人外周血白细胞处理后CD分子的变化及免疫原性

Variation of CD molecules and immunogenicity in treated human peripheral blood leukocytes

目的检测人外周血中白细胞培养后主要CD分子的变化,验证其作为免疫原制剂的可行性。方法以白细胞滤盘为原料,对比分析白细胞滤盘中淋巴细胞培养前后CD2、CD3、CD4、CD8、CD44、CD45、CD98、CD99、CD107a、CD38、CD25、CD56、CD147、CD50、CD49d、CD82、HLA-ABC共17种CD分子的变化,并与胸腺细胞、Jurkat细胞和外周血细胞进行对比。用培养的淋巴细胞进行免疫原的制备,进行E玫瑰花环抑制试验和淋巴细胞毒试验,制定并验证免疫程序。结果白细胞滤盘中的淋巴细胞培养后,检测的17种CD分子均未发生丢失,其中CD3+、CD3+CD4+、CD3+CD8+阳性率分别从培养前的59.81%、32.73%、22.15%变为98.80%、18.32%、75.31%。免疫原性分析结果显示,第3次免疫后6~15 d血浆抗体的E玫瑰花环抑制试验100%合格,9 d时淋巴细胞毒试验合格率最高。另用10头猪进行了免疫程序的验证,结果显示效价100%合格。结论人外周血白细胞处理后作为免疫原制剂可行。

Objective To verify the changes of main CD molecules in human peripheral blood leukocytes after treatment and applicability for immunogen preparations.Methods Using discarded leukocyte filter discs as raw materials,the variation of 17 main CD molecules(CD2,CD3,CD4,CD8,CD44,CD45,CD98,CD99,CD107a,CD38,CD25,CD56,CD147,CD50,CD49d,CD82,HLA-ABC)were analyzed and compared in leukocyte filter plate lymphocytes before and after culture.Variation of CD molecules in cultured lymphocytes,thymocytes,Jurkat cells and peripheral blood cells were compared.The immunogen was prepared with cultured lymphocytes,and classical E rosette inhibition test as well as complement-mediated lymphotoxicity test were used to evaluate the formulation and verification of the immunization paradigm.Results All 17 CD molecules were retained after culture in lymphocytes from leukocyte filter plate.The CD3+,CD3+CD4+and CD3+CD8+rates changed from 59.81%,32.73%,22.15%to 98.80%,18.32%,and 75.31%,respectively.The results of immunogenicity analysis showed that the qualified rate of E rosette inhibition test of 6-15 d plasma antibody after the third immunization was 100%,and the qualified rate of lymphotoxicity test was the highest at 9 d.Another 10 pigs were used to verify the immunization paradigm,and the qualified rates of titers were 100%.Conclusion Human peripheral blood leukocytes can be used for immunogen preparation after treatment.