Li J L, Chen J L, Ouyang M H, Chao S, Peng Y B, Lin L X, Chang W S, Li Y J
Department of Malaria Immunology, First Medical University of PLA, Guangzhou, P.R. China.
J Immunol Methods. 1991 Aug 28;142(1):15-20. doi: 10.1016/0022-1759(91)90287-p.
Anti-idiotypic antibodies are powerful reagents for the study of immunoregulation, and have potential interest as vaccines against tumors and infectious diseases. Three immunization strategies for the production of rat monoclonal anti-idiotope antibodies have been compared in this paper. Male Wistar rats were immunized i.p. and at multiple subcutaneous sites with 750 micrograms of purified monoclonal antibody against Plasmodium falciparum for three times and subsequently boosted by (1) intraperitoneal injection with 750 micrograms of the immunogen, (2) intravenous inoculation with 400 micrograms of the IgG, and (3) intrasplenic immunization with 200 micrograms of the idiotype. With the intraperitoneal boost method, the frequency of hybrids with anti-idiotope activity was 0.3-0.9% with 62.8-85.2% of the seeded wells containing hybrids. In the intravenous boost group, the percentage of hybrids demonstrating anti-idiotope activity increased to 11.0-13.3% with 80.2-97.9% of the hybrid efficiency. When immunized by the intrasplenic boost route, the frequency of anti-idiotope hybrids generated rose to 12.9-16.4% with 82.3-96.6% of the hybrid efficiency. There was no obvious effect of the boost immunizing methods on the generation of rat monoclonal anti-mouse IgG antibodies. These results indicated that the multiple-site immunization followed by intravenous or intrasplenic boost injection was an appropriate immunizing method for the production of monoclonal anti-idiotope antibodies.
抗独特型抗体是研究免疫调节的有力试剂，作为抗肿瘤和传染病的疫苗具有潜在的研究价值。本文比较了三种制备大鼠单克隆抗独特型抗体的免疫策略。雄性Wistar大鼠经腹腔注射和在多个皮下部位注射750微克抗恶性疟原虫纯化单克隆抗体，共免疫三次，随后分别采用以下方法进行加强免疫：（1）腹腔注射750微克免疫原；（2）静脉接种400微克IgG；（3）脾内注射200微克独特型。采用腹腔加强免疫法时，具有抗独特型活性的杂交瘤频率为0.3-0.9%，接种孔中含有杂交瘤的比例为62.8-85.2%。在静脉加强免疫组中，显示抗独特型活性的杂交瘤百分比增至11.0-13.3%，杂交瘤效率为80.2-97.9%。当采用脾内加强免疫途径时，产生的抗独特型杂交瘤频率升至12.9-16.4%，杂交瘤效率为82.3-96.6%。加强免疫方法对大鼠单克隆抗小鼠IgG抗体的产生没有明显影响。这些结果表明，多部位免疫后进行静脉或脾内加强注射是制备单克隆抗独特型抗体的合适免疫方法。
