Dunnick W A, Bach F H
J Immunol. 1977 Jun;118(6):1944-50.
A transfer factor-like activity was prepared by Sephadex G-25 chromatography of immune guinea pig leukocyte lysates. This isolated material leads to antigen-dependent migration inhibition and thymidine uptake by nonimmune lymphoid cells. Tests of the "transfer factor" from guinea pigs immunized to either ovalbumin or bovine gamma-globulin demonstrated the donor specificity of the in vitro activity. The activity is susceptible to heat (56 degrees C), alkali (0.5 M sodium hydroxide), pronase, and phosphodiesterase. The pronase susceptibility is blocked by traysylol, a protease inhibitor; the phosphodiesterase susceptibility is not bocked by traysylol. The guinea pig factor was purified further by alkaline phosphatase treatment. Sephadex G-25 chromatography, and DEAE-cellulose chromatography. The final product, active in vitro, represents about 0.03% of the cellular material absorbing 260 nm light, and contains polymerized amines and phosphate. Gel electrophoresis of the fluram-reactive components suggests a limited heterogeneity of the DEAE-cellulose-purified material. These data are consistent with the active "transfer factor" molecule including both peptide and phosphate-containing components.
通过对免疫豚鼠白细胞裂解物进行葡聚糖凝胶G - 25层析制备了一种类转移因子活性物质。这种分离得到的物质可导致非免疫淋巴细胞出现抗原依赖性迁移抑制和胸腺嘧啶核苷摄取。对用卵清蛋白或牛γ球蛋白免疫的豚鼠的“转移因子”进行的测试表明了体外活性的供体特异性。该活性对热（56℃）、碱（0.5M氢氧化钠）、链霉蛋白酶和磷酸二酯酶敏感。链霉蛋白酶敏感性可被蛋白酶抑制剂抑肽酶阻断；磷酸二酯酶敏感性不能被抑肽酶阻断。豚鼠因子通过碱性磷酸酶处理、葡聚糖凝胶G - 25层析和二乙氨基乙基纤维素层析进一步纯化。最终产物在体外具有活性，约占吸收260nm光的细胞物质的0.03%，并含有聚合胺和磷酸盐。对氟胺反应性成分的凝胶电泳表明二乙氨基乙基纤维素纯化物质的异质性有限。这些数据与活性“转移因子”分子同时包含肽和含磷酸盐成分的情况一致。
