Document Detail

Endotoxin neutralization with rabbit antisera to Escherichia coli J5 and other gram-negative bacteria.
MedLine Citation:
PMID:  3298063     Owner:  NLM     Status:  MEDLINE    
To study the mechanisms of protection against endotoxin challenge offered by antisera to smooth and rough gram-negative organisms, we have developed an assay to quantitate endotoxin neutralization based on inhibition of the Limulus amoebocyte lysate test. Dilutions of different bacterial lipopolysaccharides (LPSs) were incubated with hyperimmune rabbit sera against Escherichia coli O113, E. coli O18, and rough mutants E. coli J5 and Salmonella minnesota Re595 and were then combined with limulus lysate. The gelation reaction induced by LPS in the lysate was monitored spectrophotometrically, and the concentration of LPS resulting in a 50% lysate response was determined and correlated with antibody titers measured by enzyme-linked immunosorbent assay. Antisera to smooth organisms neutralized homologous LPS markedly and heterologous LPSs only minimally relative to neutralization by preimmune serum. Neutralization of homologous LPS occurred immediately without preincubation of serum and LPS. Antisera to rough mutants neutralized more heterologous LPS than did antisera to smooth organisms. However, this heterologous neutralization required preincubation of serum and LPS and did not appear to be correlated with antibody concentrations. We conclude that antisera to LPS rapidly neutralize the biological activity of the homologous LPS, as detected by limulus lysate, and that neutralization is at least in part antibody mediated. Antisera to rough-mutant organisms slowly neutralized the activity of heterologous LPSs, but this effect appeared not to be correlated with concentrations of antibody to the LPS of the rough mutant, as measured by enzyme-linked immunosorbent assay.
H S Warren; T J Novitsky; A Bucklin; S A Kania; G R Siber
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Infection and immunity     Volume:  55     ISSN:  0019-9567     ISO Abbreviation:  Infect. Immun.     Publication Date:  1987 Jul 
Date Detail:
Created Date:  1987-07-29     Completed Date:  1987-07-29     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0246127     Medline TA:  Infect Immun     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1668-73     Citation Subset:  IM    
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MeSH Terms
Antibodies, Bacterial / immunology*
Antibody Specificity
Antigens, Bacterial / immunology*
Dose-Response Relationship, Immunologic
Endotoxins / immunology*
Enzyme-Linked Immunosorbent Assay
Escherichia coli / immunology*
Gram-Negative Bacteria / immunology*
Immunoglobulin G / immunology
Immunoglobulin M / immunology
Limulus Test
Lipopolysaccharides / immunology*
Neutralization Tests
Salmonella / immunology
Time Factors
Grant Support
Reg. No./Substance:
0/Antibodies, Bacterial; 0/Antigens, Bacterial; 0/Endotoxins; 0/Immunoglobulin G; 0/Immunoglobulin M; 0/Lipopolysaccharides

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