Jay Prakash Gupta

Global gene expression profile of peripheral blood mononuclear cells challenged with Theileria annulata in crossbred and indigenous cattle

Bovine tropical theileriosis is an important haemoprotozoan disease associated with high rates of morbidity and mortality particularly in exotic and crossbred cattle. It is one of the major constraints of the livestock development programmes in India and Southeast Asia. Indigenous cattle (Bos indicus) are reported to be comparatively less affected than exotic and crossbred cattle. However, genetic basis of resistance to tropical theileriosis in indigenous cattle is not well documented. Recent studies incited an idea that differentially expressed genes in exotic and indigenous cattle play significant role in breed specific resistance to tropical theileriosis. The present study was designed to determine the global gene expression profile in peripheral blood mononuclear cells derived from indigenous (Tharparkar) and cross-bred cattle following in vitro infection of T. annulata (Parbhani strain). Two separate microarray experiments were carried out each for cross-bred and Tharparkar cattle. The cross-bred cattle showed 1082 differentially expressed genes (DEGs). Out of total DEGs, 597 genes were down-regulated and 485 were up-regulated. Their fold change varied from 2283.93 to -4816.02. Tharparkar cattle showed 875 differentially expressed genes including 451 down-regulated and 424 up-regulated. The fold change varied from 94.93 to -19.20. A subset of genes was validated by qRT-PCR and results were correlated well with microarray data indicating that microarray results provided an accurate report of transcript level. Functional annotation study of DEGs confirmed their involvement in various pathways including response to oxidative stress, immune system regulation, cell proliferation, cytoskeletal changes, kinases activity and apoptosis. Gene network analysis of these DEGs plays an important role to understand the interaction among genes. It is therefore, hypothesized that the different susceptibility to tropical theileriosis exhibited by indigenous and crossbred cattle is due to breed-specific differences in the dealing of infected cells with other immune cells, which ultimately influence the immune response responded against T. annulata infection.

Study on Genetic Variation of Microsatellite and Their Association with Mastitis Occurrence in Crossbred Cattle

In present study, the polymorphism at five microsatellite loci BM302, BM4505, BMS2684, CYP21 and DIK20 were investigated for finding their association with the somatic cell counts (SCC) in crossbred cattle. Total 130 animals were tested using california mastitis test (CMT) and somatic cell count (SCC) to screen animals for mastitis. Total 83 alleles were found in all five microsatellites where 23 alleles (9 in BM302, 3 in BM4505, 2 in BMS2684, 8 in CYP21 and 1 in DIK20) were differing significantly in case-control animals. Total number of genotypes for BM302, BM4505, BMS2684, CYP21 and DIK20 were 148, out of which 11 were differing significantly in case-control animals. The allele size ranges for BM302, BM4505, BMS2684, CYP21 and DIK20 were 146 bp to 166 bp, 212 bp to 252 bp, 82 bp to 108 bp, 180 bp to 228 bp and 164 bp to 196 bp, respectively. The polymorphism information content/heterozygosity/allelic diversity for BM302, BM4505, BMS2684, CYP21 and DIK20 were 0.88/0.92/0.89, 0.91/0.53/0.91, 0.81/0.56/0.82, 0.91/0.65/0.92 and 0.82/0.60/0.84. The genotypes ‘146/158’, ‘148/156’, ‘148/158’, ‘150/164’, ‘152/164’, and ‘152/166’ at BM302, ‘238/238’ at BM4505, ‘186/186’ and ‘196/196’ at CYP21 and ‘174/186’ and ‘180/192’ were differing significantly in case-control animals. It revealed that the information observed in the present study was valuable and it may be helpful for improving mastitis resistance in crossbred cattle.

Expression Profile of CXCL3 Gene in Peripheral Blood Mononuclear Cells Challenged in vitro with Theileria annulata in Crossbred Cattle

Bovine tropical theileriosis is a major haemoprotozoan disease associated with high rates of morbidity and mortality particularly in exotic and crossbred cattle. Recent studies suggest that a number of immune response genes, expressed differentially in exotic and indigenous breeds play an important role in breed specific resistance to tropical theileriosis. In the present study, expression of CXCL3 gene which has chemotactic activity for neutrophils, controls migration and adhesion of monocytes and ultimately mediates its effects on target cells by interacting with a cell surface chemokine receptor called CXCR2 was studied in crossbred cattle. The in vitro experimental result revealed significant difference in CXCL3 gene expression in Theileria annulata challenged peripheral blood mononuclear cells of crossbred animals as compared to healthy controls and a 2.53 fold increase (p < 0.05) was recorded. The results of current study indicate that CXCL3 may be involved in host-pathogen interaction during tropical theileriosis.