Journal: Nature Communications
Article Title: Candida albicans evades NK cell elimination via binding of Agglutinin-Like Sequence proteins to the checkpoint receptor TIGIT
doi: 10.1038/s41467-022-30087-z
Figure Lengend Snippet: a Cytotoxicity assay of C. albicans yeast cells using the YTS NK cell line either expressing TIGIT (YTS TIGIT) or not expressing it (YTS Eco). The YTS cells were either blocked using anti-TIGIT antibodies (black bars) or not blocked (gray bars). n = 2–4 independent experiments. Data are presented as mean values ± SD. b Cytotoxicity assay of C. albicans yeast cells using primary NK cells isolated from different human donors. The NK cells were blocked or not using anti-TIGIT antibodies. Each line represents an independent human donor. c A Diagram depicting the in-vitro T-cell activation model used in ( d ). d T cell activation by C. albicans model. CD4 + TIGIT + T cells were isolated from human donors. The T cells were activated using anti-CD3-antibody-coated P815 cells in the presence or absence of C. albicans cells and the presence (black dots) or absence (white dots) of a TIGIT-blocking antibody. A quantification of 3 independent experiments is presented. e C. albicans cytotoxicity assay using the YTS NK cell line expressing either WT TIGIT (YTS TIGIT) or a mutated version of TIGIT (YTS TIGIT Y231A or YTS TIGIT Y231Stop). The YTS cells were either blocked using anti-TIGIT antibodies or not blocked. A quantification of 3 independent experiments is presented. f TIGIT activation was assayed using the murine thymoma cell line BW expressing a chimeric TIGIT-ζ-chain receptor. The BW cells were co-incubated for 48 h in the presence of the WT C. albicans lab strain SC5314 (red bar) or clinical C. albicans strains isolated from a cohort of human invasive candidiasis patients (black bars). TIGIT activation was measured using ELISA for quantification of IL-2 secreted by the activated BW cells. n = 104 biologically independent samples containing 2–3 technical repeats examined over 2 independent experiments. Data are presented as mean values ± SD. For ( a , b , d , e )) significance was tested using Student’s t test (two-tailed and unpaired for ( a ), two-tailed and paired for ( b , e ), and one-tailed and paired for ( d )). For ( f ), one-way ANOVA with correction for multiple comparisons using Dunnett’s test was used. ns = not-significant, * = p < 0.05, ** = p < 0.01.
Article Snippet: Antibodies used for TIGIT blockade (anti-mouse TIGIT clone 1G9, BioXCell, 100 μg/mouse), NK cell depletion (anti-mouse NK1.1 PK136, BioXCell, 25 μg/mouse) or T cell depletion (anti-mouse CD3 17A2, BioXCell, 100 μg/mouse) were diluted in 1xPBS for a final volume of 200 μl/mouse and injected intraperitoneally.
Techniques: Cytotoxicity Assay, Expressing, Isolation, In Vitro, Activation Assay, Blocking Assay, Incubation, Enzyme-linked Immunosorbent Assay, Two Tailed Test, One-tailed Test
