[Interferon-α mediating the functional damage of CD56CD57natural killer cells in peripheral blood of systemic lupus erythematosuss].

Objective: To investigate the regulatory effect of interferon-α (IFN-α) on the apoptosis and killing function of CD56CD57 natural killer (NK) cells in systemic lupus erythematosus (SLE) patients, and to explore the specific mechanism.

Methods: A total of sixty-four newly treated SLE patients and sixteen healthy controls (HC) enrolled in the Second Hospital of Dalian Medical University were selected as the research subjects. And the gene expression levels of molecules related to NK cell-killing function were detected by real-time quantitative polymerase chain reaction. CD56CD57 NK cells were co-cultured with the K562 cells, and the apoptotic K562 cells were labeled with Annexin-Ⅴ and 7-amino-actinomycin D. Peripheral blood mononuclear cells were treated with 20, 40, and 80 μmol/L hydrogen peroxide (HO), and treated without HO as control, the expression level of perforin (PRF) was detected by flow cytometry. The concentration of IFN-α in serum was determined by enzyme linked immunosorbent assay. The expression levels of IFN-α receptors (IFNAR) on the surface of CD56CD57 NK cells were detected by flow cytometry, and were represented by mean fluorescence intensity (MFI). CD56CD57 NK cells were treated with 1 000 U/mL IFN-α for 24, 48 and 72 h, and no IFN-α treatment was used as the control, the apoptosis and the expression levels of mitochondrial reactive oxygen species (mtROS) were measured by flow cytometry and represented by MFI.

Results: Compared with HC(=3), the expression levels of gene in peripheral blood NK cells of the SLE patients (=3) were decreased (1.24±0.41 . 0.57±0.12, =0.05). Compared with HC(=5), the ability of peripheral blood CD56CD57 NK cells in the SLE patients (=5) to kill K562 cells was significantly decreased (58.61%±10.60% . 36.74%±6.27%, < 0.01). Compared with the control (=5, 97.51%±1.67%), different concentrations of HO treatment significantly down-regulated the PRF expression levels of CD56CD57 NK cells in a dose-dependent manner, the 20 μmol/L HO PRF was 83.23%±8.48% (=5, < 0.05), the 40 μmol/L HO PRF was 79.53%±8.56% (=5, < 0.01), the 80 μmol/L HO PRF was 76.67%±7.16% (=5, < 0.01). Compared to HC (=16), the serum IFN-α levels were significantly increased in the SLE patients (=45) with moderate to high systemic lupus erythematosus disease activity index (SLEDAI≥10) [(55.07±50.36) ng/L . (328.2±276.3) ng/L, < 0.001]. Meanwhile, compared with HC (=6), IFNAR1 expression in peripheral blood CD56CD57 NK cells of the SLE patients (=6) were increased (MFI: 292.7±91.9 . 483.2±160.3, < 0.05), and compared with HC (=6), IFNAR2 expression in peripheral blood CD56CD57 NK cells of the SLE patients (=7) were increased (MFI: 643.5±113.7 . 919.0±246.9, < 0.05). Compared with control (=6), the stimulation of IFN-α (=6) significantly promoted the apoptosis of CD56CD57 NK cells (20.48%±7.01% . 37.82%±5.84%, < 0.05). In addition, compared with the control (=4, MFI: 1 049±174.5), stimulation of CD56CD57 NK cells with IFN-α at different times significantly promoted the production of mtROS in a time-dependent manner, 48 h MFI was 3 437±1 472 (=4, < 0.05), 72 h MFI was 6 495±1 089 (=4, < 0.000 1), but there was no significant difference at 24 h of stimulation.

Conclusion: High serum IFN-α level in SLE patients may induce apoptosis by promoting mtROS production and inhibit perforin expression, which can down-regulate CD56CD57 NK killing function.