用人血型糖蛋白 A 和人红细胞分别免疫 BALB/C 小鼠。用被免疫的 BALB/C 小鼠脾细胞与 SP2/0骨髓瘤细胞融合,在37℃、5%二氧化碳培养箱中孵育10～15天,然后用 OM 和 ON 型指示红细胞分别检测培养上清液的凝集情况,筛选出能分泌高特异性...用人血型糖蛋白 A 和人红细胞分别免疫 BALB/C 小鼠。用被免疫的 BALB/C 小鼠脾细胞与 SP2/0骨髓瘤细胞融合,在37℃、5%二氧化碳培养箱中孵育10～15天,然后用 OM 和 ON 型指示红细胞分别检测培养上清液的凝集情况,筛选出能分泌高特异性和高效价抗 M 和抗 N 抗体的细胞株,并建立了 GM_4H3、GM_4H_4、N_2A_3和 N_2D_(10)细胞株。这些细胞株可持续分泌免疫球蛋白 G 类抗 M、抗 N 抗体。应用这些抗体,通过血凝法、解离法和 ELISA 斑点法,可进行血及血痕的 MN 分型。在血型检验中,优于多克隆抗 M、抗N 血清。展开更多
Bat SARS-Iike coronavirus （SL-CoV） has a genome organization almost identical to that of SARS-CoV, but the N-terminus of the Spike （S） proteins, which interacts with host receptor and is a major target of neutrali...Bat SARS-Iike coronavirus （SL-CoV） has a genome organization almost identical to that of SARS-CoV, but the N-terminus of the Spike （S） proteins, which interacts with host receptor and is a major target of neutralizing antibodies against CoVs, of the two viruses has only 63-64% sequence identity. Although there have been reports studying the overall immunogenicity of SsL, knowledge on the precise location of immunodominant determinants for SSL is still lacking. In this study, using a series of truncated expressed SsL fragments and SsL specific mouse sera, we identified two immunogenic determinants for SSL. Importantly, one of the two regions seems to be located in a region not shared by known immunogenic determinants of the SSARS. This finding will be of potential use in future monitoring of SL-CoV infection in bats and spillover animals and in development of more effective vaccine to cover broad protection against this new group of coronaviruses.展开更多
文摘用人血型糖蛋白 A 和人红细胞分别免疫 BALB/C 小鼠。用被免疫的 BALB/C 小鼠脾细胞与 SP2/0骨髓瘤细胞融合,在37℃、5%二氧化碳培养箱中孵育10～15天,然后用 OM 和 ON 型指示红细胞分别检测培养上清液的凝集情况,筛选出能分泌高特异性和高效价抗 M 和抗 N 抗体的细胞株,并建立了 GM_4H3、GM_4H_4、N_2A_3和 N_2D_(10)细胞株。这些细胞株可持续分泌免疫球蛋白 G 类抗 M、抗 N 抗体。应用这些抗体,通过血凝法、解离法和 ELISA 斑点法,可进行血及血痕的 MN 分型。在血型检验中,优于多克隆抗 M、抗N 血清。
基金funded by the State Key Program for Basic Research Grant (2010CB530100,2011CB504700)special project for infectious diseases(2009ZX10004-109) from the Chinese Ministry of Science and Technology
文摘Bat SARS-Iike coronavirus （SL-CoV） has a genome organization almost identical to that of SARS-CoV, but the N-terminus of the Spike （S） proteins, which interacts with host receptor and is a major target of neutralizing antibodies against CoVs, of the two viruses has only 63-64% sequence identity. Although there have been reports studying the overall immunogenicity of SsL, knowledge on the precise location of immunodominant determinants for SSL is still lacking. In this study, using a series of truncated expressed SsL fragments and SsL specific mouse sera, we identified two immunogenic determinants for SSL. Importantly, one of the two regions seems to be located in a region not shared by known immunogenic determinants of the SSARS. This finding will be of potential use in future monitoring of SL-CoV infection in bats and spillover animals and in development of more effective vaccine to cover broad protection against this new group of coronaviruses.
