Document Detail


Production and characterization of monoclonal antibodies to IgM of Pacific herring (Clupea pallasii).
MedLine Citation:
PMID:  22771742     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Pacific herring (Clupea pallasii) have a central role in the North Pacific ecosystem as a forage fish species and are natural reservoirs of several important finfish pathogens, including Viral hemorrhagic septicemia virus (VHSV). Here, we report the identification of the gene encoding the immunoglobulin mu (IgM) heavy chain, as well as the development and characterization of monoclonal antibodies (MAbs) that specifically react with Pacific herring IgM. Pacific herring immunoglobulin was purified and consisted of heavy and light chains of approximately 80 and 25 kDa. Three hybridoma clones were initially identified by ELISA as reactive with purified immunoglobulin but only one clone was able to detect an 80 kDa protein in Pacific and Atlantic herring (C. harengus) whole plasma by denaturing western blot. However, all three MAbs were able to precipitate an 80 kDa protein from Pacific herring and LCMS sequencing of peptide fragments derived from this protein matched the predicted amino acid sequence of the cloned, heavy chain gene. In addition, two of the MAbs stained cells within the putative lymphocyte gates for the spleen, anterior kidney and posterior kidney but were not reactive for myeloid/granulocyte gates, which is consistent with these Mabs reacting with surface IgM(+) B-cells. To our knowledge, this is the first report of IgM-related gene sequences and anti-IgM monoclonal antibodies from any member of the family Clupeidae. The antibodies produced in this study are critical for achieving our long-term goal of conducting serological surveillance to assess pathogen exposure in natural populations of Pacific herring.
Authors:
Maureen K Purcell; Erin S Bromage; Jessica Silva; John D Hansen; Samantha M Badil; James C Woodson; Paul K Hershberger
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-7-5
Journal Detail:
Title:  Fish & shellfish immunology     Volume:  -     ISSN:  1095-9947     ISO Abbreviation:  -     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-7-9     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9505220     Medline TA:  Fish Shellfish Immunol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012. Published by Elsevier Ltd.
Affiliation:
US Geological Survey-Western Fisheries Research Center, 6505 NE 65(th) Street, Seattle, WA 98115, USA.
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