Demonstration of a correlation between tumor cell H-2 antigen content, immunogenicity, and tumorigenicity using lectin-resistant tumor variants.
Demonstration of a correlation between tumor cell H-2 antigen content, immunogenicity, and tumorigenicity using lectin-resistant tumor variants.
复制标题
使用凝集素抗性肿瘤变体证明肿瘤细胞 H-2 抗原含量、免疫原性和致瘤性之间的相关性。
作者：
J. Dennis;Donaghue Tp;D. A. Carlow;R. Kerbel
摘要对高度恶性的小鼠肿瘤MDAY-D2的四个小麦胚抗凝集素(WGA R)变异体进行了检测，以观察其细胞膜的变化，这些变化可能与前面所述的它们的致瘤性降低有关。注射剂S.C.WGA R变异体MDW1或MDW3的5×106个细胞被正常同基因DBA/2宿主排斥，但它们在裸鼠体内生长迅速并呈进行性增长，提示在免疫活性宿主中强烈的T细胞介导的免疫反应是该变异体被排斥的原因。相比之下，接种10~2MDW4、MDW5或MDAY-D2细胞可导致正常DBA/2小鼠肿瘤的进行性生长。因此，MDW1和MDW3突变体似乎比MDW4、MDW5或MDAY-D2亲本肿瘤更具免疫原性。两条证据表明，事实上，所有WGA R变异体和亲本肿瘤都共享一个共同的肿瘤相关抗原：(A)DBA/2小鼠对MDW1肿瘤细胞的排斥使它们能够抵抗后续MDAY-D2或MDW4细胞的攻击；(B)在4小时51Cr释放试验中检测到了抗肿瘤细胞毒性T淋巴细胞(CTL)活性；(C)从荷瘤小鼠的脾组织中提取51Cr，并用丝裂霉素C处理的肿瘤细胞进行体外再刺激。MDAY-D2和WGA R变异体具有交叉反应，既可作为再刺激阶段的免疫原，又可作为CTL靶标。与致瘤系相比，MDW1和MDW3是更有效的肿瘤特异性CTL刺激因子，在CTL检测中是更好的靶点。同样，用三硝基苯酚修饰的肿瘤细胞刺激同基因的DBA/2脾细胞表明，MDW1和MDW3是三硝基苯特异性反应的更有效的刺激因子，并且比其他WGA R变体或MDAY-D2更容易被三硝基苯定向的CTL裂解。十二烷基硫酸钠对微粒体膜蛋白的分离：聚丙烯酰胺凝胶电泳显示，在致瘤性差的变异体MDW1和MDW3的微粒体中存在两种糖蛋白水平升高的糖蛋白，表观分子量分别为43,000和47,000。这些糖蛋白分别鉴定为H-2D和H-2K。肿瘤细胞对同种异体血清的吸收实验表明，WGA R变异体和MDAY-D2的细胞表面有相似数量的H-2。相反，使用肿瘤细胞内质网和质膜部分的吸收测试，以及用十二烷基硫酸钠：聚丙烯酰胺凝胶电泳法对膜部分的分析，显示出致癌能力差的MDW1和MDW3变种的内质网中H-2水平增加。结果表明，不同肿瘤细胞系的致瘤性与免疫原性和细胞H-2水平呈负相关。
Abstract Four wheat germ agglutinin-resistant (WGA R ) variants of the highly malignant murine tumor MDAY-D2 were examined for membrane alterations which might correlate with their decrease in tumorigenicity described previously. Injection s.c. of as many as 5 × 10 6 cells of the WGA R variants MDW1 or MDW3 was rejected by the normal syngeneic DBA/2 host, but they grew rapidly and progressively in athymic nude mice, suggesting that a vigorous T-cell-mediated immune response was responsible for rejection of the variants in the immunocompetent host. In contrast, an inoculum of as few as 10 2 MDW4, MDW5, or MDAY-D2 cells gave rise to progressively growing metastatic tumors in normal DBA/2 mice. Hence, the MDW1 and MDW3 variants appeared more immunogenic than did the MDW4, MDW5, or MDAY-D2 parent tumor. Two lines of evidence suggested that all of the WGA R variants and parental tumor in fact shared a common tumor-associated antigen; ( a ) rejection of MDW1 tumor cells by DBA/2 mice immunized them against a subsequent challenge of either MDAY-D2 or MDW4 cells; ( b ) antitumor cytotoxic T-lymphocyte (CTL) activity was detected in a 4-hr 51 Cr release assay in spleens taken from tumor-bearing mice and restimulated in vitro with mitomycin C-treated tumor cells. MDAY-D2 and WGA R variants were cross-reactive, both as immunogens at the restimulation stage and as CTL targets. Compared to the tumorigenic lines, MDW1 and MDW3 were more effective stimulators of tumor-specific CTL and were superior targets in the CTL assay. Similarly, stimulation of syngeneic DBA/2 spleen cells with trinitrophenol-modified tumor cells showed MDW1 and MDW3 were more effective stimulators of the trinitrophenyl-specific response and were more readily lysed by trinitrophenyl-directed CTL than were the other WGA R variants or MDAY-D2. Separation of microsomal membrane proteins on sodium dodecyl sulfate:polyacrylamide gel electrophoresis revealed an elevated level of two glycoproteins with apparent molecular weights of 43,000 and 47,000, present in the microsomes of the poorly tumorigenic variants MDW1 and MDW3. The glycoproteins were identified as H-2D and H-2K, respectively. Absorption of alloantiserum by whole tumor cells indicated that the WGA R variants and MDAY-D2 had similar amounts of H-2 on their cell surface. In contrast, absorption tests, using tumor cell endoplasmic reticulum and plasma membrane fractions, as well as analysis of the membrane fractions by sodium dodecyl sulfate:polyacrylamide gel electrophoresis, showed an increased level of H-2 in the endoplasmic reticulum of the poorly tumorigenic MDW1 and MDW3 variants. The results demonstrate an inverse correlation between the tumorigenicity and both the immunogenicity and cellular H-2 levels of the variant tumor cell lines.