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Journal of Clinical Microbiology, July 2002, p. 2459-2465, Vol. 40, No. 7
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.7.2459-2465.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Cross-Reactivity among Several Recombinant Calicivirus Virus-Like Particles (VLPs) with Monoclonal Antibodies Obtained from Mice Immunized Orally with One Type of VLP

Noritoshi Kitamoto,1 Tomoyuki Tanaka,2 Katsurou Natori,3 Naokazu Takeda,3 Shuji Nakata,4 Xi Jiang,5,{dagger} and Mary K. Estes6*

School of Humanities for Environmental Policy and Technology, Himeji Institute of Technology, Hyogo 670-0092,1 Sakai Institute of Public Health, Sakai 590-0953,2 Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640,3 Department of Pediatrics, Sapporo Medical University School of Medicine, Sapporo 060-8543, Japan,4 Center for Pediatric Research, Eastern Virginia Medical School, Children's Hospital of The King's Daughters, Norfolk, Virginia 23510,5 Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas 770306

Received 17 August 2001/ Returned for modification 18 November 2001/ Accepted 28 April 2002

Human caliciviruses (HuCVs) are classified into the Norwalk-like viruses (NLV) and Sapporo-like viruses (SLV) as genera within the family Caliciviridae. The NLV genus is further classified into genogroups I and II, based on sequence similarities. To study the antigenic determinants on the HuCV capsid protein and develop new diagnostic tools for field samples, we established and characterized monoclonal antibodies (MAbs) against baculovirus-expressed recombinant HuCV virus-like particles (VLPs). Hybrid clones producing MAbs were obtained from cultures of PAI myeloma cells fused with spleen or mesenteric lymph node cells from mice immunized orally with either a single type of recombinant Norwalk virus (rNV), Kashiwa 47 virus (rKAV), Snow Mountain agent (rSMA), or Sapporo virus (rSV) VLP or with mixtures of two types of VLPs from different genogroups. Twenty MAbs, obtained as mouse ascites, were characterized and classified into six groups according to their enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB) cross-reactivity patterns to VLPs. Five groups of MAbs reacted by both WB and ELISA and were classified as follows: common cross-reactive MAbs for four genogroup I and six genogroup II VLPs (group A), genogroup I-specific MAbs (group B), genogroup II-specific MAbs (group C), and strain-specific MAbs (groups D and E). One MAb group (group F) reacted only by ELISA. The group A MAbs, which showed broad cross-reactivity with VLPs of both NLV genogroups, were obtained from mice immunized orally with a single type of VLP (either rNV or rKAV). Two MAbs, which were obtained from mice immunized with rSV, reacted with rSV but not with any NLV VLP. These are the first MAbs to be reported for any SLV. These strain-, genogroup-, and genus-reactive MAbs will be useful tools for further study of the antigenic and structural topography of the HuCV virion and for diagnostic assays for HuCVs.

* Corresponding author. Mailing address: Department of Molecular Virology and Microbiology, Mailstop BCM-385, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX 77030-3498. Phone: (713) 798-3585. Fax: (713) 798-3586. 8552. Fax: (858) 410-8622. E-mail: mestes{at}bcm.tmc.edu.

{dagger} Present address: Department of Pediatrics, University of Cincinnati, Cincinnati, OH 45229.

Journal of Clinical Microbiology, July 2002, p. 2459-2465, Vol. 40, No. 7
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.7.2459-2465.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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