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目的:研究CD86嵌合抗体ch1D1对天然高表达CD86分子的人恶性B淋巴瘤细胞株Raji增殖抑制及杀伤效应。方法:经流式细胞术检测ch1D1对Raji细胞表面CD86分子的识别;经ch1D1(终浓度10μg/ml)处理Raji细胞,MTT法检测对Raji细胞增殖的抑制效应;流式细胞术检测ch1D1诱导Raji细胞表面FasL及Fas的表达;MTT法检测ch1D1介导的外周血单个核细胞(PBMCs)对Raji为靶细胞的ADCC效应。结果:ch1D1可特异性识别Raji细胞表面CD86分子,阳性结合率为97.5%。ch1D1对Raji细胞增殖的抑制率为27.1%,较鼠源性亲本抗体1D1组(抑制率为38.8%)略下降,但两者统计学比较无差异(P>0.05)。与CD80嵌合抗体ch4E5联合应用抑制率可达56.3%,明显高于ch1D1单独作用的抑制率(P<0.05)。ch1D1作用4小时后,Raji细胞表达FasL水平开始升高,处理72小时阳性表达率为6.8%,较亲本抗体1D1组(阳性表达率3.2%)升高,但统计学比较无显著差异(P>0.05);经ch1D1作用24小时,Raji细胞Fas表达水平开始上调,阳性率为6.1%,较1D1组(阳性表达率4.8%)升高,但无统计学差异(P>0.05)。与ch4E5联合应用均较ch1D1单独作用Fas及FasL的表达水平升高,但统计学比较仍无显著差异(P>0.05)。经ch1D1处理4、24、48、72小时,介导PBMC对Raji细胞杀伤率分别为25.0%、52.0%、71.3%、72.7%,与人IgG1及鼠源亲本抗体对照组相比均明显升高,具有显著差异(P<0.05)。结论:抗人CD86嵌合抗体可抑制高表达CD86分子的肿瘤细胞增殖并诱导其凋亡,与嵌合抗体Fc段介导的ADCC效应具有协同抗肿瘤作用。
AIM: To investigate the inhibitory and cytotoxic effects of CD86 chimera antibody ch1D1 on Raji proliferation in human malignant B lymphoma cell line with high expression of CD86. Methods: The expression of CD86 on the surface of Raji cells was detected by flow cytometry. Raji cells were treated with ch1D1 (final concentration 10μg / ml), the inhibitory effect on Raji cells was detected by MTT assay, and the effect of ch1D1 on Raji cells was analyzed by flow cytometry Cell surface expression of FasL and Fas; MTT assay of chlD1-mediated peripheral blood mononuclear cells (PBMCs) on Raji target cells ADCC effect. Results: ch1D1 could specifically recognize CD86 molecules on the surface of Raji cells. The positive binding rate was 97.5%. The inhibitory rate of ch1D1 on Raji cell proliferation was 27.1%, which was slightly lower than that of murine 1D1 antibody (38.8%), but there was no statistical difference between the two groups (P> 0.05). The inhibitory rate with chimeric antibody ch4E5 was 56.3%, which was significantly higher than that of ch1D1 alone (P <0.05). After 4 hours of ch1D1 treatment, the level of FasL expression in Raji cells began to increase, the positive expression rate of Raji cells after 72 hours treatment was 6.8%, which was higher than that of the parent antibody 1D1 group (positive rate was 3.2%), but there was no significant difference (P> 0.05). After ch1D1 treatment for 24 hours, the expression level of Fas in Raji cells began to increase, the positive rate was 6.1%, which was higher than that in 1D1 group (4.8%), but there was no significant difference (P> 0.05). In combination with ch4E5, the expression of Fas and FasL increased compared with that of ch1D1 alone, but there was no significant difference between the two groups (P> 0.05). The cytotoxicity of PBMC to Raji cells was 25.0%, 52.0%, 71.3% and 72.7% at 4, 24, 48 and 72 hours after ch1D1 treatment, respectively, which were significantly higher than those of human IgG1 and mouse parental antibody control groups , With significant difference (P <0.05). Conclusion: The anti-human CD86 chimeric antibody can inhibit the proliferation and induce apoptosis of CD86 molecule-rich tumor cells, which has a synergistic anti-tumor effect with the ADCC-mediated ADCC effect of the chimeric antibody.