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Oral Vaccination with Hepatitis E Virus Capsid Protein and Immunobiotic Bacterium-Like Particles Induce Intestinal and Systemic Immunity in Mice

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Abstract

The hepatitis E virus (HEV) genotype 3 (GT3) is an emergent pathogen in industrialized countries. It is transmitted zoonotically and may lead to chronic hepatitis in immunocompromised individuals. We evaluated if the major antigen of HEV, the capsid protein, can be used in combination with immunobiotic bacterium-like particles (IBLP) for oral vaccination in a mouse model. We have cloned and expressed the RGS-His5-tagged HEV GT3 capsid protein (ORF2) in E. coli and purified it by NiNTA. IBLP were obtained from two immunobiotic Lactobacillus rhamnosus strains acid- and heat-treated. ORF2 and the IBLP were orally administered to Balb/c mice. After three oral immunizations (14-day intervals), blood, intestinal fluid, Peyer´s patches, and spleen samples were drawn. IgA- and IgG-specific antibodies were determined by ELISA. Mononuclear cell populations from Peyer’s patches and spleen were analyzed by flow cytometry, and the cytokine profiles were determined by ELISA to study cellular immunity. Orally administered recombinant ORF2 and IBLP from two L. rhamnosus strains (CRL1505 and IBL027) induced both antigen-specific humoral and cellular immune responses in mice. IBLP027 was more effective in inducing specific secretory IgA in the gut. IFN-γ, TNF-α, and IL-4 were produced by Peyer’s plaques lymphocytes stimulated with ORF2 ex vivo suggesting a mixed Th1/Th2-type adaptive immune response in immunized mice. Oral vaccines are not invasive, do not need to be administered by specialized personal, and elicit both systemic and local immune responses at the port of entry. Here, we present an experimental oral vaccine for HEV GT3, which could be further developed for human and/or veterinary use.

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Acknowledgments

We thank Dr. Roque Carrero Valenzuela for sharing his facilities with us.

Funding

This study received financial support from the Agencia Nacional de Ciencia y Técnica (PICT-2016-0853 and PICT-2016-0410), CONICET (PIP057), and Fundación Allende.

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Author notes

  1. J. Villena and M. G. Vizoso Pinto contributed equally to this work.

Authors and Affiliations

  1. Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET-UNT, and Facultad de Medicina, UNT, Av. Kirchner 2100, (4000) San Miguel de Tucumán, Tucumán, Argentina

    L. P. Arce, M. F. Raya Tonetti, M. F. Müller & M. G. Vizoso Pinto

  2. Laboratorio de Ciencias Básicas. OR. Genética. Facultad de Medicina de la Universidad Nacional de Tucumán, Tucumán, Argentina

    L. P. Arce, M. F. Raya Tonetti, M. P. Raimondo, M. F. Müller & M. G. Vizoso Pinto

  3. Laboratorio de Inmunobiotecnología, CERELA (CONICET), Chacabuco 145, (4000) San Miguel de Tucumán, Tucumán, Argentina

    S. Salva, S. Álvarez & J. Villena

  4. LGL, Bavarian Health and Food Safety Authority, Oberschleissheim, Germany

    A. Baiker

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Correspondence to J. Villena or M. G. Vizoso Pinto.

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Statement on the Welfare of Animals

This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the Guidelines for Animal Experimentation of the Reference Centre for Lactobacilli (CERELA-CONICET, Tucuman, Argentina). The CERELA Institutional Animal Care and Use Committee prospectively approved this research under the protocol BIOT-CRL-17.

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The authors declare that they have no conflict of interest. LA, FRT, and MM hold CONICET scholarships. SGA, JV, and MGVP are members of CONICET.

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Arce, L.P., Raya Tonetti, M.F., Raimondo, M.P. et al. Oral Vaccination with Hepatitis E Virus Capsid Protein and Immunobiotic Bacterium-Like Particles Induce Intestinal and Systemic Immunity in Mice. Probiotics & Antimicro. Prot. 12, 961–972 (2020). https://doi.org/10.1007/s12602-019-09598-7

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