Cytokines expression in milk somatic cells during mastitis in cattle and buffaloes
Bhupal, Gramsci (2007) Cytokines expression in milk somatic cells during mastitis in cattle and buffaloes. Masters thesis, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand.
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The present study was carried out to investigate the role of inflammatory cytokines viz. interleukins (IL)-6, IL-8, IL-12, granulocyte macrophage-colony stimulating factor GM-CSF, interferon IFN–γ and tumour necrosis factor TNF-α in immune response of clinical cases of mastitis in HF crossbred cattle (n=4) and indian buffaloes (n=6) and subclinical cases of mastitis in HF crossbred cattle (n=14), Indian buffaloes (n=6) and Kankrej cattle (n=6) using quantitative real time PCR. The mRNA abundance of these target genes was calibrated with that of a reference gene (GAPDH) and expressed as fold of induction over the unstimulated bovine WBCs. Only IL-8, IL-12 and IFN-γ genes were transcribed in unstimulated WBCs in low amounts. All cytokines were up regulated in concavalin- A stimulated, cultured bovine polymorph-nuclear cells which served as positive control. In clinical mastitis all genes were more transcribed than subclinical mastitis except TNF–α, which was seen more in subclinical mastitis in HF crossbreds. In buffaloes the overall expression of cytokines was lower than HF crossbreds. IL-8 gene was down regulated in subclinical mastitis in buffaloes and IFN-γ gene was observed to be down regulated in both clinical and subclinical mastitis in buffaloes. In Kankrej, the expression of cytokines was significantly higher in subclinical stages than the expression of cytokines in other breeds. When Pearson`s correlation coefficient was determined between cytokine transcripts and SCC, it revealed correlation between SCC and transcriptional activity of IL-6, IL-8 and TNF-α in HF crossbreds during subclinical mastitis. The transcriptional activity of IL-8 was found to be correlated with GMCSF and IFN-γ in HF crossbreds. Transcriptional activities of IFN-γ were observed to be correlated with that of GM-CSF in buffaloes during clinical mastitis and in Kankrej cattle during subclinical mastitis. In Kankrej cattle, IL-6 and TNF-α activity were found to be correlated with each other during subclinical mastitis. The findings of this study indicated the distinguishable pattern of cytokine transcription during clinical and subclinical stages of mastitis as well as breed and species variation in cattle and buffalo. Specific preventive and therapeutic strategies are required for each type of mastitis. Understanding the inflammatory responses elicited by these cytokines is fundamental to developing such strategies. This study may form the basis for the development of interventions that can mimic the cytokine response that leads to eradication of intramammary pathogens.
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