检测免疫低下小鼠抗体形成功能的方法比较

Comparison on two methods for detection antibody forming cell function

目的:比较溶血空斑试验(PFC)和微量溶血分光光度法(QHS)检测免疫低下小鼠抗体形成细胞功能的优缺点。方法:BALB/C种小鼠用环磷酰胺(CP)造成免疫低下,分成四个80 mg/kg CP组和四个60 mg/kg CP组,其下均设模型对照组及蛹虫草子实体高、中、低三个剂量组。另设正常小鼠组。30 d后进行PFC和微量QHS。另外用80 mg/kg CP模型对照组及60 mg/kg CP模型对照组小鼠不同脾细胞浓度做微量QHS。结果:环磷酰胺80 mg/kg BW和60 mg/kg BW制造免疫低下小鼠模型成立,蛹虫草子实体高、中、低三个剂量组溶血空斑数和吸光度与CP模型对照组比较差异均无显著性意义,两种检测方法所得的结果一致,且溶血空斑数与吸光度值呈正相关。PFC法显示免疫低下80 mg/kg CP模型对照组和60 mg/kg CP模型对照组小鼠的溶血空斑数仅为正常小鼠的11%和12%,而微量QHS测得的A值则为正常小鼠组的55%和58%,表明两种检测方法存在差异。本研究为明确这一差异的产生原因,对CP模型对照组不同脾细胞浓度进行微量QHS检测,结果显示脾细胞数在1×107-4×107之间时与A值线性关系良好。结论:两种方法均能有效地检测抗体形成细胞的功能,但在抗体形成细胞功能组间有很大差异时,PFC能更直观准确的表现其差异,较微量QHS法优越。

Objective：To compare the two methods of plaque forming cell assay and quantitative hemolysis spectrophotometry for detection of antibody forming cell function of immunocompromised mice.Methods： Model of immunocompromised mice were developed with cyclophosphamide,then were divided to four 80 mg/kg CP groups and four 60 mg/kg CP groups respectively including model control group and high,middle,low dose groups of cordyceps,and the normal mice group was also setup.30 days later,the PFC and QHS were carried out.Used different spleen cell density of cyclophosphamid 80 mg/kg and 60 mg/kg model control mice to detect antibody forming cell function by QHS.Results： Model of immunocompromised mice was successfully setup with 80 mg/kg Bw and 60 mg/kg BW cyclophosphamide.There were no significant difference upon haemolysis barren spot value and A value among four CP groups,and also there was a correlation between the results by using the two methods.However The haemolysis barren spot value of 80 mg/kg BW and 60 mg/kg BW cyclophosphamide immunocompromised model mice group were only 11% and 12% of normal group,and the A vaule were 55% and 58% instead,suggesting the difference between these two method.The result of QHS with different spleen cell density shows that the linear relationship with A value were great when the density were between 1×107 to 4×107.Conclusion： Two methods can detect antibody forming cell function effectively,but when there are huge differences among the groups,PFC will be more intuitive and accurate to perform the differences than QHS.