Developmental control of CD8+ T cell–avidity maturation in autoimmune diabetes
J. Clin. Invest. Bingye Han, et al. 115:1879 doi:10.1172/JCI24219 [
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Figure 1Avidity maturation of NRP-A7–reactive CD8
+ cells is associated with changes in Vα17 usage. (
A) Changes in Vα17 usage by islet-derived, NRP-A7–reactive CD8
+ cells with age (data from Supplemental Figure 1). (
B) Predicted amino acid sequences of the 3 Vα17 elements expressed by NOD mice. Gray boxes highlight sequence differences compared with Vα17.6, the least used Vα17 element by 20 weeks. (
C) Average mfi for Vβ8 and CD8 on TCR-αβ transfectants expressing the 8.3 TCR-β chain and the 8.3 –TCR-α (Vα17.4
+) or 8.3-like TCR-α chains (using Vα17.6 or Vα17.5) (
n = 3 clones each). (
D) Tetramer binding to transfectants. (
E) Flow cytometric analysis of 17.4α/8.3β and 17.5α/8.3β transfectants stained with FPLC-purified, Cy5-labeled NRP-V7/K
d tetramers. Numbers on the upper right corner correspond to average mfi values. (
F) Production of IL-2 by Vα17.6
+, Vα17.5
+, and Vα17.4
+ transfectants in response to anti-CD3–coated microbeads or IGRP
206–214–pulsed NOD splenocytes (average ± SEM for 1, 4, and 4 clones, respectively). The peptide concentrations that give the half-maximum response (SD50) for IL-2 secretion corresponding to Vα17.6
+, Vα17.5
+, and Vα17.4
+ cells were as follows: 173 ± 54 (Vα17.6
+), 13 ± 8 (Vα17.5
+), and 73 ± 2 (Vα17.4
+) μg/ml (for NRP-V7), and greater than 1000 (Vα17.6
+), 1 ± 1 (Vα17.5
+), and 135 ± 17 (Vα17.4
+) μg/ml (for IGRP
206–214), respectively (
P < 0.001) (data are from a separate experiment employing 3 clones of each transfectant type). (
G) Flow cytometric analysis of Vα17.5 and Vα17.6 transfectants coexpressing another Vβ8.1
+ TCR-β rearrangement cloned from tetramer
+ cells of 9-week-old NOD mice (cDNA clone NC28, with the CDR3 sequence CASSDPENTL). Numbers on the graphs correspond to average mfi values of 2–6 clones per group.
