Function study of the anti⁃infection effects for an anti⁃SraPL⁃Lectin antibody in mouse macrop⁃hages infected by Staphylococcus aureus
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    Abstract:

    Objective:To determin the functions of an anti-SraPL-Lectin monoclonal antibody in the process of phagocytosis and killing Staphylococcus aureus(S.aureus)in the mouse macrophages. Methods:Differernt gene sequence of sraP was amplied by PCR and specific amplification products were inserted into pET28a plasmid. The rpET28a-SraPL-Lectin plasmid was transferred into E.coli.BL21 and induced by 0.1 mmol/L IPTG at 25 ℃. The recombinant protein was purified by nickel column and the specificity of this antibody was detected by Western blot. The expression level of inflammatory factors in macrophages was detected by qPCR. CCK8 assay was carried out to assess the inhibition rate of S.aureus proliferation. The number of S.aureus colonies in the supernatant and lysate of macrophages was counted on the coated plate. Results:Anti-SraPL-Lectin monoclonal antibody could specifically bind to recombinant SraP truncated proteins and cell wall protein of S.aureus. The co-incubation of monoclonal antibody with S.aureus could induce the down-regulation of pro-inflammatory cytokine(TNF-α,IL-1β,IL-12p40)and up-regulation of anti-inflammatory cytokine(IL-10)in macrophages. The proliferation of S.aureus USA300 LAC was obviously inhibited in the co-effect of gentamicin and antibody. Anti-SraPL-Lectin reduced the amount of S.aureus in macrophage supernatant. Conclusion:The immune complex of anti-SraPL-Lectin antibody and S.aureus can alleviate the immune response of macrophages and promote the clearance of macrophages to S.aureus.

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许秀华,郑 峰,岳 岩,汪春晖,周婷婷,胡龙华.抗金黄色葡萄球菌富丝氨酸重复蛋白SraPL⁃Lectin抗体在小鼠巨噬细胞抗感染中的功能研究[J].南京医科大学学报(自然科学版英文版),2019,(4):472-477.

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  • Received:January 29,2019
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  • Online: May 07,2019
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