Study Of Cytotoxicity And Expansion Of CD3AK Cells From Different Origin

Background and ObjectiveTumor's biological therapy is one of the methods of suppressing or removing tumor's growth through tumor's defense mechanism or the biological pharmaceutics and it has become the focus and front line of tumor's therapy, now. Tumor's biological therapy presented by adoptive cellular immunotherapy (ACI/AIT) has become the fourth model of tumor's treatment following operation, chemotherapy and radiotherapy. ACI/AIT is one of the methods of tumor's biological therapy, which is to inject activated immunized cells, which can kill tumor cells directly or activate immunity effect, to deal with tumor. Now, tumor's biological therapy has been thought one of the effective treatments of malignant tumor with such advantages.LAK (lymphokine activated killer) cells and TIL (tumor infiltrating lymphocyte) cells is one of ACI / AIT. But LAK and TIL cells all were limited to be used because of their obvious multi-system-side-effect, or complex process of separating original cells, or lone time of culturine. CD3Ak cells, which is induced bv anti-CD3 monoclonalantibody and rIL-2, is attaching importance based on it's rapid expansion, long time of living, strong anti-tumor activity and low side -effect. But there is few research on which origin cells is the best choice to obtain rapider speed of expansion, and stronger activity of killing tumor cells, nor does what is the difference of killing activity among different tumor cells line killed by one kind of CDSAk cells, and nor does the killing mechanism. This experiment plans to compare the expansion speed, cytotoxicity between two-origin CD3AK cells, from umbilical cord blood and peripheral blood, and the difference of umbilical cord blood CDSAk cells'cytotoxicity.Material and Methods16 pieces of umbilical cord blood and peripheral blood come from normal healthy fetus' peripheral and healthy adult respectively. Obtain cord blood mononuclear cells (CMNC) and peripheral blood mononuclear cells (PMNC) by density gradient centrifugation, culture and harvest CDSAK cells induced by CDSMcAb and rIL-2, calculate and compare the expansion times of CDSAK cells from two origins. Put CDSAK cells and different tumors cells line together cultured, and compare the cytotoxicity of them with MTT. Observe super-microstructure and changes of tumor cells killed. Process the data using f-test and analysis of variance of repetive metrical data with SPSS 10.0 statistical software, a (equals 0.05) was considered significant test level.Results(l).CDSAK cells' dynamics effect comparison: CDSAK cells from two origins, umbilical cord blood CD3AK cells expansion speed is higher, and there is no significant difference (P>0.05)on the fourth day of culturing. With culture time passing by, umbilical cord blood CDSAK cells' expansion speed is obviously higher than peripheral blood CD3AK cells', and there is significant difference (8d, 12d, 16d, 21d,), (P<0.05). The expansion effect can maintain at least three weeks.(2).Comparison of CDSAK cells' cytotoxicity; put two kinds of CDSAK cells andtumor cells line Ecal09 together cultured, the fourth day's CD3Ak cells shows some cytotoxicity and reachs the highest level at the twelfth day of culturing, umbilical cord blood CD3AK cells' cytotoxicity are higher than peripheral blood CD3AK cells at 8d, 12d, 16d, 2Id, and there are significant difference (P<0.05). Two kinds of CD3AK cells can maintain their cytotoxicity at least three weeks(3).Comparison of cytotoxicity mechanism: perhaps umbilical cord blood CD3AK cells kill tumor cells to necrosis mainly, and peripheral blood CD3AK cells induced tumor cells to apoptosis mainly. And there is significant difference (P<0.01).(4).Umbilical cord blood CD3AK cells' cytotoxicity to different tumor cells(esophageal cancer cells line Ecal09,stomach cancer cells line BGC823, lung cancer cells line A549, and leukemia cells line HL-60): To these tumor cells, the cytotoxicity of umbilical cord blood CD3AK cells were different, there is significant difference between leukemia cells line HL-60's...