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Theses (Ph.D.)
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ThesisItem Open Access EVALUATION OF NUCLEIC ACID-BASED MOLECULES FOR STIMULATION OF TYPE I IFN PATHWAYS IN BUFFALO CELL CULTURE SYSTEM(ICAR-NDRI, KARNAL, 2020) ASHUTOSH VATS; SACHINANDAN, DEInnate immune system is activated upon virus invasion of a host cell by recognizing viral component, such as dsRNA through specific receptors, resulting in the production of type- I IFNs. This provides an antiviral state to the host cell and the neighboring cells. In the present study the immunostimulatory effect of synthetic ribonucleotides based RLRs ligands were evaluated. Fibroblast, monocyte and macrophage cells derived from water Buffalo (Bubalus bubalis) were exposed to a synthetic dsRNA analogue, poly I:C to mimic viral invasion in each cell type. Recognition of poly I:C through cytosolic helicase receptors RIG-I and MDA5 molecule lead to the activation of MAVS-IRF3/7 cascade and the production of antiviral effector molecule like IFNβ and ISGs. Within these different cell types, we identified variability in RLR receptor and IFNβ expression after poly I:C administration. Fibroblasts responded quickly and strongly with IFNβ production, followed by macrophages and monocytes. Despite absolute expression variability among different cell types the expression trend of RLRs pathway genes were similar. Length of poly I:C molecule also influence IFNβ expression. Short (LMW) poly I:C induce stronger IFN-β expression in myeloid (macrophage and monocyte) cells. In contrast, long (HMW) poly I:C preferably elicit higher IFNβ expression in non-myeloid (fibroblast) cell. Therefore, MDA5 and RIG-1 plays an indispensable role in eliciting antiviral response in non- immune (fibroblast) host cell. Thus, stimulation of RLR pathway with suitable and potentially cell-type specific agonist molecules successfully elicit antiviral state in the host cell, with fibroblasts revealed a stronger antiviral state than monocytes and macrophages. Seven short RNA motifs of various length, composition and structure were evaluated that specifically target RIG-I and MDA5. The short RNA motifs successfully elicit type I interferon primarily IFNβ and Interferon Stimulated Genes (ISGs). A 90 nucleotides long 5’ppp-RNA motif with stem loop named C5A were exclusively stimulate highest RLRs as well as IFNβ response. Three more RNA motif (P5A, F3A and P3A) also effectively induced IFNβ higher than poly I:C. A RNA motif of 30 nucleotides, 13% uridine and single stem loop structured, F5B failed to induce IFNβ response. Therefore, these results showed that 5’ppp containing short RNA motifs can be rationally designed to achieve an optimum RLRs-mediated antiviral response against in buffalo fibroblast cells.ThesisItem Open Access Studies on production of cloned embryos using somatic cell nucleus transfer in buffalo.(NDRI, Karnal, 2006) Singh, Birbal; Singla, S.K.ThesisItem Open Access Folate receptors and differential expression of folate regulated developmental genes in Goat Embryos.(ICAR-NDRI, Karnal, 2017) Saini, Sikander; Malakar, D.ThesisItem Open Access Effect of pulsed electromagnetic field on the developmental competence quality and gene expression in Buffalo Embryos(ICAR-NDRI, Karnal, 2017) Sharma, Aditya Kumar; Palta, P.
