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IFNγ expression by an attenuated strain of Salmonella enterica serovar Typhimurium improves vaccine efficacy in susceptible TLR4-defective C3H/HeJ mice

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Abstract

C3H/HeJ mice carry a mutated allele of TLR4 gene (TLR4 d) and thus are hyporesponsive to the lethal effects of lipopolysaccharide (LPS). Characteristically, however, the mice are also hypersusceptible to infections, particularly by Gram-negative bacteria such as Salmonella enterica serovar Typhimurium (S. typhimurium) and are known to be difficult to vaccinate against virulent exposure. This is observed despite the expression of wild-type allele of Nramp1 gene, another important determinant of Salmonella susceptibility. In contrast, C3H/HeN mice (TLR4 n Nramp1 n) express a functional TLR4 protein and are resistant to infection, even by virulent strains of S. typhimurium. In the present study, we describe the immune system-enhancing properties of an attenuated strain of S. typhimurium engineered to express murine IFN-γ. This strain (designated GIDIFN) was able to modulate immune responses following systemic inoculation by upregulating the production of inflammatory mediators (IL-6 and IL-12) and anti-bacterial effector molecules (nitric oxide; NO). Consequently, this led to a more effective control of bacterial proliferation in systemic target organs in both C3H/HeJ and C3H/HeN mice. Although evidence for the enhancement in immune responses could be observed as early as few hours post-inoculation, sustained improvements required 2–3 days to manifest. Vaccination of C3H/HeJ mice with GIDIFN strain, even at low doses, conferred a significantly higher degree of protection against challenge with virulent Salmonella in susceptible C3H/HeJ mice. Our data demonstrate that IFNγ-expressing Salmonella are immunogenic and confer excellent protection against virulent challenge in susceptible C3H/HeJ mice; in addition they may be used as an effective mucosal delivery vectors against virulent infection and for boosting immune responses in immunodeficient hosts.

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Acknowledgments

We thank Drs F.Y. Liew and D. Xu (University of Glasgow, Western Infirmary, UK) for generously providing the BRD509 and GIDIFN Salmonella strains. We wish to thank Mohamed El-Wasilah and Arshad Khan for animal care and husbandry. This work was funded by a PhD fellowship from the Ministry of higher education, Government of Libya (to SMA) and by grants from the Research Grants Committee of the Faculty of Medicine and Health Sciences, UAE University (to BKA).

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The authors declare no competing interests.

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Authors and Affiliations

  1. Department of Medical Microbiology and Immunology, Faculty of Medicine and Health Sciences, United Arab Emirates University, P.O. Box 17666, Al Ain, United Arab Emirates

    Samia M. Al-Ojali & Basel K. al-Ramadi

  2. School of Biomedical Science, University of Ulster, Coleraine, Northern Ireland, UK

    Samia M. Al-Ojali & C. B. Tara Moore

  3. Department of Biochemistry, Faculty of Medicine and Health Sciences, United Arab Emirates University, Al Ain, United Arab Emirates

    Maria J. Fernandez-Cabezudo

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  1. Samia M. Al-Ojali
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  2. C. B. Tara Moore
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  3. Maria J. Fernandez-Cabezudo
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  4. Basel K. al-Ramadi
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Correspondence to Basel K. al-Ramadi.

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Al-Ojali, S.M., Tara Moore, C.B., Fernandez-Cabezudo, M.J. et al. IFNγ expression by an attenuated strain of Salmonella enterica serovar Typhimurium improves vaccine efficacy in susceptible TLR4-defective C3H/HeJ mice. Med Microbiol Immunol 202, 49–61 (2013). https://doi.org/10.1007/s00430-012-0248-z

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  • DOI: https://doi.org/10.1007/s00430-012-0248-z

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