Chinese Journal of Dermatology ›› 2009, Vol. 42 ›› Issue (7): 481-483.

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Effect of culture supernatant of CpG ODN-stimulated neutrophils from patients with psoriasis vulgaris on the proliferation of keratinocytes

  

  • Received:2008-06-11 Revised:2009-04-09 Online:2009-07-15 Published:2009-07-08

Abstract:

Objective To investigate the role of neutrophils in the pathogenesis of psoriasis vulgaris. Methods Neutrophils were isolated from venous blood samples of 25 patients with psoriasis vulgaris (including 13 cases of active psoriasis and 12 cases of inactive psoriasis) as well as 25 normal human controls, and cultured. Then, these neutrophils were grouped and treated with lipopolysaccharide (LPS, 100 g/L), CpG-A (50 mg/L), CpG-B (50 mg/L), and RPMI 1640 culture medium, respectively, for 24 hours followed by the collection of culture supernatants. Human keratinocytes (HaCaT) were cultured in the presence of supernatants of treated or untreated neutrophils for 72 hours followed by the detection of cell proliferation with MTT assay. To determine the role of proinflammatory factors, SOD/CAT and monoclonal antibody to IL-8 and TNF-alpha of 400 u/mL were used to pretreat HaCaT cells 1 hour prior to the stimulation with supernatants of neutrophils. Results Compared with culture medium, the supernatant of unstimulated neutrophils from normal controls or patients with inactive psoriasis had no significant effect on the proliferation of HaCaT cells (P > 0.05), but that from patients with active psoriasis markedly promoted the proliferation of HaCaT cells (t = 2.41, P < 0.05). After stimulation by LPS, CpG-A and CpG-B, the supernatant of active patient- derived neutrophils significantly promoted the proliferation of HaCaT cells compared with that of normal control-derived neutrophils (t = 3.11, 2.89, 2.29, respectively, all P < 0.05). In comparison with unstimulated neutrophils, the supernatant from LPS- and CpG-A stimulated neutrophiles significantly accelerated the proliferation of HaCaT cells. Furthermore, the proliferation of HaCaT cells induced by the supernatants of LPS-, CpG-A-, CpG-B-stimulated neutrophils from psoriatic patients was statistically suppressed by the pretreatment with the monoclonal antibody to IL-8, TNF-alpha and SOD/CAT (all P < 0.05). Conclusions In patients with psoriasis vulgaris, there is an abnormal secretion of IL-8, TNF-alpha and superoxide by neutrophils in peripheral blood, and these proinflammatory factors could promote the proliferation of HaCaT cells.