ABSTRACT
<p><b>BACKGROUND AND OBJECTIVE</b>Cytokine-induced killer (CIK) cells and autologous dendritic cells-CIK (DC-CIK) cells co-cultured with autologous dendritic cells (DCs) and CIK cells are commonly used for immunotherapy recently. We compared the anti-tumor immune response of CIK cells, autologous DC-CIK cells, and semi-allogeneic DC-CIK cells to explore a more effective anti-tumor adoptive immunotherapy approach.</p><p><b>METHODS</b>Peripheral monocytes were isolated from patients with renal carcinoma, lung cancer, or maxillary squamous cell carcinoma and their healthy adult children. Isolated cells were cultured and induced as DCs and CIK cells in vitro. CIK cells from patients were co-cultured with autologous DCs and DCs from their children respectively, generating DC-CIK cells and semi-allogeneic DC-CIK cells. The anti-tumor activities of autologous CIK cells, autologous DC-CIK cells, and semi-allogeneic DC-CIK cells were measured by LDH assay. Intracellular staining was used to test the secretion of cytokines. Flow cytometry was applied for detecting the phonotype changes of these three types of cells. Cell proliferation and cell apoptosis were detected by 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) and Annexin V/PI respectively.</p><p><b>RESULTS</b>Compared with autologous CIK cells and DC-CIK cells, semi-allogeneic DC-CIK cells significantly enhanced the anti-tumor activity and IFN-gamma secretion, reduced IL-4 secretion, increased the ratio of CD3(+)CD56(+) cells and CD3(+)CD8(+) cells, decreased the number of CD4(+)CD25(+) cells, promoted cell proliferation, and lessened cell apoptosis.</p><p><b>CONCLUSIONS</b>Semi-allogeneic DC-CIK cells had a stronger anti-tumor effect than did autologous CIK cells and DC-CIK cells. Our results provided experimental evidence for clinical application of DC-CIK cells.</p>
Subject(s)
Humans , Apoptosis , Carcinoma, Squamous Cell , Metabolism , Pathology , Cell Line, Tumor , Cell Proliferation , Cells, Cultured , Coculture Techniques , Cytokine-Induced Killer Cells , Cell Biology , Allergy and Immunology , Metabolism , Cytokines , Metabolism , Cytotoxicity, Immunologic , Dendritic Cells , Cell Biology , Allergy and Immunology , Metabolism , Hep G2 Cells , Immunotherapy, Adoptive , Interferon-gamma , Bodily Secretions , Interleukin-4 , Bodily Secretions , K562 Cells , Kidney Neoplasms , Metabolism , Pathology , L-Lactate Dehydrogenase , Metabolism , Lung Neoplasms , Metabolism , Pathology , Maxillary Neoplasms , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate whether dendritic cells fused with tumor cells could elicit in vitro antitumor responses against renal cell carcinoma (RCC) cells.</p><p><b>METHODS</b>Renal carcinoma cells were purified from tumor tissue excised from patients with metastatic RCC through tumor cell purifying technique and cultured in RPMI-1640 medium containing 10% FCS. Monocyte-derived DCs generated from peripheral blood mononuclear cell of RCC patients were cultured in the presence of human recombinant granulocyte-macrophage colony stimulating factor and interleukin-4. Tumor cells and DCs were cocultured in the presence of polyethylene glycol (PEG) to generate cell fusion. The phenotype of tumor cells, DCs and fusion cells were detected by flow cytometry. MTT was used to measure the ability of fusion cells to stimulate T cell proliferation. T cell-mediated antitumor responses were measured by lactate dehydrogenase release (LDH) assay for lysis of autologous tumor cells.</p><p><b>RESULTS</b>The DCs expressed MHC class I, MHC class II and costimulatary molecules (CD80 and CD86), while the renal carcinoma cells expressed a high molecular glycoprotein MUC-1. The DC/tumor fusion cells coexpressed MUC-1 and the phenotype of DCs, and could stimulate T cell proliferation effectively. CTLs stimulated by the fusion vaccine showed distinct lytie activity in vitro to autologous tumor cells.</p><p><b>CONCLUSION</b>Dendritic cells fused with tumor cells can elicit distinct antitumor responses in vitro against tumor cells from patients with metastatic RCC, providing a basis for further research on the clinical application of fusion vaccine in treatment for renal cancers.</p>
Subject(s)
Humans , B7-2 Antigen , Metabolism , Cancer Vaccines , Allergy and Immunology , Carcinoma, Renal Cell , Allergy and Immunology , Metabolism , Pathology , Cell Fusion , Cell Proliferation , Coculture Techniques , Cytotoxicity, Immunologic , Allergy and Immunology , Dendritic Cells , Cell Biology , Allergy and Immunology , Metabolism , Hybrid Cells , Cell Biology , Allergy and Immunology , Metabolism , Kidney Neoplasms , Allergy and Immunology , Metabolism , Pathology , Mucin-1 , Metabolism , T-Lymphocytes , Allergy and Immunology , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Tumor Cells, CulturedABSTRACT
<p><b>OBJECTIVE</b>To analyze the allelic loss of heterozygosity (LOH) in the region of chromosome 4p15.1-4q12 in nasopharyngeal carcinoma (NPC) patients with family history.</p><p><b>METHODS</b>Tumor cells and lymphocytes were obtained from paraffin-embedded biopsied tissue section by microdissection. LOH detections were carried out on 25 NPC patients with family history by PCR-based microsatellite polymorphism analysis using 7 pairs of microsatellite markers primers. The microsatellite loci located in 4p15.1-4q12 region. Genescan software was used to analyse LOH at each locus.</p><p><b>RESULTS</b>Ninety-two percent of NPC cases (23/25) with family history was showed at least one microsatellite marker of LOH. Higher frequencies of LOH were found at three loci: D4S238 (56%), D4S350 (50%), D4S1547 (50%). The minimal common region of deletion might be defined between D4S350 and D4S1547.</p><p><b>CONCLUSION</b>The higher incidence of LOH at D4S350 and D4S1547 suggests that there may be a potential tumor suppressor gene located in the two regions.</p>