Wiegers K J, Uhlig H, Dernick R
J Gen Virol. 1986 Sep;67 ( Pt 9):2053-7. doi: 10.1099/0022-1317-67-9-2053.
In vivo immunization of BALB/c mice with poliovirus type 1, strain Mahoney, or with its purified polypeptides resulted in 0.2 to 0.5 antigen-specific hybridoma microcultures per 10(6) spleen cells. Stimulation of spleen cells from mice immunized with poliovirus or with its polypeptides in vitro with poliovirus 6 days prior to fusion with the myeloma cells led to a six- to 20-fold increase in the number of positive microcultures, i.e. after stimulation the yield of poliovirus-specific hybridomas was up to 3.8 antigen-specific microcultures per 10(6) spleen cells. The in vitro stimulation of spleen cells primed in vivo was demonstrated by the detection of poliovirus-specific antibody-producing cells 6 days after in vitro cultivation in the presence of poliovirus as antigen. Only spleen cells stimulated under these conditions in vitro gave rise to specific antibody-producing cells and yielded antigen-specific hybridomas after somatic hybridization.
用1型脊髓灰质炎病毒Mahoney株或其纯化多肽对BALB/c小鼠进行体内免疫，每10⁶个脾细胞可产生0.2至0.5个抗原特异性杂交瘤微培养物。在与骨髓瘤细胞融合前6天，用脊髓灰质炎病毒体外刺激经脊髓灰质炎病毒或其多肽免疫的小鼠的脾细胞，可使阳性微培养物数量增加6至20倍，即刺激后脊髓灰质炎病毒特异性杂交瘤的产量可达每10⁶个脾细胞3.8个抗原特异性微培养物。在以脊髓灰质炎病毒为抗原进行体外培养6天后，通过检测脊髓灰质炎病毒特异性抗体产生细胞，证明了体内致敏的脾细胞的体外刺激。只有在这些条件下体外刺激的脾细胞才能产生特异性抗体产生细胞，并在体细胞杂交后产生抗原特异性杂交瘤。
