Betton G R, Dekkers-Bijma A M, Lansdorp P M
J Immunol Methods. 1980;32(2):157-66. doi: 10.1016/0022-1759(80)90069-1.
In order to produce long term lymphoid cell cultures from canine lymphocytes of known histocompatibility antigen specificities, mitogenic responses to staphylococcal protein A (SpA) were examined and compared with those of phytohaemagglutinin (PHA) and concanavalin A (Con A). SpA was found to be the strongest mitogen tested with significant responses to concentrations as low as 31 ng/ml. There was a decrease in responsiveness above optimal mitogen concentrations with SpA and PHA. Peak responses were observed at lower concentrations for longer incubation times. PHA showed a rapid fall off in thymidine uptake below optimal concentrations whereas the SpA dose-response curve was less steep and a shoulder or secondary peak of activity was observed at low SpA concentrations in some cases. Continuous SpA stimulation of lymphocyte cultures resulted in an initial period of cell proliferation followed usually by a second period of cell proliferation around week 7 of culture. To date, viable cell cultures have been maintained for up to 12 weeks in vitro. SpA lymphoblast cultures behave normally in microcytotoxicity tests for serologically defined DLA histocompatibility antigens and remain functional in natural killer (NK) and PHA induced cell mediated cytotoxic reactions against 51Cr-labelled tumour target cells but were not themselves susceptible as target cells for NK activity.
为了从已知组织相容性抗原特异性的犬淋巴细胞中产生长期淋巴细胞培养物，检测了对葡萄球菌蛋白A（SpA）的促有丝分裂反应，并与植物血凝素（PHA）和刀豆球蛋白A（Con A）的促有丝分裂反应进行了比较。发现SpA是所测试的最强有丝分裂原，对低至31 ng/ml的浓度有显著反应。SpA和PHA在最佳有丝分裂原浓度以上时反应性降低。在较低浓度下，较长孵育时间可观察到峰值反应。PHA在低于最佳浓度时胸苷摄取迅速下降，而SpA剂量反应曲线较平缓，在某些情况下，在低SpA浓度下观察到活性肩峰或次级峰。对淋巴细胞培养物进行持续SpA刺激会导致细胞增殖的初始阶段，随后通常在培养第7周左右出现第二个细胞增殖阶段。迄今为止，活细胞培养物已在体外维持长达12周。SpA淋巴母细胞培养物在针对血清学定义的DLA组织相容性抗原的微量细胞毒性试验中表现正常，并且在自然杀伤（NK）和PHA诱导的针对51Cr标记肿瘤靶细胞的细胞介导细胞毒性反应中保持功能，但它们本身作为NK活性的靶细胞并不敏感。
