从蜂蜇患者中筛选变应原。
Biopanning of allergens from wasp sting patients.
机构信息
Emergency Department, Taihe Hospital, Shiyan 442000, China.
Department of Pediatrics, Shiyan Maternal and Child Health-Care Hospital, Shiyan 442000, China.
出版信息
Biosci Rep. 2018 Oct 17;38(5). doi: 10.1042/BSR20181113. Print 2018 Oct 31.
OBJECTIVE
Wasp venom is a potentially important natural drug, but it can cause hypersensitivity reactions. The purpose of the present study was to systematically study the epitopes of wasp venom.
METHODS
Using a random 12-peptide phage library, we performed antibody-binding epitope panning on ten serum samples from wasp sting victims at 3 h and 4 days after the sting. The panning epitopes were identified by high-throughput sequencing and matched with wasp venom proteins by BLAST. The panned antibody-binding epitopes were verified by ELISA.
RESULTS
A total of 35 specific potential wasp venom epitopes in 4 days were identified. Amongst them, twelve peptide epitopes were matched with nine wasp venom proteins, namely, vitellogenin precursor, hexamerin 70b precursor, venom carboxylesterase-6 precursor, MRJP5, major royal jelly protein 8 precursor, venom acid phosphatase Acph-1 precursor, phospholipase A2, venom serine protease 34 precursor, and major royal jelly protein 9 precursor. The changes in serum IgM antibodies induced by wasp venom were confirmed by ELISA based on the 12 peptide epitopes.
CONCLUSION
The nine wasp venom proteins are potential allergens, which should be excluded or modified in the potential biomedical applications of wasp venom.
目的
黄蜂毒液是一种潜在的重要天然药物，但它会引起过敏反应。本研究旨在系统研究黄蜂毒液的抗原表位。
方法
采用随机 12 肽噬菌体文库，在蜇伤后 3 小时和 4 天，对 10 例黄蜂蜇伤患者的血清样本进行抗体结合表位淘选。通过高通量测序鉴定淘选表位，并通过 BLAST 将其与黄蜂毒液蛋白相匹配。通过 ELISA 验证淘选的抗体结合表位。
结果
在 4 天内共鉴定出 35 个潜在的黄蜂毒液特异性表位。其中，12 个肽表位与 9 种黄蜂毒液蛋白相匹配，分别为卵黄原蛋白前体、六聚体 70b 前体、毒液羧酸酯酶-6 前体、MRJP5、蜂王浆蛋白 8 前体、毒液酸性磷酸酶 Acph-1 前体、磷脂酶 A2、丝氨酸蛋白酶 34 前体和蜂王浆蛋白 9 前体。通过 ELISA 基于 12 个肽表位证实了黄蜂毒液诱导的血清 IgM 抗体的变化。
结论
这 9 种黄蜂毒液蛋白可能是过敏原，在黄蜂毒液的潜在生物医学应用中应将其排除或修饰。
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