Vaccines for Chronic Myeloid Leukemia CML

CML is characterized by a t(9 22) translocation that results in the expression of chimeric bcr abl fusion oncoproteins necessary for oncogenesis. Like AML cells, CML-derived DCs show potential as tools for therapy. Leukemia cells of patients with CML will undergo substantial differentiation toward DCs and may be used to drive autologous T cells to acquire antileukemic cytotoxicity (for a review, see ref. 126). Early studies showed that both CD34+ bone marrow cells (127) and peripheral blood...

Correlation Between Antibody Induction and Tumor Growth Inhibition Increased Survival in Vaccinated Mice

Several studies in mice have demonstrated a correlation between antibody induction and tumor growth inhibition in vaccinated mice. Early studies have shown a correlation between antibody formation to Moloney sarcoma virus and regression of virus-induced sarcomas (45). Tumor growth inhibition was mediated by ADCC. Similarly, cytotoxic antibodies to feline sarcoma virus have been implicated in tumor regression of virus-induced sarcomas in dogs (46). Anti-idiotypic antibodies binding to the...

Exosomes and their potential application in immunotherapy

Active immunotherapy with mature DCs is being extensively tested for treatment of malignancies. DC-based immunotherapy, however, remains difficult to handle for scale up, definition of quality control parameters, and long-term storage. Indeed, many investigators have shown that one leukopheresis enables up to 4-10 DC injections, whereas there is evidence that a larger number of injections, perhaps spanning over a year or more, appear to be required for long-lasting antitumor protections....

Critical developments for adoptive immunotherapy using autologous t cells

Clinical applications of adoptive immunotherapy will likely be most effective and least toxic if they use an autologous source of tumor-reactive T cells. T cells recognize tumor antigens in the form of peptide fragments of proteins presented by MHC molecules. Thus, each tumor antigen that is targeted must be matched for the relevant MHC molecule. Additionally, humans have a large number of alleles in the HLA gene complex and a mismatch of donor and recipient at either class I or class II loci...

Clinical trials of chaperone protein vaccines

In the first published study, Janetzki et al. (101) reported on 16 patients with various advanced malignancies, vaccinated with GRP94 gp96 that was prepared by the Srivastava group's purification procedure. Significant toxicities or autoimmune reactions were not observed. Immune responses were evaluated using ELISPOT assays of peripheral blood T cells. Immunization with autologous GRP94 gp96 elicited MHC class I-restricted, tumor-specific CD8+ T cells in 6 of the 12 immunized patients....

Antiid antibodies and human cancer

Carcinoembryonic Antigen (CEA) A number of anti-Id antibodies (Table 1) have been generated that mimic CEA (1523). CEA is one of the first TAAs to be identified and is one of the most widely investigated human TAAs. CEA is a 180 kD glycoprotein and was originally detected in colonic carcinoma and fetal gut. However, it has since been detected at high density in 95 of colorectal carcinoma, 70 of lung adenocarcinoma, and 50 of breast cancer. In order to generate the...

Tumor antigens expressed by lung cancer

Numerous tumor antigens have been described for lung cancers. Among the most frequently expressed are the MAGE antigens with MAGE-1 reported in 11-36 , MAGE-2 in 30 , MAGE-3 in 38 -60 , and MAGE-4 in 13 of NSCLC (28-31). Using tissue microarray technology, expression of tumor-associated antigens (TAAs) of the MAGE family have been reported in 50 of squamous cell carcinomas of the lung and in 38 of large-cell carcinomas of the lung (32). In another study, MAGE-A10 was expressed only by SCLC, and...

Ganglioside Vaccines

We have been refining our ability to induce antibodies against GM2 in melanoma patients for 15 yr, since it was first demonstrated that patients immunized with irradiated melanoma cells occasionally produced antibodies against GM2, and that vaccines containing purified GM2 could be more immunogenic than vaccines containing tumor cells expressing GM2 (72). Initially GM2 adherent to bacille Calmette-Guerin (BCG) was selected as optimal, inducing immunoglobulin M (IgM) antibody responses in 85 of...

Adjuvants for tumor vaccines

Regardless of the model for T-cell activation, because tumors express self molecules, to which the immune system may be tolerant, and because the tumor environment may contain immunosuppressive molecules such as interleukin-10 (IL-10) and TGF-P, additional danger signals will be needed to properly activate the immune response. The most frequent manipulation to tumor cell vaccines is irradiation to render them unable to propagate in the recipient of the vaccine. Although irradiation causes...

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Expression of EphA3 in normal tissues appeared to be confined to tissues that do not constitutively express HLA class II molecules (70). Expression of EphA3 in normal tissues appeared to be confined to tissues that do not constitutively express HLA class II molecules (70). single point mutation resulting in the substitution of a leucine for a serine residue was also identified as the target of HLA-DRp1*0401-restricted, tumor-reactive CD4+ TIL (50). Transfectants expressing the normal CDC27...

Plasmids With Genes for Fusion Proteins

Another approach to multifunctional DNA vaccines is based on covalent linking of two or more proteins (Fig. 2C). A number of DNA vaccines containing genes encoding fusion proteins have been generated. The array of possible applications of such vaccines ranges from inhibition of allergic reactions (87), through suppression of autoimmunity (88,89), to boosting vaccine-induced immunity (90). Here we present only a few out of many promising applications of this technology. The access of antigens to...

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....

Active tumormediated immunosuppression mechanisms of tgfP action

As discussed above, secretion of TGF-P by tumors and the surrounding stroma cells is perhaps the most widely used method by which tumors actively suppress the immune system. The immunosuppressive effects of TGF-P have been demonstrated in vitro and in vivo, with exogenous TGF-P 1 producing immunosuppression in animal models (14). How does TGF-P mediate its immunosuppressive effects Although TGF-P has diverse effects on virtually every cell type in the immune system, for the purposes of this...

Initial Clinical Trials With Frozen Lysates

In our phase I and II trials (comprising 19 and 25 patients, respectively) with frozen lysates, lysates and DETOX adjuvant were given sc. Nine of the 44 patients (5 of 19 and 4 of 25, respectively) (20.5 ) with measurable metastatic disease had major responses, 3 complete response (CR) (6.8 ), and 6 partial response (PR) (13.6 ). One additional patient in the phase II trial (2.3 ) had 17 mo stability of lung nodules developing brain metastases at that juncture which we felt was also a response...

Advantages of heterologous primeboost immunizations

Heterologous prime-boost immunizations have the following advantages over repeated immunizations with the same vector. First, vector specific immune responses are not boosted (see Fig. 1). Second, the best immunostimulatory properties of each strategy may be combined. Third, any toxicities of a vector are limited by the lower frequency with which any one vector (particularly those that induce significant inflammation) need be administered. Regarding vector-specific immunity, antivector immunity...

Vaccine Draining LN T Cells

LNs draining a site of vaccine inoculation are an enriched source of T lymphocytes that have been sensitized to tumor antigens. Preclinical kinetic studies have clearly demonstrated that the optimal number of preeffector T cells is present between 6 and 14 d with subsequent waning of activity. These findings recommend a strategy of placing a tumor vaccine in a region that is uninvolved with tumor so that newly sensitized T cells can be obtained by removal of the vaccine draining LNs at the peak...

Replication Defective Pox Viral Vectors

The avipox viruses represent potentially attractive vectors for use in cancer vaccines. Though the immunogenicity of the inserted transgene may not be as potent as a transgene in a vaccinia virus, avipox viruses such as fowlpox and ALVAC can be administered numerous times to enhance immunogenicity (26-28). Since they are replication defective, induction of any host immune responses should be relatively inconsequential. Avipox viruses are also distinguished from vaccinia in that the inserted...

Minigenes Based Polyepitope Vaccines

Revealing the molecular mechanism of MHC peptide recognition by aP T-cell receptors (TCRs) (104) allowed the identification of a large number of immunogenic epitopes (105). Synthetic peptide combinatorial libraries proved to be a powerful tool for screening millions of peptide candidates, thus greatly speeding up discovery of new immunogenic epitopes (106), with immediate consequences for the design of vaccines. Numerous immunogenic epitopes have now been identified in many potential targets of...

Role of primeboost for polyepitope vaccines

Polyvalent vaccines are being developed to induce broad CTL responses against multiple tumor antigens and epitopes restricted by more than one major histocompatibility complex (MHC) class I molecule. Poly-epitope vaccines aiming to induce such polyvalent responses have been tested in combination with the heterologous prime-boost approach. Instead of engineering single epitopes or antigens into heterologous vectors, poly-epitope and or poly-protein expressing constructs have been used to drive...

Molecular Mechanisms of DC Dysfunction in Cancer

Cell differentiation is a complex process, which involves multiple genes controlled by multiple transcription factors. One of the transcription factors, NF-kB, is especially important for DC biology. NF-kB regulates the transcription of many genes involved in immune responses (reviewed in refs. 121 and 122). NF-kB is composed of 50- and 65-kDa subunits (p50, p52, p65 RelA , cRel, and RelB), which are bound to a 10-bp motif in the promoter of responsive genes. These subunits form both homo- and...

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...

Replication Competent Pox Viral Vectors

One of the most thoroughly investigated pox viral vectors is vaccinia. This virus has been used since 1796, when Jenner first demonstrated the ability of vaccinia to protect against subsequent variola inoculation and thus protect against smallpox. This technique was widely applied with over a billion doses given worldwide, thereby leading to the eradication of smallpox (4). The intensity of this vaccination program has led to the compilation of a highly accurate safety profile of vaccinia (see...

Cytokinemodified tumor vaccines

The ability of bacterial products to elicit inflammatory cytokines underlies the antitumor activity observed with the administration of bacterial extracts as immunologic adjuvants. The ability of locally delivered BCG or recombinant cytokines to enhance antigen-specific immunity led to the testing of cytokine-secreting tumor vaccines as a strategy for delivering relevant tumor antigens in the context of immune-activating cytokines. A number of cytokines, including IL-2 (76-80), IL-4 (81-83),...

Other uses of rcc vaccines

Vaccine-Primed Lymph Node Cells Lymph nodes that drain malignancies or sites of tumor vaccination presumably harbor preeffector cells that can be activated and expanded ex vivo using anti-CD3 antibody and low concentrations of IL-2. In an early-phase clinical trial, 12 patients with metastatic RCC were treated with vaccine-primed lymph node (VPLN) cells and high-dose bolus IL-2 (44). There were two complete and two partial responses. In a similar study, 20 patients with metastatic RCC were...

Natural killer cell defects in cancer

NK cells are cytotoxic effector cells that are involved in the lysis of a tumor (134). These cells produce IFN-y and TNF-a, which further stimulate both T cells and NK cells. Three main mechanisms are responsible for the cytotoxic function of NK cells against tumor targets. These mechanisms include the CD95L-CD95 (Fas ligand-Fas) pathway (135,136), TNF-related apoptosis (137,138), and granule exocytosis. The latter involves the release of a pore-forming protein, perforin (139), and apoptotic...

DCDerived Exosomes Bear Functional MHC Class I and Class IIPeptide Complexes

Exosomal MHC class I molecules can be efficiently loaded directly, after acid elution of purified exosomes (32 Dee Shu, unpublished data). In contrast, exosomal MHC class II molecules can present peptides that have been pulsed onto whole DC culture (25,26 Dee Shu, unpublished data). We are currently reporting that MHC class I and II peptide complexes harbored by mouse or human exosomes activate specific CD8+ or CD4+ T lymphocytes but require DCs (29,31). Exosomal HLA-A2 Mart1 peptide complexes...

Human Papillomavirus

Carcinomas of the anogenital tract, particularly cancer of the cervix, account for almost 12 of all cancers in women, and so represent the third most frequent gynecological malignancy in the world (44). It is well established that chronic infection of cervical epithelium by human papilloma viruses (HPVs) is necessary for the development of cervical cancer. HPV DNA has been demonstrated in more than 99 of all tumor biopsy specimens, with high-risk HPV16 and HPV18 being most prevalent (45). HPVs...

Tumor Infiltrating Lymphocytes

T lymphocytes obtained from tumor deposits are presumably highly enriched for tumor-reactive effector memory cells that preferentially migrated into tumors and have been retained there. Enthusiasm for clinical use of TILs was engendered by murine tumor models that demonstrated tumor-specific cytolytic activity and cytokine release in vitro and regression of established tumors following adoptive transfer of relatively low cell numbers when compared with LAK cells (13,27,82). The most extensive...

Cytotoxic T Cells and NKMediated Immunity

In the L1210 GZL murine model, Raychaudhuri et al. (71) demonstrated the induction of tumor-specific cytotoxic T lymphocytes (CTLs) following immunization with the anti-Id antibody 2F10 mimicking an epitope of the mouse mammary tumor virus-encoded envelop glycoprotein gp52. The frequency of L1210 GZL tumor cell-reactive CTLs was similar in mice immunized with the anti-Id antibody or irradiated tumor cells when examined at the precursor level. Rather than using a classic Ab2 approach, Ruiz et...

Presence of Antitumor Immunity in Lung Cancer Patients

Despite immune defects, antitumor immune responses can be detected in some lung cancer patients. For example, more than half of SCLC and NSCLC patients have been described to have circulating antibodies reactive with autologous tumor proteins (9). Presence of these antibodies predicts better response to therapy and survival in some studies (9-11), whereas others, such as those specific for p53, are associated with a survival effect in some but not other studies (12-20). Detecting in vivo...

Summary and challenges to vaccines for lung cancer

Lung cancers, like other malignancies, can escape immune recognition by downregulation of HLA class I molecules. Administration of IFN-gamma has been shown to upregulate HLA class I on NSCLC and thus may be a useful adjunct (37). Fas-ligand (Fas-L), frequently detected in lung carcinoma cell lines and resected tumors, can cause apoptosis of T cells. In fact, lung carcinoma cells were capable of killing the Fas-sensitive human T-cell line Jurkat in coculture experiments (65). Inhibiting Fas-L...

Immunotherapy of experimental tumors with haptenmodified vaccines

There is considerable evidence that the failure of immunotherapy to eradicate cancers, whether spontaneous human cancers or experimental transplantable tumors, is a result of immunological tolerance. The most striking illustration of this hypothesis remains the work of Mullen et al. (25). They produced two variants of the fibrosarcoma 1591 a regressor tumor that was highly immunogenic and always rejected by normal mice and a progressor tumor that had become nonimmunogenic as a result of losing...

Conclusion

The purification process used for exosome isolation from DC cultures and ascites fluid allows harvesting of reproducible yields of 40- to 100-nm vesicles that are highly immunogenic in vitro. Exosomes are currently defined by their morphology (electronic microscopy), by their physical properties (stable at high temperature, floating at a density of 1.13-1.210 g mL in a sucrose D2O gradient), by their proteic patterns (endocytic markers, i.e., tetraspanins and hsp73), and by their enrichment in...

Peptide vaccines

Tumors express peptides, often oncogenes, that are unique to the malignant cells. Typically, these proteins are mutated forms of the native protein. Though variation in the specific mutations seen would make immunologic targeting difficult across a patient population, several oncogenes share significant homology among patients. For example, K-ras commonly is mutated in codon 12 and could serve as a potential target for vaccine development. Several investigators are testing this hypothesis in...

Dendritic cell vaccines

Dendritic cell vaccines for colon cancer have generally been loaded with MAGE (a cancer antigen) or CEA. In one study, MAGE was expressed by one-third of colon cancers (48). In a larger study, MAGE expression was exclusive to the tumor tissue with at least one of the 10 MAGE antigens tested being present on 70 of the 80 samples and MAGE-3 expression was associated with increased metastatic potential (49). Clinical studies are ongoing using MAGE as a vaccine target in GI cancers. In one,...

Le and Globo H Vaccines

The development of Ley and Globo H vaccines was previously limited by the lack of sufficient quantities of antigen for vaccine construction and testing. Over the last 6 yr, Dr. Samuel Danishefsky in our group has successfully synthesized both antigens (76-78). We have immunized groups of mice with Globo H-ceramide plus or minus adjuvants QS-21 and Salmonella Minnesota mutant R595, and with Globo H covalently attached to KLH or bovine serum albumin (BSA) plus immunological adjuvants QS-21 or...

Vaccines for Acute Myeloid Leukemia AML

AML cells can be made into cell vaccines for the disease. A strategy that has been extensively pursued is the differentiation or maturation of leukemia cells into DCs, which reflects the knowledge of DC development from primitive myeloid cells (32-34). A representative study by Choudhury and coworkers (113) demonstrated that the cytokine combination of GM-CSF and IL-4 together with CD40L or TNF-a induced primary AML cells from 18 of 19 patients to differentiate into DC-like cells that were able...

Use of autologous or allogeneic vaccines

One question in the development of whole-cell vaccines is whether autologous or allogeneic vaccine should be used. Although autologous (and thus personalized vaccines) are appealing, they are difficult to produce. The small size of many tumor specimens obtained by biopsy can make it difficult to obtain sufficient cells for therapy, especially when multiple immunizations are part of the clinical protocol. In some instances, it is possible to propagate tumor cells in vitro to increase the number...

Idiotype Based Protein Vaccines for Myeloma

Our group at the Karolinska Institute, Stockholm, Sweden, was the first to introduce active immunization of myeloma patients with Id proteins (72,73). Having considered that immunotherapy may work better in immunocompetent patients with a low tumor burden, we targeted untreated patients with early disease. In our first pilot study, we recruited and immunized five previously untreated patients with stages I-III MM with the autologous Id protein precipitated in an aluminum phosphate suspension...

Nonspecific active immunotherapy

Although the purpose of this chapter is to describe the results for vaccines in the treatment of lung cancer, it is important to consider that nonspecific inflammatory mediators have demonstrated activity in lung cancer and although experimentally, they are giving way to the more specific tumor vaccines, they may be combined with vaccines in the future. Initial studies utilizing the bacille Calmette-Guerin (BCG) strain of Mycobacterium tuberculosis, bovis BCG, administered by the intrapleural,...

Abnormal Dendritic Cell Differentiation and Accumulation of Immature Myeloid Cells in Cancer

The induction of an effective antitumor immune response requires antigen presentation by host APCs (94). DCs are the most potent APCs. DCs, macrophages, and granu-locytes are differentiated from common myeloid progenitors. Only mature DCs can efficiently prime CD8+ CTL precursors (95). Impaired differentiation of the myeloid lineage may result in the accumulation of cells able to inhibit CD8+ T cells (96). Tumors severely affect differentiation of myeloid cells, which results in the decreased...

Delayed Type Hypersensitivity

In the delayed-type hypersensitivity test (DTH), an intradermal injection of antigen in the form of soluble protein alone or as antigen loaded onto antigen-presenting cells is administered and the diameter of any resulting erythema or induration after 48-72 h is measured. CD4+ Th cells that recognize the antigen presented on local antigen-presenting cells mediate the DTH response by releasing inflammatory, Th1 cytokines that increase vascular permeability and recruit monocytes and other...

And Kenneth A Foon MD

Anti-Id Antibodies in Murine Tumor Models Anti-Id Antibodies and Human Cancer Network Antigens in Lymphoma and Leukemia Formats of Anti-Id Antibodies Immune Pathways Induced by Anti-Idiotypic Antibodies Conclusion and Future Directions of Anti-Id Cancer Vaccines References Active specific immunotherapy (ASI) is an attractive approach to cancer therapy, especially in an adjuvant setting. ASI is intended to boost or induce a host antitumor response, in contrast to passive immunotherapy, where...

AntiId DNA Vaccines

Our anti-Id antibody, 1A7 mimicking GD2, induced both Ab3 and Ab1' humoral responses in injected small animals (54), primates (38), as well as melanoma patients (55). However, presumed cellular responses could not be detected in the injected patients. Various plasmid constructs have been reported to invoke combined humoral and cellular immune responses in injected hosts against the transgene product. To investigate the induction of cellular immune responses we constructed two plasmids...

Tumorassociated antigen vaccines

The demonstration that some RCC lines express antigenic determinants that can be recognized by MHC-restricted cytotoxic T lymphocytes (CTLs) has led to efforts aimed at identifying tumor-associated antigen (TAA) in human RCC. One recently cloned candidate RCC-TAA is G250 (32). DCs loaded with G250 peptide and cultured with autologous T cells can be used to generate human CTLs capable of lysing G250-express-ing targets (33). This suggests that TAA-based immunotherapeutic strategies may be...

Breaking tolerance with haptenmodified proteins

Weigle extended these observations to proteins that were not immunogenic in their native state. Rabbits that had been rendered tolerant to bovine serum albumin (BSA) by neonatal injections of this protein failed to produce anti-BSA antibody even after injection with Freund's adjuvant. In contrast, unresponsive rabbits injected with BSA conjugated to sulfanilic acid (SA) produced antibody not only to SA-conjugated BSA but to native BSA as well (3). Thus immunization with a hapten-modified...

Advantages of Vaccines That Induce Antibodies Over Vaccines Designed to Augment TLymphocyte Immunity

The concept of a vaccine that consistently augments T-lymphocyte immunity against cancer cells is exciting and offers enormous potential for clinical benefit. However, demonstration that this has been achieved has proven to be more difficult than initially appreciated, for a variety of reasons 1. Ideally, autologous cancer cells are required for testing and these are rarely available as cell lines or in frozen samples in sufficient quantities for a thorough analysis of the immune response and...

Dendritic Cell Based Vaccines for BCell Lymphoma

DCs are the most potent APCs and are ideally suited to serve as natural adjuvants for purposes of vaccination and immunotherapy for cancers (30,31). Methods have been developed to obtain substantial numbers of DCs from proliferating CD34+ progenitors in bone marrow and peripheral blood, as well as from nonproliferating precursor cells, such as CD14+ monocytes, in human blood (32-34). Induction of antitumor immune responses by injection of antigen-loaded DCs has been extensively studied in...

Future development of recombinant pox virus vaccines

The future development of recombinant pox virus vaccines holds considerable promise for cancer management. Virtually all of the clinical studies described above have been carried out in patients with relatively advanced cancers. These patients most likely have subtle but depressed immune responses, which have been exhibited in two ways (a) via downregulation of the Z chain of the T-cell receptor and (b) in the cytokine profile of T cells obtained from advanced cancer patients (type 2 profile)...

Th1 vs Th2Type Response in Cancer

In animal studies, the production of Th1 cytokines IFN-y and IL-2 by CD4+ T cells is linked to the generation of therapeutically efficient immune responses that reject tumors (84). IL-2 is required for T-cell proliferation and the acquisition of cytotoxic effector function. On the other hand, type 2 cytokines, IL-4 and IL-10, promote Th2 responses that are necessary for the induction of a humoral immune response. In animal models of cancer and cancer patients, the tumor microenvironment can...

TF Tn and sTn Antigens

Mucins are major cell-surface antigens in a variety of epithelial cancers. They are primarily large extracellular molecules made up of multiple copies of serine- and threo-nine-rich tandem repeats (30-33). Though mucins (including carbohydrate and peptide epitopes) are also expressed on some normal tissues, they have proved to be excellent targets for anticancer attack for two reasons First, expression on normal tissues is largely restricted to the ductal border of secretory cells (31-33), a...

Early studies of adoptive immunotherapy of tumors

Two of the fundamental concepts that underlie tumor immunotherapy, namely that cellular elements could confer immunity upon na ve recipients and that unique tumor antigens were the relevant targets, were established empirically long before T lymphocytes were defined or the nature of antigen recognition was understood at a molecular level. Billingham observed that immunity to skin grafts could be transferred to na ve recipients by lymph node (LN) cells and introduced the term adoptive immunity...

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...

Recombinant viral vaccines

As a means to increase antigen processing and expression in APCs and improve the expression of costimulatory molecules, recombinant viral-based vaccines have been developed. The pox virus family has been most commonly used (vaccinia, fowlpox) but others such as adenovirus have been used. Genes encoding TAAs are genetically recom-bined to the virus and then administered. The virus serves as a vector, infecting cells including APCs the passenger gene is then transcribed and translated into a...

Experimental evidence supporting a role of tgfP in active tumormediated immunosuppression

The evidence presented above supporting TGF-P as a major mechanism for tumormediated immunosuppression is primarily circumstantial. If TGF-P is the major mechanism for tumor-mediated immunosuppression, one would predict that blocking the TGF-P-signaling pathway would result in more effective tumor vaccines, and conversely, that increased TGF-P signaling would allow tumors to escape the immune system. These predictions have been experimentally verified. In terms of increasing TGF-P signaling,...

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 (>...

Drug Induced Autoimmunity

Ingested drugs can act as haptens, which combine with normal tissue proteins forming immunogenic complexes that are recognized by T cells. Although drugs tend not to be highly chemically reactive molecules, their metabolites may well be. An example is penicillin, which can be associated with autoimmune hemolytic anemia. Although not likely to conjugate proteins itself, in solution penicillin spontaneously forms penicillenic acids with a highly reactive oxazolone group (22) that modifies...

Models of hapteninduced autoimmunity

As described above, in his studies on cross-reactivity of hapten-modified and native proteins, Weigle (3) serendipitously found that mice immunized with hapten-modified thyroglobulin developed histological evidence of autoimmune thyroiditis. More recently haptens have been intentionally applied to develop animal models of what are presumed to be human autoimmune diseases. Neurath et al. (20) applied small amounts of the hapten, trinitrobenzene sulfonic acid (TNBS) (a derivative of TNP), to the...

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...

Toxicity of DNPModified Autologous Vaccine

In most patients, the toxicity was limited to the reaction at the vaccine injection site. All patients developed pruritic papules that progressed to pustules, sometimes with small ulcerations, that resolved into small white or pink scars. The intensity of the local reactions was ameliorated by reducing the dose of bacille Calmette-Gu rin (BCG). As expected, about one-third of patients developed nausea, sometimes with vomiting, following administration of cyclophosphamide, but systemic toxicity...

And W Martin Kast PhD

Oncogenic Viruses Conclusion Acknowledgments References Viruses implicated in the development of human cancer include hepatitis B (HBV) and hepatitis C (HCV) viruses, human papilloma virus (HPV), Epstein-Barr virus (EBV), human T-cell lymphoma virus, and human herpes virus 8. Together they contribute significantly to the total incidence of cancer worldwide. Current work in each of these virus systems seeks to understand the mechanisms of viral action and identify strategies of immune...

1

IP rIL2 or rIFNy with or without expanded TIL ip intraperitoneal TIL tumor-infiltrating lymphocyte CR complete response PR partial response rlL-2 recombinant interleukin-2 rIFNy recombinant interferon-y MHC major histocompatability complex. ip intraperitoneal TIL tumor-infiltrating lymphocyte CR complete response PR partial response rlL-2 recombinant interleukin-2 rIFNy recombinant interferon-y MHC major histocompatability complex. clinical activity observed. Thus, after initial encouraging...

Immunogenicity of Peritoneal Ascitis Derived Exosomes

Tumor-derived exosomes are not simply released in vitro by tumor cell lines in culture supernatants. We examined malignant effusions for the presence of tumor-derived exosomes and analyzed exosome immunogenicity on autologous peripheral T lymphocytes. Ultracentrifugation on sucrose and D2O gradients of11 malignant effusions allowed isolation of abundant amounts of exosomes. Malignant effusions accumulate high amounts of membrane vesicles with a mean diameter of 60-90 nm. These vesicles bear...

Multiple chaperone vaccines

If individual chaperone protein vaccines are effective against tumors, would one vaccine containing numerous chaperone protein family members be better The perceived advantages would be the probability of increasing the overall number of peptides available for antigen presentation because of multiple chaperone proteins, as well as the odds of increasing peptide diversity if members of the different chaperone families tend to escort distinct sets of peptides. Although their utilization has been...

Info

Sc subcutaneous Th1 T-helper-1 Th2 T-helper-2. antigen-presenting DCs to the vaccination site and thus encourage epitope spreading by creating a microenvironment conducive to optimal antigen presentation and T-cell stimulation. An impressive 14 of 18 patients generated T-cell proliferative responses, and 28 of patients demonstrated epitope spreading. Additional studies confirmed that DTH responses correlated with in vitro T-cell activity in 32 patients with HER-2-overexpressing breast, ovarian,...

TCell Responses at the Tumor Site

A rather surprising observation was made early into the first clinical trials of DNP-modified autologous vaccine the development of inflammatory responses in metastatic sites (50). These responses were initially observed in superficial (nodal or subcutaneous) metastases, and consisted of marked erythema, warmth, and tenderness of the tumors and the overlying skin. Responding metastatic lesions varied in size from 5-mm diameter skin metastases to 10-cm lymph node masses. The number of inflamed...

References

Allogeneic and autologous hematopoietic cell transplantation. In Beutler E, Lichtman MA, Coller BS, Kipps TJ, Seligsohn U, eds. Williams hematology, 5th ed. New York McGraw-Hill Medical Publishing Division, 2001 209-235. 2. Press OW, Leonard JP, Coiffier B, Levy R, Timmerman J. Immunotherapy of non-Hodgkin's lymphoma. Hematology (Am Soc Hematol Educ Program) 2001 221-240. 3. Countouriotis A, Moore TB, Sakamoto KM. Cell surface antigen and molecular targeting in the...

Flegnevgkty

Use of a T-cell clone that recognized a squamous cell carcinoma to screen a cDNA library resulted in the identification of a mutated Caspase-8 gene product (37). A nucleotide substitute in the normal stop codon resulted in extension of the normal open reading frame and generation of the T cell epitope. This mutation was observed in only one out of a large number of tumor cell lines, however, indicating that this was a relatively rare event. A mutated CDK4 gene...

Oil Based Adjuvants

Oil-based adjuvants are formulated as emulsions, which are dispersions of an antigen-containing aqueous phase with an oil phase. Water-in-oil (W O) emulsions contain higher concentrations of oil, typically 30-80 , consist of water droplets immersed in an oil milieu, and are quite viscous. In contrast, oil-in-water (O W) emulsions are composed of oil droplets immersed in an aqueous phase, and are predominantly aqueous in nature, with oil concentrations of only 2-10 . W O emulsions are more apt...

Phase I Trial Using Human DCDerived Exosomes

These data were the rationale for launching vaccination with DC-derived exosomes in metastatic tumor-bearing patients. A GMP has been set up to harvest large amounts of MD-DC-derived exosomes (ultrafiltration followed by ultracentrifugation on a high-density sucrose D2O cushion) and load exosomal MHC class I molecules with synthetic tumor peptides. Phenotypical analysis of clinical-grade exosomes is allowed by FACS-beads-assay, i.e., DC-derived exosomes, are fixed onto 4.5-micron beads stained...

Nuclear Localization of Plasmid DNA

Once released from endosomes into the cytoplasm, plasmids are facing another barrier the nuclear membrane. Generally, higher levels of transgene expression are observed in actively dividing cells where the nuclear membrane disappears during mitosis (34). However, unless tumor or bone marrow cells are being targeted for transfection most cells, including muscle and dendritic cells, are nondividing. To overcome this barrier plasmids should contain specific nuclear localization sequences. The...

In Vitro Measures of Functional Antigen Specific Immune Responses

In addition to phenotypic assays, in vitro assays of T-cell function play an important role in detecting and quantitating antigen-specific immune responses. Functional assays measure a T-cell activity in response to a specific antigenic stimulus, include proliferation, cytokine secretion, and cytolytic function. These assays can be performed on specimens stimulated in vitro with antigen and cytokines, or on PBMC samples, without any preceding in vitro stimulation. The capacity of T cells to...

Peptide vaccine strategies

Over the last decade, at least 20 HLA class I-restricted peptide epitopes have been defined from multiple melanoma differentiation and cancer-testis antigens. They are reviewed in ref. 15. Peptide vaccine trials have been conducted with individual or multiple peptides in patients with stage IV disease in aqueous solution or using different adjuvants, with different cytokines, or via DNA plasmid delivery. Some trials have already been conducted in high-risk resected patients, which would appear...

Pcr

Reverse transcriptase real-time quantitative cytokine mRNA, 535-536 PCR-based cDNA subtraction. See Subtractive hybridization (SSH) PCTA-1, 453 PEL, 571 Penicillin, 279 Peptide(s) binding site modification, 131 improving immunogenicity, 129-132 Peptide-based vaccines, 121-132 Peptide immunization human clinical trials, 129-131 Peptide vaccine, 382-384 gastrointestinal cancer vaccines, 499-500 Peptide vaccine adjuvants dendritic cells (DC), 387-388 Peripheral blood lymphocytes (PBL), 367,...

Including the Exogenous Pathway

The presentation of vaccine antigens on MHC class I is needed to activate CD8+ CTLs, which are critical for protective antitumor immunity. The classical, endogenous pathway for presenting peptides on MHC class I products begins when endogenously synthesized proteins are clipped by the proteasome, and peptide fragments are transported via transporters associated with antigen presentation (TAPs) into the rough endoplasmic reticulum (13). There, the resulting peptides bind the peptide-binding...

TF Tn and sTn Vaccines

Patients with various epithelial cancers have been immunized with unclustered TF-KLH and sTn-KLH vaccines plus various adjuvants (84,85). High-titer IgM and IgG antibodies against TF and sTn antigens resulted. In our hands the majority of the reactivity was against antigenic epitopes present in the vaccine that were not present on naturally expressed mucins (porcine or ovine submaxillary mucins PSM or OSM ) or tumor cells (84,86). Based on previous studies with Tn antigen (87), Kurosaka and...

Malcolm S Mitchell MD

Allogeneic Melanoma Lysates for the Treatment of Disseminated Melanoma Adjunctive Therapy With Lysates in Resected Stage II and III Disease Melacine vs Observation for Resected Stage II Intermediate Thickness (1.5-4.0 mm) Melanoma Southwest Oncology Group Trial Melacine Followed by Interferons (IFN-a) in Disseminated Melanoma Immunological and Immunohistochemical Observations Other Clinical Trials With Allogeneic Lysates Conclusions and Future References

Discovery of haptens

Haptens are tiny lights that illuminate the dark recesses of the immune system. They were discovered by Karl Landsteiner, who used haptens to explore the breadth and fine sensitivity of antibody responses. Landsteiner (1) worked with a variety of simple chemicals, including nitrophenyls and phenyl arsonates, that were incapable of inducing an immune response by themselves, but became immunogenic when they were attached covalently to a protein carrier. He coined the term hapten from the Greek...

Antibody Responses in Vaccinated Cancer Patients

Table 3 summarizes humoral immune responses in vaccinated cancer patients including only those studies in which the binding of vaccine-induced antibodies to the surfaces of tumor cells was demonstrated. Antibodies binding to the tumor cell surface may destroy the cells via ADCC or CDC mechanisms or induce apoptosis, whereas antibodies binding to intracytoplasmic structures are most likely of no benefit to the patient. Thus, there are numerous reports on the induction of antibodies to isolated...

TCell Anergy Tolerance

T-Cell Anergy in Cancer Patients Selective T-cell nonresponsiveness to a particular antigen, as shown by defective antigen-specific IFN-y production and proliferation, has been repeatedly reported using murine tumor models. T-cell anergy has been shown to be an early event in tumor progression (30). Naive CD4+ T cells specific for a tumor-associated antigen, transferred into tumor-bearing mice, had a significantly diminished response to the peptide antigen (30). In clinical settings, the...

Alphavirus replicons and related vector systems general characteristics

Some efforts have been made to generate vaccine vectors from fully replication-competent alphaviruses. In these cases, attenuated strains are used, and the subgenomic 26S promoter is duplicated to allow expression of both the viral structural proteins and a heterologous gene (74). However, such vectors are limited with respect to both the size and stability of the heterologous gene, and in addition, efficiently induce antivector immune responses. Thus, replicon vectors have proven to be more...

Alphavirus characteristics applicable to vector development

Alphaviruses have several characteristics that are potentially advantageous for the derivation of vaccine vector systems. Within the general population, preexisting anti-alphavirus immunity is not widespread. Alphaviruses generally exhibit diverse cellular tropism within a given host, allowing for a broad range of administration routes for antitumor immunotherapeutics, although some alphaviruses such as Venezuelan equine encephalitis (VEE) show a natural and pronounced initial tropism for...

DC Dose Frequency Route and Schedule of Injections

In most studies thus far, the DCs were usually given at 2- to 4-wk intervals, and at doses between 4 and 40 million without striking differences in results. In vitro studies on human T-cell activation by DCs would predict that higher doses of DCs given more frequently should provide more intense and durable TCR triggering and thus promote T-cell priming and polarization. However, frequent stimulation might also cause activation-induced death of T cells. Also, the induced CTLs may kill booster...

Introduction

The success of cancer immunotherapy is dependent on the discovery of tumor-associated antigens (TAAs) for vaccine targets, but equally important is the need for effective delivery strategies and potent adjuvants to induce an appropriate immune response. Although there is evidence that many cancer patients do mount an immune response against their tumor, the affinity and frequency of responding T cells appears to be low. Additionally, the natural immune response to some cancers is a T helper 2...

MPLContaining Adjuvants

Ribi et al. (36,40-43) carried out systematic studies on the antitumor activities of cell wall components from both mycobacteria and Gram-negative Salmonella species. The findings from these studies led to the development of two adjuvants currently in advanced clinical studies with cancer vaccines Corixa Corporation's monophosphoryl lipid A (MPL adjuvant) and Enhanzyn immunostimulant. LPS is the major component of Gram-negative bacterial cell walls. Evaluation of different LPS preparations for...

Carbohydrate Cell Surface Cancer Antigens The Mskcc Experience

We have screened a variety of malignancies and normal tissues with a series of 40 monoclonal antibodies against 25 antigens that were potential target antigens for immunotherapy (18-21). Results for the 12 defined antigens expressed strongly in 50 or more of biopsy specimens of breast, ovary, prostate cancer, melanoma, sarcoma, and small-cell lung cancer (SCLC) are shown as examples in Table 2. The 13 excluded antigens (including CEA carcinoembryonic antigen and HER2 neu) were expressed in 0-2...

John Fikes PhD

The Challenge of Overcoming Immunological Tolerance 1. the challenge of overcoming immunological tolerance For all cancer vaccine strategies, a major challenge facing efforts to induce a clinically effective T-cell response is the necessity to break tolerance to normal, self antigens. To control auto-reactivity, some T cells with high avidity for tumor-associated antigen TAA epitope-major histocompatibility MHCs complexes are deleted in the thymus and the remaining T cells are controlled by...

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

A potential pitfall of the use of single class I binding peptides is illustrated in two similar studies vaccinating patients with an HLA-A2-binding peptide, p369-377, derived from the protein sequence of HER-2 neu, a well-defined tumor antigen. In an initial clinical study, HLA-A2-positive patients with metastatic HER-2 neu-overexpressing breast, ovarian, or colorectal carcinomas were immunized with 1 mg of p369-377 admixed in incomplete Freund's adjuvant IFA every 3 wk 38 . Peptide-specific...

Humana Press

2004 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512 All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permission from the Publisher. All articles, comments, opinions, conclusions, or recommendations are those of the author s , and do not necessarily reflect the views of the publisher. Due...

Paul F Robbins PhD

Antigens Recognized by HLA Class I-Restricted T Cells Antigens Recognized by Tumor-Reactive, Class II-Restricted T Cells Clinical Applications References Studies first carried out in the early 1980s demonstrated that incubation of tumor-infiltrating lymphocytes TILs with high doses of interleukin-2 IL-2 resulted in the generation of CD8 cytotoxic T cells CTL that recognized tumor cells in a major histocompatibility complex MHC -restricted manner. This procedure resulted in the generation of...

James L Gulley MD PhD Philip M Arlen MD and Jeffrey Schlom PhD

Vaccinia Vector

Dendritic Cell Vaccines With Pox Vectors Cytokine Expression Driven by Pox Vectors Future Development of Recombinant Pox Virus Vaccines Tumor-associated antigens TAAs are by definition either weakly immunogenic or functionally nonimmunogenic. Vaccine strategies must be developed in which the presentation of these TAAs to the immune system results in far greater activation of T cells than is being achieved naturally in the host. One approach to create an inflammatory milieu designed to trigger a...

Adenovirus use in cancer vaccine strategies

With so much known about Ad biology, it was a natural step to use recombinant Ad for vaccination purposes. As a first example, E1- Ad vectors have been engineered that express genes encoding epitopes or whole proteins derived from a number of pathogens, including but not limited to malaria, bovine herpesvirus type 1 , foot-and-mouth disease virus FMDV , measles virus, and HIV human immunodeficiency virus 5-7 . Vaccination with these Ad vectors has shown positive efficacy in preventing viral...

Practical Issues of Peptide Immunization Assessed in Early Human Clinical Trials

One of the first considerations addressed in initial clinical trials of peptide-based vaccines was the route of administration. The primary routes of administration that have been used are the intramuscular, subcutaneous, and intradermal routes. The dose and volume of the vaccine itself, the choice of adjuvant, and the desired immune response have largely determined immunization route. There have been few human clinical peptide vaccine studies comparing the routes of administration. For...

Melacine vs observation for resected stage ii intermediate thickness 1540 mm melanoma southwest oncology group trial

The results of this trial, recently analyzed for the first time after 9 yr of study, have been published by Sondak and collaborators 11 . A group of 689 patients was randomly assigned to receive Melacine over the course of a year, at the schedule used in our one-arm studies but unfortunately by the im route preceded by cyclophosphamide. Patients were required to have no palpable regional lymph nodes, but no formal axillary or inguinal node dissections were required. The sentinel node procedure...

Endosome Escape Sequences

The acidic environment of endosomes is detrimental to DNA, thus it is imperative for the plasmid to exit the endosome as quickly as possible. Unlike several viral peptides that facilitate escape of the virus from the endosome, such sequences are probably missing in most plasmids. Apart from providing a component of cellular targeting, cationic polymers can also protect naked DNA from degradation in the endosome. This might be another reason why transfection with cationic lipid-plasmid DNA...

Production and purification of exosomes

Since exosomes mediate MHC class I- and class II-restricted T-cell stimulatory capacity 29-31 and efficiently substitute for whole DC cultures 5 , good manufacturing process GMP laboratory procedures for exosome harvesting and purification have been set up for clinical implementations 32 . Exosomes derived from DC and tumor cell culture supernatants can be readily purified within 4-5 h starting from 2-3 liters of culture supernatant based on their physical properties. Exosomes float on sucrose...

Systemic and Local Suppression of T Cells

Immunosuppressive mechanisms that promote tumor growth include suppressive factors directly produced by tumor cells and those that are produced by other cells, although induced by the tumor. The most studied suppressive factor that can be produced by both the tumor and immunocompetent cells is the transforming growth factor-P TGF-P . TGF-P inhibits differentiation of both CD4 and CD8 naive T cells via different transcriptional activators 16 . Blocking of TGF-P signals in T cells in mice leads...

Biological effects of exosomes

The primary aim of exosomal release for a cell might be to discard membrane proteins. This role was suggested for reticulocyte-derived exosomes that carry transferring receptors TfRs useless in erythrocytes 36 . Thus, exosomes could be an alternative to lysosomal degradation, for example, to eliminate proteins that resist degradation by lysosomal proteases. However, based on their protein composition, i.e., enrichment for MHC complexes, hsp, and some targeting molecules, it is conceivable that...

Bacterial Extracts in Cancer Vaccines

Over a century ago, a New York physician, William B. Coley, noted that some cancer patients experienced tumor regression following episodes of acute bacterial illness. Dr. Coley hypothesized that the two events were linked and treated an inoperable cancer patient with a viable bacterial culture to induce a commotion in the blood. Remarkably, the patient recovered from both the bacterial infection and the tumor. From 1900 to 1936, Coley went on to successfully treat many tumor-bearing patients...

Contributors

Andrea Amalfitano, do, phd Departments of Pediatrics, Molecular Genetics and Microbiology, and Pathology, Duke University Medical Center, Durham, NC Fabrice Andre, md Department of Clinical Biology, Institut Gustave Roussy IGR , Villejuif, France Philip M. Arlen, md Laboratory of Tumor Immunology and Biology, Center for Cancer Research, National Cancer Institute, Bethesda, MD Jory R. Baldridge, phd Corixa Corporation, Hamilton, MT David Berd, md Division of Medical Oncology, Department of...