Figure 3.

Enhanced function of educated mouse uNK cells. (A) Gating strategy for uterine group 1 ILC and delineation into three subsets including uterine ILC1 (uILC1), conventional NK cells (cNK) and tissue-resident CD49a⁺ NK. (B) Representative staining of all three group 1 ILC subsets for self-receptors in C57BL/6 mice. cNK, trNK or uILC1 subsets were stained for Ly49C/I, and NKG2A. Coloured line shows proportion of each subset positive for one or more of these educating inhibitory receptors. Grey line indicates staining with isotype matched negative control antibody. (C) Assessment of the functional responsiveness (intracellular IFN-γ and surface CD107a) after crosslinking by plate bound anti-NK1.1 antibody in cells expressing inhibitory NK receptors for self MHC compared to those that do not. Shown are the % of cells in each quadrant (grey text in corners) as well as the relative percentage of responders among cells expressing receptors for self and responders that do not have self-receptors (in bold). The relative percentage in the top left (Q1) and right (Q2) quadrants were calculated from the raw values as follows: Q1/(Q1+Q3) and Q2/(Q2+Q4). (D, E) Enumeration of IFN-γ producing (D) and CD107a⁺ NK cells (E) among cells expressing at least one receptor for self MHC class I (educated subset, self-receptor +) and those that do not (uneducated, self-receptor -). uILC1 are not depicted due to the paucity of uneducated cells in this subset. Data representative of 3 (A, B) or 2 (C-E) experiments with n = 6 mice per group. P-values in (D, E), comparing cells in cNK or CD49a⁺ NK subsets expressing an educating self-receptor with those that do not, within each mouse using paired two-tailed Student’s t-tests.