Browsing by Author "Rabindra Prasad Singh"

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Foot-and-mouth disease virus alters MUC genes expression and function of goblet cells in the intestines of naturally infected cattle calves
(Academic Press, 2025) Monalisa Sahoo; Rajendra K. Singh; Sagar M. Patel; Susen Kumar Panda; A. P. Acharya; Ritun C. Patra; Saminathan M; Vinay Kumar Sd; Rajesh Kumar Vandre; Jigyasa Rana; Mamata Pasayat; Jagannath Prasad Tripathy; N. R. Sahoo; Jitendra Kumar Biswal; Rabindra Prasad Singh
Foot-and-mouth disease (FMD), the “Risk Group 4” animal pathogen, causes devastating economic losses world over. The mechanism of the disease process is still not fully explored despite extensive research work going on. The present work highlights the intestinal pathology with mucin gene expression in 41 calves died from FMD virus infection in two separate outbreaks. A total of 41 intestinal (both small and large intestines) and mesenteric lymph node tissue were collected from these cases and tested for the presence of the FMD virus/antigen and various host cell responses using patho-molecular techniques. For the comparison, intestine and mesenteric lymph nodes from 2 apparently healthy calves were collected. Grossly, the small and large intestines showed marked congestion, thickening of the mucosa and the presence of catarrhal exudates within the lumen. The mesenteric lymph nodes (MLNs) were swollen and congested (21 cases). Microscopically, the intestine showed villous atrophy, crypt elongation, crypt cell proliferation, reduced number of goblet cells, necrosed villi, and lymphocytic infiltration in the lamina propria and sub-mucosa. The MLNs showed depleted cortical follicles and apoptotic bodies (19 cases). The lesions were predominant in the small intestine as compared to the large intestine. The affected intestine showed higher apoptosis cell counts, increased levels of proinflammatory cytokine genes (TNF-α, IL-1β), and reduced number of goblet cells containing sulphated mucins. In multiplex-PCR, the viral genome was detected in the intestine, and lymph nodes of 32 and 21 cases, respectively. Immunohistochemically, abundant cytoplasmic immunoreactivity was noticed mostly in the crypt epithelium of the affected intestine suggesting the association of the virus with intestinal pathology. The MLNs showed moderate immunoreactivity for viral antigen with higher apoptosis counts in 9 cases, suggesting the association of the virus with lymphocytolysis. The reduced expression of MUC1, MUC2, MUC4 mRNAs, while increased. expression of MUC5AC and MUC20 mRNAs was observed in the affected intestine. However, no significant difference in the expression level of MUC15 mRNA was observed. A similar distribution of MUC1, MUC2, and MUC5AC antigens was observed in the paraffin embedded tissue sections of the intestine of FMDV infected calves. The histopathological lesions, immunodetection of viral antigens, increased levels of pro-inflammatory cytokines, reduced goblet cell numbers with altered mucin gene expression likely lead to a defective absorptive function of the intestinal epithelium, which contributes to FMDV pathogenesis. © 2025
SLAM (CD150) receptor homologous peptides block the peste des petits ruminants virus entry into B95a cells
(John Wiley and Sons Inc, 2024) Aditya Agrawal; Rajat Varshney; Anil Gattani; Perumalraja Kirthika; Rohini Gupta; Deepak Kumar; Rabindra Prasad Singh; Praveen Singh
The fusion of haemagglutinin-neuraminidase (HN) protein of peste des petits ruminant (PPR) virus with signaling lymphocyte activation molecules (SLAM) host cell receptor consequences the virus entry and multiplication inside the host cell. The use of synthetic SLAM homologous peptides (i.e., molecular decoy for HN protein of PPR virus) may check PPR infection at the preliminary stage. Hence, the predicted SLAM homologous peptides using bioinformatics tools were synthesized by solid phase chemistry with standard Merrifield's 9-fluorenylmethoxycarbonyl (Fmoc) chemistry and were purified by reverse phase high performance liquid chromatography. The secondary structures of synthesized peptides were elucidated by circular dichroism spectroscopy. The in vitro interactions of these peptides were studied through indirect Enzyme Linked Immuno Sorbent Assay (ELISA) and visual surface plasmon UV-visible spectroscopy. The SLAM homologous peptides were able to interact with the peste des petits ruminant virus (PPRV) with varying binding efficiency. The interaction of SLAM homologous peptide with the PPR virus was ascertained by the change in the plasmon color from red wine to purple during visual detection and also by bathochromic shift in absorbance spectra under UV-visible spectrophotometry. The cytotoxic and anti-PPRV effect of these peptides were also evaluated in B95a cell line using PPR virus (Sungri/96). The cytotoxic concentration 50 (CC50) value of each peptide was greater than 1000 μg mL−1. The anti-PPRV efficiency of SLAM-22 was relatively high among SLAM homologous peptides, SLAM-22 at 25 μg mL−1 concentration showed a reduction of more than log10 3 virus titer by priming of B95a cell line while the use of SLAM-15 and Muco-17 at the same concentration dropped virus titer from log10 4.8 to log10 2.5 and log10 3.1 respectively. The concentration of SLAM homologous peptide (25 μg mL−1) to exert its anti-PPRV effect was much less than its CC50 level (>1000 μg mL−1). Therefore, the synthetic SLAM homologous peptides may prove to be better agents to target PPRV. © 2023 Wiley Periodicals LLC.