Evaluation of Fc Receptor Gene Variants in Bovine Neutrophils

Mulumebet Worku1, Jamie S. Williams1, Ralph Noble1, Antoine Alston1

Abstract


The objectives of this study were to detect FcR I, FcR II and their sub isoforms in cows, evaluate genetic variation, the impact of IgG binding on transcription of FcγR and secreted levels of phosphorylated Syk.  Neutrophils contain a heterogeneous population of Fc receptors (FcR).   The FcγRs (I, II and sub isoforms) bind specifically to IgG molecules and stimulate effector cells to destroy pathogens by phagocytosis and intracellular killing.  The SYK kinase is critical for FcγR-mediated phagocytosis and signal transduction.  Blood was collected from Holstein-Friesian (n=6) and JerseyXHolstein (n=4) lactating cows at the North Carolina A&T State University dairy farm.  The somatic cell count and packed cell volume were used to evaluate overall health. DNA was isolated from blood stored on FTA cards. Specific primers for FcRI (CD64), FcRII (CD32) and FcRII sub-isoforms were used to amplify Fc receptor genes. The housekeeping gene GAPDH was used to normalize the data. Amplified products were separated on a 1% agarose gel and observed following staining with ethidium bromide. Polymorphisms were evaluated by SSCP analysis.  Neutrophils treated with whole bovine IgG molecules were used for q-PCR. FcRI and FcRIIc were detected in all cows. Packed cell volume remained within the range of a healthy cow. All genes were transcribed.  An increase was observed in the transcription of FcγRII in Jersey X Holstein cows, and a significant increase was observed in the Syk protein expression of Jersey X Holstein cows (p<0.05) when compared to Holstein-Friesian cows.  Heterogeneity in FcγR gene expression may be related to breed differences in cows and impact downstream cellular function following binding of IgG to Fc receptors.  The activating (FcRIIa) and inhibitory (FcRIIb) respectively were detected only in Holstein × Jersey crossbred cows regardless of SCC. Further studies are needed to evaluate the effect of FcR gene polymorphism on the cellular response in cattle. These results have implications for control of inflammatory diseases such as mastitis in cattle.


Keywords


Fc Receptor; Crossbreed; Genetic Variation

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