TY - JOUR
T1 - Rational design of a hydrolysis-resistant mycobacterial phosphoglycolipid antigen presented by CD1c to T cells
AU - Reijneveld, Josephine F.
AU - Marino, Laura
AU - Cao, Thinh-Phat
AU - Cheng, Tan Yun
AU - Dam, Dennis
AU - Shahine, Adam
AU - Witte, Martin D.
AU - Filippov, Dmitri V.
AU - Suliman, Sara
AU - van der Marel, Gijsbert A.
AU - Moody, D. Branch
AU - Minnaard, Adriaan J.
AU - Rossjohn, Jamie
AU - Codée, Jeroen D.C.
AU - van Rhijn, Ildiko
N1 - Funding Information:
Acknowledgments—Biotinylated CD1c monomers were obtained through the NIH Tetramer Core Facility. This work was supported by the Medical Research Council of Australia and the Australian Research Council (ARC) (CE140100011).
Funding Information:
Funding and additional information—J. C., G. A. v. d. M., and A. J. M. were supported by Nederlands Wetenschappelijk Onderzoek (NWO) Toppunt grant 15.002. D. B. M. was supported by National Institutes of Health grant R01 AI049313. D. B. M. and I. V. R. were supported by and NIH Tuberculosis Research Unit Network, Grant U19 AI111224. A. S. was supported by an ARC DECRA Fellowship (DE210101031). J. R. is supported by an Australian ARC Laureate Fellowship. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2021 THE AUTHORS.
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/10
Y1 - 2021/10
N2 - Whereas proteolytic cleavage is crucial for peptide presentation by classical major histocompatibility complex (MHC) proteins to T cells, glycolipids presented by CD1 molecules are typically presented in an unmodified form. However, the mycobacterial lipid antigen mannosyl-β1-phosphomycoketide (MPM) may be processed through hydrolysis in antigen presenting cells, forming mannose and phosphomycoketide (PM). To further test the hypothesis that some lipid antigens are processed, and to generate antigens that lead to defined epitopes for future tuberculosis vaccines or diagnostic tests, we aimed to create hydrolysis-resistant MPM variants that retain their antigenicity. Here, we designed and tested three different, versatile synthetic strategies to chemically stabilize MPM analogs. Crystallographic studies of CD1c complexes with these three new MPM analogs showed anchoring of the lipid tail and phosphate group that is highly comparable to nature-identical MPM, with considerable conformational flexibility for the mannose head group. MPM-3, a difluoromethylene-modified version of MPM that is resistant to hydrolysis, showed altered recognition by cells, but not by CD1c proteins, supporting the cellular antigen processing hypothesis. Furthermore, the synthetic analogs elicited T cell responses that were cross-reactive with nature-identical MPM, fulfilling important requirements for future clinical use.
AB - Whereas proteolytic cleavage is crucial for peptide presentation by classical major histocompatibility complex (MHC) proteins to T cells, glycolipids presented by CD1 molecules are typically presented in an unmodified form. However, the mycobacterial lipid antigen mannosyl-β1-phosphomycoketide (MPM) may be processed through hydrolysis in antigen presenting cells, forming mannose and phosphomycoketide (PM). To further test the hypothesis that some lipid antigens are processed, and to generate antigens that lead to defined epitopes for future tuberculosis vaccines or diagnostic tests, we aimed to create hydrolysis-resistant MPM variants that retain their antigenicity. Here, we designed and tested three different, versatile synthetic strategies to chemically stabilize MPM analogs. Crystallographic studies of CD1c complexes with these three new MPM analogs showed anchoring of the lipid tail and phosphate group that is highly comparable to nature-identical MPM, with considerable conformational flexibility for the mannose head group. MPM-3, a difluoromethylene-modified version of MPM that is resistant to hydrolysis, showed altered recognition by cells, but not by CD1c proteins, supporting the cellular antigen processing hypothesis. Furthermore, the synthetic analogs elicited T cell responses that were cross-reactive with nature-identical MPM, fulfilling important requirements for future clinical use.
UR - http://www.scopus.com/inward/record.url?scp=85116816207&partnerID=8YFLogxK
U2 - 10.1016/j.jbc.2021.101197
DO - 10.1016/j.jbc.2021.101197
M3 - Article
C2 - 34536421
AN - SCOPUS:85116816207
SN - 1083-351X
VL - 279
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 4
M1 - 101197
ER -
