Localization of the regions on the C-terminal domain of the heavy chain of botulinum toxin A recognized by T lymphocytes and by antibodies after immunization of mice with pentavalent toxoid
Localization of the regions on the C-terminal domain of the heavy chain of botulinum toxin A recognized by T lymphocytes and by antibodies after immunization of mice with pentavalent toxoid
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DOI：
10.3109/08820139709022704
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发表时间：
1997-01-01
影响因子：
2.8
通讯作者：
Atassi, MZ
中科院分区：
文献类型：
--
作者：
Rosenberg, JS;Middlebrook, JL;Atassi, MZ
我们用五价环免疫两个近交系小鼠(BoNTs A、B、C、D和E)后，在A型肉毒神经毒素重链的C-末端区域(H-c)定位了T和/或B细胞(Abs)识别的区域。使用一组覆盖整个H-c结构域(残基855-1296)的合成重叠多肽，我们证明了Balb/c(H-2(D))小鼠的T细胞，用一次环状注射启动，识别残基897-915和939-957中的两个主要区域。经多次环状免疫后，Balb/c小鼠T细胞的识别能力增强，对1261-1279肽的攻击反应良好，对1249-1167肽刺激的反应中等。与Balb/c T细胞不同的是，环状免疫SJL(H-2(S))小鼠的T细胞表现出更复杂的特征，并以大量重叠的多肽响应挑战。然而，在一次环状注射后，897-925、939-957/953-971和1052-1069三个肽是最有效的T细胞刺激因子。环状注射3次后，897-915和1051-1069仍为免疫优势区，而第三区上移至925-943/939-957重叠。897-915肽中的免疫优势表位仅被T细胞识别，因为没有检测到针对该区域的抗体。两个品系小鼠的抗体结合谱非常相似，只显示出很小的数量差异。Balb/c和SJL抗环抗体主要与2177-1195肽结合，其次是869-887/883-902重叠多肽和2275-1296多肽。此外，相当数量的Balb/c抗环抗体与多肽1235-2153结合。与与肽995-1013和1052-1069弱相互作用的Balb/c抗体不同，SJL小鼠的抗环抗体与这两种多肽表现出较强的结合。结果表明，在给定的菌株中，抗环抗体和T细胞识别的区域可能重合，也可能是唯一的B细胞或T细胞决定簇。
We have mapped the regions recognized by T and/or B cells (Abs) on the C-terminal domain (H-c) of the heavy chain of botulinum neurotoxin serotype A (BoNTIA) after immunization of two inbred mouse strains with pentavalent toroid (BoNTs A, B, C, D and E). Using a set of synthetic overlapping peptides, encompassing the entire H-c domain (residues 855-1296), we demonstrated that T cells of Balb/c (H-2(d)) mice, primed with one injection of toroid, recognized two major regions within residues 897-915 and 939-957. After multiple inoculations with toroid, T cells of Balb/c expanded their recognition ability and responded very well to challenge with peptide 1261-1279 and moderately to stimulation with peptide 1249-1167. Unlike Balb/c T cells, those of toroid-primed SJL (H-2(S)) mice exhibited a more complex profile and responded to challenge with a large number of overlapping peptides. After one toroid injection, however, three peptides, 897-925, 939-957/953-971 overlap and 1052-1069, were the most potent T cells stimulators. After three toroid injections, peptides 897-915 and 1051-1069 remained immunodominant while the third region was shifted upstream to 925-943/939-957 overlap. The immunodominant epitope within peptide 897-915 was recognized exclusively by T cells, since no Abs were detected against this region. The Ab binding profiles of the two mouse strains were quite similar, showing only small quantitative differences. Both, Balb/c and SJL anti-toroid Abs displayed strong binding mainly to peptide 2177-1195, followed by peptides 869-887/883-902 overlap and 2275-1296. In addition, a significant amount of Balb/c anti-toroid Abs was bound to peptide 1235-2153. Unlike Balb/c Abs, that interacted weakly with peptides 995-1013 and 1052-1069, the anti-toroid Abs of SJL mice exhibited strong binding toward both peptides. The results showed that, in a given strain, the regions recognized by anti-toroid Abs and T cells may coincide or may be uniquely B or T cell determinants.