为研制广谱性禽流感口服疫苗,将4种禽流感病毒特异的基质蛋白2胞外区(matrix protein 2ectodomain,M2e)多肽与黏膜免疫佐剂CTA1-DD串联,原核表达并纯化CTA1DD-AVI4M2e融合蛋白。以200μg融合蛋白CTA1DD-AVI4M2e口服免疫BALB/c小鼠,结果...为研制广谱性禽流感口服疫苗,将4种禽流感病毒特异的基质蛋白2胞外区(matrix protein 2ectodomain,M2e)多肽与黏膜免疫佐剂CTA1-DD串联,原核表达并纯化CTA1DD-AVI4M2e融合蛋白。以200μg融合蛋白CTA1DD-AVI4M2e口服免疫BALB/c小鼠,结果显示实验组肠液IgA抗体效价和血清IgG抗体效价比对照组显著升高,血清中特异性IgG抗体效价达约360倍。分割的3段肠样中:第1段浸出液IgA抗体效价约为360倍,第2段约为120倍,第3段约为9 720倍。结果表明,本研究制备的CTA1DDAVI4M2e具有显著口服免疫效果,为后续研究提供了基础。展开更多
The matrix protein 2 (M2) is a preferred target for developing a universal vaccine against the influenza A virus (IAV). This study aimed to develop a method for assessing antibody-dependent cell-mediated cytotoxicity ...The matrix protein 2 (M2) is a preferred target for developing a universal vaccine against the influenza A virus (IAV). This study aimed to develop a method for assessing antibody-dependent cell-mediated cytotoxicity (ADCC) associated with M2-based immunization in mice. We first established a stable cell line derived from mouse lymphoma cells (YAC-1) expressing M2 of H3N2. This cell line, designated as YAC-1-M2, was generated using a second-generation lentiviral tricistronic plasmid system to transduce the M2 gene into YAC-1 cells. The ADCC effect induced by polyclonal antibodies targeting matrix protein 2 ectodomain (M2e) was demonstrated by YAC-1-M2 cell lysis by natural killer cells (NK) derived from mice, in the presence of anti-M2 antibodies obtained from mice immunized with an mRNA vaccine based on M2e. This ADCC effect was found to be stronger compared to the effect induced by monoclonal antibodies (14C2) against M2. Moreover, the ADCC effect was enhanced as the effector-to-target ratio of NK to YAC-1-M2 cells increased. In conclusion, we established a novel method to detect ADCC of M2 of IAV, which paves the way for the development of an M2-based universal vaccine against IAV and an in-depth analysis of its mechanism of broad-spectrum immune protection in mice.展开更多
季节性流感疫苗需每年接种,且保护型别有限,因此需研发一种针对多种毒株的通用流感疫苗。国外已研发出基于流感病毒血凝素(hemagglutinin,HA)茎部、基质蛋白2(matrix protein 2,M2)离子通道的胞外域(M2e)、多部位重组蛋白及合成多肽的...季节性流感疫苗需每年接种,且保护型别有限,因此需研发一种针对多种毒株的通用流感疫苗。国外已研发出基于流感病毒血凝素(hemagglutinin,HA)茎部、基质蛋白2(matrix protein 2,M2)离子通道的胞外域(M2e)、多部位重组蛋白及合成多肽的通用疫苗,且在动物实验及早期临床试验中显示出较好的安全性及免疫原性,但目前均尚未批准上市。本文就不同病毒靶点的通用流感疫苗的研究进展作一综述,为进一步的研发提供理论依据。展开更多
文摘为研制广谱性禽流感口服疫苗,将4种禽流感病毒特异的基质蛋白2胞外区(matrix protein 2ectodomain,M2e)多肽与黏膜免疫佐剂CTA1-DD串联,原核表达并纯化CTA1DD-AVI4M2e融合蛋白。以200μg融合蛋白CTA1DD-AVI4M2e口服免疫BALB/c小鼠,结果显示实验组肠液IgA抗体效价和血清IgG抗体效价比对照组显著升高,血清中特异性IgG抗体效价达约360倍。分割的3段肠样中:第1段浸出液IgA抗体效价约为360倍,第2段约为120倍,第3段约为9 720倍。结果表明,本研究制备的CTA1DDAVI4M2e具有显著口服免疫效果,为后续研究提供了基础。
基金the National Key Research and Development Program of China(2021YFC2300101).
文摘The matrix protein 2 (M2) is a preferred target for developing a universal vaccine against the influenza A virus (IAV). This study aimed to develop a method for assessing antibody-dependent cell-mediated cytotoxicity (ADCC) associated with M2-based immunization in mice. We first established a stable cell line derived from mouse lymphoma cells (YAC-1) expressing M2 of H3N2. This cell line, designated as YAC-1-M2, was generated using a second-generation lentiviral tricistronic plasmid system to transduce the M2 gene into YAC-1 cells. The ADCC effect induced by polyclonal antibodies targeting matrix protein 2 ectodomain (M2e) was demonstrated by YAC-1-M2 cell lysis by natural killer cells (NK) derived from mice, in the presence of anti-M2 antibodies obtained from mice immunized with an mRNA vaccine based on M2e. This ADCC effect was found to be stronger compared to the effect induced by monoclonal antibodies (14C2) against M2. Moreover, the ADCC effect was enhanced as the effector-to-target ratio of NK to YAC-1-M2 cells increased. In conclusion, we established a novel method to detect ADCC of M2 of IAV, which paves the way for the development of an M2-based universal vaccine against IAV and an in-depth analysis of its mechanism of broad-spectrum immune protection in mice.
文摘季节性流感疫苗需每年接种,且保护型别有限,因此需研发一种针对多种毒株的通用流感疫苗。国外已研发出基于流感病毒血凝素(hemagglutinin,HA)茎部、基质蛋白2(matrix protein 2,M2)离子通道的胞外域(M2e)、多部位重组蛋白及合成多肽的通用疫苗,且在动物实验及早期临床试验中显示出较好的安全性及免疫原性,但目前均尚未批准上市。本文就不同病毒靶点的通用流感疫苗的研究进展作一综述,为进一步的研发提供理论依据。
