A Natural Skin Cancer Cure

How To Prevent Skin Cancer

How To Prevent Skin Cancer

Complete Guide to Preventing Skin Cancer. We all know enough to fear the name, just as we do the words tumor and malignant. But apart from that, most of us know very little at all about cancer, especially skin cancer in itself. If I were to ask you to tell me about skin cancer right now, what would you say? Apart from the fact that its a cancer on the skin, that is.

Get My Free Ebook

How I Survived Malignant Melanom

By The Time You've Finished Reading How I Survived Melanoma Skin Cancer Seven Survivors Tell Their Stories. You'll Feel Like A New Person, with A New, More Positive Outlook! You will learn: 1. How do I know if I have melanoma? What are the signs and symptoms? I wanted to know why the doctor was so concerned when she looked at that little mole on my forearm. What was it that looked so sinister? How worried should I be? Was the doctor over-reacting? 2. What tests will the doctor carry out to see if I have melanoma? Will they be able to tell me on the spot if there is a problem? Or will I have to wait for days, fretting about whats going on? 3. How curable is melanoma? If they do tell me its melanoma, what exactly does that mean? Is it a death sentence? Will they tell me You have 12 months to live. Get your life in order and prepare for the worst.? 4. What are the stages of the disease? The reading Id done said that there were different stages of melanoma. What are the symptoms of each stage? What are the survival rates of each stage? If I had a later stage melanoma, wouldnt I know about it? Wouldnt I actually feel like I was sick? 5. How quickly does the disease progress or spread? Should I have gone to the doctor sooner? Id noticed the mole changing over about 3 months. Was this delay critical? 6. How is melanoma normally treated? Would I have to go through chemotherapy and radiation treatment? If so, for how long? What are the odds of curing the disease using these treatments? How extensive is any surgery likely to be? How big will the scars be? 7. What are the common side effects of the treatments? Would I lose my hair? Would I become sterile? What else could I expect? 8. What alternative treatments are available? Id heard of people going on special macro-biotic diets. Id seen lots of herbal remedies on the internet. Which of these are proven and documented, and which ones are snake oil? Is it possible to combine alternative treatments with surgical other western treatments? How do I find a doctor that is open to using both alternative and western treatments? 9. What are the latest treatments being developed, and who is carrying out clinical trials of these new treatments?

How I Survived Malignant Melanom Summary


4.6 stars out of 11 votes

Contents: EBook
Author: Daryl Grant
Price: $39.00

My How I Survived Malignant Melanom Review

Highly Recommended

All of the information that the author discovered has been compiled into a downloadable book so that purchasers of How I Survived Malignant Melanom can begin putting the methods it teaches to use as soon as possible.

When compared to other ebooks and paper publications I have read, I consider this to be the bible for this topic. Get this and you will never regret the decision.

Download Now

Is There an Association Between Levodopa Therapy and Melanoma

The connection between levodopa therapy, PD, and malignant melanoma has been a matter of debate for three decades. It originally derived from the biochemistry of the drug. Levodopa is a substrate for the development of dopamine, which, in turn, develops into neuromelanin in CNS nigral neurons. It is the dopaquinones derived from levodopa that are oxidized to form neuromelanin in these cells (126). Hence, it has been proposed that levodopa may also affect the activity of melanocytes in the skin, possibly promoting malignant transformation, although this connection has never been proven. In addition, it is now known that 70 of melanoma cases in the general population appear to be connected with a genetic mutation unrelated to PD. Thus, it would seem unlikely that there would be a connection between PD, lev-odopa, and melanoma. Nevertheless, reports of melanoma in patients treated in the 1970s led the FDA to require language in the package insert that cautions against the use of levodopa...

Mucosal Melanomasspecial Considerations

Mucosal melanomas of the head and neck are rare, with only about 1000 cases reported. Of more than 84,000 melanoma cases in the National Cancer Data Base, only 1.3 were mucosal, and 55 of that subgroup arose in the head and neck (15). The majority of head and neck mucosal melanomas are sinonasal in origin, and the others are primarily from the oral mucosa. Among the sinonasal tumors, 81 arise in the nose and 19 in the sinuses, although the size of some of these tumors makes it difficult to be sure of the site of origin. Most of the oral lesions arise from the hard palate and maxillary alveolar ridge. Melanomas arising in other head and neck mucosal sites such as the larynx, pharynx, and cervical esophagus have been reported, but are quite rare. A melanoma arising in the oropharynx is shown in Figure 3. The oral lesions are often detected by healthcare personnel as a dark spot in the mouth, but the sinonasal lesions are often only apparent after symptoms such as epistaxis or nasal...

Allogeneic melanoma lysates for the treatment of disseminated melanoma

We began our studies with allogeneic melanoma lysates late in 1985, in patients with disseminated melanoma. This was done primarily for ethical reasons, because we did not know what the toxicity of this form of treatment would be. That would preclude its use in patients with minimal residual disease the group that might best benefit from vac-cination until phase I data showed that toxicity was acceptable. We should note that the use of this admittedly crude whole-cell vaccine, together with an adjuvant that may further have obscured the effects of the tumor preparation itself, was a decision we made deliberately after some experience with extracts of autologous cells given without adjuvant our initial attempt to use a better defined, individual-specific antigen. The failure to generate either an immune response or a clinical response in a small group of patients led us to the diametrically opposite approach whole-cell lysates with antigens that gave the patients the choice of which to...

Other genetic factors and skin cancer

An increasing number of studies have recently reported on associations between various candidate genes and skin cancer. Most plausible are reports of associations between genes involved in DNA repair and skin cancer. Patients with xeroderma pigmentosum (OMIM 278730), an autosomal recessive disorder characterized by mutation in one of a number of genes involved in nucleotide excision repair, have a grossly elevated risk of both melanoma and NMSC (often presenting in early childhood with multiple tumors). Various studies have recently reported associations between some of the genes implicated in xeroderma pigmento-sum and skin cancer (Dybdahl et al., 1999 Tomescu et al., 2001 Winsey et al., 2000), whilst other studies have reported on associations between polymorphisms in pathways that may play a role in oxidative damage control in skin in response to UVR (such as the glutathione transferases) (Lear et al., 1997 Ramsay et al., 2001) or skin immunity (tumour necrosis factor a) (Hajeer et...

Uveal melanoma of choroid and ciliary body

Malignant melanoma of choroid. W Figure 37.1. Malignant melanoma of choroid. W Figure 37.2. Malignant melanoma of choroid. W Figure 37.2. Malignant melanoma of choroid. W Figure 37.3. Malignant melanoma of choroid. W Figure 37.3. Malignant melanoma of choroid. W Figure 37.4. Malignant melanoma of choroid. W Figure 37.4. Malignant melanoma of choroid. W

The MC1R and human pigmentation and skin cancer

In 1995 we showed that sequence variation at the MC1R was common in man and that particular variants were associated with red hair and ''pale'' skin (Valverde et al., 1995). Such individuals are characterized clinically by a tendency to burn rather than tan in response to UVR, and pale skin on sites protected from the sun (such as the buttock). This clustering of phenotypic characteristics is known to be associated with an elevated risk of most forms of skin cancer (Rees, 2002c). Red hair approximates to an autosomal recessive trait although the penetrance depends on which MC1R variants are present (typically penetrance is around 0.85). This bold statement ignores some of the subtleties of phenotype hair color changes with age from childhood through early and late adulthood, and different body sites frequently have different colored hair. Given that the MC1R is a determinant of pigmentary phenotype and that pigmentary phenotype is a risk factor for both melanoma and NMSC it is not...

Glioma and Melanoma

Yamanaka and colleagues have used SFV replicons for multiantigen immunotherapy in the 203 glioma and B16 melanoma models derived from C57BL 6 mice (54). Tumor cDNA was generated from tumor mRNA and cloned into the SFV replicon constructs yielding SFVcDNA-RNA (54). This mixture of SFV replicons expressing a wide array of tumor-derived heterologous proteins was incubated with syngeneic, Day 6, bone marrow-derived DCs that were then used as the immunogen via three intraperitoneal injections at 1-wk intervals. Splenocytes from immunized animals were evaluated for CTL responses and the SFV replicon-loaded DCs yielded substantial tumor-specific CTL responses, approximately twofold greater specific cytolysis at each effector to target ratio compared to splenocytes from animals immunized with tumor RNA or tumor lysate-pulsed DCs. In tumor protection models, tumor outgrowth was delayed with the SFV-cDNA replicon-loaded DC immunizations and 50 full protection was demonstrated using intracranial...


Rates of melanoma vary widely geographically, with the major determinant of risk being skin color. The relative incidence of subtypes of melanoma differs across racial groups with one rare subtype termed ''acral lentiginous melanoma'' occurring in all racial groups (Hudson and Krige, 1993 Stevens et al., 1990 Jimbow and Kukita, 1984) but the commoner subtypes (superficial spreading melanoma, nodular melanoma and lentigo maligna) occur essentially only in white persons. For these latter subtypes and among white populations, the highest incidence is in countries nearest to the equator, leading to the hypothesis that sun exposure is the major environmental determinant of melanoma risk (Armstrong, 1988). Australia and New Zealand have the highest rates worldwide (Jones et al., 1999 Parkin et al., 1997). Variation in skin color explains much of the variation within and between geographical locations. For instance, in Hawaii, the melanoma incidence rates for people of European descent are...

Skin cancers

There are two main types of skin cancer melanoma, arising from the neural crest derived melanocytes and basal cell carcinoma and squa-mous cell carcinoma derived from keratinocytes (the latter two referred to as non-melanoma skin cancer, NMSC) (Rees, 2002b). This broad distinction into two types is useful NMSC is extremely common and case-fatality extremely low (< 0.5 ) melanoma by contrast is less common but with a case fatality of -20 (Rees, 1998). Melanomas usually present as pigmented lesions with a darkening and increase in size of a preexisting melanocytic nevus (a benign collection of immature melanocytes) or de novo, as a lesion resembling a mole that is growing and getting darker. Melanoma may metastasize early and fatality is usually due to secondary spread. Because therapy of the tumor once spread has occurred is largely palliative, emphasis has been on primary prevention, early detection and excision of malignant lesions, and identification of high risk groups (Rees,...

Skin Cancer

Balch CM, Buzaid AC, Soong SJ, Atkins MB, Cascinelli N, Colt DG, Fleming ID, Gershenwald JE, Houghton A Jr, Kirkwood JK, McMasters KM, Mihn MF, Morton DL, Reintgen DS, Ross MI, Sober A, Thompson JA, Thompson JF. Final version of the American Joint Committee on cancer staging system for cutaneous melanoma. J Clin Oncol 2001 19 (16) 3635-3648. Banerjee SS, Harris M. Morphological and immunophenotypic variations in malignant melanoma. Histopathology 2000 36 387-402. Breslow A. Thickness, cross-sectional areas and depth of invasion in the prognosis of cutaneous melanoma. Ann Surg 1970 172 902-908. Clark WH Jr. A classification of malignant melanoma in man correlated with histogenesis and biological behaviour. In Montagna W, Hu F (eds).Advances in Biology of the Skin. Vol. 8. Pergamon Press. London 1967 621-627. Cochran AJ, Bailly C, Cook M, Crotty K, Mihm M, Mooi W, Sagebiel R. Recommendations for the reporting of tissues removed as part of the surgical treatment of cutaneous melanoma....

Non Skin Cancers

Significantly increased relative risk not only for non-melanoma skin cancer but also for buccal cancer and primary liver cancer. Whether this increased relative risk was correlated with alcoholism and or smoking could not be ascertained in this study. The data suggest a co-occurrence of HS and cancer but these observations await confirmation in other HS populations. If confirmed it may give direction to further aetio-logical and pathogenic research in HS.

Michael A Morse MD Timothy M Clay PhD and H Kim Lyerly MD

Interest in activating the immune system to control tumors dates to more than 100 yr ago and is often attributed to William B. Coley who used the inflammatory response to bacterial products as a form of immunotherapy (1). This initiated the era of attempts to nonspecifically activate the immune system with the hope that tumors would be targeted as part of the overall, predominantly inflammatory, response. Following the abandonment of the crude bacterial products in Coley's toxin, other bacteria or their cell wall products such as bacille Calmette-Guein (BCG), Corynebacterium parvum, nocardia rubra, OK-432, and others have been tested. In some circumstances, these inflammatory activators have had success as antitumor agents. For example, BCG has activity in superficial bladder cancer in situ (2). Later, cytokines such as interleukin (IL)-2 that could activate T and natural killer (NK) cells were extensively tested and demonstrated activity in renal cell carcinoma and melanoma. In the...

Ancillary Techniques in Histopathology Specimens

Immunohistochemistry The surgical pathologist's second H and E is invaluable in assessing tumour type, prognosis and treatment, e.g., carcinoma (cytokeratins) versus malignant lymphoma (CD45) and malignant melanoma (S100), or better prognostic and hormone responsive breast cancer (oestrogen receptor positive). Tumour antigenic profile is also of use in specifying the site of origin for a metastasis, e.g., prostate carcinoma (PSA positive). Electron microscopy Valuable in renal biopsy diagnosis, and tumours where morphology and immunohistochemistry are inconclusive, e.g., malignant melanoma (pre- melanosomes) and neuroendocrine carcinoma (neurosecretory granules). Quantitative methods Prognostic indicators include the Breslow depth of invasion in malignant melanoma, muscle fibre typing and diameter in myopathies and the mitotic activity index in breast carcinoma.

Molecular Correlates Of Germ Cell Tumor Development And Function In Humans

C-Kit is a transmembrane glycoprotein (molecular weight, 145Kd) produced by the c-kit gene and is a member of the receptor tyrosine kinases. It is related to the receptors for platelet-derived growth factor, macrophage colony-stimulating factor, and the flt 3 ligand.70 In mice, its natural ligand is the Steel factor or the stem cell factor, and murine studies suggest that the function of Kit tyrosine kinase is important in the development of the interstitial cells of Cajal, mast cells, and normal hematopoiesis71 and in the development and maturation of spermatogonia.43 Curiously, c-Kit expression or mutation has been observed in a range of human malignancies, including mast cell leukemia, gastrointestinal stromal tumors, melanoma, and seminoma.7,72,73 Tian and colleagues7 found activating mutations of c-kit in about 10 of seminomas but in no instances of NSGCT, but it should be noted that these workers studied only exons 11 and 17. The commonality of c-kit mutations in melanoma, GCTs,...

Antiepileptic Drugs In Neurooncology Patients

Approximately 25-30 per cent of the adults and 10-15 per cent of the children with primary or metastatic brain tumors will experience a seizure during the course of their illness 1-4 . Seizure incidence is even higher in patients with primary glioma, particularly low-grade glioma and in patients with metastases from melanoma 3,5-10 . Although routine prophylaxis with AEDs is not recommended for patients with a brain tumor who have not had a seizure, patients in the community are often on an AED, usually phenytoin 2 . This practice can lead to

Experimental evidence supporting a role of tgfP in active tumormediated immunosuppression

Strategies to block the TGF-P-signaling pathway include decreasing expression of TGF-P ligand with antisense to TGF-P, decreasing TGF-P ligand binding to responsive cells with antibodies to TGF-P ligand or expressing the soluble extracellular domain of TPRII, or expressing a mutant form of the TPRII that lacks the entire kinase domain and most of the juxtamembrane region, TBRIIAcyto, which acts to block TGF-P signaling in a dominant-negative fashion (29). Initial studies performed using antibodies to TGF-P ligand were able to enhance tumor-specific immune responses and reduce tumorigenic-ity, but not alter the ability of the immune system to eradicate the tumor (30,31). However, neutralizing antibodies to TGF-P (prevents immunosuppression) combined with IL-2 therapy (to stimulate the immune system) was able to decrease the number of metastases in a murine B16 melanoma model (32). Subsequent studies, performed by inserting an antisense construct to TGF-P ligand in resected tumors and...

Neoplastic Conditions

Extrahepatic bile duct carcinoma there is an increased incidence in various disorders including ulcerative colitis, sclerosing cholangitis, gall stones and congenital bile duct anomalies. The majority (50-75 ) arise in the upper third (including the hilum) with lesser numbers in the middle and distal thirds (10-25 each) or even diffuse and multifocal. Sometimes polypoid but often nodular, ulcerated, sclerotic or strictured, prognosis relates to the stage of disease, location and histological grade. There is characteristic perineural invasion often with involvement of regional lymph nodes, peritoneum or the liver (upper third tumours) at presentation. Other rare cancers are carcinoid tumour, malignant melanoma, lymphoma leukaemia and in childhood, embryonal rhabdomyosarcoma.

Frequency of HIT in Surgical Patients Treated with Porcine Mucosal UFH

Isolated limb perfusion (ILP) with melphalan employs extracorporeal circulation (and thus high-dose UFH) to treat melanoma or unresectable sarcoma limited to an extremity. In one study, HIT occurred in three of 108 patients (2.8 ), who also received sc UFH prophylaxis following ILP (Masucci et al., 1999). The occurrence of arterial thrombosis and partial limb amputation in two of these patients with HIT led the investigators to discontinue routine UFH prophylaxis post-ILP. The hypothesis that ILP is a high-risk situation for HIT was supported by a follow-up prospective study by these same investigators showing that heparin-dependent antibodies were formed in all nine patients who underwent ILP (despite not receiving postoperative UFH prophylaxis), with eight having strong antibodies that effected serotonin release in vitro (Masucci et al., 1999).

Chemotherapy And Boron Neutron Capture In Rat Brain Tumor Models

Neutron flux at the Brookhaven National Laboratory Medical reactor appeared curative for rats bearing 9L gliosarcoma. On the other hand, the F98 tumor was refractory to therapy. Barth et al., 49 went on to determine whether the efficacy of boron neutron capture therapy could be increased by combination with standard radiation therapy. F98 glioma-bearing rats treated with intracarotid or intravenous sodium borocaptate (30 mg kg) plus p-boronophenylalanine (250 mg kg followed 2.5 h later by exposure to a neutron flux and 7-10 days later by standard radiation therapy to a total dose of 15 Gray had survival times (61-53 days) similar to those of rats that did not receive the standard radiation therapy (52-40 days). However, animals in the same study bearing MRA 27 melanoma showed a significantly enhanced survival when boron neutron capture therapy was followed with conventional radiation therapy.

Role of primeboost for polyepitope vaccines

Of CTLs with a range of specificities (12). However, we and others have shown that, during the boosting step, competition of CTLs around the APCs is a major factor in reducing the breadth of a primed response (4,13). By simultaneously monitoring CTLs of several different specificities ex vivo we have shown that after boosting a poly-epitope-primed response, CTLs of a single specificity expand, leaving those of other specificities behind (Fig. 2 4). The phenomenon observed is termed immundomination (4). Recently, in this setting, a number of groups have investigated the underlying mechanism of immundomination (4,13-15). It has been shown that competition of CTLs around APCs (in particular DCs) can cause the inhibition of subdominant CTLs by other CTLs specific for the dominant determinant. It is possible to eliminate the effect of immundomination either by increasing the number of APCs displaying a set of epitopes, or by expressing single epitopes antigens on separate APCs....

Ordering of prime and boost immunizations

Although murine models have demonstrated the efficacy of prime-boost immunizations against tumor antigens, the best combination of platforms for immunization and the best order for delivery are still being evaluated. Schlom's group at the National Cancer Institute has demonstrated in mice transgenic for the human CEA gene that primary immunization with a vaccinia vector encoding CEA (rV-CEA) and costimulatory molecules followed by boosts with a fowlpox vaccine encoding the same proteins could generate potent CEA-specific T-cell responses that reduced tumor progression and increased survival (16). When used with DNA vaccines, it appears that attenuated vaccinia vectors should be administered as a boost (at least in infectious-disease models) (2). Similarly in a murine model of pre-B-cell leukemia, priming with a DNA plasmid vaccine followed by boosting with a vaccinia-based vaccine (both expressing a peptide derived from the tumor-specific immunoglobulin) conferred the greatest...

Risks of Other Cancers in BRCA1BRCA2 Carriers

In a parallel study based on 173 BRCA2 families, the risk of other cancers was approximately twofold in both male and female carriers (42). The largest excess risk was for prostate cancer, with an estimated 4.7-fold relative risk, increasing to sevenfold in men below age 65. A 3.5-fold risk of pancreatic cancer was also found, and significant excesses were also seen for cancers of the stomach, buccal cavity and pharynx, gallbladder and bile duct, and fallopian tube and for melanoma. A more recent study from the Netherlands also found increased risks of prostate, pancreatic cancer, and head and neck cancer (43).

Identification of Class I Peptides

Once a class I specific peptide has been identified, one must determine whether that peptide can be used to generate T cells capable of responding to and lysing HLA-matched tumor that expresses the antigen of interest. These studies may be performed by using peptide to sensitize T cells in vitro. However, in vitro propagation of human lymphocytes is labor intensive and operator dependent. In addition, establishing T-cell lines using lymphocytes derived from patients with cancer may be made more difficult due to potential inherent defects in the T-cell response in tumor-bearing hosts. For these reasons, another approach has been developed to rapidly evaluate the utility of predicted class I epitopes, active immunization of HLA-A2 transgenic mice. Vaccinating mice with class I peptides allows a more robust immune response to be generated to a particular tumor antigen than the response found endogenously in cancer-bearing patients. Moreover, initiating in vitro culture with a robust...

Patients With Cancer Can Be Immunized With Class I Peptide Based Vaccines

Clinical trials performed vaccinating cancer patients with class I binding peptides derived from a variety of tumor antigens have demonstrated that patients can be immunized to boost immunity to self-tumor antigens. Peptide-based vaccines offer an excellent model for assessing the ability to immunize by measuring immunity generated with assays that can specifically measure class I T-cell responses. Lee et al. (13) used MHC tetramer flow cytometry to evaluate in vivo response to vaccination with gp100209_217 and tyrosinase368-376 peptides in 44 HLA-A2 patients with advanced-stage melanoma. Tetramer assays performed before, during, and following the period of vaccination demonstrated appearance of gp100-specific T cells as early as 4 wk, i.e., after two vaccinations. At 18 wk at least 20 of CD8+ T cells were specific for gp100209-217 in four of six patients. Overall, 37 of 42 patients developed an immune response to gp100209-217 following vaccination as measured by MHC tetramer assay....

Gene Expression Patterns In Normal And Diseased Tissues

Relevant tumor subtypes, including B-cell lymphoma 38 , leukemia 33 , melanoma 39 , and breast cancer 40 . In relation to cancer, DNA microarrays have also been used to distinguish between patients who respond to therapy versus those who do not, despite having identical histological appearance. In addition, it has been useful to identify molecular correlates and molecular signatures of tumors of the same and different lineages. Within the field of neuro-oncology, several examples now are emerging in the literature describing differential gene expression by microar-rays. DNA microarrays have been used to profile tumor subtypes, most notably in astrocytomas 41,42 and medulloblastomas 41-43 . As each group of tumors had significantly different prognoses, treatment can then be optimized for tumor subtypes based on their molecular phenotypes.

From The Laboratory To The Clinic Oncolytic Herpes Simplex Viruses Appear Safe In Trials But Just How Effective Will

Adverse events, such as acute toxicity, viral shedding, clinically evident reactivation or encephalitis attributable to virus inoculation or replication were observed in either study at any of the doses, which escalated from 103-105 pfu with 1716 and 1 x 1063 x 109 with G207 (16,17). Patient deaths were primarily the reuslt of progressive cancer, with one report of radiation necrosis. Two of the G207 treated patients were alive as of December, 2003 (77). In addition, both studies were unable to establish a maximum tolerated dose (16,17). More recently, evidence has been obtained that is reportedly consistent with replication of 1716 within tumor samples over time, supporting the concept of tumor destruction by viral oncolysis (78). Finally, multiple injections of 1716 directly into skin lesions of a limited number of melanoma patients have proceeded without any harmful effects (79). A phase Ib II study involving G207 is currently underway open to patients with recurrent glioblastoma...

Gangliosides GM2 GD2 GD3 and Fucosyl GM1

Gangliosides are sialic acid-containing glycolipids that are expressed at the cell surface with their lipid (ceramide) moiety incorporated into the cell-surface lipid bilayer. Most gangliosides considered as potential targets for cancer therapy are expressed primarily in tissues and tumors of neuroectodermal origin. This is true for the melanoma, sarcoma, neuroblastoma, and SCLC antigens GM2, GD2, and GD3, and the SCLC antigen fucosyl GM1. The structures of these antigens are shown in Fig. 2. Surprisingly, however,

Intratumoral Gene Delivery By Alphavirus

Alphavirus vectors have been applied for several studies in tumor animal models. In this context, SFV particles expressing the p40 and p35 subunits of the murine inter-leukin-12 (IL-12) gene were injected into mice bearing B16 melanoma tumors (32). The tumor development was followed by Doppler ultrasonography and demonstrated a significant tumor regression as well as inhibition of tumor blood vessel formation. It was also shown, that repeated injections improved the tumor regression efficacy. In another study, significant regression of P185A tumor growth was achieved after intra-tumoral injections with SFV-IL-12 vectors (33). The size of the tumor played an important role in relation to efficacy as treatment of large tumors was less successful. The intratumoral injections also presented prophylactic efficacy, as long-term immunity and absence of reoccurrence of tumors were achieved.

Tumor Targeting Of Alphavirus Vectors

Yet another approach to obtain tumor selective transfection was attained by encapsulation of recombinant SFV particles in liposomes (40). By this procedure, targeted gene delivery to human LnCaP prostate tumors implanted in severe combined immunodeficiency (SCID) mice was achieved after systemic delivery of encapsulated SFV-LacZ particles. Systemic administration of encapsulated SFV particles expressing the p40 and p35 subunits of IL-12 to SCID mice with human Panc-1 pancreatic tumors resulted in statistically significant reduction in tumor growth after a single injection (41). Furthermore, an initial phase I study on advanced melanoma and kidney carcinoma patients demonstrated the safe use of this SFV vector in humans. In this study the maximum tolerated dose (MTD) for encapsulated SFV-IL-12 particles was 3 x 109 particles m2, which might seem relatively low compared with doses for other viruses, usually administered in the range of 1011 particles. However, in this case the dose is...

Cytotoxic T Cells and NKMediated Immunity

In cancer patients, evidence that anti-Id antibody can stimulate a CTL response was provided by immunization of melanoma patients with two different anti-Id antibodies mimicking high-molecular-weight proteoglycan antigen (73). Human leukocyte antigen (HLA)-A2-specific CTL responses were observed in 43 of the patients who appeared to express the HLA-A2 allele. Anti-Id antibody ACA 125, which mimics the ovarian tumor-associated antigen CA 125, also induced cellular immune responses that resulted in lysis of antigen-positive ovarian tumor cells (43). Although CTL assays in these two studies were performed with HLA-matched tumor cells, autologous tumor cells were not available and could not be used. Thus, other mechanisms of cytotoxicity could be involved in the lysis of the tumor cells. Durrant et al. (74) treated six patients with rectal cancer with the anti-Id antibody, 105AD7. PBMCs or lymph node cells from three of these patients caused significant killing of autologous tumor cells,...

Genetically Engineered VSV as a Gene Therapy Tool Against Cancer

Oncolytic studies indicated that wild type VSV exhibited considerable potential as an anticancer agent. However, the ability to modify VSV through genetic engineering obviously affords the prospect of creating new generations of custom-made VSV vectors that contain immunomodulatory and or suicide cassettes designed to increase their antitumor activity. In order to begin evaluating whether genetically engineered VSV carrying tumor-killing cassettes could be created and whether such viruses were more efficacious in tumor therapy than the wild-type VSV, we developed VSV vectors carrying, as models, the herpesvirus TK suicide cassette or the cytokine gene interleukin-4 (IL-4). The foreign genes were cloned as additional transcription units between the VSV G and L genes and all viruses were grown to exceptionally high titers (71,103). TK protein was synthesized to extremely high levels and was functional, being able to phosphorylate ganciclovirs (GCV). Recombinant viruses also retained...

Cytokine expression driven by pox vectors

Both orthopox (vaccinia) and avipox (fowlpox and canarypox) vectors have been used for the expression of GM-CSF. Previous studies have shown that direct injection of rV-GM-CSF into tumor lesions will enhance antitumor effects both in experimental animals (73) and in clinical studies in melanoma patients (74). On the other hand, fowlpox expresses transgenes for 14-21 d. A recent study in mice has compared the biological activity of recombinant GM-CSF with that of fowlpox expressing GM-CSF (40). A single injection of rF-GM-CSF led to significantly higher numbers of APCs, particularly DCs, in regional lymph nodes when compared with four daily injections of GM-CSF. In

Proteasome Inhibitors

The first in vivo demonstration of the antitumor activity of proteasome inhibitors involved treatment of Burkitt's lymphoma xenografts with PS-341, since renamed Bortezomib 173 . A screen of tumor cell lines isolated from 9 different cancer types (brain, colon, melanoma, prostate, lung, breast, renal, and ovarian) showed that Bortezomib had broad antitumor activity both in vitro and in vivo 174,175 . Additional studies found that transformed cells were selectively sensitive to the cytotoxic effects of protea-some inhibition when compared to normal cells in culture 173,176-179 . In vivo effects were detected in prostate tumor xenograft and murine mammary carcinoma models 180 . trials evaluated the use of bortezomib in the treatment of both chemoresistant and recurring solid tumors (hepatocellular carcinoma, melanoma, metastatic breast cancer, metastatic colorectal carcinoma, meta-static renal cell carcinoma, neuroendocrine tumors, ovarian carcinoma, non-small cell lung carcinoma, and...

Molecular Chemotherapy With rAAV

Consideration of molecular chemotherapy strategies for selective killing of tumor cells suggests that long-term expression of transgenes is not an imminent requirement hence, AAV-based vectors are less preferred over adenoviral vectors. Further, the efficacy of adenoviral infection in different tumor cells has been reported to be significantly higher than many other available gene therapy vectors. However, it has recently been reported that the efficiency of rAAV transduction of primary tumor material, derived from malignant melanoma and ovarian carcinoma, is significantly higher (> 90 ) than that seen in established tumor cells of the same derivation in culture (86). This observation suggests that it is possible to utilize rAAV in direct targeting of tumor cells for an effective killing by approaches such as molecular chemotherapy, cytokine gene transfer, and inactivation of protooncogene expression. In addition, studies by Su et al. using an AAV-TK-IL-2 vector reported...

Melanocortin 1 Receptor

The melanocortin 1 receptor (MC1R) is a major determinant of hair and skin pigmentation. Extensive case-control studies and studies on familial melanoma showed that genetic variations in MC1R play crucial roles in the development of both familial and sporadic melanoma.5152 More importantly, certain variants of the MC1R were associated with development of melanoma, independent of pigment synthesis. Therefore, apart from pigment synthesis, the MC1R is involved in hitherto unknown tumorigenic signal transduction pathways. Whether the genetic variants of MC1R harbor constitutively active properties, thus providing a causative link to aberrant proliferation, is yet to be determined.

Is achieving extreme old age worthwhile the centenarian phenotype

Independently functioning at age 92 (Hitt et al., 1999). Most subjects experienced a decline in their cognitive function only in the last three to five years of their lives (Perls, 1997 Silver et al., 1998). Upon further examination of the ages of onset for ten common age-associated diseases (hypertension, heart disease, diabetes, stroke, non-skin cancer, skin cancer, osteoporosis, thyroid condition, Parkinson's disease, chronic obstructive pulmonary disease and cataracts) among 424 centenarians (323 males and 101 females), the subjects were noted to fit into three morbidity profiles ''survivors,'' ''delayers'' and ''escapers'' (Evert et al., 2003). Survivors, individuals who were diagnosed with age-related illness prior to age 80, accounted for 24 of the male and 43 of the female centenarians (p 0.0009). Delayers, individuals who delayed the onset of age-related diseases until at least age 80, accounted for 44 of the male and 42 of female centenarians. Escapers, individuals who...

Global Analysis of Gene Expression

In a few cases, it has been possible to confirm functional hypotheses first suggested by expression analysis. For example, the gene encoding RhoC (ARHC) was identified as an expression correlate of tumor metastasis in a melanoma model critically, blockade of RhoC diminished metastasis and activation enhanced metastasis in this model.57 As yet, few such functional hypotheses have been validated, due to the relative mismatch between our ability to rapidly generate such hypotheses using expression microarrays and our more limited ability to test them in the laboratory. However, methods for systematically testing gene function on a global scale are rapidly evolving and are discussed briefly in the next section.

Metabotropic Glutamate Receptors

Studies in a new melanoma mouse model recently implicated the ectopic expression of the Gq 11-coupled metabotropic glutamate receptor 1 (mGluRl, Grml), a member of the C family of GPCRs, in melanomagenesis and metastasis.8889 Alterations in the expression of Grml are sufficient to promote mouse melanocyte transformation in vivo.88 In addition to the coupling to Gq 11 proteins, Grml may act to activate Gs proteins in transfected cells.90

Summary And Conclusions

Our studies and those of others (11, 25) indicated that IL-2R and endogenous IL-2 are ubiquitously expressed in human carcinomas both in culture and in situ. Data available from various laboratories provide evidence for the presence of IL-2R on melanoma cell lines and other tumour cell lines. Furthermore, the IL-2 IL-2R pathway appears to be operative in normal non-hematopoietic tissues, e.g., fibroblasts and keratinocytes (5, 26). Thus, this pathway appears to be active in vivo and in 6. Rimoldi D, Salvi S, Hartmann F, Schreyer M, Blum S, Zografos L, Plaissance S, Azzarone B, Carrel S. Expression of IL-2 receptors in human melanoma cells. Anticancer Res 1993 13 555-564 12. Alileche A, Plaisance S, Han DS, Rubinstein E, Mingari C, Bellomo R, Jasmin C, Azzarone B. Human melanoma cell line M14 secretes a functional interleukin-2. Oncogene 1993 8 1791-1796 29. Atkins MB. Interleukin-2 in metastatic melanoma establishing a role. Cancer J Sci Am 1997 3 57-58

Venous Thromboembolism

Venous thromboembolism (VTE) has long been known as a complication ofcancer. Venous thromboembolisms are usually divided into deep venous thrombosis (DVT) and pulmonary embolism (PE). When DVTs affect the proximal veins of the lower extremities they are usually treated as a PE. We discuss VTE with all brain tumor patients at the initial consultation. Mechanism of action for VTE includes, venous stasis (immobility), intimal injury, and alterations in coagulation. In brain tumor patients, Sawaya et al. (46) has shown alterations in the fibrinolytic system and an underlying coagulopathy as causes of VTE. Other associations include age, prior DVT, smoking, oral contraceptives, and obesity. During surgery, brain tumor patients often have induced dehydration and hyperosmolality that increase the VTE risk. Malignant brain tumor patients have also been shown to display an increased risk of VTE with reports of upward of 28 of patients having symptomatic events (47). At our institution we have...

Antineoplastic And Chemopreventative Effects

Many studies have reported antineoplastic effects of both oil and water soluble allyl sulfur compounds from garlic, but the effect is generally greater for the Iipid-soluble compounds (Knowles & Milner 2001). Diallyl disulfide, one of the most studied oil-soluble organosulfur compounds in garlic, has demonstrated antineoplastic activity against both hormone-dependent and hormone-independent breast cancer cell lines (Nakagawa et al 2001). It also inhibits the proliferation of human tumour cell lines for colon, lung and skin cancer (Sundaram & Milner 1996). Garlic derivatives inhibit proliferation of human prostate cancer cell lines and human breast cancer cell lines (Pinto & Rivlin 2001). In vitro results also show ajoene induces apoptosis in human leukaemic cells (Dirsch et al 2002), whereas allien, but not its precursor alliin, inhibits proliferation of human mammary, endometrial, and colon cancer cells (Hirsch et al 2000).

Environment as determinant of i genotype and ii disease

First, within a long evolutionary sense, various regional differences in the biological attributes of human populations have arisen as a result of natural selection pressures exerted by local environments on human biology. In this sense, environment acts as a determinant of the local human genotype. Many examples are well known in the anthropological and biomedical literature. They include (i) variation in skin pigmentation, which (especially in today's world in which there has been considerable redistribution of regional populations over recent centuries) contributes to variations in local population risks of skin cancer and vitamin D insufficiency (ii) various enzymatic polymorphisms which are associated with altered risks of various diseases (e.g. the N-acetylation pathway enzymes, various of the cytochrome P450 oxidative enzymes, the activity of alcohol dehydrogenase, and lactase activity) many of which such polymorphisms are presumed to reflect the allele-selecting impacts of...

Use of autologous or allogeneic vaccines

Could provide the danger signals needed to activate an immune response. For example, Ward (8) observed that in a murine melanoma model, vaccination of B6 mice with allogeneic K1735 melanoma cells protected against challenge with syngeneic B16 melanoma. This protection could not be improved on by cytokine transfection of the vaccine cells (9). In contrast, K1735 cells administered to syngeneic mice offered no protection against autologous challenge unless the K1735 were transfected with granulocyte-mac-rophage colony-stimulating factor.

Oncogenic Capacity Of The Jakstat Signaling Pathway

Extensive data describe activated Stat3 and Stat5 in tumors. Activating mutations of Stat3 and Stat5 and specific inhibition of Stat3 and Stat5 by gene deletion, antisense oligonucleotides, or dominant negative approaches have highlighted the importance of Stat3 and Stat5 in tumor formation. Mutations of Stat3 that allow spontaneous dimer-ization of the monomers in the absence of interactions between phosphorylated tyrosines and SH2 domains are sufficient to cause transformation and induce tumor formation in nude mice (98). Inhibition of Stat3 signaling using antisense Stat3 or by a Jak-selective tyrosine kinase inhibitor, AG490, restored the sensitivity of cells from patients with large granular lymphocyte (LGL) leukemia to Fas-mediated apoptosis (99). Downregulation of Fas correlates with an increase in metastatic potential and resistance of tumors to chemically and physically induced apoptosis. This effect is mediated, at least in part, by an interaction between Stat3 and c-jun,...

The CD40CD40L Pathway

CD40 is a member of the tumor necrosis factor receptor (TNFR) family (62-65) broadly expressed on antigen-presenting cells (APCs), B cells, epithelial cells, and endothelial cells (66). Engagement of CD40 on APCs by its ligand CD40L (CD154) on responding T cells augments the resulting immune response (66). Exogenous activation of the CD40 pathway by agonist antibodies is capable of substituting for T-cell help (67), and augments both humoral and CTL responses (68). Manipulation of the CD40 pathway by engineering chronic lymphocytic leukemia (CLL) cells to overexpress CD40L by adenoviral gene transfer induces the expression of multiple costimulatory molecules. This augments the antigen-presenting capacity of both CD40L-modified CLL cells and unmodified bystander CLL cells in vitro, enabling both of them to prime CTL specific for autologous CLL (69). CD40L modification also augments the antigen-presenting capacity of human multiple myeloma cells in vitro (70,71). The ability of...

Cytokinemodified tumor vaccines

IL-12-secreting tumor vaccines elicit antitumor immunity in a variety of preclinical models, including mammary carcinoma, colon carcinoma, and melanoma. IL-12-transduced autologous tumor vaccines elicit CD8+ T-cell-dependent antitumor immunity capable of curing metastatic disease in the C26 model of colon cancer (103). The antitumor effect of these vaccines, which secreted pg levels of IL-12, was enhanced by CD4+ T-cell depletion (104). In the absence of IFN-y (i.e., IFN-y knockout mice), this immunity was dependent on CD4+ T cells, neutrophils, and GM-CSF (105,106). Interestingly, a direct comparison of mechanisms of antitumor immunity mediated by IL-2-secreting C26 cells as compared to IL-12-secreting C26 cells revealed that only IL-12-modified C26 vaccination was curative despite the equivalent induction of CTL (107). This was attributed to the ability of IL-12 to shift antibody production to complement-fixing IgG2a antibodies capable of lysing C26 target cells. gens in the context...

Cancers Other Than Breast and Ovarian Cancer

Carriers families rectal Stomach bladder Pancreas Liver duct Uterine Pertonium tube Cervix Prostate Lymphoma cell skin Leukemia Melanoma combined Malignant Melanoma An increased risk of cutaneous malignant melanoma associated with having an affected family member was quantified (OR 2.69) in a study by Holman and Armstrong (124). It has been estimated that 8 to 12 of cases are attributable to inherited factors (11). There is a known association of malignant melanoma with a mutation in CDKN2 on chromosome 9 that codes for p16, another important regulator of the cell division cycle (125). Several studies have demonstrated an association of BRCA mutations and malignant melanoma. In 3728 individuals in 173 breast-ovarian cancer families with BRCA2 mutations, the BCLC (36) estimated a statistically significant RR for malignant melanoma of 2.58. However, a study by van Asperen et al. (126) noted no significant excess risk of malignant melanoma associated with BRCA mutations in first-degree...

Extent Of Local Tumour Spread

Carcinomas with a pushing border and prominent lymphocytic reaction are regarded as having a better prognosis than those with a diffusely irregular infiltrating margin and sparse lymphocytic reaction, e.g. col-orectal carcinoma, head and neck carcinoma, malignant melanoma, medullary carcinoma of breast, advanced gastric carcinoma. Malignant melanoma. Direct linear measurement (mm) and anatomical level of invasion of the vertical component are strong prognostic indicators. The TNM classification is applied to carcinoma only in the majority of tissues. Other qualifying malignant tumours are malignant mesothe-lioma, malignant melanoma, gestational trophoblastic tumours, germ cell tumours and retinoblastoma.

Dose and Schedule of Administration

Five vaccine dosage schedules have been tested and are summarized in Table 2. The animal tumor models, particularly the paper of Fujiwara et al. (32), prompted us to presensitize patients with DNFB by topical application of a 1 solution in acetone-corn oil, and this was done in schedules A, B, and C. Subsequently, the presensitization was found to be unnecessary (but not deleterious) for the induction of maximum DTH to autologous melanoma cells and it was omitted for schedules D and E. Melanoma 1) Measurable metastases 97a

Development of Delayed Type Hypersensitivity Responses 861 Method for DTH Testing

Patients were tested for DTH by modification of standard methodology (45). Cryopreserved melanoma cell suspensions and PBLs were thawed, washed, and irradiated (2500 cGy). DNP modification of melanoma cells and PBLs was performed as described above. Melanoma cells (1 x 106) and PBLs (3 x 106), each either DNP modified or unmodified, were suspended in Hanks balanced salt solution without serum, phenol red, or antibiotics and injected intradermally into the ventral forearm. The mean diameter of induration was measured after 48 h. A positive response was defined as maximum diameter of induration > 5 mm. Patients were also skin tested with intermediate strength purified protein derivative (PPD)(5 TU). DTH testing was performed before the treatment program was initiated and at various times post-treatment. Analyses were performed by determining the maximum DTH response exhibited by each patient to each of the test reagents. Pretreatment positive DTH responses to autologous melanoma...

Histological Type

Malignant melanoma in situ Lentigo maligna melanoma face Hutchinson's melanotic freckle a lentiginous single and nested basal layer proliferation of melanocytes with cytological atypia (enlarged, hyperchromatic angular nuclei and cytoplasmic vacuola-tion) architectural atypia (expanded junctional nests) on a background of dermal solar elastosis. Expansion and spindling of junctional nests and any clinically nodular areas should raise a suspicion of invasion. The clinical term lentigo maligna encompasses any degree of proliferation that is confined to the epidermis (i.e. it includes Clark level I or melanoma in situ), while lentigo maligna melanoma implies the presence of dermal invasion (at least Clark level II). Superficial spreading melanoma Nodular melanoma Acral mucosal lentiginous melanoma 1Radial growth phase includes melanoma in situ (i.e. intraepidermal) microinvasion of the papillary dermis. The radial phase may be indolent with no metastatic potential and 95-100 survival...

Studies to investigate genes and environment

The Melanoma Genetics Consortium (Bishop et al., 2002) examined the risk (penetrance) for melanoma among CDKN2A mutation carriers. Carriers of particular germline mutations in CDKN2A are known to be at increased risk of melanoma. In this analysis, based on families with multiple persons with melanoma and a germline CDKN2A mutation, the overall risk of melanoma was estimated to be 62 by age 75 years. However, there was statistical evidence of a difference with carriers in Australia having a risk higher than that of the USA which, in turn, was higher than that in Europe. In the general population, for comparable time periods, the risk of melanoma by age 75 was at its highest in Australia in which it approached 2 . On an age-specific basis, the population rates in Australia were approximately 7 times higher than in Europe, while the rates in USA were 5 times higher than those in Europe. With the limited precision of the estimates of the penetrance of CDKN2A in the published analysis, the...

Clinical Effects of DNPModified Autologous Vaccine

We have reported the results of a series of clinical trials of patients with surgically incurable metastatic melanoma with measurable metastases (48) there were 97 patients, of whom 83 were evaluable (Group 1 in Table 3). Among the 83 evaluable patients there were 11 responses 2 complete, 4 partial, and 5 mixed 2 patients were judged to have stable disease. Both complete responses and two of the four partial responses occurred in patients with lung metastases. Response durations were as follows partial responses 5, 6, 8, and 47+ mo complete responses 12, 29 mo. Two examples of antitumor responses are provided below Patient no. 20254, a 77-yr-old man, presented simultaneously with a regional lymph node metastasis in the neck and 2-cm diameter mass in the lung adjacent to the cardiac border that increased in size over 2 mo observation. Five months after beginning DNP-vaccine treatment, the same mass was thought to be slightly smaller. The mass continued to slowly regress and by the 2-yr...

Importance of Dosage Schedule

Our demonstration that the development of DTH to unmodified melanoma cells was critical to its clinical effectiveness has allowed us to study the dose response of the vaccine using DTH as the response. An analysis of 284 patients (Groups 2-4 in Table 3) who were treated following resection of regional or distant metastases showed no significant association between the magnitude of DTH and the number of live (trypan blue-excluding) melanoma cells administered per dose over a dosage range of 0.5-25.0 x 106 It has become apparent that the variation in intensity of DTH responses to autologous melanoma cells among patients receiving different treatment schedules was likely to be explained by the timing of what we have called an induction dose of vaccine (62). This is defined as an intradermal injection of autologous melanoma cells without BCG, which was intended as a baseline test of DTH reactivity. Surprisingly, retrospective analysis showed that patients who received the induction dose...

Environment and high penetrance genes

Genes associated with high penetrance of cancer have been identified for breast cancer susceptibility (genes BRCA1, BRCA2 and TP53) (Chapters 15,16), colorectal cancer (APC germline mutations and mismatch repair genes, predominantly hMSH2 and hMLH1) (Chapter 17) and melanoma (CDKN2A) (Eeles et al., 2004). The risks of malignancy at one or more anatomical site are 0.60 or higher by age 75 years for mutations in each of these genes. Population-based studies suggest that approximately 1 in 500 of the general population carries a germline mutation in BRCA1 or BRCA2 (although some populations, such as Ashkenazim, have notably higher carrier frequencies. Carrier frequencies for germline mutations in the other predisposing genes may be marginally lower than for BRCA1 and BRCA2. Persons carrying such mutations represent a challenge for clinical management and an important group in which to examine the effects of environmental exposures as any exposures which are modifiable would be targets...

Studies in Ovarian and Renal Cell Carcinoma

There are no practical impediments to trials of the DNP-modified, autologous vaccine in other cancers. However, given melanoma's reputation as being particularly immunogenic among human malignancies, some would speculate that the applicability of this immunological trick would be limited. We have conducted two phase I trials in patients with ovarian cancer. In the first trial, 9 evaluable patients (10 total) with newly diagnosed adenocarcinoma of the ovary underwent standard debulking surgery plus six cycles of chemotherapy (taxol + platinum) prior to receiving DNP-vaccine. In the second trial, 13 evaluable patients (20 total) with bulky, chemotherapy-refractory disease were treated. We chose dosage schedule D (see Table 2) because it appeared to induce optimal DTH responses in the melanoma studies. Positive DTH (5-mm induration) to autologous DNP-modified tumor cells was elicited in 19 of 22 patients (median 14-mm induration). More importantly DTH to unmodified tumor cells was...

Other Clinical and Preclinical In Situ Cytokine Gene Therapy Approaches

Been inserted into a vaccinia virus that coexpressed the MUC-1 gene, which was then delivered as an intramuscular injection to men with advanced prostate cancer (73). No toxicity was observed and one patient that received three doses had some evidence of systemic immune effects. IL-15 was shown to contribute to the development of NK and antitumor response to prostate cancer in a xenograft model with PC-3 tumors (74). IL-18 may synergize with IL-12 (75) and although it has not been used in gene therapy strategies for prostate cancer the recombinant protein has been used in combination with IL-12 gene therapy in a bladder cancer model (76). The IL-24 gene also known as a melanoma differentiation associated gene 7 (MDA-7) has been used in preclinical studies for prostate cancer gene therapy (77).

Non Cancer Health Care and Health Maintenance

The transition off of primary cancer treatment is also a second opportunity to consider whether genetic assessment might be necessary. During an initial consultation, when taking the family history, a potential genetic predisposition may be detected. However, the patient may not pursue referral to a genetic counselor at that time because they are so overwhelmed by the new diagnosis of cancer and dealing with the treatment they will have to embark upon. The completion of treatment is another opportunity to review this issue and consider making a referral. The genetics of breast, ovarian, and colorectal cancers are best understood, but increasingly associations with other cancers such as pancreatic cancer and melanoma are being

Cytokine Modified Dendritic Cells

DCs are extremely efficient APCs that are widely dispersed in tissues and peripheral blood. Because they can be manipulated ex vivo and are perhaps the most specialized APCs they have been considered the prime candidates for cell mediated cancer therapy. A number of clinical trials using DC have been performed for prostate cancer (109-116) as well as other cancers (see ref. 117 for a review of the first 1000 trials). Although melanoma is the most commonly treated cancer using DC immunotherapy, GU cancers, notably, renal and prostate, are also being evaluated (118). Most of these studies involve DC primed with specific polypeptides that will bind MHC Class I or II molecules or with tumor lysates. Tumor derived mRNA transfected into DCs is being used to circumvent the challenge of lack of identity of TAAs. In current clinical trials DCs are most often injected intradermally or subcutaneously and less commonly intravenously or into a lymph node (118). A challenge with current clinical...

Chaperoned peptides as antitumor antigens multiplicity of antigenicity and crosspriming

From the above discussion it should be apparent that when purifying chaperone proteins from tumors (or any other tissue, for that matter), one is catching them in the act of transporting peptides. At least some of the intracellular peptides remain bound by the purified chaperones, and those peptides that are antigenic can now be utilized in the generation of an immune response. Thus, chaperone protein purification may be envisioned as preserving a peptide snapshot of the cellular protein pool. This leads to the doctrine of multiple antigenicity, whereby presumably numerous peptide antigens are escorted by tumor-derived chaperone proteins. One need not even know the identity or quality of the antigens, as that is a choice the immune system will make. In a vaccine setting, such antigenic multiplicity would in theory prevent the escape of specific individual antigen-negative cells. Though such escape variants have been demonstrated to exist in some melanoma peptide vaccine trials...

Clinical trials of chaperone protein vaccines

Antigenics Inc. (New York, NY) has developed a clinical GRP94 gp96 vaccine termed HSPPC-96, or Oncophage (102). Data generated from clinical trials evaluating HSPPC-96 Oncophage against a variety of cancers have been reported at recent American Society of Clinical Oncology (ASCO) and American Association of Cancer Research (AACR) conferences. The major limiting factor for producing Oncophage from a patient's tumor is the ability to purify enough GRP94 gp96 from the autologous tissue. Oncophage is manufactured in a 10-h process from autologous tumors. The minimum tissue required to produce enough vaccine for a course of treatment is between 1 and 3 g, and patients have been immunized either subcutaneously or intradermally weekly for 4 wk and then every two to four weeks for another three to five vaccinations. In the reported trials, HSPPC-96 vaccine consisted of 2.5, 5, 10, 25, 50, or 100 g of purified GRP94 gp96. Generally, adequate protein was purified to complete the vaccination...

The Role Of Il11 In Tumour Metastasis

Human melanoma (A375M) and human breast cancer (MDA-MB-231) cells formed osteolytic bone metastasis in vivo when these tumour cells were injected into the left ventricles of BALB c nude mice (Figure 2) (17). These tumour cells promoted bone resorption in the in vitro neonatal murine calvaria organ culture system by indirectly stimulating the production of a bone resorption-inducing factor (or factors) from human osteoblast-like Saos-2 cells. In the course of examining the factors, we found that Saos-2 cells, in response to the interaction with tumour cells, produced a factor that promoted the proliferation of T10 cells. Because the T10 cell growth was IL-6 and IL-11 dependent (24), we checked the concentrations of both in the Saos-2-conditioned medium. We found that this conditioned medium stimulated the production of IL-11, but not IL-6, from Saos-2 cells. Furthermore, we found that a specific anti-IL-11, but not anti-IL-6, antibody could neutralize the abilities to induce T10...

Gene Replacement Therapy

The protein product of the mda-7 gene, Mda-7 interleukin (IL)-24, is a novel cytokine that belongs to the IL-10 family of cytokines (40). Gene delivery using Ad-mda-7 results in growth suppression and apoptosis in a broad-spectrum of cancer cell types including those of the lung, prostate, mesothelioma, pancreatic, breast, gliomas, renal, and human melanoma (41-48). In contrast, Ad-mda-7 does not elicit deleterious effects in normal cells, including those of epithelial, fibroblast, astrocyte, melanocyte, or endothelial origin (49). Based on these distinctive properties and reports of antitumor and antiangiogenic activities in human tumor xenograft animal models, a The cyclin-dependent kinase inhibitor (CDK-I), p16, was identified in a yeast two-hybrid screen as a protein that inhibits the ability of cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6) to regulate the phosphorylation status of phos-pho retinoblastoma (pRb), thereby controlling progression of cells into...

In Vivo Evidence for DCs As Strong Adjuvants

There is a large body of literature involving animal models of tumor immunity in which DCs loaded with tumor-associated antigens (TAAs) are able to induce protective antitumor responses. When tested, DCs can be superior to other vaccination strategies (24). There also are reports in which DC immunization produces significant therapeutic immunity to established tumors (reviewed in ref. 25). A number of trials have now utilized TAA-loaded DCs as vaccines in humans. Initial pioneering studies involved patients with lymphoma and melanoma (26,27). Some clinical and immune responses (T-cell proliferation and delayed-type hypersensitivity DTH ) without any major toxicity have been observed. More recent DC vaccination studies put further emphasis on the elicited immune responses and have included control antigens for CD4+ and CD8+ T-cell responses (28-31). Several of these studies are reviewed elsewhere in this handbook. Inclusion of control antigens helps to verify that the DCs are...

Assay Techniques 41 Identification of VSMCs

It may also be appropriate to confirm that cultures are von Willebrand factor negative if there is concern about endothelial cell contamination (for example, when isolating VSMCs from small blood vessels). Separate markers have also been described for microvascular SMCs (pericytes) such as 3G5 43 , but there is debate about the specificity of this marker 44 . The high molecular weight melanoma-associated antigen is also a marker for pericytes in vivo 28 .

Cancer Surveillance And Prevention

Unfortunately there are no defined guidelines for screening or prevention for most LFS cancers. The most frequent tumors would be soft tissue sarcoma, osteosarcoma and brain tumors, none of which lend themselves to presymptomatic screening. New imaging techniques or serum proteomic discriminants are clearly needed. Given the lack of specificity for a distinct cancer type within a family, with the possible exception of adrenal cortical tumors, it is difficult to recommend aggressive screening. Patient education regarding symptoms, regular physical examinations with a physician aware of LFS risk and tumor spectrum, and application of noninvasive screening for melanoma and perhaps colon cancer (91) at an early age seem reasonable.

Management of the Unaffected Carrier

Carriers of BRCA2 mutations are also at risk of developing melanoma. Patients who have a family history of melanoma should be examined annually and advised to report any suspicious skin lesions. Those with a previous personal history of melanoma should be followed up regularly by a dermatologist.

Alkylating Agent Resistance

The use of MGMT point mutants to differentially protect the hematopoietic compartment while sensitizing tumors has also been reported in animal xenograft models (126-128). Clinical trials using gene transfer of MGMT have been proposed by several investigators, and one phase I trial in patients with advanced malignancies such as melanoma, sarcoma and other solid tumors is in progress (129). The objective of this trial is to protect bone marrow stem cells from the toxic effects of chemotherapy and select for MGMT-G156A transduced cells during treatment. This strategy is expected to result in less toxicity to bone marrow and blood cells while enriching for the number

Table 231 Different characteristics of benign and malignant tumours

Malignant disease accounts for 1 in 8 deaths of people under 35 years in Australia and 1 in every 4 (25 ) of deaths in those over 45 years. 1 Cancer is the only major cause of death in Australia that is increasing in both sexes. At current rates about 1 in 3 males and 1 in 4 females will develop a cancer, excluding non-melanoma skin cancers, by the age of 75. 1 Neoplasia, especially malignancy of the silent areas, can present as undifferentiated illness and be a real masquerade. The so-called 'silent' malignancies that pose a special problem include carcinoma of the ovary, kidney, caecum and ascending colon, liver (hepatoma), melanoma and haematological tissue.

Genetic Vaccines for Cancer Therapy

In patients with compromised immunity resulting from the progression of cancer and chemotherapy. Furthermore, the common requirement of multiple vaccinations can be problematic using viral vectors. Recently, it was shown that restimulation with adeno-virally transduced DCs actually decreased the antigen-specific immune response in favor of strong antiadenovirus specific immune reactions in melanoma patients (18). Pre-existing immunity can also inhibit delivery using viral vectors. Attenuated bacteria has also shown promise for vaccine delivery, but are associated with safety concerns as well (19).

Methods Of Dna Vaccine Delivery 31 Electroporation

Studies on the delivery of TAA antigens using electroporation have yielded promising results. Mendiratta et al. reported vaccination using electroporation with both plasmid encoded human GP100 and mouse TPR2 antigen elicited complete protection from melanoma challenge (21). Lohr et al. demonstrated that introduction of plasmid encoding IL-2 and IL-12 (inflammatory cytokine signals) by electroporation at tumor sites caused transduction and inhibition of murine melanoma without the systemic cytokine levels experienced after adenoviral gene transfer (22). Further investigations have shown that electroporation can be used to facilitate the discovery of novel antigen encoded plasmid constructs. For example, Kalat et al. used electroporation methods to optimize tyrosinase related protein-2 antigens to elicit CD8+ responses and inhibition of melanoma growth in two challenge models (23). This same group later demonstrated that electroporation was capable of inducing immune responses...

Cytokines for Cancer Therapy

Cytokines such as interferons and interleukins are administered for cancer because of their broad-based immunostimulatory effects including generation of tumor-reactive lymphocytes (11). Interleukin-2 (IL-2), or aldesleukin, which is indicated for the treatment of adults with metastatic renal cell carcinoma and melanoma, is the most studied cytokine. IL-2 dose-dependently activates cellular immunity and causes release of other immune-boosting cytokines in vivo (11). Systemic cytokine therapy is generally limited by rapid degradation and elimination of the cytokine, the inability to achieve optimal concentrations in the tumor microenvironment, and dose-dependent toxicity, including life-threatening side effects such as vascular leak syndrome and orthostatic hypotension (12-15). Cytokine gene therapy, in which a cytokine gene (such as that for IL-2) is introduced into tumor cells, is being explored to overcome some of these limitations of systemic cytokine administration. However,...

AntiGD2IL2 Immunocytokine

The anti-GD2 immunocytokine hu14.18-IL-2 (EMD 273063) is currently in phase I clinical trials (18). This immunocytokine consists of recombinant human IL-2 and a humanized monoclonal antibody directed against human GD2, which is a disialogang-lioside antigen expressed by tumors of neuroectodermal origin including malignant melanoma, neuroblastoma, some sarcomas, and some small-cell lung carcinomas. Each immunocytokine molecule is composed of two cytokine molecules fused to each of the two heavy chains of a monoclonal antibody via a peptide linkage at the carboxyl terminus (Fig. 2). Preclinical studies with the anti-GD2-IL-2 fusion protein were conducted with a chimeric form (i.e., ch14.18-IL-2) as well as the humanized form that is EMD 273063 (i.e., hu14.18-IL-2). Early studies focused on establishing the biological functionality of the immunocytokine and on determining whether the fused protein retained full antibody and cytokine functions. That ch14.18-IL-2 retains its functionality...

Virus Induced Changes In Mhc Class I Presentation Of Hostderived Peptides

Heat shock protein production is amplified during a variety of cellular stresses including viral infection. Therefore, an increase in intracellular HSP concentration may lead to increased MHC class I presentation of HSP-derived peptides. Infection of immortalized human T cells expressing HLA B*0702 with HIV increases intracel-lular levels of Hsp27. These increased intracellular levels of Hsp27 lead to increased presentation of the Hsp27-derived peptide APAYSRAL by the HLA B*0702 class I molecule (Hickman et al., 2003). The Hsp90-derived HLA B*1801 ligand EEVETFAF has also been observed following HIV infection (H.D. Hickman-Miller and W.H. Hildebrand, unpublished). Increased class I presentation of HSP-derived peptides is not limited to cells stressed through HIV infection. Hsp90 production is increased in HLA A*0201 homozygous cells following Epstein-Barr virus transformation while the class I HLA of melanoma cell line MelJuSo also present Hsp90p peptides when infected with measles...

Increasing Gene Expression

Replicons have proven to be powerful enhancements to DNA vaccination, and are capable of eliciting antibody and tumor protective responses at up to 1000 times lower titers than conventional naked DNA vaccines in a P-gal expressing tumor model (132). Vaccination with replicons has also induced protective immunity to melanoma challenge in a TRP-1 expression system, unlike conventional DNA vaccines (133). Although it is

Tumor Induced Changes In Mhc Class I Presentation Of Hostderived Peptides

Alterations in gene expression can induce changes in peptide presentation, including the expression of germ cell genes, often referred to as cancer-testis (CT) genes. Normally, these genes are expressed in the trophoblast and in testicular germ cells, which do not express MHC class I (Van Der Bruggen et al., 2002). Peptides novel to the mature immune system are presented on the MHC class I upon processing of the protein products of these genes. The function of most CT gene products is unknown but they are putatively involved in spermatogenesis. Their activation is likely due to genome wide demethylation seen in many tumors (Loriot et al., 2006). This extensive demethylation may be due to the loss or downregulation of DNA methyltransferases, as a result of transcriptional disregulation or mutation. These events open up the promoter regions of numerous genes that are normally transcriptionally repressed. The MAGE, BAGE, and GAGE families of CT genes were first identified in melanoma but...

Clinical Manifestations Primary Lesion

A primary lesion is the most common manifestation of malignant melanoma of the head and neck region. This lesion can vary in appearance from the classical black-pigmented, raised lesion to an enlarging, skin-colored (amelanotic) mole (Fig. 1A and B). Melanoma can arise from a preexisting nevus or normal skin. The appearance of melanoma may be FIGURE 1 Primary melanoma has a wide range of appearances. (A) Superficial-spreading melanoma. (B) Amelanotic nodular melanoma of the scalp. (C) Lentigo maligna melanoma of the cheek. (D) Advanced cutaneous melanoma with satellite lesions of the scalp. Source Courtesy of Dr. Mark Faries. FIGURE 1 Primary melanoma has a wide range of appearances. (A) Superficial-spreading melanoma. (B) Amelanotic nodular melanoma of the scalp. (C) Lentigo maligna melanoma of the cheek. (D) Advanced cutaneous melanoma with satellite lesions of the scalp. Source Courtesy of Dr. Mark Faries. similar to other generally noninvasive skin cancers such as basal cell and...

Biological effects of exosomes

In addition to transferring preprocessed antigens in MHC molecules, exosomes might also transfer hsp-associated peptides or cytosolic whole candidate tumor antigens. We could demonstrate that melanoma cells release exosomes containing whole tumor proteins such as MART-1 melanA. Following DC uptake of such melanoma-derived exosomes, cross-presentation of MART-1 peptides by DC MHC class I molecules could be observed. In vivo, DCs pulsed with texosomes induced potent CD8+ T-cell-dependent antitumor effects against established mouse tumors (6). We recently reported that high amounts of tumor exosomes accumulate in the malignant effusions of patients bearing different types of tumors (melanoma, breast, lung, ovarian cancer, mesothelioma) (4). Exosomes harvested from ascitis of melanoma patients efficiently transport MART-1 tumor antigen to monocyte-derived dendritic cells (MD-DCs) for cross-presentation to MART-1-specific CTL clones. Moreover, in in vitro stimulation assays aimed at...

Cloning With Cancer Cells

Cloning with cancer cells recently has been reported. Hochedlinger et al. (2004) reported that cloning with several types of cancer cells (leukemia, lymphoma, breast cancer) led to the formation of blastocysts, but not to development of inner cell masses (ICMs), the early progenitor of the embryo proper and ESCs. However, ESCs were established from clones made with nuclei from a RAS-inducible melanoma cell line, and these were subsequently used to produce chimeric mice using the tetraploid complementation method. The chimeric mice were predisposed to cancer, with an expanded spectrum of tumorigenesis. This result reveals that, although developmental pluripotency can be conferred upon cancer cell nuclei by the cloning method, the genetic factors that originally induced carcinogenesis predispose the clone to develop tumors of the same type as the donor, as well as other cell types that can respond to that genetic defect.

Loco Regional Disease Nodal Assessment

After initial pathological assessment by punch or excisional biopsy, the next step is treatment of the primary site and staging of the regional lymph nodes. Treatment for the primary tumor consists of wide local excision. Local recurrence can be as high as 40 for primary melanomas that are not reexcised after biopsy (3). Melanoma-in-situ is treated by reexcision with at least 0.5-cm margins around the primary lesion or biopsy scar. When primary invasive melanomas are < 1 mm thick or Clark's level II, adequate margins are 1 cm circumferentially. Melanomas that are 1.0 to 4 mm thick or extend to Clark's level III IV are treated with 2-cm margins circumferentially. Because of the increased risk of microsatellites, melanomas thicker than 4 mm or extending to Clark's level V should be excised with 2- to 3-cm margins that include the underlying fascia (3). Wound closure is achieved primarily with local flaps or more rarely with skin grafting. Some primary sites have special...

Complications And Prognosis

The most important prognostic factors are primary tumor thickness, ulceration, and lymph node status. This is reflective of the AJCC TNM staging for melanoma (Table 1), where advancing stage corresponds with worsening prognosis (7). After biopsy and SNB, most patients can be given general prognostic counseling.

Nucleotide Excision Repair

Marked sensitivity to sunlight (ultraviolet) with subsequent formation of multiple skin cancers and premature death. The risk of developing skin cancer is increased 1000- to 2000-fold. The inherited defect seems to involve the repair of damaged DNA, particularly thymine dimers. Cells cultured from patients with xero-derma pigmentosum exhibit low activity for the nu-cleotide excision-repair process. Seven complementation groups have been identified using hybrid cell analyses, so at least seven gene products (XPA-XPG) are involved. Two of these (XPA and XPC) are involved in recognition and excision. XPB and XPD are helicases and, interestingly, are subunits of the transcription factor TFIIH (see Chapter 37).

Dongchul Kang Zaozhong Su Habib Boukerche and Paul B Fisher

Expression occurring during induction of terminal differentiation in human melanoma cells treated with interferon-beta (IFN-P) plus mezerein (MEZ) (Figure 5). In RaSH, cDNA samples are restriction-digested with frequent cutters such as DpnII to an average size of 256 bp to improve hybridization efficiency. Moreover, restriction digestion of tester cDNA only prior to hybridization enables selection and cloning of tester-unique cDNAs by direct ligation of hybridization mixtures to corresponding sites in the plasmid vector. Thus, RaSH reduces the amount of starting materials required for conventional subtraction hybridization by one tenth by using PCR and obviates the need to separate and clone subtracted cDNA species into plasmid vector or bacteriophage, a nontrivial, technically demanding step in subtractive cDNA library construction (Jiang and Fisher, 1993 Sagerstrom et al., 1997). In addition, RaSH uses reverse Northern Blot hybridization for further rapid confirmation of...

Lymphokine Activated Killer LAK Cells

LAK cells are derived from peripheral blood lymphocytes cultured in the presence of high concentrations of IL-2 for 3-4 d. They demonstrate preferential lysis of tumor cells but not normal cells. Promising data in animal tumor models and preclinical in vitro testing raised the prospect of a readily available source of autologous cells with broad reactivity (12,53,54). Analysis of the relevant precursors of LAK cells indicated a natural killer (NK) origin and the treatment effect was also independent of host T cells (55-57). The first clinical studies of LAK cells were performed at the National Cancer Institute (NCI) surgery branch and used total cell doses ranging from 1.8 to 18.4 x 1010 cells combined with bolus high-dose IL-2 (720,000 U kg). This study showed the greatest efficacy for patients with malignant melanoma or renal cell carcinoma with a 44 overall response rate (58). Based on this promising initial study, several phase II clinical trials of LAK cells were performed in the...

Peripheral Blood Lymphocytes PBLs

Antigen-experienced T cells with a memory phenotype preferentially circulate in the peripheral blood. One of the theoretical advantages of using PBLs is that they are readily accessible and can be safely harvested in large quantities through standard leukapheresis procedures. Several studies tested the clinical efficacy of PBL activated with anti-CD3 and IL-2 termed autolymphocyte therapy (ALT) in patients with metastatic RCC (72,73) or metastatic melanoma (74). Simultaneous CD3 and CD28 stimulation by mAb coupled to magnetic beads in the presence of IL-2 (100-400 IU mL) has also been tested on PBL from cancer patients resulting in 10- to 15-fold numerical expansion after 14 d of culture (75). The PBL stimulated with anti-CD3 alone had equivalent to slightly improved effector function in cytolytic and cytokine release assays compared with combined anti-CD3 anti-CD28 stimulation. Other studies also raise the issue of whether costimulation through CD28 is required to generate superior...

Targets for Antiangiogenic Antilymphangiogenic Gene Therapy 321 Inhibition of Proangiogenic Lymphangiogenic Growth

Simultaneously targeting VEGF production with antisense oligonucleotide and VEGF receptor signaling with receptor tyrosine kinase inhibitors enhances the anticancer efficacy of either therapy alone (96). Some focus has been given to the preclinical gene therapy with soluble truncated forms of VEGFR-2 (sVEGFR-2). This molecule functions in two ways by sequestering VEGF and, in a dominant-negative fashion, by forming inactive heterodimers with membrane-spanning VEGFRs (91,92). Recently, murine fibrosarcoma and melanoma cells transduced with a retrovirus expressing sVEGFR-2, showed tumor growth reduction by inhibition of angiogenesis however, it did not show

Tumor Infiltrating Lymphocytes

Of TIL therapy has been in patients with melanoma or renal cell carcinoma where unequivocal durable responses have been documented in patients with progressive bulky metastatic disease that was previously unresponsive to IL-2 alone. One consideration for the clinical application of TIL therapy is that it requires considerable expertise and effort to successfully obtain therapeutic doses of TIL. Even the most experienced investigators are not uniformly successful. In a large series from the surgery branch at NCI describing the preparation of TIL from melanoma, renal, breast, or colon carcinoma, therapeutic numbers of greater than 1011 cells were successfully grown from 45 of the samples (83). In the initial methodology, TILs were derived from tumor biopsies that were enzymatically disaggregated and cultured in the presence of high concentrations of IL-2 (7200 IU mL) and 10 autologous LAK cell culture supernatant. Among a variety of tumors, melanoma yielded the highest percentage of...

Systemic and Local Suppression of T Cells

The indirect influence of tumors on the development of a systemic immunosuppresion can be attributed to hyperproduction of IL-10, oxygen metabolism intermediates, and some enzymes. IL-10 is a type 2 cytokine that is produced by APCs and Th2 cells. It is involved in the development of T-cell anergy, promotion of Th2 responses, and inhibition of Th1 responses, which are important for the generation of efficient antitumor responses. Increased production of oxygen metabolites by macrophages isolated from the metastatic lymph nodes of patients with malignant melanoma was found to be responsible for decreased CD3-mediated stimulation of T cells and the reduction of CTL and NK cell activity (22). The inhibitory effect of macrophages on melanoma-specific CTL lines and NK cells was abrogated in the presence of catalase, a scavenger for H2O2. The mechanism of H2O2-induced immunosuppression by monocytes macrophages derived from the blood of cancer patients was attributed to the inhibition of Th1...

Vaccine Draining LN T Cells

In an initial study of VDLN cells, subjects with metastatic melanoma or RCC were treated. Irradiated autologous tumor cells admixed with bacille Calmette-Guerin (BCG) was used as a vaccine and 7 d later draining LNs were removed. The T cells were stimulated with immobilized anti-CD3 (OKT3) for 2 d followed by culture in IL-2 to reach a mean treatment dose of 8.4 x 1010 cells. These activation conditions generated a mixture of CD4 and CD8 that demonstrated specific release of GM-CSF and IFN-y against autologous tumor targets. Eleven melanoma patients and 12 RCC patients were treated with T-cell infusion and concomitant IL-2. One patient with melanoma experienced a partial response (PR) two patients with RCC had a CR and two had a PR. Immune monitoring studies indicated a trend correlating development of a positive delayed-type hypersensitivity (DTH) response to autologous tumor with clinical response (100).

Tumor Targeting 411 Transcriptional Targeting

Transcriptional targeting can be achieved by the use of an expression cassette which is activated by tissue specific promoters (TSPs) (167). The options are to use either a promoter with a high activity in tumor cells tumor ECs or to use inducible promoters to achieve therapeutic transgene expression. Examples of promoters with high tumor-selective activity (minimal expression in normal cells) are, CXCR4, cyclooxygenase-2 (COX-2), survivin (a member of the inhibitor of apoptosis protein family) and pre-proendothelin-1 (PPE-1) a precursor protein for endothelin-1. The human CXCR4 gene is expressed at high levels in many types of cancers, but is repressed in the liver. Thus, the CXCR4 promoter has a tumor-on and liver-off status in vitro and in vivo, which make it a good candidate TSP for targeted cancer gene therapy approaches, (i.e., for melanoma and breast cancers) (168). COX-2, a key enzyme in the synthesis of prostaglandins and thromboxanes, is highly up-regulated in tumor cells,...

Roberta P Glick Terry Lichtor Henry Lin and Edward P Cohen

The prognosis for patients with malignant glioma is poor. Conventional treatments such as surgery, radiation therapy, and chemotherapy have done little to affect long-term survival, and new methods of treatment are urgently needed. In this report, approaches involving cytokine gene therapy in treatment of malignant brain tumors are reviewed and contrasted with a strategy developed in this laboratory involving the use of allogeneic cells which have been genetically modified to secrete cytokines. In our studies, mice with an intracerebral glioma, melanoma, or breast carcinoma treated solely by intratumoral injections with allogeneic cells genetically modified to secrete interleukin-2 were found to survive significantly longer than mice in various control groups. The anti-tumor response was mediated predominantly by T-cell subsets (CD8+ and NK LAK cells). The injections resulted in the killing of only the neoplastic cells non-neoplastic cells were unaffected. Experiments involving...

Preliminary Evaluation Of Fnab Smears

Markedly cellular Favor malignancy Lymphoma Carcinoma Melanoma Sarcoma (rare) Moderately cellular Malignant (low cell yield) Lymphoma Carcinoma Melanoma Sarcoma (rare) Reactive Pediatric small round blue cell tumors Melanoma Sarcoma (rare) Malignant Lymphoma Carcinoma Melanoma Sarcoma (rare)

Adjunctive therapy with lysates in resected stage ii and iii disease

In 1990 and 1991, we attempted to study Melacine with the Southwest Oncology Group in a phase III randomized trial vs no treatment in resected stage III melanoma, to see whether relapse-free and overall survival could be improved. This was at a time when the ECOG 1684 study with IFN-a-2b (INTRON-A) had not yet been completed. When the SWOG joined the Intergroup IFN-a study for stage III and deeply invasive (> 4 mm) melanoma, retaining Melacine only for resected intermediate thickness (> 1.5-4.0 mm) stage II disease (SWOG 9085), our plans had to be revised. To obtain data that might support a subsequent randomized study of our own, we treated 44 consecutive patients with resected stage III melanoma from January 1992 to December 1993 over a period of 48 wk with frozen lysates + DETOX (first 17 patients) or Melacine (final 27 patients), preceded by 350 mg m2 of cyclophosphamide. An additional 23 patients referred to us with resected stage II melanoma were also treated in a separate...

Association of Specific HLA Class I Molecules With Response to Melacine

If cytolytic T cells were important for achieving a clinical response, we speculated that responding patients might have an overrepresentation of certain specific HLA class I molecules known to restrict CTL responses (20). Seventy patients with metastatic melanoma were studied first. Those patients who had HLA-A2, or the closely related HLA-A28 (now called HLA-A68), HLA-B12 split (including HLA-B44 and HLA-B45) or HLA-C3 had a far greater likelihood of responding to Melacine than those who lacked those alleles. Patients with all three alleles had a 40 response rate, whereas those who lacked all three had nearly 0 response. The group as a whole had a 20 response rate. In a confirmation and extension of these data, Sosman and colleagues (21), analyzing a Southwest Oncology Group study of 689 patients with resected stage II melanoma, found that those with HLA-A2 or HLA-C3 had a longer disease-free survival than those with other HLA phenotypes. Among patients who matched at least two of...

Obtaining Family History Information

Additionally, the pedigree should include information on all surgical procedures in affected and unaffected family members such as bilateral oophorectomy performed either for prophylaxis or for a benign condition or removal of skin lesions. Such surgeries may impact that individuals risk for cancer (as would be the case with oophorectomy) or may indicate another possible unsuspected cancer in the pedigree, such as a melanoma. An example of a cancer pedigree is presented in Figure 1 and important details of the family history are listed in Figure 2.

Immunohistological Studies

We also studied the immunohistology of lesions that were undergoing rejection after vaccine therapy in a group of seven patients, compared with a group of six untreated melanoma patients, and other control subjects, including a patient who had a DETOX-induced granuloma at the site of injection, which allowed us to compare the rejection site Fig. 3. Immunohistology of a melanoma lesion undergoing rejection after treatment with allogeneic lysates after staining with specific monoclonal antibodies to cluster determinants. (A) CD8+ T cells, (B) CD4+ T cells, (C) Macrophages, (D) HLA class I antigens (w6 32), (E) HLA class II antigens. Note the predominance of CD4+ T cells over CD8+ T cells in the lesion, the peripheral and perivascular distribution of T cells in general, and the predominance of macrophages over all. HLA class I molecules (by beta-2 microglobulin staining) were present in all 11 lesions examined from six patients, whereas HLA class II molecules were absent from the tumor...

Other clinical trials with allogeneic lysates

Other investigators have also concluded that allogeneic materials are potentially of greater benefit to patients than autologous tumor-derived immunogens. Among the earliest of these studies were those of Cassel and associates, who utilized viral oncolysates of melanoma cell lines as immunogens (24). These studies were extremely promising, with a number of remissions lasting a decade or more, but unfortunately may have been ahead of their time. Attempts to reproduce the results in large-scale cooperative trials were unsuccessful, but those were hampered by failure of attention to important details, such as viability of the cells before viral treatment. Wallack immunized patients with allogeneic viral oncolysates created with vaccinia virus, and did one of the first double-blind, randomized, multicenter studies in 250 patients perhaps too small a number to show a small difference between the groups with resected stage III melanoma. Protein representing 5 million melanoma cells in...

Conclusions and future

Our personal experience with allogeneic melanoma lysate vaccines, given to several hundred patients, has convinced us that although the response rates in disseminated disease are at most in the range of 15-20 , there is indeed activity of these preparations even when billions of tumor cells are present. There have been a number of patients with a gratifying dramatic increase in survival lasting 5-10 yr or more. Allogeneic lysates may perhaps be effective in preventing recurrence of melanoma in patients with resected stage III disease, and have been equivalent to toxic four-drug chemotherapy even when given by a suboptimal route of administration in a large multicenter trial. With the same caveat, suboptimal (im) route of administration, a large Southwest Oncology Group phase II study of Melacine vs observation in resected intermediate thickness stage II melanoma was initially positive at a follow-up of 1 yr (p 0.04), although further follow-up showed no difference. Melacine was...

Production and purification of exosomes

II concentration of the starting clarified supernatant. This methodology was extended to a miniscale process with comparable results. Likewise, the classical sedimentation technique is a tedious and less productive process carrying over along with exosomes contaminants of the culture medium. Furthermore, the development of quality control assays allowed qualitative and quantitative standardization of exosome preparations. Immunocapture assays assessing exosomal contents in MHC class I and II molecules as well as FACS determination of exosomal protein patterns after coupling onto macroscopic beads are currently used to calibrate exosome dosages in the first phase I trial (24). Exosomes are secreted from monocyte-differentiating DCs from Day 5 to Day 6 using culture conditions devoid of maturing agents (granulocyte-macrophage colony-stimulating factor GM-CSF interleukin-4 IL-4 ). The amounts of exosomal MHC class I molecules recovered in a 24-h DC culture supernatant using such a...

Other uses of rcc vaccines

Gene-modified tumors have also been used as the lymph node-activating vaccine. For example, investigators primed subjects with autologous tumor cells modified ex vivo to express HLA-B7 and P2-microglobulin (48). Lymphocytes from lymph nodes draining the vaccination site were obtained and expanded ex vivo with anti-CD3 and IL-2, and adoptively transferred to 9 patients with melanoma and 11 patients with metastatic RCC. In this phase I II study no objective responses were observed.