Fig 2: CD155 surface expression by APCs. Pooled flow cytometry analysis of PBMCs ex vivo, of healthy controls (n = 7, symbol with cross), psoriasis (n = 7, open symbol), and psoriatic arthritis (n = 7, filled symbol) patients. Shown data are from six APC subsets: CD14+CD16- classical monocyte (cM), CD14+CD16+ intermediate monocyte (iM), CD14-CD16+ non-classical monocyte (ncM), CD141+ myeloid DC (mDC), CD1c+ mDC, and CD303+ plasmacytoid DC (pDC). (A) Proportion of APC subset in PBMCs (gate ‘Single Cells’ Supplementary Fig. S1A). (B) Proportion of CD155-positive cells in APC subset. (C–H) Gating strategy of the selection of CD155-positive cells in cM (C), iM (D), ncM (E), CD141+ mDC (F), CD1c+ mDC (G), and pDC (H).

Fig 3: High TNF production by CD155 expressing APCs. Flow cytometry analysis of psoriatic disease patients PBMCs stimulated for 4 hours with LPS (100 ng/ml) in the presence of Brefeldin A (1:1000). (A) Proportion of TNF producing cells within CD155-positive and CD155-negative APC subsets of psoriasis patients (n = 13, open symbol) and psoriatic arthritis patients (n = 15, filled symbol): CD14+CD16- classical monocyte (cM), CD1c+ myeloid dendritic cell (mDC), and CD123+ plasmacytoid dendritic cell (pDC) (detailed gating strategy shown in Supplementary Fig. S1B). (B–D) Representative flow plots of TNF production by CD1c+ mDC: CD155-positive TNF FMO control (B), CD155-negative CD1c+ mDC (C), and CD155-positive CD1c+ mDC (D). *Significant P-value MWU. NA: not applicable.

Fig 4: KSHV infection drives CD56-CD16+ NK cell differentiation with expression of CD38, CD39, CD69, and CXCR6 in huNSG mice(A) UMAP visualization of single, live, human CD45+Lin- (CD3, CD19, CD14, CD4) CD7+ cells isolated at 4 weeks p.i. from spleen and liver of mock-, EBVzko-, and EBVzko+KSHV-infected mice depicted as contour plot.(B) UMAP plot depicting relative marker expression from samples in (A). (C) Heatmap depicting median fluorescent intensity (MFI) of surface markers on CD56-and CD16-stained NK cell subsets (pre-gated on human CD45+NKp46+Lin-) and on human CD45- cells of EBVzko+KSHV co-infected animals.(D) Heatmap depicting the median frequency of cells positive for indicated surface markers among different CD56-and CD16-stained NK cell subsets. In (C) and here, composite data are from 1–4 independent experiments with n = 6–20 animals. Data are normalized per column. Statistical analysis was performed by multiple comparison tests following paired one-way ANOVA or Friedmann’s test (q values). See also Figures S3A and S3B.(E and F) Expression of (E) CD38 and CD2 or (F) CXCR6 and CD62L among splenic and hepatic NK cells. Colors in the pie charts indicate CD56/CD16 subsets. Pie arc colors depict the expression of (E) CD38 and CD2 or (F) CXCR6 and CD62L. In (E), scatterplots depict subset frequencies within NKp46+ cells of individual mice (symbol color: black, spleen; clear, liver). Median.(G) Frequencies of CXCR6+ splenic or hepatic NK cells.(H and I) Frequencies of cells expressing (H) CD69 or (I) CD39 among CXCR6+ and CXCR6- NK cells from EBVzko+KSHV mice. Wilcoxon test.In (G)–(I), three independent experiments. Median (IQR). In (C)–(I), NK cells were pre-gated on huCD45+, live, Lin- (CD3, CD19, CD14, CD4) NKp46+ cells. *p < 0.05, **p < 0.01, ***p < 0.001. Unless otherwise stated, statistical analysis was performed by Dunn’s test following KW test (q values).See also Figures S2 and S3.

Fig 5: CD56-CD16+ NK cells are enriched in the NK cell compartment of KSHV-infected huNSG mice, as well as in Kenyan KSHV-sero-positive children(A) Representative flow cytometry dot plots of human CD45+NKp46+Lin- splenocytes of EBVzko and EBVzko+KSHV mice.(B) Frequency of peripheral blood CD56-and CD16-stained NK cell subsets over time in mock (n = 5), EBVzko (n = 5), and EBVzko+KSHV (n = 6) mice. REML followed by Fisher’s LSD test, q values, mean ± SEM. (C and D) Frequency of CD56-and CD16-stained NK cell subsets among splenic (C) or hepatic (D) CD45+CD3-NKp46+ cells of mice from four and two independent experiments, respectively, relative to the mean of the respective EBVzko group. Mean ± SEM, MWU.(E) Correlations between splenic KSHV DNA load and the frequency of CD56+CD16- (red) and CD56-CD16+ (yellow) NK cells are depicted for EBVzko+KSHV co-infected mice from four independent experiments. Solid lines represent trend lines obtained by linear regression, and shaded areas indicate 95% confidence interval (CI) of each trend line. Mice with values below 0.1 ORF26 copies per 106 cells were excluded.(F) Cytokine concentration was measured in the serum of EBVzko (n = 15) and EBVzko+KSHV (n = 16) mice at 4 weeks p.i. The median (IQR) is shown with minimum and maximum ranges as whiskers. Composite data are from four independent experiments, MWU. Dashed lines indicate lower level of quantification (LLOQ). (G) Frequency of CD56-CD16+ NK cells in KSHV-seropositive and KSHV-negative (KSHVab + and -) pediatric healthy donors (HDs). Mean ± SD. MWU.(H) Correlation between CD56-CD16+ NK cell frequencies and serum IL-18 in pediatric HDs and eBL patients.*p < 0.05, **p < 0.01, ***p < 0.001. rS, Spearman’s correlation. See also Figure S1.