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Class-I Restricted T Cell-Associated Molecule (CRTAM) is a protein that is expressed after T cell activation. The interaction of CRTAM with its ligand, nectin-like 2 (Necl2), is required for the efficient production of IL-17, IL-22, and IFN&gamma, by murine CD4 T cells, and it plays a role in optimal CD8 T and NK cell cytotoxicity. CRTAM promotes the pro-inflammatory cytokine profile, therefore, it may take part in the immunopathology of autoimmune diseases such as diabetes type 1 or colitis. Thus, antibodies that block the interaction between CRTAM and Necl2 would be useful for controlling the production of these inflammatory cytokines. In this work, using bioinformatics predictions, we identified three short disordered epitopes (sDE1-3) that are located in the Ig-like domains of murine CRTAM and are conserved in mammalian species. We performed a structural analysis by molecular dynamics simulations of sDE1 (QHPALKSSKY, Ig-like V), sDE2 (QRNGEKSVVK, Ig-like C1), and sDE3 (CSTERSKKPPPQI, Ig-like C1). sDE1, which is located within a loop of the contact interface of the heterotypic interaction with Nectl2, undergoes an order&ndash, disorder transition. On the contrary, even though sDE2 and sDE3 are flexible and also located within loops, they do not undergo order&ndash, disorder transitions. We evaluated the immunogenicity of sDE1 and sDE3 through the expression of these epitopes in chimeric L1 virus-like particles. We confirmed that sDE1 induces polyclonal antibodies that recognize the native folding of CRTAM expressed in activated murine CD4 T cells. In contrast, sDE3 induces polyclonal antibodies that recognize the recombinant protein hCRTAM-Fc, but not the native CRTAM. Thus, in this study, an exposed disordered epitope in the Ig-like V domain of CRTAM was identified as a potential site for therapeutic antibodies.

Type 1 diabetes (T1D) is an autoimmune disease that is characterized by the specific destruction of insulin-producing pancreatic β cells. Invariant natural killer T (iNKT) cells have been associated with development of T1D. Class I MHC-restricted T cell-associated molecule (CRTAM) is expressed on activated iNKT, CD8(+), and CD4(+) T cells, and it is associated with the pro-inflammatory profiles of these cells. Crtam gene expression in CD3(+) lymphocytes from non-obese diabetic (NOD) mice is associated with T1D onset. However, expression of CRTAM on T cells from patients with T1D has not yet been evaluated. We compared iNKT cell (CD3(+)Vα24(+)Vβ11(+)) numbers and CRTAM expression in a Mexican population with recent-onset T1D and their first-degree relatives with control families. Remarkably, we found lower iNKT cell numbers in T1D families, and we identified two iNKT cell populations in some of the families. One iNKT cell population expressed high iTCR levels (iNKT(hi)), whereas another expressed low levels (iNKT(lo)) and also expressed CRTAM. These findings support a probable genetic determinant of iNKT cell numbers and a possible role for these cells in T1D development. This study also suggests that CRTAM identifies recently activated iNKT lymphocytes.

Background Class-I MHC-restricted T cell-associated molecule (CRTAM) is a protein expressed by activated natural killer T (NKT) cells, natural killer (NK) cells, CD8 T cells, and certain CD4 T lymphocytes. It is also expressed in Purkinje neurons and epithelial cells. However, no studies have examined the expression of CRTAM in peripheral blood cells during homeostasis or disease. Therefore, we explored whether CRTAM expression is influenced by the presence of allergic asthma. Methods We collected whole peripheral blood cells from non-asthmatic control subjects (n = 17) and patients with asthma (n = 17). All patients with asthma tested positive in allergen skin prick tests. We analyzed CRTAM expression in CD4+ and CD8+ T lymphocyte populations. CRTAM expression was also analyzed in CD177+ neutrophils and IL5Rα+ eosinophils. Findings The percentage of CD4+CRTAM+ and CD8+CRTAM+T lymphocytes in peripheral blood was higher in allergic asthma patients compared with healthy controls. Furthermore, the percentage of CD177+CRTAM+ neutrophils in peripheral blood was also elevated in patients with allergic asthma. However, the percentage of IL5Rα+CRTAM+ eosinophils in peripheral blood was not significantly different in patients with allergic asthma compared with healthy controls. Conclusions CRTAM expression on T cells, eosinophils, and neutrophils may be involved in bronchial inflammation in allergic asthma. Determination of CRTAM expression in peripheral blood may be useful for the diagnosis of bronchial inflammation and/or to identify recently activated immune cells.