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J. Microbiol. Biotechnol. 2024; 34(3): 1-11

Published online March 28, 2024 https://doi.org/10.4014/jmb.2307.07040

Copyright © The Korean Society for Microbiology and Biotechnology.

Mucosal Administration of Lactobacillus casei Surface-Displayed HA1 Induces Protective Immune Responses against Avian Influenza A Virus in Mice

Dung T. Huynh, W.A. Gayan Chathuranga, Kiramage Chathuranga, Jong-Soo Lee*, and Chul-Joong Kim*

College of Veterinary Medicine, Chungnam National University, Daejeon 34314, Republic of Korea

Correspondence to:Jong-Soo Lee,          jongsool@cnu.ac.kr
Chul-Joong Kim,       cjkim@cnu.ac.kr

Received: July 28, 2023; Revised: October 12, 2023; Accepted: October 18, 2023

Abstract

Avian influenza is a serious threat to both public health and the poultry industry worldwide. This respiratory virus can be combated by eliciting robust immune responses at the site of infection through mucosal immunization. Recombinant probiotics, specifically lactic acid bacteria, are safe and effective carriers for mucosal vaccines. In this study, we engineered recombinant fusion protein by fusing the hemagglutinin 1 (HA1) subunit of the A/Aquatic bird/Korea/W81/2005 (H5N2) with the Bacillus subtilis poly γ-glutamic acid synthetase A (pgsA) at the surface of Lactobacillus casei (pgsAHA1/L. casei). Using subcellular fractionation and flow cytometry we confirmed the surface localization of this fusion protein. Mucosal administration of pgsA-HA1/L. casei in mice resulted in significant levels of HA1-specific serum IgG, mucosal IgA and neutralizing antibodies against the H5N2 virus. Additionally, pgsA-HA1/L. casei-induced systemic and local cell-mediated immune responses specific to HA1, as evidenced by an increased number of IFN-γ and IL-4 secreting cells in the spleens and higher levels of IL-4 in the local lymphocyte supernatants. Finally, mice inoculated with pgsAHA1/L. casei were protected against a 10LD50 dose of the homologous mouse-adapted H5N2 virus. These results suggest that mucosal immunization with L. casei displaying HA1 on its surface could be a potential strategy for developing a mucosal vaccine against other H5 subtype viruses.

Keywords: Avian influenza, HA1, Lactobacillus casei, poly &gamma,-glutamic acid synthetase A, surface display, mucosal delivery