Document Detail


Bovine whole-blood culture as a tool for the measurement of endotoxin activities in Gram-negative bacterial vaccines.
MedLine Citation:
PMID:  23465356     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
In order to analyze bovine immune reactions against the Gram-negative bacterial vaccine, bovine whole-blood culture was used to investigate the pro-inflammatory cytokine responses stimulated with lipopolysaccharides (LPS) extracted from Escherichia coli, Salmonella enterica, Pseudomonas aeruginosa, and Klebsiella pneumoniae. We also examined the interaction between LPS and aluminum hydroxide gel for endotoxin activity and pro-inflammatory cytokine responses of whole bovine blood. Alteration in the mRNA concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-10 in whole-blood culture at 4h after stimulation with different doses of LPS was observed and determined by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The mRNA concentrations of TNF-α and IL-1β changed in a dose-dependent manner and differed depending on the type of LPS. Limulus test revealed that endotoxin activity was remarkably reduced when aluminum hydroxide gel was added to LPS. In contrast, the mRNA concentration of TNF-α in whole bovine blood was enhanced by LPS mixed with aluminum hydroxide gel. These results suggest that bovine whole-blood culture can be utilized to detect endotoxin activity of Gram-negative bacterial vaccines. In addition, whole-blood culture offers several advantages, such as ease of performance, few preparation artifacts, and a physiological cell environment, for investigating bovine immune response compared with the Limulus test.
Authors:
Saiki Imamura; Mari Nakamizo; Michiko Kawanishi; Nao Nakajima; Kinya Yamamoto; Mariko Uchiyama; Fumiya Hirano; Hidetaka Nagai; Mayumi Kijima; Ryoyo Ikebuchi; Hirohisa Mekata; Shiro Murata; Satoru Konnai; Kazuhiko Ohashi
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-2-8
Journal Detail:
Title:  Veterinary immunology and immunopathology     Volume:  -     ISSN:  1873-2534     ISO Abbreviation:  Vet. Immunol. Immunopathol.     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-3-7     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8002006     Medline TA:  Vet Immunol Immunopathol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2013 Elsevier B.V. All rights reserved.
Affiliation:
National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1, Tokura, Kokubunji, Tokyo 185-8511, Japan. Electronic address: saiki_imamura@nval.maff.go.jp.
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