| Part 1 A Elenmental Experimental Study Objective: To probe the immunobiological features of CD3 monoclonal antibody activated killer cells(CD3AK) and its anti-tumor mechanism Methods: CD3 monoclonal antibody (CD3McAb) and recombinant interleukin-2(rIL-2) costimulated human peripheral blood mononeuclear cells (PBMC) for inducing CD3 AK. APAAP and MTT methods were separately used to investigate the phenotype and cytotoxicity of CD3AK against tumor cell lines K562~ A2780 and H7402. ELISA method was adopted to detect the dynamic changes of TN7F ?a IFN ?y secreting levels before and after CD3AK culturing with A2780 and H7402. Morphological change of tumor cells apoptosis was observed by Transmision Electronic Microscope. Results: 1 The proliferative ability of CD3AK cells: The proliferative multiple of CD3AK and LAK were similar at the first 4 days(P>O.05). With time going on and CD3McAb concentration increasing,the proliferative multiple of CD3AK was significantly higher than that of LAK (P0.05). 6~ Detection of ThTF ?cx IFN ?y of CD3AK and LAK supemant before and after culturing with A2780 and H7402: TNF ?a~ IFN ?y secreting levels of group CD3AK and group LAK had a significant difference (P<0.0 1 ).The postculture concentration of TNF ?cx and IFN ?y were markedly higher than that of preculture.The time that got to highest levels for Th4F ?a was separatly 8h( group LAK) and 1 2h(group CD3 AK). But the time that two groups reached to highest concentration for IFN ?y were at 8h postculture. 7~ The morphological changes about A2780 and H7402 apoptosis: Under transmission electronic microscope, there were typical apoptotic cell features, such as: nucleus shrinks, chromatin accumulates under nuclear membrane and taking the shape of star-moon, cell membrane swells, apoptotic bodies,etc. Conclusion: CD3AK cell is easily to be induced and proliferated, 6 and has a strong cytotoxicity against tumor cell lines. It is a new immunoeffect cell better than LAK. |
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