Panickan, S. | Search bioRxiv

bioRxiv2026

Immunogenicity and protective efficacy of a Brucella abortus L7/L12 DNA vaccine delivered via chitosan modified PLGA nanoparticles in mice

The present study evaluated the immunogenicity and protective efficacy of the chitosan (CS)-modified poly-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) delivering Brucella abortus L7/L12 DNA in a mouse model. The NPs were prepared by solvent displacement method and characterized for size, charge, morphology, cellular uptake and cytotoxicity. The cationic CS-PLGA NPs were spherical with a mean size of [~]165 nm with a positive zeta potential (+20 mV). DNA loading efficiency of 1.2% and DNA adsorption shifted zeta potential to -45 mV. In vitro studies in RAW 264.7 cell line demonstrated efficient uptake of the DNA loaded cationic NPs and expression of L7/L12 protein. Intramuscular immunization of the L7/L12 DNA vaccine loaded CS-PLGA NPs elicited both humoral and cell-mediated immunity with upregulation of Th1 and Th2 cytokines along with induction of IgG antibodies in mice. IFN-{gamma}, IL-2, and IL-4 levels were significantly (P < 0.001) higher than control group. The protective efficacy of the DNA loaded NPs against virulent B. abortus 544 infection (105 CFU) was significantly higher than that of the naked DNA (P<0.001). These findings suggest that the CS-PLGA NPs were efficiently delivered L7/L12 DNA and exhibited adjuvant potential, conferring protection against experimental murine brucellosis.

microbiology↗

bioRxiv2026

Pathology of dose dependent inocula of H5N8 avian influenza viruses in experimentally infected chicken

In the present study, we assessed the pathogenicity of H5N8 avian influenza viruses belongs to the clade 2.3.4.4b in chicken. Birds of three different dose groups, 102, 104, and 106 EID50 were used in the study. No mortality was observed in 102 EID0 group. Percent cumulative mortality of 104 and 106 EID50 group was 66.67 and 100 %, respectively. Varying duration of MDT of 3.2 and 2 days was observed in 104 and 106 EID50 group, respectively. The CID50 of virus was found to be 104.5 EID50. High no. of viral RNA copies were found both in oropharyngeal and cloacal swabs and in various organs of birds infected in 104 and 106 EID50 group. Significant gross and histological changes and presence of viral antigen in various organs were observed in 104 and 106 EID50 group. So, the study concludes that Indian HPAI, H5N8 isolates are highly pathogenic in nature to chicken by affecting most organs systemically. CID50 of this H5N8 virus indicates poor adaption in chicken and it implies poor transmission possibility of this virus for host species in field condition. Though this virus is highly pathogenic in nature as that of HPAI, H5N1 viruses, absence of endothelial staining in most organ attributes variation in replication process and pathogenesis from HPAI, H5N1 viruses. Hence, further studies need to be done to elucidate the pathobiology of this virus in various bird species. HighlightsO_LIH5N8 virus belong to the clade 2.3.4.4b, Indian isolate is highly pathogenic in nature as that of HPAIV, H5N1. C_LIO_LIThe dose inocula, 102 EID50 is noninfectious to chicken. C_LIO_LIThe dose inocula, 104 and 106 EID50 had caused significant mortality in the inoculated chicken with MDT of 2 and 3.2 days, respectively. C_LIO_LIH5N8 virus was detected with high viral titres in clocal and oral shedding and in multiple organ with the dose inocula, 104 and 106 EID50. C_LIO_LI104 and 106 EID50 of H5N8 inocula virus caused significant gross and histological changes in multiple organs and viral antigens were detected in respective organs. C_LI

microbiology↗