Lamb J R, Ivanyi J, Rees A D, Rothbard J B, Howland K, Young R A, Young D B
EMBO J. 1987 May;6(5):1245-9. doi: 10.1002/j.1460-2075.1987.tb02360.x.
Two complementary approaches were used to determine the epitope specificity of clonal and polyclonal human T lymphocytes reactive with the 65-kd antigen of Mycobacterium leprae. A recombinant DNA sublibrary constructed from portions of the 65-kd gene was used to map T cell determinants within amino acid sequences 101-146 and 409-526. Independently, potential T cell epitopes within the protein were predicted based on an empirical analysis of specific patterns in the amino acid sequence. Of six peptides that were predicted and subsequently synthesised, two (112-132 and 437-459) were shown to contain human T cell epitopes. This corroborated and refined the results obtained using the recombinant DNA sublibrary. Both of these regions are identical in M. leprae and M. tuberculosis and are distinct from the known B cell epitopes of the 65-kd protein. This combination of recombinant DNA technology and peptide chemistry may prove valuable in analysis of the cellular immune response to infectious agents.
采用两种互补方法来确定与麻风分枝杆菌65-kd抗原发生反应的克隆性和多克隆性人T淋巴细胞的表位特异性。从65-kd基因部分构建的重组DNA亚文库用于绘制氨基酸序列101 - 146和409 - 526内的T细胞决定簇。另外,基于对氨基酸序列中特定模式的经验分析预测了该蛋白内潜在的T细胞表位。在预测并随后合成的六种肽中,有两种(112 - 132和437 - 459)被证明含有人类T细胞表位。这证实并完善了使用重组DNA亚文库获得的结果。这两个区域在麻风分枝杆菌和结核分枝杆菌中是相同的,并且与65-kd蛋白已知的B细胞表位不同。重组DNA技术和肽化学的这种结合可能在分析对感染因子的细胞免疫反应中被证明是有价值的。