Vitiligo is a disease in which melanocytes are selectively destroyed. The disease is thought to be an autoimmune process being there are antibodies to pigment cells in the sera of patients and animals with vitiligo. In the present study, sera from vitiligo patients were examined for reactivity with the human melanoma cell line, SK-Mel-28, by Western blot analysis of solubilized membrane antigens of these cells to identify the pigment cell antigens defined by antibodies in the patients with vitiligo. Antibody reactivity to human melanoma cells (SK-Mel-28) was investigated in 14 patients with vitiligo, and 16 with normal control individuals. Antibodies to the 116-113, 60, 40 KD antigens were associated with vitiligo being present in 79%, 86%, and 43% respectively of the patients with vitiligo, but in only 6%, 38% and 6% of the normal controls. In contrast, antibodies to the 160-155, 78 and 64 KD antigens were equally common in vitiligo and in normal individuals. The results suggest that autoreactivity to pigment cells occurs more commonly in patients with vitiligo than in the normal control and high autoreactivity to pigment cells in the vitiligo sera might be an impertinent epiphemenon to destroyed pigment cell.
Antibodies, Neoplasm/*blood ; Antigens, Neoplasm/immunology ; Autoantibodies/blood ; Blotting, Western ; Human ; Melanoma/*immunology ; Vitiligo/*immunology

This report gives a better emphasis on the role of targeted effectors (e.g. a combination of 5-FC with CD-NSPCs as compared to the application of NSPCs alone) and how such delivery of pro-drug activating enzymes and other tumor-killing substances may overcome melanocytic defence system, interact with and promote the host defence and immune response modulations not only in melanoma but, potentially, in other highly-metastatic cancers.
Brain Neoplasms ; immunology ; surgery ; Humans ; Immunity, Innate ; immunology ; Melanoma ; immunology ; secondary ; surgery ; Models, Immunological ; Stem Cell Transplantation ; methods

Listeria monocytogenes (LM), a Gram-positive facultative intracellular bacterium, can be used as an effective exogenous antigen expression vector in tumor-target therapy. But for successful clinical application, it is necessary to construct attenuated LM stain that is safe yet retains the potency of LM based on the full virulent pathogen. In this study, attenuated LM and recombinants of LM expressing melanoma inhibitory activity (MIA) were constructed successfully. The median lethal dose (LD(50)) and invasion efficiency of attenuated LM strains were detected. The recombinants were utilized for immunotherapy of animal model of B16F10 melanoma. The level of MIA mRNA expression in tumor tissue was detected by using real-time polymerase chain reaction (PCR) with specific sequence, meanwhile the anti-tumor immune response was assayed by flow cytometric analysis and enzyme-linked immunosorbent spot (ELISPOT) assay. The results showed the toxicity and invasiveness of attenuated LM were decreased as compared with LM, and attenuated LM expressing MIA, especially the double-genes attenuated LM recombinant, could significantly induce anti-tumor immune response and inhibit tumor growth. This study implicates attenuated LM may be a safer and more effective vector for immunotherapy of melanoma.
Animals ; Cancer Vaccines ; genetics ; immunology ; Extracellular Matrix Proteins ; genetics ; immunology ; Listeria monocytogenes ; immunology ; Male ; Melanoma ; genetics ; immunology ; Mice ; Mice, Inbred C57BL ; Vaccines, Attenuated ; genetics ; immunology

Chronic inflammatory responses have long been observed to be associated with various types of cancer and play decisive roles at different stages of cancer development. Inflammasomes, which are potent inducers of interleukin (IL)-1β and IL-18 during inflammation, are large protein complexes typically consisting of a Nod-like receptor (NLR), the adapter protein ASC, and Caspase-1. During malignant transformation or cancer therapy, the inflammasomes are postulated to become activated in response to danger signals arising from the tumors or from therapy-induced damage to the tumor or healthy tissue. The activation of inflammasomes plays diverse and sometimes contrasting roles in cancer promotion and therapy depending on the specific context. Here we summarize the role of different inflammasome complexes in cancer progression and therapy. Inflammasome components and pathways may provide novel targets to treat certain types of cancer; however, using such agents should be cautiously evaluated due to the complex roles that inflammasomes and pro-inflammatory cytokines play in immunity.
Animals ; Carcinoma ; immunology ; pathology ; therapy ; Gastrointestinal Neoplasms ; immunology ; pathology ; therapy ; Humans ; Inflammasomes ; metabolism ; Melanoma ; immunology ; pathology ; therapy ; Neoplasms ; immunology ; pathology ; therapy ; Skin Neoplasms ; immunology ; pathology ; therapy

OBJECTIVE: To determine the effect of activation of specific anti-tumor cytotoxic T lymphocytes (CTL) and the ability of cross-presentation in vitro by fusion of HLA-A2+ human dendritic cells (DCs) with HLA-A2- melanoma cells. METHODS: The HLA-A2+ human dendritic cells and HLA-A2- melanoma cells were fused by PEG and were cultivated in complete RPMI1640 media containing FCS (10%) and GM-CSF for 24-48 h, and then co-cultured fusion cells with Melan-A specific T cells. HLA-A2- melanoma cells were negative control,While T2 cells and DC+Pts were positive control. The activation of anti-tumor CTL elicited by the fusion cells was detected by intracellular cytokine staining. RESULTS: The immature DC could express CD80, CD83, CD86, HLA-DR, and HLA-ABC,but the mature DC induced by TNF-alpha, PGE-2, and CD40L further highly expressed above molecules. The rate of specific CTL cells primed by the fusion cells was 16.72%+/-4.26%, negative control was 0.21%+/-1.84%,and positive control was 28.60%+/-5.67%. The CTL from vaccine by fusing DC and LAR6 induced lysis of HLA-A2+ LAR1 cells. CONCLUSION The HLA-A2 restricted specific anti-tumor CTL can be induced in vitro by fusion of HLA-A2+ human dendritic cells with HLA-A2- melanoma cells.
Antigen Presentation ; immunology ; Antigens, Neoplasm ; immunology ; Cancer Vaccines ; immunology ; Cell Fusion ; Cell Line, Tumor ; Dendritic Cells ; cytology ; immunology ; HLA-A2 Antigen ; immunology ; Humans ; MART-1 Antigen ; immunology ; Melanoma ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo explore mechanisms of the augmented anti-tumor immunity observed in reconstituted lymphopenic mice (RLM) receiving melanoma vaccination.

METHODSThe study is to investigate the anti-tumor immunity of tumor vaccination during early immune reconstitution period following irradiation and cyclophosphamide (CY)-induced lymphopenia. Lymphopenic mice were subsequently reconstituted with naive splenocytes from syngeneic mice and immunized with irradiated melanoma cells F10 (irradiation experiment) and GM-CSF-modified D5 melanoma cells (D5-G6) (CY experiment). Controls included normal C57BL/6 mice receiving the corresponding vaccination, un-immunized naive mice and RLM. 8 - 10 days after vaccination, tumor vaccine draining lymph nodes (TVDLN) were harvested and phenotyped by FACS analysis. T cells purified from TVDLN were stimulated with anti-CD3 and anti-TCRbeta and proliferation was assessed by [(3)H]-TdR incorporation and FACS assay was performed for CD69 expression.

RESULTSThe augmented anti-tumor immunity correlated with a significant increase in the percentage of T cells with activation/memory phenotype in the TVDLN of vaccinated RLM, compared to that of the controls. There was also a significant increase in the density of DCs in TVDLNs. The activation threshold of T cells generated from vaccinated RLM was significantly decreased, resulting in markedly enhanced proliferating capability upon anti-CD3 stimulation.

CONCLUSIONThis study suggests that the augmented anti-tumor immunity observed in vaccinated RLM is due to down regulated activation threshold of T cells during lymphopenia-driven T cell proliferation, which may in turn facilitate the breaking down of immune tolerance to weak tumor antigens upon vaccination with tumor cell vaccines.

Animals ; Cancer Vaccines ; administration & dosage ; Cyclophosphamide ; Lymph Nodes ; immunology ; Lymphopenia ; etiology ; immunology ; Male ; Melanoma, Experimental ; immunology ; therapy ; Mice ; Mice, Inbred C57BL ; T-Lymphocytes ; immunology ; Whole-Body Irradiation

OBJECTIVETo investigate the immunotherapy efficacy of both helper T lymphocytes (Th) and cytotoxic T lymphocytes (CTL) epitopes augmented dendritic cells (DCs) tumor vaccine on gastric cancer.

METHODSNaïve spleen T cells were stimulated by mixed peptides (a mixture of Th epitope MAGE-3 (22-36)) primed DCs per week in vitro. After 4 cycles of restimulation, peptide specific T cells were harvested and subgroups of which were determined with flow cytometry. Cytokines secreting profiles by CD4+ T cells and cytotoxicities of CD8+ T cells on tumor cells were assessed. The protective immunity by referred DCs tumor vaccines was also monitored.

RESULTSBoth Th and CTL epitopes primed DCs could elicit both CD4+ T cells and CD8+ T cells in vitro,of which CD4+ T cells released high amount of Th1 type cytokines (IFN-gamma, IL-2) on recognizing specific antigen, as well as CD8+ T cells exhibited efficient tumor-killing capacity. The effects induced by DCs pulsed with single epitope (Th or CTL epitope) in vivo were less effective than those induced by DCs pulsed with mixture epitopes.

CONCLUSIONSBoth Th and CTL epitopes augmented DCs tumor vaccine can induce CD4+ Th1 and CD8+ CTL mediated immune responses to eradicate gastric cancer cells.

Animals ; Cancer Vaccines ; immunology ; therapeutic use ; Cell Line ; Cell Line, Tumor ; Dendritic Cells ; immunology ; Epitopes, T-Lymphocyte ; immunology ; Immunotherapy ; Melanoma, Experimental ; Mice ; Peptides ; immunology ; Stomach Neoplasms ; therapy ; T-Lymphocytes, Cytotoxic ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology

OBJECTIVETo explore the in vitro immune response to taxol resistance associated antigen 3 (TRAG-3)-derived cytotoxic T lymphocyte (CTL) epitope-pulsed dendritic cell (DC).

METHODSThe HLA-A2.1 restricted CTL epitope of TRAG-3 was previously identified to be a nonameric peptide sequence from 58 to 66 amino acid residues (TRAG-3(58-66)). The peptide was synthesized according to Fmos procedure, purified and molecular weight determined. Peripheral blood mononuclear cells (PBMC) of normal HLA-A2.1(+) individuals were used to isolate DC and DCs' phenotype identified by flow cytometry. Induction of CTL was achieved by in vitro stimulation of HLA-A2.1(+) PBMC with peptide-pulsed DC. CTL activity against HLA-A2.1(+)/TRAG-3(+) melanoma cell line LB373-MEL was assessed by (51)Cr release assay and IFN-gamma released by ELISA.

RESULTSThe synthetic nonameric peptide was above 90% pure and the determined values of molecular weight were conformed to their theoretical values. The phenotype of the isolate DCs was characteristic for mature ones. PBMC stimulated in vitro by TRAG-3-derived epitope-pulsed DCs for three times could specifically kill the target cells and secreted high concentration of IFN-gamma.

CONCLUSIONTRAG-3-derived epitope-pulsed DC can elicit specific anti-tumor immune response in vitro. Clinical trials using it as tumor vaccine may be feasible as specific immunotherapy for patients with TRAG-3 expressing cancers.

Cancer Vaccines ; immunology ; Cell Line, Tumor ; metabolism ; Cells, Cultured ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Drug Resistance, Neoplasm ; Epitopes, T-Lymphocyte ; immunology ; Humans ; Interferon-gamma ; metabolism ; Melanoma ; pathology ; Neoplasm Proteins ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Taxoids ; pharmacology

OBJECTIVETo investigate the possibility of using melanoma antigen-1 (MAGE-1) peptide as a tumor vaccine to treat hepatocellular carcinoma (HCC).

METHODSThe expressions of MAGE-1 in 8 HCC cell lines and in liver cancer tissue from a patient were detected using RT-PCR. The type of human leucocyte antigen I(HLA I) of both 8 HCC cell lines and peripheral blood mononuclear cells of the patient was detected using a microcytotoxicity method to screen out target cell lines for the cytotoxicity assay. Peripheral blood mononuclear cells from the HCC patient pulsed with an MAGE-1 peptide (NYKCRFPEI) were used as antigen presenting cells. Autogenous peripheral blood mononuclear cells were stimulated with antigen presenting cells every 7 days for 4 times to elicit cytotoxic T lymphocytes. The phenotype of effector cells was analyzed using flow cytometry. The cytotoxicity of effector cells was detected with a lactate dehydrogenase releasing assay.

RESULTSThe expressions of both MAGE-1 and HLA-A24 were detected in BEL7405 cell line which were used as the positive target cell line in the cytotoxicity assay. The expression of MAGE-1 alone was detected in HLE, BEL7402, BEL7404, QGY7703 and SMMC7721 cell lines, and the expression of neither MAGE-1 nor HLA-A24 was shown in QGY 7701 and HpG2 cell lines. The last 7 cell lines could be used as negative target cell lines in the cytotoxicity assay. Peripheral blood mononuclear cells expanded 32 folds during 28-day culture. The ratio of CD3(+) T cells increased by 16% (from 54% to 70%), and the ratio of CD8(+) T cells increased by 20% (from 36% to 56%) during 28-day culture. When the ratio of effector cells to target cells was 10:1, effector cells exhibited 62.5% cytotoxicity against autogenous lymphoblasts pulsed with the peptide (NYKCRFPEI) of MAGE-1 antigen, 40.25% cytotoxicity against BEL7405 cells, compared with 17.88% cytolysis observed against autogenous lymphoblasts, 19.55% against HLE cells, and 1.6% against QGY7701 cells. When the ratio of effector cells to target cells was 3.3:1, the cytotoxicity of effector cells against the peptide pulsed autogenous lymphoblasts was 53.6%, which was much higher against autogenous lymphoblasts, HLE cells and QGY7701 cells at 15.6%, 13% and 1%, respectively.

CONCLUSIONThe results demonstrate that cytotoxic T lymphocytes with the ability to specifically lyse target cells expressing both MAGE-1 and HLA-A24 could be successfully induced by the MAGE-1 peptide NYKCRFPEI in vitro. This indicates that a good result might be anticipated if this peptide is used as a tumor vaccine to treat HLA-A24 HCC patients.

Adult ; Antigens, Neoplasm ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; immunology ; HLA-A Antigens ; analysis ; HLA-A24 Antigen ; Humans ; Liver Neoplasms ; immunology ; Male ; Melanoma-Specific Antigens ; Neoplasm Proteins ; genetics ; immunology ; RNA, Messenger ; analysis ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

OBJECTIVETo observe the inhibitory effect of dendritic cells (DCs) activated cytotoxicity T lymphocyte (CTL) combined with MAGE-1 nonapeptide on transplanted human hepatocyte carcinoma (HCC) in nude mice.

METHODSA model of HCC transplanted tumor was established by injecting BEL-7402 cell line HCC cells subcutaneously on the back of nude mice. Successful transplantation rate was 73%. Specific CTLs (1 x 10(6)), which were activated by DCs combined with MAGE-1 nonapeptide, were injected into the site of transplanted tumor (group A, n = 5). Another group of 17 mice were treated with same amounts of different kinds of cells, and they were divided into groups B, C, D, E, and F. The growth of tumor was observed, and pathological examination was also done.

RESULTS(1) The activated lymphocytes induced by DCs combined with MAGE-1 nonapeptide could suppress the growth of tumor and reduce the tumor size. In group A, 5/5 mice survived for at least two weeks, while the tumors grew rapidly and the majority of the mice died within two weeks in other groups (groups B, C, D, E, F) (P < 0.01). (2) Extensive necrosis and apoptosis were found in transplanted tumors in group A.

CONCLUSIONSThe DCs combined with MAGE-1 nonapeptide could not only inhibit the growth of HCC, but also result in produce death and apoptosis of HCC, hence preventing tumor metastasis and recurrence. The mechanism underlying tumor immunization resulted from DCs might be enhanced in apoptosis of tumor cells. MAGE-1 nonapeptide combined with DCs might be a potential novel tumor vaccine for the treatment of HCC.

Animals ; Antigens, Neoplasm ; Apoptosis ; Cancer Vaccines ; immunology ; Dendritic Cells ; immunology ; Humans ; Liver Neoplasms, Experimental ; immunology ; pathology ; therapy ; Melanoma-Specific Antigens ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Proteins ; administration & dosage ; genetics ; Neoplasm Transplantation ; T-Lymphocytes, Cytotoxic ; immunology ; Transplantation, Heterologous