Buus S, Colon S, Smith C, Freed J H, Miles C, Grey H M
Proc Natl Acad Sci U S A. 1986 Jun;83(11):3968-71. doi: 10.1073/pnas.83.11.3968.
The binding of 125I-labeled immunogenic peptides to purified Ia molecules in detergent solution was examined by equilibrium dialysis. We used the chicken ovalbumin peptide ovalbumin-(323-339)-Tyr, which is immunogenic in the BALB/c mouse and restricted to I-Ad. 125I-labeled ovalbumin-(323-339)-Tyr was shown to bind to I-Ad but not to I-Ed, I-Ek, or I-Ak. This binding was inhibited by unlabeled ovalbumin-(323-339) but not by ovalbumin-(329-339), which is the longest N-terminally truncated peptide that fails to stimulate any of the I-Ad-restricted hybridomas that have been raised to ovalbumin-(323-339)-Tyr. As a further specificity control, we also used the chicken egg lysozyme peptide Tyr-(46-61), which has recently been studied by similar methods [Babbitt, B. P., Allen, P. M., Matsueda, G., Haber, E. & Unanue, E. R. (1985) Nature (London) 317, 359-361]. We have confirmed that it bound to I-Ak but not to I-Ek, I-Ad, or I-Ed. Thus, a specific interaction between Ia and antigen that correlates with the major histocompatibility complex restriction was demonstrated, strongly arguing in favor of a determinant selection hypothesis for such restriction.
通过平衡透析法检测了去污剂溶液中125I标记的免疫原性肽与纯化的Ia分子的结合。我们使用了鸡卵清蛋白肽卵清蛋白-（323 - 339）-酪氨酸，它在BALB/c小鼠中具有免疫原性且受I-Ad限制。结果显示，125I标记的卵清蛋白-（323 - 339）-酪氨酸可与I-Ad结合，但不与I-Ed、I-Ek或I-Ak结合。未标记的卵清蛋白-（323 - 339）可抑制这种结合，而卵清蛋白-（329 - 339）则不能，卵清蛋白-（329 - 339）是N端截短最长的肽段，无法刺激任何针对卵清蛋白-（323 - 339）-酪氨酸产生的I-Ad限制的杂交瘤细胞。作为进一步的特异性对照，我们还使用了鸡卵溶菌酶肽酪氨酸-（46 - 61），最近用类似方法对其进行了研究[巴比特，B.P.，艾伦，P.M.，松枝田，G.，哈伯，E. & 乌纳纽，E.R.（1985年）《自然》（伦敦）317，359 - 361]。我们已证实它可与I-Ak结合，但不与I-Ek、I-Ad或I-Ed结合。因此，证实了Ia与抗原之间存在与主要组织相容性复合体限制相关的特异性相互作用，有力地支持了这种限制的决定簇选择假说。
