Recognition of non-lipid antigens by Type 2 NKT cells — ASN Events

Recognition of non-lipid antigens by Type 2 NKT cells (#148)

Catarina Filipa dos Santos Sa e Almeida 1 2 , Dylan Smith 3 , Tamara Hilmenyuk 1 2 , Adam P Uldrich 1 2 , Ildiko van Rhijn 4 , David B Moody 4 , Daniel G Pellicci 1 2 , Spencer J Williams 3 , Dale I Godfrey 1 2
  1. Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, The University of Melbourne , Melbourne, VIC, Australia
  2. Australian Research Council Centre of Excellence in Advanced Molecular Imaging, The University of Melbourne, Melbourne, VIC, Australia
  3. School of Chemistry , Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC, Australia
  4. Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA

Natural Killer T (NKT) cells recognise lipid antigens presented by the MHC Class I-like molecule CD1d. Following activation, they rapidly secrete a broad range of immunoregulatory cytokines that can influence other mediators of immune responses and therefore they represent a promising therapeutic target for cancer and other diseases. The most extensively studied are type 1 NKT cells, which recognise a derivative of a marine sponge glycolipid α-galactosylceramide (α-GalCer), express a semi-invariant T cell receptor (TCR), and have a well-established role in the immune system. Much less is known about type 2 NKT cells, which do not recognise α-GalCer and express a diverse TCR repertoire. An early study showed that a non-lipid molecule – phenyl 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonate (PPBF) – can also be detected by type 2 NKT cells in the context of CD1d. The structure of PPBF shares similarities with several sulfonamide drugs that have been reported to cause hypersensitivity in humans suggesting that type 2 NKT cells may be involved in such hypersensitivity reactions. Through investigation of various PPBF analogues, we identify 3-chlorophenyl 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonate (ClPPBF) as a stronger activator of type 2 NKT cells than the original PPBF compound. Importantly, we demonstrate that type 2 NKT cell modulation by PPBF is due to TCR recognition of PPBF-CD1d complexes. Using CD1d-ClPPBF tetramers we identified a novel population of type 2 NKT cells. Single cell sequencing of these cells revealed a polyclonal TCR repertoire distinct from type 1 NKT cells. Through the generation of retrovirally TCR-transduced cell lines we validated the reactivity of sorted cells. Thus, we provide evidence that a polyclonal repertoire of type 2 NKT cells are capable of reacting with PPBF-loaded CD1d. This study provides valuable insight into the diversity of antigens recognized by CD1d-restricted NKT cells, suggesting that NKT cell activation can be modulated by non-lipid molecules.

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