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Application of Long-term cultured Interferon-γ Enzyme-linked Immunospot Assay for Assessing Effector and Memory T Cell Responses in Cattle
Published on: July 11, 2015
Candidate gene promoter polymorphisms and antibody response kinetics in chickens: interferon-gamma, interleukin-2,
H Zhou1, A J Buitenhuis, S Weigend
1Department of Animal Science, Iowa State University, Ames 50011-3150, USA.
Abstract:
An F2 population was produced from mating G0 highly inbred (>99%) males of two MHC-congenic Fayoumi lines with G-B1 Leghorn hens. The F2 population was essentially a full-sibship with the F1 sire line reflecting MHC effect. Adult F2 hens (n = 158) were injected twice with SRBC and whole fixed Brucella abortus (BA). Agglutinating antibody titer at 7 d after primary immunization and mean titer of the final three samples (Days 18, 32, and 63 after the second immunization) were used as parameters for primary and equilibrium phases, respectively. Secondary phase parameters of minimum (Ymin), maximum titers (Ymax) and time needed to achieve minimum (Tmin) and maximum (Tmax) titers were estimated from seven postsecondary titers with a nonlinear regression model. Three candidate genes, interferon-gamma (IFN-gamma), interleukin-2 (IL-2), and immunoglobulin G light chain (IgL) were studied. Primers for the promoter regions were designed from EMBL chicken genomic sequences. Polymorphisms between parental lines were detected by direct sequencing. Polymerase chain reaction-restriction fragment length polymorphism methods were then developed to directly detect the polymorphism. There were significant main effects (P < 0.05, general linear model analysis) of IFN-gamma polymorphism on Ymax of BA antibody and interaction of IFN-gamma by IgL on primary antibody response to SRBC and BA, and on Tmin and Ymin of antibody response to SRBC in F2 offspring of M5.1 grandsires. There were significant main effects of IFN-gamma polymorphism on Tmax of BA and interaction of IFN-gamma by IL-2 on Ymin to SRBC in F2 offspring of M15.2 grandsires. The results suggest that IFN-gamma genes play an important role in chicken primary and secondary antibody response to SRBC and BA antigens, and there exists interaction among genes for antibody production.
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