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
To explore the significance of cancerous immunoglobulin(Ig)in cancer cell growth,HeLa cervical cancer cells were stably transfected with small interfering RNA(siRNA)that specifically,efficiently and consistently silen...To explore the significance of cancerous immunoglobulin(Ig)in cancer cell growth,HeLa cervical cancer cells were stably transfected with small interfering RNA(siRNA)that specifically,efficiently and consistently silences the expression of heavy chain genes of all immunoglobulin isotypes.This stable cell line was used to examine cell viability,colony formation and tumor growth in athymic nude mice.The results of these experiments indicated that siRNA-mediated knockdown of cancerous Ig inhibited cell growth in vitro and suppressed tumor cell growth in immune-deficient nude mice in vivo.Similarly,this siRNA also inhibited the growth of MGC gastric cancer cells and MCF-7 breast cancer cells.Furthermore,the presence of cancerous Ig specifically reduced antibody-dependent cell-mediated cytotoxicity(ADCC)induced by an anti-human epithelial growth factor receptor(EGFR)antibody in a dose-dependent manner,suggesting that the cancerous Ig-Fc receptor interaction inhibits natural killer cell(or NK cell)effector function.The prevalent expression of Ig in human carcinomas and its capacity to promote growth and inhibit immunity might have important implications in growth regulation and targeted therapy for human cancers.展开更多
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.
基金the Major Program of National Natural Science Foundation of China(39830410)National High Technology Research and Development Program(863)of China(2006AA02A404)CMB project(99665)and National Natural Science Foundation of China(30772465).
文摘To explore the significance of cancerous immunoglobulin(Ig)in cancer cell growth,HeLa cervical cancer cells were stably transfected with small interfering RNA(siRNA)that specifically,efficiently and consistently silences the expression of heavy chain genes of all immunoglobulin isotypes.This stable cell line was used to examine cell viability,colony formation and tumor growth in athymic nude mice.The results of these experiments indicated that siRNA-mediated knockdown of cancerous Ig inhibited cell growth in vitro and suppressed tumor cell growth in immune-deficient nude mice in vivo.Similarly,this siRNA also inhibited the growth of MGC gastric cancer cells and MCF-7 breast cancer cells.Furthermore,the presence of cancerous Ig specifically reduced antibody-dependent cell-mediated cytotoxicity(ADCC)induced by an anti-human epithelial growth factor receptor(EGFR)antibody in a dose-dependent manner,suggesting that the cancerous Ig-Fc receptor interaction inhibits natural killer cell(or NK cell)effector function.The prevalent expression of Ig in human carcinomas and its capacity to promote growth and inhibit immunity might have important implications in growth regulation and targeted therapy for human cancers.
