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Online Journal of Veterinary Research©
Volume 23 (10):977-988, 2019.
Quantitative analysis of toll like receptor 9 mRNA expression by different tissues of buffalo calves (Bubalus bubalis)
A. Manuja, M. Dhingra, S. Kumar, S. Sarkar and B. Kumar.
Central Institute for Research on Buffaloes, Sirsa road, Hisar-125001, Haryana, India
Manuja A, Dhingra M, Kumar S, Sarkar S, Kumar B., Quantitative analysis of toll like receptor 9 mRNA expression by different tissues of buffalo calves (Bubalus bubalis). Onl J Vet Res., 23 (10):977-988, 2019. Toll-like receptors (TLR9) recognize unmethylated CpG dinucleotides in bacterial or viral DNA and trigger innate immune response. Synthetic CpG oligodeoxynucleotides (ODN) mimic the effect of naturally occurring CpG motifs in activation of mammalian innate immune responses through TLR9. The potential of synthetic CpG-ODN as therapeutic agents and vaccine adjuvants has been demonstrated in animal models of infectious diseases, allergy and cancer. Proper targeting of CpG ODN for the induction of innate immune defenses requires an understanding of TLR9 expression by different immune compartments. In the present study, transcriptional expression of TLR9 was studied in variety of lymphoid tissues (mesenteric lymphnode, tonsil and spleen), blood and mucosal tissues (lungs and intestine) of buffalo calves. The abundance of TLR9 in different tissues was determined by normalizing the integrated density values (IDV) of TLR9 expression by that of GAPDH, a housekeeping gene. TLR9 mRNA expression levels were also determined and validated by real-time PCR. All tissues and peripheral blood mononuclear cells (PBMCs) exhibited variable mRNA expression of TLR9. The higher expression of TLR9 was exhibited by lymphoid tissues as compared to PBMCs, lungs and the intestine.
Key words: TLR9, mRNA expression, Buffalo, Real time PCR.