NHC Key Lab of Reproduction Regulation, Shanghai Engineering Research Center of Reproductive Health Drug and Devices, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Shanghai, China.
Department of Histo-Embryology Genetics and Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Med Virol. 2024 Jan;96(1):e29388. doi: 10.1002/jmv.29388.
The use of precise epitope peptides as antigens is essential for accurate serological diagnosis of viral-infected individuals, but now it remains an unsolvable problem for mapping precise B cell epitopes (BCEs) recognized by human serum. To address this challenge, we propose a novel epitope delimitation (ED) method to uncover BCEs in the delineated human IgG-reactive (HR) antigenic peptides (APs). Specifically, the method based on the rationale of similarities in humoral immune responses between mammalian species consists of a pair of elements: experimentally delineated HR-AP and rabbit-recognized (RR) BCE motif and corresponding pair of sequence alignment analysis. As a result of using the ED approach, after decoding four RR-epitomes of human papillomavirus types 16/18-E6 and E7 proteins utilizing rabbit serum against each recombinant protein and sequence alignment analysis of HR-APs and RR-BCEs, 19 fine BCEs in 17 of 22 known HR-APs were defined based on each corresponding RR-BCE motifs, including the type-specificity of each delimited BCE in homologous proteins. The test with 22 known 16/20mer HR-APs demonstrated that the ED method is effective and efficient, indicating that it can be used as an alternative method to the conventional identification of fine BCEs using overlapping 8mer peptides.
利用精确的表位肽作为抗原对于准确诊断病毒感染个体的血清学至关重要，但目前仍无法确定人类血清中识别的精确 B 细胞表位 (BCE)。为了解决这一挑战，我们提出了一种新的表位界定 (ED) 方法，以揭示已划定的人类 IgG 反应性 (HR) 抗原肽 (AP) 中的 BCE。具体来说，该方法基于哺乳动物种间体液免疫反应相似性的原理，包括一对要素：实验划定的 HR-AP 和兔识别 (RR) BCE 基序以及相应的序列比对分析。使用 ED 方法后，利用针对每个重组蛋白的兔血清解码人乳头瘤病毒 16/18-E6 和 E7 蛋白的四个 RR-表位，并对 HR-AP 和 RR-BCE 进行序列比对分析，根据每个相应的 RR-BCE 基序，在 22 个已知的 HR-AP 中的 17 个中定义了 19 个精细 BCE，包括同源蛋白中每个界定 BCE 的特异性。对 22 个已知的 16/20mer HR-AP 的测试表明，该 ED 方法是有效和高效的，表明它可以替代使用重叠 8mer 肽的传统精细 BCE 识别方法。
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