title: A hemolysis-hemagglutination assay for characterizing constitutive innate humoral immunity in wild and domestic birds.
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.
authors.affiliation: Department of Biology, University of Missouri, 8001 Natural Bridge Road, St Louis, MO 63121-4499, USA. kdm50c@umsl.edu
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Matson
authors.forename: Robert E
authors.initials: RE
authors.lastname: Ricklefs
authors.forename: Kirk C
authors.initials: KC
authors.lastname: Klasing
mesh_terms: D000367:Age Factors; D000818:Animals; D000917:Antibody Formation / Q000276:immunology*; D001717:Birds / Q000276:immunology*; D019411:Clinical Laboratory Techniques; D005260:Female; D006386:Hemagglutination Tests; D006461:Hemolysis / Q000276:immunology; D007113:Immunity, Innate / Q000276:immunology*; D008297:Male; D011817:Rabbits