Table 1.
Patient characteristics.
Figure 1.
CD8+ T-cells from IFN-OFF patients have an increased central memory compartment.
Memory status of CD8+ T-cells was determined by FACS-analysis using anti-CD45RA and –CCR7 antibodies. A) Examples of 3 IFN-OFF, 3 IFN-ON and 3 healthy controls. TCM, T central memory cells; TEM, T effector memory cells; TEMRA, T CD45RA+ effector memory cells B) The proportion of naïve CD8+ T-cells (CD45RA+CCR7+), C) CD8+ CD45RA+ effector memory T-cells (CD45RA+CCR7−), D) CD8+ central memory T-cells (CD45RA−CCR7+), and E) CD8+ effector memory T-cells (CD45RA−CCR7−) from all CD8+ T-cells. White dots represent pregnant patients in IFN-ON group (Table 1).
Figure 2.
CD4+ T-cells from IFN-OFF patients have an increased effector memory compartment.
Memory status of CD4+ T-cells was determined by FACS-analysis using anti-CD45RA and –CCR7 antibodies. A) Examples of 3 IFN-OFF, 3 IFN-ON and 3 healthy controls. TCM, T central memory cells; TEM, T effector memory cells; TEMRA, T CD45RA+ effector memory cells. B) The proportion of naïve CD4+ T-cells (CD45RA+CCR7+), C) CD45RA+ effector memory CD4+ T-cells (CD45+CCR7−), D) CD4+ central memory T-cells (CD45RA−CCR7+), and E) CD4+ effector memory T-cells from all CD8+ T-cells. White dots represent pregnant patients in IFN-ON group (Table 1).
Figure 3.
CD4+ T-cells from IFN-OFF patients are more prone to secrete TNF-α/IFN-γ than CD4+ T-cells from the healthy.
T-cells were stimulated with anti-CD3, –CD28 and -CD49d. After 6 h incubation, cell surface markers and intracellular cytokines were stained and analyzed with flow cytometry. A) Non-stimulated and stimulated TNF-α/IFN-γ secretion of CD3+ T-cells. Shown are 3 representative patients from each group (IFN-OFF, IFN-ON and healthy). Percentages are calculated from CD4+ T-cell population. B) TNF-α/IFN-γ secretion of CD4+ T-cells in all patients C) TNF-α/IFN-γ secretion of CD8+ T-cells in all patients. White dots represent pregnant patients in IFN-ON group (Table 1).
Figure 4.
Function of T-cell subsets in IFN-OFF patients.
A) T-cells were stimulated with anti-CD3, –CD28 and -CD49d and after 6 h incubation, cell surface markers and intracellular cytokines were stained and analyzed with flow cytometry. The cytokine secretion of CD4+ T-cell subsets (effector and central memory, naïve and temra cells) was analyzed separately. The percentage values present the proportion of cytokine secreting cells from all CD4+ cells showing that the effector and central memory T-cells are the main cytokine producers. B) The proportion of cytokine secreting effector and central memory CD4+ T-cells were also analyzed separately and the % values show the proportion of positive cells from a subset in question. C) Cytokine secretion of CD8+ T-cell subsets and D) the proportion of cytokine secreting effector and central memory CD8+ T-cells separately. E) Degranulation of CD8+ T-cells was analyzed by CD107a/b expression with flow cytometry after 6 h incubation with anti-CD3, -CD28 and -CD49d. Unstimulated MNCs were used as a control.
Figure 5.
NK-cells from IFN-OFF patients have a mature phenotype.
NK-cell proportions and surface markers CD62L, CD57 and CD27 were analyzed with flow cytometry and absolute NK-cell counts were counted from total lymphocyte numbers. A) The proportion of NK-cells in IFN-OFF and IFN-ON patients B) Absolute amount of NK-cells in IFN-ON and IFN-OFF patients C) The proportion of CD56DIM NK-cells from CD56 NK-cells in IFN-ON and IFN-OFF patients D) Absolute amounts of CD56DIM NK-cells in IFN-ON and IFN-OFF patients. E) CD57 expression in CD56DIM NK-cells. F) CD62L expression in CD56DIM NK-cells G) CD27 expression in CD56DIM NK-cells. White dots represent pregnant patients in IFN-ON group (Table 1).
Figure 6.
The cytotoxicity of NK-cells from IFN-OFF and IFN-ON patients is impaired.
MNCs were used as effector cells and K562 cells as target cells. The NK-cell percentage was determined by flow cytometry and the number of effector MNCs was counted accordingly. Cells were co-incubated for 6 h at +37°C at effector:target ratios 4∶1 and 8∶1. The graphs present alive K562 cells after the co-incubation with effector cells.
Figure 7.
Degranulation and cytokine secretion of NK-cells.
MNCs were incubated for 6 hours with and without K562 cells at +37°C and the degranulation (CD107a/b expression) and cytokine secretion of NK-cells was measured by flow cytometry. A) Degranulation of NK-cells without stimulation B) Degranulation of NK-cells after the stimulation with K562-cells C) IFN-γ/TNF-α secretion by NK-cells without stimulation D) IFN-γ/TNF-α secretion by NK-cells after the stimulation with K562-cells. White dots represent pregnant patients in IFN-ON group (Table 1).