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A EXPERIMENTAL AND CLINICAL PRIMARY APPLIED STUDY ON CD3AK TREATING MALIGNANT TUMOR

Author: KuangZhiPeng
Tutor: LiangAnMin
School: Guangxi Medical University
Course: Oncology ( biological treatment )
Keywords: CD3AK cells Biological characteristics Cytokines
CLC: R73-36
Type: Master's thesis
Year: 2000
Downloads: 46
Quote: 0
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Abstract


, Part of the purpose of the basic experiment: Understanding activated killer cells in cord blood CD3AK (anti-CD3 monoclonal antibody-induced method of, cord blood CD3AK) cells and immune biological characteristics, further proved its anti-tumor mechanism. Methods: anti-CD3 monoclonal antibody (anti-CD3 monoclonal antibody, CD3McAb) and recombinant interleukin-2 (recombinant interleukine-2, rIL-2) and other co-stimulation of human peripheral blood mononuclear cells (peripheral blood mononuclear cells PBMC) to induced CD3AK cell with the alkaline phosphatase bridge-linked immunosorbent enzyme linked immunosorbent assay (APAAP method) analysis CD3AK cells phenotype, tetrazolium salt colorimetric test (MTT assay) detection of K 562 cells the most appropriate effector to target ratios, and observed tumoricidal activity of dynamic change and K 562 A 2780 , H , 7402 tumor cell lines cytotoxic active comparators. Enzyme-linked immunosorbent assay CD3AK with A 2780 H 7402 the total sterility before and after the tumor necrosis factor (TNF)-α, interferon (IFN)-γ dynamic changes in A 2780 , H 7402 apoptotic morphological changes in transmission electron microscope. Results: 1, of CD3AK cell proliferation kinetics observations: CD3AK and simple rIL-2-induced LAK cells induced CD3McAb and rIL-2 costimulatory close in the first four days of amplification culture (P> 0.05), but after With the incremented of time and CD3McAb dose, the amplification factor of CD3AK cells is much larger than LAK cells (P <0.01). CD3AK cells maintained for at least 16 days of growth trends. 2, cord blood CD3AK LAK cells phenotype analysis: CD3AK cells in CD3 , the the CD4 CD8 cell-based heterogeneity of cell populations which CD3 , CD4 , CD8 , were significantly higher than LAK cells (P <0.01). 3, CD3AK cells K;;; killing effect of the cell line is the most suitable concentration ③ North cells with target cells 10:1 effector to target ratio of total sterility tumoricidal activity was significantly lower than 20:1 and 40 BU and 20 * and 40J the two groups, no significant differences, suggesting CD3AK cells K; ; Most suitable cell effector to target ratio of 20: l. Tumoricidal activity CD3AK cells dynamic changes observed results: CD3AK cells with K;;; cells sterile results found that higher tumoricidal activity in cultured to four days CD3AK-cells to 10 Telsda peak . 5 cord blood CD3AK, LAK and NK cells tumoricidal activity * compared with the results: of CD3AK cells K;;;, H; . ; Cytotoxic activity of the cells was significantly higher than LAK and NK cells scared 0.05). 6, CD3AK, LAK and AnM H Nakagawa bite co-culture supernatant cytokines TNF-a, IFN-Y secretion test results: cord blood CD3AK LAK cells of another with A;;; , H;;; 0:1,4 0:1 effector to target ratio of sterile 2 4 h before and after results showed that the role of TNF-a, IFN-Y secretion with a significant difference p <0.01 plant a total of sterile after the H secretion levels were significantly higher than the total sterility before. LAK, cord blood CD3AK the two groups, the secretion of TNF-a level reached the peak time, respectively, for a total of infertility sh, 12h, while the level of secretion of IFN-Y of the two groups in the role after sh reach secretion peak. 7, CD3AK cells induced by A;;; , H,; ; Apoptosis morphological observations: CD3AK cells with A;;; H;; ; Were incubated for 24 hours in the transmission electron microscope under visible tumor cells showed typical morphological features of apoptosis: nucleus pyknosis was Xingyue-like accumulation of chromatin along the nuclear membrane, membrane swelling blistering apoptotic body. Conclusion CD3AK cells easy to induce amplification cytotoxic survive a long time, is a novel anti-tumor immune effector cells better than LAK cells.

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