PMC3568014_01
HIV and HCV viral loads, hepatic functions and CD4 count. A: Hepatic functions determined by liver enzyme assays (ASAT: L-aspartate aminotransferase (light grey circles) (U/L); ALAT: L-alanine aminotransferase (grey squares) (UL/) and gammaGT: gamma-glutamyltransferase (triangles)(U/L)) and HCV viral load (black crosses, number of HCV UI/mL) over time. B: Absolute numbers of CD4+ T-cells (grey line, number of CD4/mm3) and HIV viral load (black line, number of copies/mL; limit of detection: 40 copies/ml) over time.
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F1
Moreover his hepatic assessment, as observed with L-aspartate aminotransferase (ASAT), L-alanine aminotransferase (ALAT) and gammaGT gamma-glutamyltransferase markers is not so much disrupted (Figure 1A). In 1989 his initial CD4+ T-cells count was 196/mm3 and since 1995 remains between 5 and 59/mm3 (Figure 1B). HIV viral loads have been mostly undetectable (below 40 copies/mL) or very low (matching deliberate HAART interruptions) since 2008 (Figure 1B).
Percentages of Natural Killer (NK) cells and their subsets; lytic potential of NK cells. A: Representative dot plot of CD16lowCD56bright; CD16+CD56bright, CD16+, CD56dim and CD16brightCD56- cells. Based on forward and side scatter analysis, cells were first gated on lymphocytes (not shown) and then on CD3- cells. B: Absolute numbers of CD4+ T-cells (light grey line, number of CD4/mm3), % of CD56bright in total CD56 NK cells (dark grey line, % of CD56bright cells) and HIV viral load (black line, number of copies/mL) during one year. C: Representative dot plots of the expression of activating and inhibitory receptors on NK-cells among CD56+ CD16+/- cells. D: Representative dot plots of CD107a degranulation and IFN-gamma secretion on CD56dim and CD56bright cells, following PBMC activation by K562.
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F2
Over the past two years, the immunophenotypic and functional status of his CD3-CD56+ NK-cells were explored by multi-parameter flow cytometry (reagents from Becton Dickinson, MountainView, CA, Figures 2A-D). Surprisingly, an abnormally high percentage of NK-cells has been steadily observed, representing approximatively 40% of peripheral lymphocytes and characterized by an expansion of the CD56bright subset (~30 to 50% of NK-cells) (Figures 2A and 2B). The percentage of CD56bright cells seems to increase when the number of CD4 decreases moreover HIV viral load is below 40 copies/ml when % of CD56bright cells is the more important (Figure 2B). The inhibitory CD94/NKG2A receptor is expressed by all CD56bright cells and a majority of CD56dim NK-cells while there is a low expression of CD158b on CD56dim and CD56bright NK-cells and a faint labeling with CD158a (Figure 2C). Strikingly, these cells did not produce IFN-gamma following NK activation by K562 (Figure 2D).