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A modified method of Jerne's assay was used to determine haemolytic plaque-forming cells (PFC) in mouse spleen. By means of this method we measured the dynamic changes of the immune status in tumor-bearing mice. The tumors used were solid Sarcoma 180 maintained in Tian-jin albino mice, four ascitic tumors including Sarcoma 180 maintained in Tian-jin albino and 615 strain mice, and Hepatoma and Ehrlich carcinoma maintained in C57BL mice. The results. showed that spleen PFC decreased as the tumors progressed,... A modified method of Jerne's assay was used to determine haemolytic plaque-forming cells (PFC) in mouse spleen. By means of this method we measured the dynamic changes of the immune status in tumor-bearing mice. The tumors used were solid Sarcoma 180 maintained in Tian-jin albino mice, four ascitic tumors including Sarcoma 180 maintained in Tian-jin albino and 615 strain mice, and Hepatoma and Ehrlich carcinoma maintained in C57BL mice. The results. showed that spleen PFC decreased as the tumors progressed, and was hardly detected when the tumors were far advanced. The tumor-bearing mice commonly had striking splenomegaly with. decrease in number of small lymphocytes and. increase of 'immature' plasma cells, which might explain the poor immune responses of tumor-bearing mice. 本文应用改良的Jerne氏方法测定小鼠脾脏溶血空斑形成细胞。用此方法测定一株实体瘤(在津白小鼠上移植之肉瘤180)和四株腹水瘤(津白小鼠上移植之肉瘤180,615小鼠上移植之肉瘤180,以及C57BL小鼠上移植之肝癌和艾氏癌)带瘤动物免疫状态的动态变化。结果表明,随着肿瘤进行性生长,小鼠脾脏溶血空斑形成细胞逐渐减少,到晚期几乎不能测得。带瘤小鼠的脾脏普遍增大,病理组织学观察提示带瘤小鼠溶血空斑反应之低下与其脾脏的病理学改变相关。 It has been shown by haemolytic plaque-forming cell assay that there is an immunosuppressive factor(s) with high activity in the cell-free aseitic fluid of Sarcoma 130 maintained in 615 strain mice. The suppression is dose-dependent and becomes apparent only when the aseitic fluid is administered before immunization with sheep red blood cells. The immunosuppressive factor (s) seems partially heat-labile. The serum of the same tumor-bearing mice also possesses a strong immunosuppressive activity.... It has been shown by haemolytic plaque-forming cell assay that there is an immunosuppressive factor(s) with high activity in the cell-free aseitic fluid of Sarcoma 130 maintained in 615 strain mice. The suppression is dose-dependent and becomes apparent only when the aseitic fluid is administered before immunization with sheep red blood cells. The immunosuppressive factor (s) seems partially heat-labile. The serum of the same tumor-bearing mice also possesses a strong immunosuppressive activity. Furthermore, similar immunosuppressions were observed in normal recipient mice treated with cell-free aseitic fluids and sera of mice bearing some other strains of tumor including Hepatoma and Ehrlich carcinoma maintained in C57BL mice and Sarcoma 180 maintained in Tian-jin albino mice. Histopathologic changes of spleens induced by the cell-free aseitic fluid resembled those observed in spleens of tumor-bearing mice. 用溶血空斑试验证明,在615小鼠上移植的肉瘤180的无细胞腹水液中存在着高活性的免疫抑制因子。腹水液的免疫抑制作用与剂量相关,且仅在绵羊红血球抗原致敏前注射才能显示。此免疫抑制因子具有部分热不稳定的性质。肉瘤180带瘤小鼠的血清也有强免疫抑制活性。在C57BL小鼠上移植的肝癌和艾氏癌,在津白Ⅰ号小鼠上移植的另一株肉瘤180的腹水液和带瘤动物血清也有类似的免疫抑制作用。肿瘤无细胞腹水液引起的小鼠脾脏病理组织学变化与带瘤动物脾脏所见相类似。 The spleen cells taken from Balb/ cJ mice 3 days after immunization with PDRBC were fused with NS-1 myelo-ma cells by two different procedures in the presence of PEG.The hybridcells secreting the specific antibody were selected out in the HAT medium by means of the spot test of the culture fluid.(Table 1).The antibody secre-ting hybrid cells were then cloned by ... The spleen cells taken from Balb/ cJ mice 3 days after immunization with PDRBC were fused with NS-1 myelo-ma cells by two different procedures in the presence of PEG.The hybridcells secreting the specific antibody were selected out in the HAT medium by means of the spot test of the culture fluid.(Table 1).The antibody secre-ting hybrid cells were then cloned by either limiting dilution or micromani-pulation methods and a cell line thus obtained was designated as CBH-1.After ten months of culture in vitro up to 69 generations,the ability of this cell line to secret specific monoclonal antibody to PDRBC was not decreased. The CBH-1 cells were also ino-culated into the peritoneal cavity of Balb/cJ or Balb/cCr mice and could be kept in these mice for at least four transplantation generations.The ability of three different agents for inducing the ascitic fluid was compared.As a result,pristane was found to be superior to liquid paraffin and Arlacel A. Attempt was also made to cultureCBH-1 in serum-free medium instead of the conventional medium containing 20% newborn calf serum (NCS).When the serum-free medium was provided with 5γ transferrin per ml,the CBH-1 cells could be maintained just as well as that in DMEM+20% NCS within 15 days (Table 2) The proportion of plaque forming cells after the first cloning was found to be 97.5% and this was increased to 99.8% after second cloning (Table 3).This result implies that the cell popula-tion of the CBH-1 clone was very ho-mogeneous with respect to the secre-tion of the specific antibody to PDR-BC. 以北京鸭红血球为抗原免疫Balb/cJ小鼠,三天后取脾细胞与小鼠NS-1骨髓瘤细胞在PEG作用下,用两种方法进行细胞融合。只有淋巴细胞杂交瘤细胞能在一种选择性培液——HAT液中生长,具有分泌抗体活性的杂交瘤细胞,借助其上清液的斑点试验加以选择。对斑点试验阳性的杂交瘤细胞进行克隆化和扩大培养,得到能专一分泌抗北京鸭红血球单克隆抗体的淋巴细胞杂交瘤,定名为CBH-1。在体外经过69代培养,历时十月余,分泌能力仍未见衰减。体内可传4—5代,腹水中可检出大量分泌抗体。用降植烷、Arlacel A和石蜡油诱发动物的腹水,其量以降植烷最高、石蜡油次之。使用国产石蜡油作为诱发腹水试验似具有一定实用价值。加5γ转铁蛋白至每毫升不含牛血清的DMEM液中,在15天的培养期内,结果至少与加20%小牛血清的培液的结果相似。首次克隆化后的CBH-1细胞中,空斑形成细胞达97.5%,再次克隆化的细胞中,空斑形成细胞达99.8%,证明是均一的细胞群落。本文还就杂交瘤制备过程及细胞稳定性问题进行了讨论。
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