Bachmann A E, Pedace E A, Sperperato A M, Conway de Macario E
Instituto de Investigaciones Hematológicas de la Academia Nacional de Medicina, Melo 3081, Buenos Aires, Argentina.
Medicina (B Aires). 1965 Nov-Dec;25(6):353-9.
Immunological modifications in guinea pigs injected with homologous lymph node extracts. In studies carried out in patients with lymphomas an accentuation of the B2M-globulin line was observed; occasionally paraproteins and isoantibodies were detected. For this reason the authors decided to find out whether the injection of homologous lymph node extracts in guinea pigs would produce serum, immunological and histological modifications comparable to the ones observed in the human being. A total of 100 guinea pigs were divided into 4 groups. Group 1, 50 untrated control animals; Group 2, 10 animals injected with complete Freunds adjuvant every 10 days, for 3 times, and sacrificed 7 days after the last injection; Group 3, 10 animals injected with the antigen prepared with the lymph nodes from normal guinea pigs, in the same way as the preceding group; Group 4, 30 animals injected with the same antigens emulsionated with the adjuvant, again by the same method as the 2 preceding groups. The antigen was prepared with the lymph nodes from normal guinea pigs. The cells were "ultrasonicated" and lyophilized; the lipids were extracted with chloroform. The extract thus obtained contained an average of 2 mg/ml of total proteins. A microimmunoelectrophoretic study (MIE) against rabbit immuno serum for guinea pigs anti-serum proteins, showed that it was contaminated with various serum proteins (fig. 1, A). In the 3 injected groups, most of the guinea pigs showed a 10% decrease in serum protein (Table). Using MIE there was a re-enforcement of the B2M-globulin precipitation line (IgM), in all the animals of groups 2 and 4, as compared to the groups 1 and 3 (Table and figure 1. B and C). Using microdiffusion in agar, the serum of 6% of the guinea pigs of group 4 gave precipitation lines with the homologous antigen (Table, figure 1, D). Using passive hemagglutination, between 30 and 40% of the serum of the animals of all 4 groups gave positive results with the same antigen, with a low titer (1/40). Using complement fixation, positive results were obtained with the lymph node antigen in 10% and 20% of the animals of groups 3 and 4 respectively: from 25 to 50% fixation with a titer of 1/10 (Table). Using immunofluorescence, positive images in the cytoplasm of lymphocytes (from improntas of normal guinea pigs lymph nodes) were obtained only in the labeled gamma-globulin (IgG) of the animals of group 4 (Table and figure 2, C). Histological study of the lymph nodes of guinea pigs from groups 2 and 4 showed hyperplasia of the reticular tissue with apparent decrease in the size of the follicles. In some of the lymph nodes of the animals of group 4, giant cells were observed (Fig. 2, B), pironinophilic. In the group 3 hypertrophy of the lymphoid follicles was observed (fig. 2, A). Non-specific modifications of the serum proteins, similar to the ones observed in patients with lymphomas were detected in some of the animals belonging to groups 2 and 3 in a greater number of animals in group 4. Circulating isoantibodies could be compared to those detected in 2 cases of lymphosarcoma. In spite of definite histological alterations of the lymphatic organs, these cannot be compared with the alterations detected in the human being.
注射同源淋巴结提取物的豚鼠的免疫改变。在对淋巴瘤患者进行的研究中,观察到B2M球蛋白谱带增强;偶尔还检测到副蛋白和同种抗体。因此,作者决定研究向豚鼠注射同源淋巴结提取物是否会产生与人类观察到的类似的血清、免疫和组织学改变。总共100只豚鼠被分为4组。第1组,50只未处理的对照动物;第2组,10只动物每10天注射一次完全弗氏佐剂,共注射3次,并在最后一次注射后7天处死;第3组,10只动物注射用正常豚鼠淋巴结制备的抗原,方式与前一组相同;第4组,30只动物注射与佐剂乳化的相同抗原,同样采用与前两组相同的方法。抗原用正常豚鼠的淋巴结制备。细胞经“超声处理”后冻干;脂质用氯仿提取。由此获得的提取物平均含有2mg/ml的总蛋白。针对豚鼠抗血清蛋白的兔免疫血清进行的微量免疫电泳研究(MIE)表明,它被各种血清蛋白污染(图1,A)。在3个注射组中,大多数豚鼠血清蛋白降低了10%(表)。与第1组和第3组相比,使用MIE时第2组和第4组所有动物的B2M球蛋白沉淀线(IgM)均增强(表和图1,B和C)。使用琼脂微量扩散法,第4组6%的豚鼠血清与同源抗原产生沉淀线(表,图1,D)。使用被动血凝法,所有4组动物中30%至40%的血清与相同抗原产生阳性结果,效价较低(1/40)。使用补体结合法,第3组和第4组分别有10%和20%的动物对淋巴结抗原产生阳性结果:25%至50%的结合,效价为1/10(表)。使用免疫荧光法,仅在第4组动物标记的γ球蛋白(IgG)中,在淋巴细胞细胞质中获得了阳性图像(来自正常豚鼠淋巴结的印记)(表和图2,C)。对第2组和第4组豚鼠淋巴结的组织学研究显示网状组织增生,滤泡大小明显减小。在第4组动物的一些淋巴结中,观察到了嗜派洛宁的巨细胞(图2,B)。在第3组中观察到淋巴滤泡肥大(图2,A)。在第2组和第3组的一些动物以及第4组更多的动物中检测到了与淋巴瘤患者中观察到的类似的血清蛋白非特异性改变。循环中的同种抗体可与在2例淋巴肉瘤中检测到的相比。尽管淋巴器官有明确的组织学改变,但这些改变与人类中检测到的改变无法相比。