Experimental Studyon Anti－tumor Effect of SplenocytesInduced by Anti－CD3McAb，PHA and IL－2

The proliferation of splenocytes from healthy adults was induced by anti-CD3 McAb,PHA and IL-2.The proliferative capability and anti-tumor activity as well as phenotypes of the splenocytes cultured in different medium systems were studied.The results showed that anti-CD3 McAb and PHA not only enhanced the proliferation of splenocytes induced by IL-2,but also produced synergism if used simultaneously.The expressions of CD4 and Tac of cellular surface markers were increased after splenocytes were induced by anti-CD3 McAb and PHA.The results of anti-tumor activity of LAK cells suggested that PHA had the capability to promote anti-tumor activity of LAK cells by both direct and in direct pathways,but anti-CD3 McAb indirectly promoted anti-tumor activity of LAK cellsby enhanctng splenocyte proliferation.

PHA、抗CD_3单抗、IL-2诱导脾细胞增殖及其抗肿瘤效应的实验研究

本文应用PHA、抗CD_3单抗、IL-2体外诱导正常人脾细胞增殖,并探讨了在不同条件的诱导下脾细胞体外增殖能力和抗肿瘤效应,以及细胞表面标志的表达特征.结果显示,抗CD_3单抗、PHA不仅可增强IL-2诱导脾细胞的增殖作用,且两者合用有协同增强效应.PHA、抗CD_3单抗还可增强体外扩增细胞CD_4和Tas受体的表达.LAK细胞抗肿瘤活性测定结果提示,PHA有直接和间接增强LAK细胞抗肿瘤效应的作用,抗CD_3单抗则可通过增加细胞增殖作用,间接地增强LAK细胞的抗肿瘤活性.

抗抗CD3 ScFv单克隆抗体的制备及鉴定

目的:制备抗抗CD3 ScFv单克隆抗体并研究其生物活性。方法:分离纯化后的抗CD3 ScFv蛋白免疫BALB/c小鼠,采用传统杂交瘤技术制备抗抗CD3 ScFv单克隆抗体;采用ELISA和Western blot鉴定其亚类和抗原结合特异性;采用FACS测定抗抗CD3 ScFv单克隆抗体对Jurkat细胞特异活性。结果:成功筛选出一株能稳定分泌抗抗CD3 ScFv单克隆抗体的杂交瘤细胞株(10B7),其分泌的抗体亚类为IgG1。该抗抗CD3 ScFv单克隆抗体直标后可特异性结合抗CD3 ScFv蛋白和抗CD3抗体。结论:文中所研制的抗抗CD3 ScFv单克隆抗体具有与抗CD3抗体、抗CD3 ScFv蛋白特异结合的活性,在肿瘤导向治疗中抗CD3/抗肿瘤双特异抗体的亲和层析纯化、药代动力学监测等方面具有广泛的应用前景。

可溶性肿瘤抗原和CD3单克隆抗体共同诱导的杀瘤细胞——T-AK细胞

用可溶性肿瘤抗原和CD3单克隆抗体共同刺激外周血单个核细胞(PBMC)产生CD8^+T细胞为主的杀瘤细胞,称其为T-AK细胞。与LAK细胞和CD3-AK细胞比较,T-AK细胞扩增快、低依赖IL-2,培养14天细胞数扩增24倍,比CD3-AK细胞高8倍,比LAK细胞高15倍,并能持续生长21天。T-AK细胞中含CD3^+86％、CD4^+20％、CD8^+95％、CD16^+40％、CD25^+53％,它们是CD8^+T细胞为主的异质性细胞群。T-AK细胞对刺激它的STA来源的靶细胞高亲和性杀伤,杀伤率>90％,对NK敏感和不敏感细胞也有杀伤,表现非MHC限制杀伤。

小鼠CTL体外非特异性扩增对其杀瘤活性的影响

目的 在体外用抗CD3单抗、抗CD2 8单抗和白细胞介素 2 (IL 2 )扩增肿瘤特异性CTL ,为肿瘤过继治疗提供足够数量的、具有高度杀瘤活性的效应细胞。方法 采用两种方案培养肿瘤细胞免疫的小鼠脾细胞。 (1)抗CD3单抗刺激 48h后 ,加入抗CD3单抗和 2 0U mlrIL 2 (抗 CD3+IL 2组 ) ;(2 )抗CD3单抗和抗CD2 8单抗同时刺激 48h后 ,加入抗CD3单抗、抗CD2 8单抗和 2 0U mlrIL 2 (抗 CD3+抗 CD2 8+IL 2组 )。分别检测 2组效应细胞的增殖水平、杀瘤活性及表型。结果 抗 CD3+IL 2组细胞的3H TdR掺入量在第 6天、12天、2 0天分别为 2 2 12 6、5 42 6、2 0 72 ,抗 CD3+抗 CD2 8+IL 2组细胞的3H TdR掺入量在第 6天、12天、2 0天分别为 32 16 8、12 92 2、32 6 5 ,后者明显高于前者。在培养 12d时 ,抗 CD3+IL 2组细胞对FBL 3的最大杀伤活性为 6 6 .4% ,抗 CD3+抗 CD2 8+IL 2组细胞对FBL 3的最大杀伤活性为 77.8%。细胞表型FACS分析结果表明 ,抗 CD3+抗 CD2 8+IL 2组培养 12d后的细胞 90 %以上为Thy1.2 + 细胞 ,且CD2 5 + 细胞在抗 CD3+抗 CD2 8+IL 2组、抗 CD3+IL 2组分别为 2 3.0 0 %、8.15 %。结论 抗CD3单抗、抗CD2 8单抗和低剂量IL 2同时非特异性刺激 ,可获得大量扩增的、具有高度杀瘤活性的肿瘤特异性CTL。