Development of an autologous canine cancer vaccine system for resectable malignant tumors in dogs.

While conventional therapies exist for canine cancer, immunotherapies need to be further explored and applied to the canine setting. We have developed an autologous cancer vaccine (K9-ACV), which is available for all dogs with resectable disease. K9-ACV was evaluated for safety and immunogenicity for a variety of cancer types in a cohort of companion dogs under veterinary care.

Differential pathways regulating innate and adaptive antitumor immune responses by particulate and soluble yeast-derived β-glucans.

β-glucans have been reported to function as a potent adjuvant to stimulate innate and adaptive immune responses. However, β-glucans from different sources are differential in their structure, conformation, and thus biologic activity. Different preparations of β-glucans, soluble versus particulate, further complicate their mechanism of action.

Pilot study of 1650-G: a simplified cellular vaccine for lung cancer.

Introduction: Cancer immunotherapy is a conceptually attractive since it is highly specific and can deal with disseminated disease with minimal impact on normal tissues. Early phase clinical trials have well established the ability of a variety of immunotherapeutic approaches to induce antigen specific immune responses in lung cancer patients. Although no immunotherapy is likely to be a panacea, recent data from randomized phase IIB studies offer promise of therapeutic activity in both early and late stage lung cancer.

Generation and characterization of non-small-cell lung cancer cell lines and clones for use in the study of immunotherapy.

The in vitro study of cancer has been made easier by the use of stable tumor cell (TC) lines derived from patients to study antigen expression, immunogenicity, and response to both experimental and conventional therapeutic agents. However, the routine generation of these cell lines in some tumor histologies such as non-small-cell lung cancer (NSCLC) is difficult. In many cases, colonies of TCs do not survive, most likely due to a lack of critical growth factors in cell culture medium.

Characteristics of PBMC obtained from leukapheresis products and tumor biopsies of patients with non-small cell lung cancer.

The current study characterized peripheral blood mononuclear cells (PBMC) obtained from leukapheresis products of patients with non-small cell lung cancer (NSCLC) for cytokine release, the ability to incorporate tritiated thymidine following stimulation using PHA as well as the levels of both CD4 and CD8 regulatory T cells (Tregs) as defined by FoxP3 expression. Results were compared to normal donor PBMC obtained from buffy coat products. Heterogeneous levels of Th1 (gamma interferon and IL-2), Th2 (IL-10 and IL-13), pro-inflammatory (TNF-alpha and IL-6) and the hematopoietic inducing cytokine GMCSF were detected from both populations of PBMC as measured using ELISA.

Immunotherapy for lung cancer.

Immunotherapy is a conceptually attractive approach, because it is highly specific and can deal with disseminated disease with minimal impact on normal tissues. Ability to induce antigen-specific immune responses in patients with lung cancer is now well established in early-phase clinical trials using a variety of immunotherapeutic approaches. Although no immunotherapy is likely to be a panacea, randomized phase IIB studies offer promise of therapeutic activity in both early- and late-stage lung cancer.

Identification of HLA-CW3, GNAS and IMPA as cytotoxic T-lymphocyte (CTL) target antigens using an allogeneic mixed lymphocyte tumor cell culture (MLTC) system and subsequent cDNA library screening.

Allogeneic mixed lymphocyte tumor cell cultures (MLTCs) were established using lymphocytes from non-small-cell lung cancer (NSCLC) patient UKY-53 and HLA-A2+ NSCLC tumor cells (UKY-29). The tumor cells expressed the lymphocyte costimulatory molecule CD80 (UKY29.7).

Immunization of NSCLC patients with antigen-pulsed immature autologous dendritic cells.

Only a handful of NSCLC patients have been included in dendritic cell (DC) vaccine clinical trials. We had previously reported a series of 16 individuals with stages IA-IIIB NSCLC who received autologous DC vaccines matured with dendritic cell/T cell-derived maturation factor (DCTCMF). Here we report the results of a continuation study with similar inclusion criteria, immunization protocol, and analysis, using an immature DC vaccine.

Issues concerning the large scale cryopreservation of peripheral blood mononuclear cells (PBMC) for immunotherapy trials.

Immunotherapy of cancer is being developed as an alternative or adjuvant to conventional therapies such as: surgery, chemotherapy and/or radiation treatment. Immunotherapy laboratories routinely process and prepare for injection large numbers of anti-tumor effector cells. The process of cryopreservation is critical to the success of immunotherapy.

Vaccines for lung cancer.

Immunotherapy is based on the knowledge that the immune system can distinguish cancerous cells from normal cells. Conceptually, this is an attractive adjuvant approach because it is highly specific and can deal with disseminated disease with minimal impact on normal tissues. In this review, we focus on strategies that use host immune machinery to generate anti-tumor effects, known as active immunotherapy.

The large scale generation of dendritic cells for the immunization of patients with non-small cell lung cancer (NSCLC).

In the current study, we generated large numbers of dendritic cells (DCs) from patients with non-small cell lung cancer (NSCLC) for a vaccine trial. The DCs were generated from CD14+ cells obtained by immuno-magnetic bead column separation technique. The CD14+ cells were placed in culture in the presence of granulocyte macrophage colony stimulating factor (GMCSF) and Interleukin 4 (IL-4).

On the road to a tumor cell vaccine: 20 years of cellular immunotherapy.

Cellular immunotherapy (CI), as we now know it, began in the early 1980s with the use of lymphokine-activated killer cells (LAK) and progressed to the use of the immunologically specific, tumor-infiltrating lymphocytes (TIL). TIL were shown to be particularly effective against melanoma and it was in these trials that we learned the importance of immunologic specificity for tumor. With the identification and characterization of tumor antigens recognized by TIL, we now see the use of these antigens in various forms constituting vaccines.

Autologous dendritic cell vaccines for non-small-cell lung cancer.

Purpose: Therapeutic outcomes of definitively treated non-small-cell lung cancer (NSCLC) are unacceptably poor. A wealth of preclinical information, and a modest amount of clinical information indicate that dendritic cell (DC) vaccines have therapeutic potential. Only a handful of NSCLC patients have been included in DC clinical trials.

Growth and functional reactivity of lymphocytes obtained from three anatomic compartments in patients with non-small-cell lung cancer (NSCLC).

Non-small-cell lung cancer (NSCLC) claims the lives of both men and women worldwide. In this study, we describe the ability to expand lymphocytes from solid tumor biopsies, pleural fluid, and peripheral blood of patients with NSCLC. While lymphocytes readily expanded from both solid biopsies and pleural fluids, the incidence of obtaining tumor-specific lymphocytes was low.

Cell surface phenotyping and cytokine production of Epstein-Barr Virus (EBV)-transformed lymphoblastoid cell lines (LCLs).

Epstein-Barr Virus-transformed B lymphoblastoid cell lines (EBV-LCLs) are routinely used for the in vitro expansion of T cells. However, these cell lines are reported to produce the cytokine IL-10, which is inhibitory for T cells. We, therefore, characterized a panel of 37 EBV-LCLs for a variety of cell surface markers, for secretion of various cytokines including IL-10 and for immunoglobulin production.

Characterization of human non-small cell lung cancer (NSCLC) cell lines for expression of MHC, co-stimulatory molecules and tumor-associated antigens.

A panel of 31 long-term non-small cell lung cancer (NSCLC) cell lines was examined for the expression of protein and/or mRNA transcripts for 11 distinct immune response related molecules or tumor associated antigens (TAA). To assess whether cytokine stimulation might up-regulate expression of the genes of interest, cells were cultured in 500 U/ml of gamma-interferon (gamma-IFN) for 48-72 h prior to analysis. Major histocompatibility complex (MHC) Class I antigens were detected by indirect immunofluorescence and were constitutively expressed on all of the cell lines.

Recognition of shared melanoma antigens in association with major HLA-A alleles by tumor infiltrating T lymphocytes from 123 patients with melanoma.

A total of 123 tumor-infiltrating T lymphocyte (TIL) cultures established from patients with HLA-A1, -A2, -A3, -A24, or -A31 metastatic melanoma in the Surgery Branch, National Cancer Institute, were screened for recognition of shared melanoma antigens including five melanosomal proteins (tyrosinase, MART-1/melan-A, gp100, TRP1, TRP2) as well as peptides derived from MAGE-1 and MAGE-3. Examination of the specificity of these T cells indicated that 16% of HLA-A1 TIL, 57% of HLA-A2 TIL, 7% of HLA-A3 TIL, 13% of HLA-A24 TIL, and 27% of HLA-A31 TIL recognized shared melanoma antigens restricted by major histocompatibility complex class I. Melanosomal proteins were frequently recognized by these TIL, and MART-1(27-35), gp100(154-162), gp100(209-217), and gp100(280-288) represent highly immunogenic epitopes that were recognized by a high percentage of HLA-A2 restricted melanoma reactive TIL.

Clinical and immune responses in advanced melanoma patients immunized with an anti-idiotype antibody mimicking disialoganglioside GD2.

Purpose: To determine immune responses and toxicity to the anti-idiotype vaccine, as well as clinical responses and survival, we initiated a clinical trial for patients with advanced melanoma treated with an anti-idiotype antibody (TriGem) that mimics the disialoganglioside GD2.

Patients And Methods: Forty-seven patients with advanced melanoma received either 1-, 2-, 4-, or 8-mg doses of TriGem (Titan Pharmaceuticals Inc, South San Francisco, CA) mixed with QS-21 adjuvant (Aquila Biopharmaceuticals, Inc, Worcester, MA) 100 microg subcutaneously weekly for 4 weeks and then monthly until disease progression. Median age was 57 years, there were 32 men and 15 women, 43% of patients had undergone prior therapy for metastatic disease, 55% had disease confined to soft tissue, and 45% had visceral metastasis.

Genetic manipulation of primitive leukemic and normal hematopoietic cells using a novel method of adenovirus-mediated gene transfer.

Gene transfer into early hematopoietic cells has been problematic due to the quiescent nature of primitive cells and the lack of gene transfer vehicles with high efficiency for hematopoietic cell types. Previously, we have shown that adenoviral vectors can be used for the transduction of normal human progenitors with gene transfer efficiencies of approximately 30%. However, this approach is limited by relatively slow uptake kinetics (24-48 h) and a strong dependence on the presence of exogenous cytokines.

CD80 expression in an HLA-A2-positive human non-small cell lung cancer cell line enhances tumor-specific cytotoxicity of HLA-A2-positive T cells derived from a normal donor and a patient with non-small cell lung cancer.

To generate non-small cell lung cancer (NSCLC)-reactive lymphocytes, we transfected an HLA-A2-expressing human NSCLC line (1355) with the cDNA encoding the lymphocyte co-stimulatory molecule CD80. Following selection in G418, 1355.7 demonstrated stable cell-surface expression of CD80.

T cell-receptor V gene use by CD4+ melanoma-reactive clonal and oligoclonal T-cell lines.

Tumor-reactive CD4+ T cells can be isolated and expanded from the peripheral blood and tumor lesions of patients with melanoma. In contrast to CD8+ T cells, little is known about the antigens recognized by these CD4+ T cells. As a consequence, little is known about the diversity of the T-cell receptor (TcR) use by melanoma-reactive CD4+ T cells.

Molecular mimicry of carcinoembryonic antigen by peptides derived from the structure of an anti-idiotype antibody.

Our goal was to use carcinoembryonic antigen (CEA) as a target for immunotherapy in CEA-positive cancer patients who are all immune tolerant to the native antigen. We isolated and characterized an anti-idiotype monoclonal antibody 3H1, which mimics a distinct and specific epitope of the Mr 180,000 CEA and can be used as a surrogate for CEA. In Phase Ib clinical trials in a group of 23 advanced colorectal cancer patients, 3H1 induced both humoral and cellular anti-3H1 responses, as well as anti-CEA immunity.

Adhesion of tumour-infiltrating lymphocytes to endothelium: a phenotypic and functional analysis.

Efficacy of cancer immunotherapy with cultured tumour-infiltrating lymphocytes (TILs) depends upon infused TILs migrating into tumour-bearing tissue, in which they mediate an anti-tumour response. For TILs to enter a tumour, they must first bind to tumour endothelium, and this process depends on TILs expressing and regulating the function of relevant cell-surface receptors. We analysed the cell-surface phenotype and endothelial binding of TILs cultured from human melanoma and compared them with peripheral blood T cells and with allostimulated T cells cultured under similar conditions.

Tumor-infiltrating lymphocytes. Cytologic, phenotypic and morphometric analysis.

Objective: To characterize the cytologic, phenotypic and morphometric features of tumor-infiltrating lymphocytes (TILs).

Study Design: Morphology and cell size parameters of 50 TILs and five normal donor peripheral blood lymphocyte (PBL) specimens were compared using light microscopy, manual ocular measurements and digital image processing.

Results: Cytologically, all TILs demonstrated similar morphologic findings regardless of the initial tumor type.