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N-glycosylation modification of heat shock protein gp96 affects its immunological function / 生物工程学报
Chinese Journal of Biotechnology ; (12): 4036-4046, 2021.
Artículo en Chino | WPRIM | ID: wpr-921484
ABSTRACT
N-glycosylation modification, one of the most common protein post-translational modifications, occurs in heat shock protein gp96. The purpose of this study is to investigate the effect of N-glycosylation modification on immunologic function of the recombinant gp96 using the mutant gp96 in N-glycosylation sites. Firstly, wild-type and mutant gp96 proteins were expressed by insect expression system and their glycosylation levels were detected. To determine the effect of N-glycosylation on gp96 antigen presentation function, the IFN-γ+ CD8+ T cells in gp96-immunized mice and secretion level of IFN-γ were examined by flow cytometry and ELISA. The ATPase activity of gp96 was further detected by the ATPase kit. Finally, the effect of N-glycosylation on adjuvant function of gp96 for influenza vaccine was investigated in immunized mice. It was found that total sugar content of mutant recombinant gp96 was reduced by 27.8%. Compared to the wild type recombinant gp96, mutations in N-glycosylation sites resulted in decreased antigen presentation ability and ATPase activity of gp96. Furthermore, influenza vaccine-specific T cell levels induced by mutant gp96 as adjuvant were dramatically reduced compared to those by wild type recombinant gp96. These results demonstrate that N-glycosylation modification is involved in regulation of ATPase activity and antigen presentation function of gp96, thereby affecting its adjuvant function. The results provide the technical bases for development of gp96- adjuvanted vaccines.
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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Glicosilación / Vacunas contra la Influenza / Adyuvantes Inmunológicos / Linfocitos T CD8-positivos / Proteínas de Choque Térmico Límite: Animales Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2021 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Glicosilación / Vacunas contra la Influenza / Adyuvantes Inmunológicos / Linfocitos T CD8-positivos / Proteínas de Choque Térmico Límite: Animales Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2021 Tipo del documento: Artículo