Fig. 5: CD2 enhances T-cell sensitivity.
From: Antigen discrimination by T cells relies on size-constrained microvillar contact
a CD45 exclusion at close contacts formed by J8-GECI-CD2WT cells. The images show a J8-GECI-CD2WT cell during the scanning stage on an SLB2 presenting pMHCnull; inset shows GECI fluorescence, indicating a lack of signaling. A min/max normalized intensity line profile was taken along the direction of the white arrow and indicates CD45 exclusion at close contacts. The kymograph shows contact stabilization and persistent CD45 exclusion from contacts. See Supplementary Movies 21–23. b–e Close contact feature analysis for both signaling and non-signaling J8-GECI-CD2WT and variant cells during the scanning stage on an SLB2 presenting pMHCnull; n = 9 (J8-GECI-CD2WT, from 2 SLBs), 11 (J8-GECI-CD2ΔCYT, from 2 SLBs), and 17 (J8-GECI-CD2KO, from 3 SLBs) cells. The boxplots indicate the quartiles with a line at the median. Whiskers extend to points that lie within 1.5 IQRs of the lower and upper quartile. Conditions were compared using the Kruskal–Wallis H test, and, if p < 0.05, further compared using the pairwise Mann–Whitney U test. The p values are shown. b Mean number of close contacts per cell. c Mean single close-contact area. The analysis uses the same cells as in (b). d Mean glycocalyx exclusion at close contacts. The analysis uses the same cells as in (b). e Mean exclusion from close contacts of CD45 on the J8-GECI cell surface. The analysis uses the same cells as in (b). f Calcium response curves for J8-GECI-CD2WT, J8-GECI-CD2ΔCYT, and J8-GECI-CD2KO cell lines interacting with SLB2s presenting pMHCnull ± pMHC9V at the indicated densities. Data were fitted using a four-point dose-response curve, constrained to a minimum based on responses to pMHCnull; n = 27 (J8-GECI-CD2WT), 25 (J8-GECI-CD2ΔCYT), and 25 (J8-GECI-CD2KO) SLBs per curve with ≥139 cells analyzed per SLB. The EC50 values for J8-GECI-CD2WT, J8-GECI-CD2ΔCYT, and J8-GECI-CD2KO cells were 0.7, 3.2, and 64.5 molecules pMHC9V/μm2, respectively. g Increased Lck recruitment to close contacts for cells expressing full-length CD2 versus CD2ΔCYT on an SLB2 presenting pMHCnull + pMHC9V-lo; n = 1080 (J8-GECI-CD2WT) and 983 (J8-GECI-CD2ΔCYT) contacts analyzed from 10 FOVs. Means were compared using a two-sided Student’s t-test. Source data are provided in the Source data file.
