Identification and characterisation of conserved epitopes of E. ruminantium that activate Th1 CD4+ T cells : towards the development of a multi-epitope vaccine

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dc.contributor.author Thema, Nontobeko
dc.contributor.author Tshilwane, Selaelo I.
dc.contributor.author Pretorius, Alri
dc.contributor.author Son, L.
dc.contributor.author Smith, R.M.
dc.contributor.author Steyn, Helena C.
dc.contributor.author Liebenberg, Junita
dc.contributor.author Van Kleef, Mirinda
dc.date.accessioned 2019-03-13T11:28:50Z
dc.date.issued 2019-03
dc.description.abstract Several studies have shown that cytotoxic T lymphocytes (CTL) require CD4 + Th1 epitopes to generate strong immune responses to intracellular pathogens. However, not much is known about Ehrlichia ruminantium epitopes, particularly those that can be considered potential candidates for inclusion in a multi-epitope vaccine. In order to identify CD4+ Th1 epitopes that induce IFNγ, a number of proteins previously identified as immunogenic were first screened to determine if they induce cellular immunity in tick infected immune sheep PBMC. Significant IFN-γ production and other Th1 cytokines were evident for 10 recombinant proteins in all sheep tested. Secondly, peptides (n = 246) derived from the top 10 E. ruminantium vaccine candidate proteins were assayed using enzyme linked immunospot (ELISPOT) assay, quantitative real-time PCR and flow cytometry. Of the 246 peptides, 23 peptides, Erum0660 (p0660-42), Erum1150 (p1150-18, p1150-19), Erum2540 (p2540-6, p2540-16, p2540-19, p2540-20, p2540-21), Erum5420 (p5420-13, p5420-14), Erum7140 (p7140-6, p7140-7, p7140-12, p7140-13, p7140-20), Erum7320 (p7320-8, p7320-9, p7320-21), Erum7350 (p7350-9), Erum7360 (p7360-8), Erum7620 (p7620-2, p7620-12) and Erum8010 (p8010-8) were identified that stimulate the best and different cell mediated immune responses. Amino acid sequences of these peptides except for p7140-12, p7140-13, p7140-20, and p7350-9 were conserved between 13 different local strains. These peptides could efficiently induce memory CD4+ T cells to rapidly proliferate and significantly increase IFN-γ production in immune sheep PBMC. The upregulation of pro-inflammatory cytokines, which include, IL-1α, IL-2, IL-12p40, TNF-α, IFN-γ, inducible nitric oxide synthase (iNOS) and granulocyte-macrophage colony stimulating factor (GM-CSF) was also detected. Our results show that these peptides could serve as promising candidates for a multi-epitope vaccine against E. ruminantium. en_ZA
dc.description.department Veterinary Tropical Diseases en_ZA
dc.description.embargo 2020-03-01
dc.description.librarian hj2019 en_ZA
dc.description.sponsorship The NRF, THRIP, and RMRD-SA in collaboration with OBP and Economic Competitiveness Support Programme and the ARC-OVR, South Africa. en_ZA
dc.description.uri http://www.elsevier.com/locate/molimm en_ZA
dc.identifier.citation Thema, N., Tshilwane, S.I., Pretorius, A. et al. 2019, 'Identification and characterisation of conserved epitopes of E. ruminantium that activate Th1 CD4+ T cells : towards the development of a multi-epitope vaccine', Molecular Immunology, vol. 107, pp. 106-114. en_ZA
dc.identifier.issn 0161-5890 (print)
dc.identifier.issn 1872-9142 (online)
dc.identifier.other 10.1016/j.molimm.2018.12.012
dc.identifier.uri http://hdl.handle.net/2263/68643
dc.language.iso en en_ZA
dc.publisher Elsevier en_ZA
dc.rights © 2018 Elsevier Ltd. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Molecular Immunology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. A definitive version was subsequently published in Molecular Immunology, vol. 107, pp. 106-114, 2019. doi : 10.1016/j.molimm.2018.12.012. en_ZA
dc.subject Cytotoxic T lymphocytes (CTL) en_ZA
dc.subject Enzyme linked immunospot (ELISPOT) en_ZA
dc.subject Ehrlichia ruminantium en_ZA
dc.subject Antigens en_ZA
dc.subject CD4+T cells en_ZA
dc.subject Peptides en_ZA
dc.subject Interferon gamma (IFNγ) en_ZA
dc.title Identification and characterisation of conserved epitopes of E. ruminantium that activate Th1 CD4+ T cells : towards the development of a multi-epitope vaccine en_ZA
dc.type Postprint Article en_ZA


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