PURPOSE: Vaccine strategies utilizing dendritic cells (DCs) to elicit anti-tumor immunity are the subject of intense research. Although we have shown that DCs pulsed with heat-treated tumor lysate (HTL) induced more potent anti-tumor immunity than DCs pulsed with conventional tumor lysate (TL), the underlying molecular mechanism is unclear. In order to explore the molecular basis of this approach and to identify potential antigenic peptides from pancreatic cancer, we analyzed and compared the major histocompatibility complex (MHC) ligands derived from TL- and HTL-pulsed dendritic cells by mass spectrophotometry. MATERIALS AND METHODS: Human monocyte-derived dendritic cells were pulsed with TL or HTL prior to maturation induction. To delineate differences of MHC-bound peptide repertoire eluted from DCs pulsed with TL or HTL, nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (nLC-ESI-MS-MS) was employed. RESULTS: HTL, but not TL, significantly induced DC function, assessed by phenotypic maturation, allostimulation capacity and IFN-gamma secretion by stimulated allogeneic T cells. DCs pulsed with TL or HTL displayed pancreas or pancreatic cancer-related peptides in context of MHC class I and II molecules. Some of the identified peptides had not been previously reported as expressed in pancreatic cancer or cancer of other tissue types. CONCLUSION: Our partial lists of MHC-associated peptides revealed the differences between peptide profiles eluted from HTL-and TL-loaded DCs, implying that induced heat shock proteins in HTL chaperone tumor-derived peptides enhanced their delivery to DCs and promoted cross-presentation by DC. These findings may aid in identifying novel tumor antigens or biomarkers and in designing future vaccination strategies.
Antigens, Neoplasm/*immunology ; Cell Line, Tumor ; Dendritic Cells/*immunology ; Humans ; Pancreatic Neoplasms/*immunology

OBJECTIVETo investigate the role of B7-H1 expression in IL-10 production, the B7-H1 and IL-10 expression levels in pancreatic carcinoma tissues and to analyze the correlation between B7-H1 expression and IL-10 level.

METHODSThe mRNA and protein levels expressions of B7-H1 and IL-10 in 35 cases of pancreatic cancer and corresponding paracarcinoma tissues and 5 cases of normal pancreas tissues were detected by RT-PCR, Western blot and immunohistochemistry respectively.

RESULTSThe findings for the first time provided the evidences that there was a clear trend for B7-H1 and IL-10 expressions to be most highly expressed in carcinoma tissue, intermediately expressed in paracarcinoma tissue, and expressed at the lowest level in normal pancreatic tissue at mRNA and protein levels. Moreover, there were statistically significant differences in B7-H1 and IL-10 expression between pancreatic carcinoma tissues, corresponding paracarcinoma tissues and normal pancreatic tissues at mRNA and protein levels (P < 0.05). Furthermore, the immunohistochemistry indicated that there were high expression levels of B7-H1 (60.5% +/- 12.7%) and IL-10 (65.3% +/- 16.2%) in pancreatic carcinoma tissues while there were no significant expressions in normal pancreatic tissues. Meanwhile, correlation analysis revealed that B7-H1 expression was significant associated with IL-10 level in tumor tissues at mRNA (P = 0.008, r = 0.841) and protein levels (P = 0.007, r = 0.838).

CONCLUSIONSOver-expression of B7-H1 may be responsible for the increasing IL-10 production in pancreatic cancer, which caused reduced immune response to tumor cells and contributed to pancreatic carcinoma escape from immune attack.

Antigens, CD ; immunology ; B7-H1 Antigen ; Humans ; Immune Evasion ; Interleukin-10 ; immunology ; Pancreatic Neoplasms ; immunology

This phase I clinical trial (NCT01935843) is to evaluate the safety, feasibility, and activity of chimeric antigen receptor-engineered T cell (CART) immunotherapy targeting human epidermal growth factor receptor 2 (HER2) in patients with advanced biliary tract cancers (BTCs) and pancreatic cancers (PCs). Eligible patients with HER2-positive (>50%) BTCs and PCs were enrolled in the trial. Well cultured CART-HER2 cells were infused following the conditioning treatment composed of nab-paclitaxel (100-200 mg/m) and cyclophosphamide (15-35 mg/kg). CAR transgene copy number in the peripheral blood was serially measured to monitor the expansion and persistence of CART-HER2 cells in vivo. Eleven enrolled patients received 1 to 2-cycle CART-HER2 cell infusion (median CAR T cell 2.1 × 10/kg). The conditioning treatment resulted in mild-to-moderate fatigue, nausea/vomiting, myalgia/arthralgia, and lymphopenia. Except one grade-3 acute febrile syndrome and one abnormal elevation of transaminase (>9 ULN), adverse events related to the infusion of CART-HER2 cells were mild-to-moderate. Post-infusion toxicities included one case of reversible severe upper gastrointestinal hemorrhage which occurred in a patient with gastric antrum invaded by metastasis 11 days after the CART-HER2 cell infusion, and 2 cases of grade 1-2 delayed fever, accompanied by the release of C-reactive protein and interleukin-6. All patients were evaluable for assessment of clinical response, among which 1 obtained a 4.5-months partial response and 5 achieved stable disease. The median progression free survival was 4.8 months (range, 1.5-8.3 months). Finally, data from this study demonstrated the safety and feasibility of CART-HER2 immunotherapy, and showed encouraging signals of clinical activity.
Aged ; Biliary Tract Neoplasms ; immunology ; therapy ; Female ; Humans ; Immunotherapy, Adoptive ; Male ; Middle Aged ; Pancreatic Neoplasms ; immunology ; therapy ; Receptor, ErbB-2 ; immunology ; Receptors, Chimeric Antigen ; immunology ; T-Lymphocytes ; immunology

OBJECTIVESTo identify HLA-restricted epitope of mucoprotein 4 (MUC4) antigen as a tumor associated antigen of pancreatic ductal adenocarcinoma (PDAC), and to validate its natural presentation in PDAC patient peripheral blood.

METHODSTwo epitope prediction databases (SYFPEITHI and ProPred-I) were used to predict HLA-A*0201 restricted MUC4 epitope, T2 cell assay was used to determine the peptide binding affinity with HLA-A*0201 molecule. Dendritic cells (DCs) were induced from the HLA-A* 0201-positive healthy individuals' peripheral blood mononuclear cells (PBMC). Mature DCs were pulsed with synthesized peptides. Autologous CD8(+) T cells from the HLA-A* 0201 healthy donor were stimulated with the peptide-pulsed DCs as CTL. CTL activity was assessed by lactate dehydrogenase release assay and IFN-γ released by enzyme-linked immunospot assay. Pentamer was synthesized for HLA-A* 0201 restricted epitope P1126, then was used to detect specific CTL in PBMC of PDAC patients.

RESULTSFive candidate HLA-A*0201 epitopes were predicted, LLLGVGTFV (P1125) and LLGVGTFVV (P1126) were determined as the two with more HLA-A*0201 affinity. Mature DCs could be induced from PBMCs. CTL induced by peptide P1126 could lyses T2 cells pulsed with peptide P1126 and HCT-116 cells [MUC4(+), HLA-A2(+)]. The number of CTL induced by peptide P1126 which could secret IFN-γ (130.3 ± 6.6) was obviously higher than that in the negative group. By Pentamer assay, P1126-pentamer and CD8 double positive CTL could be detected in PBMC of PDAC patients with MUC4(+) than patients with MUC4(-), but no significant difference of CTL frequency between patients with HLA-A2(+) and with HLA-A2(-) in MUC4(+) PDAC patients.

CONCLUSIONSTumor associated antigen MUC4-derived HLA-A* 0201-restrictive cytotoxic T lymphocyte (CTL) epitope P1126 can induce CTL reaction. The CTL can secret immunologic active material to induce the specific target cells lysis. P1126 epitope can be naturally presented in PBMC of PDAC patients, but its HLA-restriction may not be perfect.

Antigens, Neoplasm ; immunology ; Cells, Cultured ; Dendritic Cells ; immunology ; Epitopes, T-Lymphocyte ; immunology ; HLA-A Antigens ; immunology ; HLA-A2 Antigen ; immunology ; Humans ; Mucin-4 ; immunology ; Pancreatic Neoplasms ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

With the advances in immunology and molecular biology, new recognition in the pathogenesis, progression, and metastasis of carcinoma have been achieved. Studies on gene therapy for pancreatic carcinoma have been attempted in different ways, such as inhibiting oncogene, activating tumor suppressor gene, inducing apoptosis, applying gene directed enzyme prodrug therapy, and immune activation. New specific target genes and further development of gene technology may bring the break-through in this field.
Animals ; Genes, Tumor Suppressor ; Genetic Therapy ; methods ; Humans ; Immunotherapy ; Pancreatic Neoplasms ; genetics ; immunology ; therapy

OBJECTIVETo verify the obtained immunogenic membrane antigens candidate of pancreatic cancer in the performed research.

METHODSPancreatic cancer cell line SW1990 membrane protein underwent immunoblot with serum IgG purified from clinically collected sera of 66 pancreatic cancer patients. Number 3 and number 8 positive dots of immunoblot were identified by MALDI-TOF mass spectrometry and peptide mass fingerprinting matching. The candidate membrane antigens were further validated in cell lines by RT-PCR, real-time PCR and Western blot, and their different expression level of gene and protein in pancreatic cancer cell lines were contrastly studied.

RESULTSNumber 3 and number 8 positive dots were identified as: voltage-dependent anion channel (VDAC3) and catechol-o-methyltransferase (COMT). RT-PCR, real-time PCR and Western blot showed that gene and protein of VDAC3 and COMT were expressed in the pancreatic cancer cell line SW1990, AsPc and P3 respectively.

CONCLUSIONVDAC3 and COMT might be the candidate immunogenic membrane antigens of human pancreatic cancer, and their gene and protein are differently expressed in the pancreatic cancer cell line SW1990, AsPc and P3.

Antigens, Neoplasm ; analysis ; Cell Line, Tumor ; Humans ; Pancreatic Neoplasms ; immunology ; Proteomics

Autoimmune Diseases ; diagnostic imaging ; immunology ; Female ; Humans ; Immunoglobulin G ; immunology ; Middle Aged ; Pancreatic Neoplasms ; diagnostic imaging ; immunology ; Pancreatitis ; diagnostic imaging ; immunology ; Tomography, X-Ray Computed

Antineoplastic Agents, Immunological ; immunology ; pharmacology ; Cell Line, Tumor ; GPI-Linked Proteins ; antagonists & inhibitors ; immunology ; Humans ; Neoplasm Proteins ; analysis ; immunology ; Pancreatic Neoplasms ; drug therapy ; immunology ; pathology

Prostate stem cell antigen (PSCA) is a cell surface antigen expressed in normal prostate and overexpressed in cancers associated with prostate, bladder and pancreas. The sensitivity of PSCA labeling is higher than PSA in prostate cancer. PSCA can be used in the preparation of protein vaccine and nucleic acid vaccine. Further studies are required to confirm its safety and efficacy as a diagnostic means.
Antigens, Neoplasm ; GPI-Linked Proteins ; Humans ; Immunotherapy ; Male ; Membrane Glycoproteins ; analysis ; genetics ; immunology ; Neoplasm Proteins ; analysis ; genetics ; immunology ; Pancreatic Neoplasms ; diagnosis ; Prostatic Neoplasms ; diagnosis ; Urinary Bladder Neoplasms ; diagnosis

OBJECTIVETo explore the adoptive immunotherapy effect of peripheral gammadeltaT cells in pancreatic cancer nude mice model.

METHODSThirty BALB/c nude mice were inoculated subcutaneously 5 x 10(5) Cap-1 cells to regularly developed hypodermal tumors, and then divided into 3 groups randomly, gammadeltaT cells, alphabetaT cells and control group. 2.5 x 10(6) gammadeltaT cells or alphabetaT cells or 100 microl RPMI-1640 were respectively injected into abdominal cavity of mice, combined with 10(4) U rhIL-2 for 3 times. Tumor volume, the survival rate and anti-carcinogenic rate of three groups were compared.

RESULTSEight control nude mice developed hypodermal tumors, which progressively increased in size, and animals had a mean survival of 88 d. Nine nude mice in gammadeltaT cells group and eight in alphabetaT cells group developed tumors (P > 0.05). Tumor growth was arrested and tumor size was reduced remarkably in gammadeltaT cells group. Mean survival was increased to 113 d with less rate of tumor metastasis and more cases of tumor necrosis in gammadeltaT cells group when compared with alphabetaT cells group and controls.

CONCLUSIONSThe anti-tumor effects of gammadeltaT cells against pancreatic cancer are better than those of alphabetaT cells and control groups, and might be promising in the adoptive immunotherapy of pancreatic cancer.

Animals ; Humans ; Immunotherapy, Adoptive ; methods ; Lymphocytes, Tumor-Infiltrating ; immunology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Pancreatic Neoplasms ; immunology ; pathology ; therapy ; T-Lymphocytes ; immunology