Epitope mapping and biological function analysis of antibodies produced by immunization of mice with an inactivated Chinese isolate of severe acute respiratory syndrome-associated coronavirus (SARS-CoV)

被引:22
作者
Chou, THW
Wang, SX
Sakhatskyy, PV
Mboudoudjeck, I
Lawrence, JM
Huang, S
Coley, S
Yang, BA
Li, JM
Zhu, QY
Lu, S
机构
[1] Univ Massachusetts, Sch Med, Dept Med, Lab Nucle Acid Vaccines, Worcester, MA 01605 USA
[2] Acad Mil Med Sci, Inst Microbiol & Epidemiol, Beijing 100071, Peoples R China
关键词
SARS-CoV; monoclonal antibody; epitope mapping; inactivated vaccine;
D O I
10.1016/j.virol.2005.01.035
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Inactivated severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been tested as a candidate vaccine against the reemergence of SARS. In order to understand the efficacy and safety of this approach, it is important to know the antibody specificities generated with inactivated SARS-CoV. In the current study, a panel of twelve monoclonal antibodies (mAbs) was established by immunizing Balb/c mice with the inactivated BJ01 strain of SARS-CoV isolated from the lung tissue of a SARS-infected Chinese patient. These mAbs could recognize SARS-CoV-Infected cells by immunofluorescence analysis (IFA). Seven of them were mapped to the specific segments of recombinant spike (S) protein: six on SI subunit (aa 12-798) and one on S2 subunit (aa 797-1192). High neutralizing titers against SARS-CoV were detected with two mAbs (1A5 and 2C5) targeting at a subdomain of S protein (aa 310-535), consistent with the previous report that this segment of S protein contains the major neutralizing domain. Some of these S-specific mAbs were able to recognize cleaved products of S protein in SARS-CoV-infected Vero E6 cells. None of the remaining five mAbs could recognize either of the recombinant S, N, M, or E antigens by ELISA. This study demonstrated that the inactivated SARS-CoV was able to preserve the immunogenicity of S protein including its major neutralizing domain. The relative ease with which these mAbs were generated against SARS-CoV virions further supports that subunit vaccination with S constructs may also be able to protect animals and perhaps humans. It is somewhat unexpected that no N-specific mAbs were identified albeit anti-NIgG was easily identified in SARS-CoV-infected patients. The availability of this panel of mAbs also provided potentially useful agents with applications in therapy, diagnosis, and basic research of SARS-CoV. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:134 / 143
页数:10
相关论文
共 47 条
[1]   A highly unusual palindromic transmembrane helical hairpin formed by SARS coronavirus E protein [J].
Arbely, E ;
Khattari, Z ;
Brotons, G ;
Akkawi, M ;
Salditt, T ;
Arkin, IT .
JOURNAL OF MOLECULAR BIOLOGY, 2004, 341 (03) :769-779
[2]   Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus [J].
Berry, JD ;
Jones, S ;
Drebot, MA ;
Andonov, A ;
Sabara, M ;
Yuan, XY ;
Weingartl, H ;
Fernando, L ;
Marszal, P ;
Gren, J ;
Nicolas, B ;
Andonova, M ;
Ranada, F ;
Gubbins, MJ ;
Ball, TB ;
Kitching, P ;
Li, Y ;
Kabani, A ;
Plummer, F .
JOURNAL OF VIROLOGICAL METHODS, 2004, 120 (01) :87-96
[3]   The coronavirus spike protein is a class I virus fusion protein: Structural and functional characterization of the fusion core complex [J].
Bosch, BJ ;
van der Zee, R ;
de Haan, CAM ;
Rottier, PJM .
JOURNAL OF VIROLOGY, 2003, 77 (16) :8801-8811
[4]   THE TIME COURSE OF THE IMMUNE-RESPONSE TO EXPERIMENTAL CORONAVIRUS INFECTION OF MAN [J].
CALLOW, KA ;
PARRY, HF ;
SERGEANT, M ;
TYRRELL, DAJ .
EPIDEMIOLOGY AND INFECTION, 1990, 105 (02) :435-446
[5]  
Che Xiao-yan, 2003, Di Yi Jun Yi Da Xue Xue Bao, V23, P640
[6]   Inactivation of the coronavirus that induces severe acute respiratory syndrome, SARS-CoV [J].
Darnell, MER ;
Subbarao, K ;
Feinstone, SM ;
Taylor, DR .
JOURNAL OF VIROLOGICAL METHODS, 2004, 121 (01) :85-91
[7]   RIBONUCLEOPROTEIN OF AVIAN INFECTIOUS-BRONCHITIS VIRUS [J].
DAVIES, HA ;
DOURMASHKIN, RR ;
MACNAUGHTON, MR .
JOURNAL OF GENERAL VIROLOGY, 1981, 53 (MAR) :67-74
[8]  
Fang Li-qun, 2003, Zhonghua Liu Xing Bing Xue Za Zhi, V24, P484
[9]   Molecular characterization of a panel of murine monoclonal antibodies specific for the SARS-coronavirus [J].
Gubbins, MJ ;
Plummer, FA ;
Yuan, XY ;
Johnstone, D ;
Drebot, M ;
Andonova, M ;
Andonov, A ;
Berry, JD .
MOLECULAR IMMUNOLOGY, 2005, 42 (01) :125-136
[10]  
He Li, 2003, Di Yi Jun Yi Da Xue Xue Bao, V23, P1128