| Objective: Study the difference of kill rate at two tumor's cells ,and the changes of CD3AK cell's imrnunoassay after activiation, through the compare of proliferate speed , cytotoxicity between two-origin CD3AK cells,from umbilical cord blood and peripheral blood, and provide the theories ground that CD3AK cells from umbilical cord blood were to be used in adoptive cellular immuno therapy.Materials & Methods : 10 pieces of umbilical cord and peripheral blood come from normal healthy feus' umbilical cord and healthy adult respectively.(1)Obtain cord blood mononuclear cells(CMNC) and peripheral blood (PMNC) by Ficoll-Paque centrifugation.culture and harvest CD3AK cells induceed by different' CD3McAb and same concetration of rIL-2(200U /ml),caculate and compare the proliferation times of CD3AK cells from two origins at different concentration and different time,and select feasible concentration induced factor to further compare the proliferation times at the different time. (2)Put two kinds of CD3AK cells and K562 cells together cultured to compare the cytotoxicity of them in different proportion with MTT,and select feasible proportion to compare the cytotoxicity of two kinds of CD3AK cells act on two kinds of tumor'cells named K562 and Hela (3) Study the characteristic of immunophenotype from two origins at 0 day and 7dJay .?Process the data using t-test and analysis of variance of repeat ive metrical data with SPSS 11.5 statistical software, a (equals 0.05 ) was considered significant test level.? Observe the cells's changes of morphologic under the light microscopy and inverted micoscopy. Results:1. CD3AK cells1 multiplication dynamic comparison: (l)when the concentrate-icn of CD3McAb is 1 u g/ml, the proliferation speed of CD3AK cells is higher than other concentration groupsC/VO.OS except at third day).at the tenth day,the proliferation speed is higer than other time groups (P<0.05).(2)At the same concentration of induce factor,the proliferation speed of two kinds of CD3AK cells attain their peak at the tenth day(f<0.05).2,The Comparison of CD3AK cells' cytotoxicity: (1) At the same concentrate-on of induce factor and cultured time, the cytotoxicity of two kinds ofCD3AK ctj^lls act on leukaemia cells (K562) in 20:1 and 40:1 proportion is higher than oijher concentration groups(P<0.05). (2)Put two kinds of CD3AK cells and two tijmor's cells together cultured respectively at the same concentration of induct factor, the same culture time and same proportion. ?The chart show thatIi iat the tenth day , the cytotoxicity of CD3AK cells from two orgins action K562 arr- ived the highest level(P<0.05). ?Under the same contion, the cytotoxicity of two kinds of CD3AK cells act on K562 cells are all higher than on Hela cells(P<0.05), the cytotoxicity of CB'CD3AK cells act on two ki nds of tumor'cells are all higher than PB' CD3AKcells (f?<0.05).3.1 Comparsion of immunoassay of CD3AK cell's from two origins at differentitime: at Oth day ; the content of CD3 ; CD4 ; CDS' in umbilical cord blood1 CD3-AJK cells are lower than in peripheral blood' CD3AK cells ; but the content ofjCD34+in umbillical cord blood CD3AK. cells is obvious higher than in periph-cM blood.At tenth day; the content of CD3+;CD44;CD8+ in two kinds of CD3-AK cells arc all increased in different degree;and it is so obvious that the content of CD3+;CD4\CD8+ in umbilical blood' CD3AK cells are all close to that in peripheral blood'CD3AK cells.At the same time; the content of CD34+ isn't aijy change. Conclusion:(l|) The proliferation speed and cytotoxicity of umbilical cord blood' CD3AK ctfclls are all higer than peripheral hlood';so umbilical cord blood can be made a.4 CD3AK cells' important organize orgins.(2) Different tumor cells have different sensitivity to AIT therapy; so we should select it's suitable ACL(3) Provide the theories ground that the superiority in the terms of low graft -versus-host disease GVHD);high graft versus leukaemia(GVL); and the function in immunotherapy and haematogenous in adoptive cellular immuno (ACI)thcrapy. |
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