Steele J K, Chu N R, Chan A, North J, Levy J G
J Immunol. 1987 Jul 15;139(2):469-75.
A T cell hybridoma (Fd11) has been produced from B10.D2 mice that secretes a putative antigen-specific T suppressor factor (TsF). The TsF is isolable from culture supernatants of Fd11 by affinity purification over columns containing either a monoclonal antibody (B16G) shown previously to be capable of binding murine TsF or ferredoxin (Fd), the nominal antigen to which the Fd11 TsF binds. Specificity of the Fd11 TsF for Fd was established by comparing it to another TsF isolated by us (A10 TsF) in a sandwich ELISA, and by demonstrating the specific reactivity to Fd of the hybridoma in calcium flux studies. The Fd11 affinity-purified TsF was shown to contain two major unique components with m.w. in the region of 80,000 and 35,000 when run on reducing polyacrylamide gels in the presence of sodium dodecyl sulphate. Specific immunosuppressive properties of Fd11 were demonstrated when Fd11 TsF (10 micrograms) was injected i.v. into Fd-primed syngeneic mice at the time of antigen boost. Fd11 TsF specifically and significantly diminished the secondary antibody response to Fd in DBA/2 mice.
已从B10.D2小鼠中制备出一种分泌假定抗原特异性T抑制因子(TsF)的T细胞杂交瘤(Fd11)。通过在含有先前已证明能够结合鼠TsF的单克隆抗体(B16G)或铁氧化还原蛋白(Fd,Fd11 TsF结合的名义抗原)的柱上进行亲和纯化,可从Fd11的培养上清液中分离出TsF。通过在夹心ELISA中将Fd11 TsF与我们分离的另一种TsF(A10 TsF)进行比较,并在钙流研究中证明杂交瘤对Fd的特异性反应性,确定了Fd11 TsF对Fd的特异性。在十二烷基硫酸钠存在下,在还原聚丙烯酰胺凝胶上运行时,Fd11亲和纯化的TsF显示含有两种主要的独特成分,分子量在80,000和35,000左右。当在抗原加强时将Fd11 TsF(10微克)静脉注射到用Fd致敏的同基因小鼠中时,证明了Fd11具有特异性免疫抑制特性。Fd11 TsF特异性且显著地降低了DBA/2小鼠对Fd的二次抗体反应。