Fig 1.
KLRC2 mRNA levels are elevated in CMV and SIV infection.
(A) Representative gating strategy showing the criteria for identifying NK cells, as well as strategy for differentiating KLRC1+ or KLRC2+ populations. (B) Data showing KLRC1+ or KLRC2+ NK cells as a percentage of all lymphocytes, and (C) as a percentage of NKG2AC+ NK cells. The horizontal bars in (B) and (C) indicate medians. Each point corresponds to a single animal: SPF (n = 10), CMV (n = 12) and SIV (n = 8). Mann-Whitney U test; *p < 0.05, **p < 0.01, ***p < 0.0001.
Fig 2.
NK cell phenotyping reveals quadrant-specific distribution and phenotypic differences among animal groups.
(A) Representative gating strategy showing identification of KLRC1±KLRC2± NK cells among NKG2AC+ NK cells as shown in Fig 1A. (B) Distribution of NK cell KLRC1±KLRC2± subpopulations in SPF and infected animals. K1+, K1+K2+, K2+ and K1-K2- correspond to KLRC1+KLRC2-, KLRC1+KLRC2+, KLRC1-KLRC2+ and KLRC1-KLRC2- populations, respectively. (C) Phenotypic markers on KLRC1±KLRC2± NK cells in SPF and infected animals. Data are show as mean ± SEM. The numbers of animals shown per group are as follows: SPF (n = 10), CMV (n = 12) and SIV (n = 8). Mann-Whitney U test was used to compare quadrant populations of different infection groups; Wilcoxon test was used to compare different quadrant populations within the same infection group; *p < 0.05, **p < 0.01, ***p < 0.0001 for panel B. Statistical data for panel C is presented in S1 Table.
Fig 3.
Increased frequencies of KLRC1-KLRC2+ NK cells are positively correlated with higher rhCMV-specific IgG.
Linear regression analysis showing correlation between rhCMV-binding antibody equivalents and (A)KLRC1+KLRC2-, (B)KLRC1+KLRC2+, (C) KLRC1-KLRC2- and (D) KLRC1-KLRC2+ NK cells.
Fig 4.
Infection increases NK cell diversity.
(A) t-SNE plots showing relationship among KLRC1±KLRC2± NK cells within SPF (n = 10), SIV+ (n = 8) or CMV+ (n = 12) animals. (B) Phenotypic histograms per group on which t-SNE plots were generated in (A). Histogram colors match their corresponding populations.
Fig 5.
KLRC2+ NK cells are functionally responsive.
Data showing CD107a expression, or production of IFN-γ and TNF-α following stimulation with PMA+ ionomycin in NK cell subpopulations from SPF, CMV+ or SIV+ animals. Means + SEM are shown. Numbers of animals per independent experiment: SPF (n = 5), CMV (n = 5) and SIV (n = 5). Mann-Whitney U test; *p < 0.05, **p < 0.01, ***p < 0.0001.