Ascher M S, Schneider W J, Valentine F T, Lawrence H S
Proc Natl Acad Sci U S A. 1974 Apr;71(4):1178-82. doi: 10.1073/pnas.71.4.1178.
The chief impediment to the precise biochemical identification of transfer factor and its mechanism of action has been the lack of a reproducible in vitro assay. We now report on a method by which dialysates containing transfer factor of proven in vivo potency can convert nonimmune lymphocytes to immune responsiveness in vitro, as reflected by antigen-triggered lymphocyte proliferation. However, "water-dialyzed transfer factor" (TF(D)) prepared for in vivo use by the conventional method of dialysis against large volumes of water exhibits diminished activity in vitro and is frequently toxic to lymphocyte cultures. This problem can be avoided by dialysis of transfer factor into tissue culture medium. When this precaution is taken, such "media-dialyzed transfer factor" (TF(DM)) causes nonimmune lymphocytes to respond to antigen by an increment of thymidine incorporation that ranges from 2 to 25 times that of such cells cultured with antigen alone. This response is generally observed only in the presence of those antigens to which the TF(DM) donor expresses delayed cutaneous reactivity and is distinguishable from nonspecific adjuvant effects.
对转移因子进行精确的生化鉴定及其作用机制的主要障碍一直是缺乏一种可重复的体外检测方法。我们现在报告一种方法,通过该方法,含有经体内效力验证的转移因子的透析液可在体外将非免疫淋巴细胞转化为免疫反应性,这可通过抗原触发的淋巴细胞增殖来反映。然而,通过常规的大量水透析方法制备用于体内的“水透析转移因子”(TF(D))在体外表现出活性降低,并且对淋巴细胞培养物常常有毒性。通过将转移因子透析到组织培养基中可以避免这个问题。采取这种预防措施后,这种“培养基透析转移因子”(TF(DM))会使非免疫淋巴细胞通过胸苷掺入量的增加来对抗原作出反应,其增加幅度是仅用抗原培养的此类细胞的2至25倍。这种反应通常仅在TF(DM)供体对其表现出迟发性皮肤反应性的那些抗原存在时才观察到,并且与非特异性佐剂效应不同。