抗CD3/CD28单克隆抗体联合PHA对T淋巴细胞活化和增殖的影响

Effect from anti-CD3/CD28 monoclonal antibody combined with PHA in activation and proliferation of T lymphocytes

目的探讨抗人CD3/CD28单克隆抗体联合植物血凝素(PHA)对人外周血T淋巴细胞活化增殖的影响。方法通过Ficoll密度梯度离心法分离健康成人外周血单核细胞后,用CD3免疫磁珠分选T淋巴细胞,抗CD3/CD28单抗、PHA及重组人白介素-2(recombinant human IL-2,rhIL-2)刺激T淋巴细胞。用CCK-8试剂盒(CCK-8 cell counting kit)检测细胞增殖情况,ELISA测定细胞上清液干扰素γ(interferon-γ,IFN-γ)的含量。结果CCK-8结果显示在增殖3 d时,CD3/CD28+PHA组A450 nm值与PHA组、IL-2组与空白组比较,差异有统计学意义(P<0.05);比较各组在6、9、12 d的增殖情况,CD3/CD28+PHA组的A450 nm值与其他各组差异均有统计学意义(P<0.05)。刺激3 d各组分泌的IFN-γ质量浓度是最高的。刺激9 d时,CD3/CD28+PHA组、CD3/CD28组、PHA组上清液中IFN-γ浓度分别为(235.27±27.80)、(189.23±24.79)和(124.66±3.39)pg/mL,CD3/CD28+PHA组上清液中IFN-γ的质量浓度高于其余2组(P<0.05)。结论抗CD3/CD28单抗联合PHA比单用抗CD3/CD28单抗、PHA或rhIL-2能更好地激活T细胞及持久地促使T细胞增殖。

Objective To investigate the effect from anti-CD3/CD28 monoclonal antibody(Anti-CD3/CD28 mAb)combined with phytohemagglutinin(PHA),in activation and proliferation for human peripheral blood T lymphocytes.Methods After peripheral blood mononuclear cells were isolated from healthy adults by Ficoll density gradient centrifugation,T lymphocytes were selected with CD3 MicroBeads,then stimulated with anti-CD3/CD28 mAb,PHA and recombinant human IL-2(rhIL-2).The proliferation of T lymphocytes was detected by CCK-8 and IFN-γin the supernatant of T lymphocytes was measured by ELISA.Results CCK-8 showed that on 3 d of proliferation,A450 nm value of CD3/CD28+PHA group was in a statistical significance(P<0.05)compared with that of PHA group,IL-2 group and blank group,respectively.The proliferation of each group at 6,9 and 12 d,A450 nm value of CD3/CD28+PHA group was in a statistical significance(P<0.05)compared with that of other groups.IFN-γconcentration was the highest in all groups after 3 d.After 9 d stimulation,IFN-γof CD3/CD28+PHA group,CD3/CD28 group and PHA group was(235.27±27.80),(189.23±24.79)and(124.66±3.39)pg/mL respectively,and IFN-γconcentration of CD3/CD28+PHA group was higher than that of other groups(P<0.05).Conclusion Anti-CD3/CD28 mAb combined with PHA could better activate T cells and prolong T cell proliferation than performanc of eanti-CD3/CD28 mAb,PHA or IL-2 alone.