Yu Xiao Fei, Liang Lin Hui, She Ming, Liao Xiao Long, Gu Jie, Li Yan Han, Han Zhong Chao
State Key Laboratory of Experimental Hematology, Institute of Hematology, Chinese Academy of Medical Sciences & Peking Union Medical College, 288 Nanjing Road, Tianjin 300020, PR China.
Immunol Lett. 2005 Sep 15;100(2):177-81. doi: 10.1016/j.imlet.2005.03.015.
Severe acute respiratory syndrome (SARS) is a highly infectious disease caused by a novel coronavirus (SARS-CoV). Specific monoclonal antibodies (mAbs) against the SARS-CoV are vital for early diagnosis and pathological studies of SARS. Direct intrasplenic inoculation of plasmid DNA encoding antigen is an effective and fast approach to generate specific mAb when the protein antigen is difficult to prepare or dangerous in use. In this study, we selected one fragment of SARS-CoV spike protein (S1-(3)) as antigenic determinant by immunoinformatics. Single intrasplenic immunization of plasmid DNA encoding S1-(3) induced anti-spike protein antibodies. We established one hybridoma cell line secreting specific mAb and evaluated this mAb with murine leukemia virus pseudotyped with SARS-CoV spike protein (MLV/SARS-CoV). The mAb could recognize the spike protein on the MLV/SARS-CoV-infected Vero E6 cells albeit with no neutralizing effect on the infectivity of the pseudotype virus. Our results show that a single-shot intrasplenic DNA immunization is efficient for the production of specific mAb against SARS spike protein, and a linear epitope of the spike protein is recognized in this study.
严重急性呼吸综合征(SARS)是一种由新型冠状病毒(SARS-CoV)引起的高传染性疾病。针对SARS-CoV的特异性单克隆抗体(mAb)对于SARS的早期诊断和病理研究至关重要。当蛋白质抗原难以制备或使用危险时,直接脾内接种编码抗原的质粒DNA是产生特异性mAb的有效且快速的方法。在本研究中,我们通过免疫信息学选择了SARS-CoV刺突蛋白的一个片段(S1-(3))作为抗原决定簇。编码S1-(3)的质粒DNA单次脾内免疫诱导产生了抗刺突蛋白抗体。我们建立了一个分泌特异性mAb的杂交瘤细胞系,并用携带SARS-CoV刺突蛋白的鼠白血病病毒假型(MLV/SARS-CoV)对该mAb进行了评估。该mAb能够识别MLV/SARS-CoV感染的Vero E6细胞上的刺突蛋白,尽管对假型病毒的感染性没有中和作用。我们的结果表明,单次脾内DNA免疫对于产生针对SARS刺突蛋白的特异性mAb是有效的,并且在本研究中识别出了刺突蛋白的一个线性表位。