TY - JOUR
T1 - Identification of antigens presented by MHC for vaccines against tuberculosis
AU - Bettencourt, Paulo
AU - Müller, Julius
AU - Nicastri, Annalisa
AU - Cantillon, Daire
AU - Madhavan, Meera
AU - Charles, Philip D.
AU - Fotso, Carine B.
AU - Wittenberg, Rachel
AU - Bull, Naomi
AU - Pinpathomrat, Nawamin
AU - Waddell, Simon J.
AU - Stylianou, Elena
AU - Hill, Adrian V.S.
AU - Ternette, Nicola
AU - McShane, Helen
N1 - Funding Information:
H.McS is a Wellcome Trust Investigator (WT 206331/Z/17/Z). H.McS, N.T. and A.V.S.H. are Jenner Investigators. S.J.W. is funded by the Wellcome Trust 204538/Z/16/Z. Mass Spectrometry data was acquired at the Target Discovery Institute Mass Spectrometry Laboratory led by Professor Benedikt M. Kessler. The authors would like to acknowledge Dr Rajko Reljic from St George's University of London, for kindly providing M. bovis BCG-GFP; Dr Matthew K. O’Shea for kindly providing PBMC samples from TB disease patients; Dr Alison Turner from the Viral Vector Facility of the Jenner Institute for producing the viral vectors; Dr Vivian Clark, Mrs Heather Grey and Mr Stephen Laird from the WTCHG in Oxford, for animal welfare during the mouse M.tb challenge experiments; Dr Giselda Bucca, Dr Andrew Hesketh and Professor Colin Smith at the Brighton Genomics Centre for RNA-seq; Dr Tom Partridge for kindly providing human primary monocytes; Dr Martin Barnardo from the Oxford University Hospitals NHS Trust for HLA genotyping. The authors would like to acknowledge Professor Todd L. Lowary from The University of Alberta, Canada, for kindly supplying the plasmid containing glfT2 that was used on initial experiments but not included in this study. BEI resources; Manassas USA, provided M.tb Erdman K01 (TMC107).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Mycobacterium tuberculosis (M.tb) is responsible for more deaths globally than any other pathogen. The only available vaccine, bacillus Calmette-Guérin (BCG), has variable efficacy throughout the world. A more effective vaccine is urgently needed. The immune response against tuberculosis relies, at least in part, on CD4+ T cells. Protective vaccines require the induction of antigen-specific CD4+ T cells via mycobacterial peptides presented by MHC class-II in infected macrophages. In order to identify mycobacterial antigens bound to MHC, we have immunoprecipitated MHC class-I and class-II complexes from THP-1 macrophages infected with BCG, purified MHC class-I and MHC class-II peptides and analysed them by liquid chromatography tandem mass spectrometry. We have successfully identified 94 mycobacterial peptides presented by MHC-II and 43 presented by MHC-I, from 76 and 41 antigens, respectively. These antigens were found to be highly expressed in infected macrophages. Gene ontology analysis suggests most of these antigens are associated with membranes and involved in lipid biosynthesis and transport. The sequences of selected peptides were confirmed by spectral match validation and immunogenicity evaluated by IFN-gamma ELISpot against peripheral blood mononuclear cell from volunteers vaccinated with BCG, M.tb latently infected subjects or patients with tuberculosis disease. Three antigens were expressed in viral vectors, and evaluated as vaccine candidates alone or in combination in a murine aerosol M.tb challenge model. When delivered in combination, the three candidate vaccines conferred significant protection in the lungs and spleen compared with BCG alone, demonstrating proof-of-concept for this unbiased approach to identifying new candidate antigens.
AB - Mycobacterium tuberculosis (M.tb) is responsible for more deaths globally than any other pathogen. The only available vaccine, bacillus Calmette-Guérin (BCG), has variable efficacy throughout the world. A more effective vaccine is urgently needed. The immune response against tuberculosis relies, at least in part, on CD4+ T cells. Protective vaccines require the induction of antigen-specific CD4+ T cells via mycobacterial peptides presented by MHC class-II in infected macrophages. In order to identify mycobacterial antigens bound to MHC, we have immunoprecipitated MHC class-I and class-II complexes from THP-1 macrophages infected with BCG, purified MHC class-I and MHC class-II peptides and analysed them by liquid chromatography tandem mass spectrometry. We have successfully identified 94 mycobacterial peptides presented by MHC-II and 43 presented by MHC-I, from 76 and 41 antigens, respectively. These antigens were found to be highly expressed in infected macrophages. Gene ontology analysis suggests most of these antigens are associated with membranes and involved in lipid biosynthesis and transport. The sequences of selected peptides were confirmed by spectral match validation and immunogenicity evaluated by IFN-gamma ELISpot against peripheral blood mononuclear cell from volunteers vaccinated with BCG, M.tb latently infected subjects or patients with tuberculosis disease. Three antigens were expressed in viral vectors, and evaluated as vaccine candidates alone or in combination in a murine aerosol M.tb challenge model. When delivered in combination, the three candidate vaccines conferred significant protection in the lungs and spleen compared with BCG alone, demonstrating proof-of-concept for this unbiased approach to identifying new candidate antigens.
UR - http://www.scopus.com/inward/record.url?scp=85077623869&partnerID=8YFLogxK
U2 - 10.1038/s41541-019-0148-y
DO - 10.1038/s41541-019-0148-y
M3 - Article
C2 - 31908851
AN - SCOPUS:85077623869
SN - 2059-0105
VL - 5
JO - npj Vaccines
JF - npj Vaccines
IS - 1
M1 - 2
ER -
