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目的探讨小细胞肺癌(SCLC)抗独特型抗体3F6和其单链抗体(3F6ScFv)诱导体液和细胞免疫应答的能力,以证明其作为抗SCLC疫苗的可行性。方法3F6和3F6ScFv(Ab2)免疫BALB/c小鼠获得抗血清,以ELISA和Westernblot方法分别检测抗血清中Ab3结合SCLC细胞膜表面特异抗原(NCI H128抗原)的能力,用竞争Westernblot检测Ab3与2F7竞争结合NCI H128抗原的能力。以迟发型超敏反应(DTH反应)和小鼠脾脏淋巴细胞增殖实验检测3F6及3F6ScFv诱发细胞免疫应答的潜能。结果ELISA和Westernblot方法均证明,3F6和3F6ScFv免疫同系小鼠所产生的Ab3能特异的与NCI H128抗原相结合,与对照血清相比,差异有统计学意义(P<0.001),且有很强的与2F7(Ab1)竞争结合靶抗原的能力。在DTH反应中,3F6和3F6ScFv所致小鼠足垫肿胀的程度均明显高于对照组(P<0.001)。小鼠脾脏淋巴细胞增殖反应试验证明,用3F6和3F6ScFv免疫的小鼠脾脏淋巴细胞对靶细胞的再次刺激有明显的增殖反应,与阴性肿瘤细胞对照组和阴性抗体对照组相比,差异有统计学意义(P<0.05)。结论抗独特型抗体3F6和3F6ScFv均有模拟SCLC细胞膜表面特异抗原的能力,成功诱导了相应的体液和细胞免疫应答,可作为抗SCLC疫苗进行深入研究。
Objective To investigate the ability of small cell lung cancer (SCLC) anti-idiotypic antibody 3F6 and its single chain antibody (3F6ScFv) to induce humoral and cellular immune responses in order to prove its feasibility as an anti-SCLC vaccine. Methods BALB / c mice were immunized with methods 3F6 and 3F6ScFv (Ab2) to obtain antiserum. The ability of Ab3 to bind specific surface antigen (NCI H128 antigen) of SCLC was detected by ELISA and Western blot respectively. Competition of Ab3 with 2F7 was detected by competitive Western blot The ability to bind to the NCI H128 antigen. The potential of 3F6 and 3F6ScFv-induced cellular immune responses was examined by delayed-type hypersensitivity (DTH) and splenic lymphocyte proliferation in mice. Results Both ELISA and Western blot showed that Ab3 produced by 3F6 and 3F6ScFv immunocompetent mice could specifically bind to NCI H128 antigen, with significant difference (P <0.001) as compared with control serum Of the ability to compete with 2F7 (Abl) for binding to the target antigen. In the DTH reaction, the degree of footpad swelling in 3F6 and 3F6ScFv mice was significantly higher than that in the control group (P <0.001). The mouse splenic lymphocyte proliferation response test demonstrated that the spleen lymphocytes immunized with 3F6 and 3F6ScFv had a significant proliferative response to the re-stimulation of the target cells compared with the negative tumor cell control group and the negative antibody control group, the difference was statistically Significance (P <0.05). Conclusions Both anti-idiotypic antibodies 3F6 and 3F6ScFv have the ability to mimic SCLC-specific cell surface antigen and successfully induced corresponding humoral and cellular immune responses, which can be further investigated as an anti-SCLC vaccine.