Dorner R W, Gantner G E, Goldschmidt J, Zuckner J, Fudenberg H H
Immunology. 1974 Nov;27(5):781-93.
lymphocyte transformation was used in an attempt to demonstrate hypersensitivity reaction to gamma-globulin in patients with rheumatoid arthritis. Peripheral blood lymphocytes from Latex-positive rheumatoid patients were cultured in TC 199 fluid for 3 days in the presence of autologous gamma-globulin preparations (concentrations of 0.01–11 mg/ml), and the percentage blast formation determined microscopically on 1000 cells. Negative controls and phytohaemagglutinin controls were included. Transformation of 10 per cent or more was considered significant. Whole gamma-globulin obtained by ammonium sulphate precipitation failed to stimulate autologous lymphocyte transformation. IgM and IgG fractions obtained by gel filtration on Sephadex G-200 at pH 4.05 stimulated transformation in five out of ten and eight out of ten autologous lymphocyte cultures. The activities of the IgM and IgG fractions could be distinguished by their specificities (homo- and auto-specificity, respectively). Controls including IgM and IgG fractions from healthy persons, Latex-negative rheumatoids, and patients with other connective tissue diseases in autologous culture and IgG from rheumatoids in homologous culture failed to stimulate transformation. Further fractionation of blasto-genically active IgG fractions on DEAE—cellulose and on goat antihuman IgG coupled to Sepharose, in order to determine if the activity was associated with IgG, was unsuccessful, since most of the activity was lost during the procedures.
采用淋巴细胞转化试验来试图证明类风湿性关节炎患者对γ球蛋白的过敏反应。将来自乳胶阳性类风湿患者的外周血淋巴细胞在含有自体γ球蛋白制剂(浓度为0.01 - 11mg/ml)的TC 199培养液中培养3天,然后在显微镜下测定1000个细胞中的母细胞形成百分比。实验设置了阴性对照和植物血凝素对照。母细胞形成率达到10%或更高被认为具有显著性。通过硫酸铵沉淀获得的全γ球蛋白未能刺激自体淋巴细胞转化。在pH 4.05条件下通过Sephadex G - 200凝胶过滤获得的IgM和IgG组分分别在十分之五和十分之八的自体淋巴细胞培养物中刺激了细胞转化。IgM和IgG组分的活性可通过其特异性(分别为同种特异性和自身特异性)来区分。包括来自健康人、乳胶阴性类风湿患者和其他结缔组织病患者的IgM和IgG组分的自体培养对照以及类风湿患者的IgG的同种培养对照均未能刺激细胞转化。为了确定活性是否与IgG相关,将具有母细胞生成活性的IgG组分在DEAE - 纤维素和偶联到琼脂糖的山羊抗人IgG上进一步分级分离,但未成功,因为在这些操作过程中大部分活性丧失了。