Table 1.
Summary of demographical data of study participants.
Figure 1.
Loss of CCR7-expressing CD56bright NK cells correlates with clinical disease markers.
(A) Representative flow cytometry plots defining CD56bright NK cells. Numbers indicate percentage of the gated population. (B) Representative CCR7, CD62L, CXCR3 and CD16 expression data and summary data all gated on CD56bright NK cells. Horizontal bars in dot plot indicate mean values. (C) Pearson’s correlation analysis between frequencies of either CD62L+ or CD16+CD56bright NK cells and CCR7+CD56bright NK cells in untreated HIV-seropositive patients. (D) Pearson’s correlation analysis between frequencies of CCR7+, CD62L+, CXCR3+ or CD16+ cells of total CD56bright NK cells with either viral load or CD4+ T cell counts in untreated HIV-positive subjects. *, P<0.05; ***, P<0.001; NS – not significant.
Figure 2.
Relative and absolute loss of CCR7+CD56bright NK cells is not attributable to apoptosis.
(A) Absolute cell numbers of CD56bright NK cells are depicted. Horizontal bars indicate means. (B) Absolute cell numbers of either CCR7+ or CCR7−CD56bright NK cells are shown. (C) Representative flow cytometry data of CD95 on gated CD56bright NK cells and respective summary data derived from untreated HIV-patients. Numbers in flow cytometry plots indicate frequencies of quadrants and horizontal bars in dot plot indicate mean values. (D) Pearson’s correlation analysis between frequencies of CCR7- and CD69-expressing CD56bright NK cells. *, P<0.05; ***, P<0.001; NS – not significant.
Figure 3.
CCR7−CD56bright NK cells exhibit phenotypic features of CD56dimCD16+ cells.
Representative expression data of CD62L, NKG2A, CD27, KIR3DL1, CD69 and CD16 on gated CCR7+ or CCR7− CD56bright cells and respective summary data including CD56dimCD16+ NK cells, from untreated HIV-seropositive individuals. Numbers represent percentages of gated events and horizontal bars in dot plots indicate mean values. *, P<0.05; **, P<0.01; ***, P<0.001.
Figure 4.
Functional differences between CCR7
− and CCR7+CD56bright NK cells from HIV-infected donors indicate high activation states. (A) Representative flow cytometry plots of granzyme B and perforin expression on gated CD56bright NK cells and summary data including CD56dimCD16+ cells, from untreated HIV-infected subjects. Horizontal bars in dot plots show mean values. Numbers in corners represent percentage of quadrant. (B) Representative histograms and summary data of CD107a degranulation in CD56bright cells from uninfected controls, untreated and HAART-treated HIV-patients. Data was generated using sorted NK cells stimulated with IL-12, IL-15 and K452 cells. (C) Representative flow cytometry plot of CD107a degranulation on gated CD56bright NK cells and summary data of degranulation in CCR7+CD56bright, CCR7−CD56bright and CD56dim NK cells from untreated HIV-infected subjects is shown. Data was generated using whole PBMCs stimulated with IL-12, IL-15 and K562 cells. Numbers in corners represent percentage of quadrant. (D) Spontaneous expression of IFN-γ in medium-only treated NK cell subsets is shown in a representative flow cytometry plot and a summary data graph. Data from untreated HIV-positive patients is shown and numbers in corners indicate percentages of quadrants. (E) Representative Ki-67-expression data and summary data on gated CCR7+ or CCR7− CD56bright cells and respective summary data including CD56dimCD16+ NK cells from untreated HIV-seropositive subjects. Numbers in flow cytometry histogram plots indicate percentage of gated events. ***, P<0.001; NS – not significant.