In vitro intracellular IFNγ, IL-17 and IL-10 producing T cells correlates with the occurrence of post-transplant opportunistic infection in liver and kidney recipients

Figure 1 FACS analysis of cultured CD3⁺ T lymphocyte. Whole blood peripheral CD4⁺ and CD8⁺ T lymphocytes from individual ESLF and ESRF patients as well as HC subjects were stain with anti-CD3, anti-CD8, anti-CD69, anti-IL-17, anti-IL-10 and anti-IFNγ monoclonal antibodies following manufacture’s guidelines upon in vitro stimulation with Io and PMA. CD4⁺ and CD8⁺ lymphocytes were gated within CD3⁺CD69⁺ population following polyclonal activation for 4 h. CD4⁺ T cells were consider to approximate CD3⁺CD8^- T cells. At least 50000 events were acquired. Io: Ionomycin; PMA: Phorbol myristate acetate.

Figure 2 Quantitative analysis of cultured CD4 ⁺ and CD8 ⁺ T lymphocytes from individual end-stage liver failure, end-stage renal failure and healthy control subjects. A: % CD4⁺CD69⁺INFγ⁺; B: CD4⁺CD69⁺IL-17⁺; C: CD4⁺CD69⁺IL-10⁺ cells in ESLF patients and HC individuals; D: % CD4⁺CD69⁺INF-γ⁺; E: CD8⁺CD69⁺IL-10⁺; F: CD4⁺CD69⁺IL-10⁺ cells in ESRF patients and HC individuals. The horizontal lines reflect median values for each group and vertical lines reflect interquartile range. ESLF: End-stage liver failure; ESRF: End-stage renal failure; HC: Healthy control.

Figure 3 Cytokine-producing CD4⁺ and CD8⁺ T cells follow-up along first year after liver and kidney transplantation. A: % of cultured IFNγ-producing CD8⁺ lymphocytes; B: % of cultured TH17 lymphocytes; C: % of cultured TH2 lymphocytes in LTr; D: % of cultured TH1 lymphocytes; E: % of cultured IL-10-producing CD8 T lymphocytes; F: % of cultured TH2 lymphocytes inKTr. The horizontal lines reflect median values for each group and vertical lines reflect interquartile range. ^aP < 0.05, ^bP < 0.01, ^dP < 0.001. Significances express the difference against baseline level.

Figure 4 Stratification analysis of the percentage for the intracellular cytokine production capacity of CD4⁺ and CD8⁺ T lymphocytes between the 1^st and 6^th month post-transplantation. A: % of CD8⁺CD69⁺IFNγ⁺ T lymphocytes in LTr with and without OI; B: % of CD4⁺CD69⁺IFNγ⁺ T lymphocytes in KTr with and without OI; C: % of CD4⁺CD69⁺IL-17⁺ T lymphocytes in LTr with and without OI; D: % of CD8⁺CD69⁺IL-10⁺ T lymphocytes in KTr with and without OI; E: % of CD4⁺CD69⁺IL-10⁺ T lymphocytes in LTr with and without OI; F: % of CD4⁺CD69⁺IL-10⁺ T lymphocytes in KTr with and without OI. LTr: liver transplant recipients; KTr: kidney transplant recipients. ^aP < 0.05, ^bP < 0.01, ^dP < 0.001.

Figure 5 Post-transplantation receiver operating characteristic curve for the intracellular cytokine production capacity and the effect of the % of intracellular cytokine production capacity in stimulated T lymphocytes cut-off values for the discrimination of liver and kidney recipients likely to develop opportunistic infection between the 1^st and 6^th month post-transplantation (Kaplan–Meier analysis). A: Post-transplant % of CD8⁺CD69⁺IFNγ⁺ in LTr; B: Post-transplant % of CD4⁺CD69⁺IFNγ⁺ in KTr; C: Post-transplant % of CD4⁺CD69⁺IL-17⁺ in LTr; D: Post-transplant % of CD8⁺CD69⁺IL-10⁺ in KTr; E: Post-transplant % of CD4⁺CD69⁺IL-10⁺ in LTr; F: post-transplant % of CD4⁺CD69⁺IL-10⁺ in KTr. LTr: Liver transplant recipients; KTr: Kidney transplant recipients.