Article

A hemolysis-hemagglutination assay for characterizing constitutive innate humoral immunity in wild and domestic birds

Department of Biology, University of Missouri, 8001 Natural Bridge Road, St Louis, MO 63121-4499, USA.
Developmental & Comparative Immunology (Impact Factor: 3.71). 02/2005; 29(3):275-86. DOI: 10.1016/j.dci.2004.07.006
Source: PubMed

ABSTRACT Methods to assess immunocompetence requiring only a single sample are useful in comparative studies where practical considerations prevent holding or recapturing individuals. The assay for natural antibody-mediated complement activation and red blood cell agglutination described here, requiring approximately 100 microl of blood, is highly repeatable. The effects of complement deactivation, 2-mercaptoethanol (2-ME), age, and lipopolysaccharide (LPS)-induced sickness response were examined to validate comparisons among diverse avian species. Complement deactivation by heating significantly reduces lysis and treatment with 2-ME reduces both lysis and agglutination. Lysis and agglutination both increase with age in chickens; LPS treatment does not influence these variables in 11-week-old chickens. In a comparison of 11 species, both lysis (0.0-5.3 titers) and agglutination (1.8-8.0 titers) vary significantly among species. Accordingly, this assay can be used to compare constitutive innate humoral immunity among species and with respect to age, sex, and experimental treatments within populations.

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    • "Half scores were recorded when SRBC precipitated partially but not to the extent of control wells. Lysis of erythrocytes was not scored since this process has been shown to be subjective (Matson et al., 2005). "
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    • "To assess the levels of circulating natural antibodies and the activity of the complement system, we used the hemolysis–hemagglutination assay as developed by Matson et al. (2005), with several minor alterations . This assay is based on the interaction of avian plasma samples and rabbit red blood cells, which results in agglutination (HA) and lysis (HL) of the cells. "
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