| Part1:Th22, But Not Th17Might Be a Good Index to Predict the Tissue Involvement of Systemic Lupus ErythematosusBackgroundT helper cells, also called auto-reactive effector CD4+T cells, were first subdivided into2groups, Thl and Th2cells, and Thl cells were believed to a main drive of autoimmune diseases. However, this notion often met challenged with some inconsistencies. The identification of newer effector T-cell subsets in recent years such as Th17and Th22cells brings new insights to these diseases. Th17cells, an independent T cell lineage distinct from classical Thl and Th2cells, which produce IL-17A, play a pro-inflammatory role in autoimmune diseases via the up-regulation of additional proinflammatory and neutrophil-recruiting cytokines and chemokines. Furthermore, increasing evidence both in humans and in mouse models suggesting that Th17cells play a role in the progression of SLE has been noted. Th22subset, a new subset of CD4+T-helper, differentiate from naive T cells in response to TNF-a and IL-6and is characterized by secretion of IL-22but not IL-17or IFN-y. Abnormal expression of Th22cells was seen in peripheral blood of some autoimmune diseases including psoriasis [19], systemic sclerosis (SSc)[20], and rheumatoid arthritis (RA)[21,22]. IL-22-positive CD+T cells were present in high quantities in psoriatic skin [15] and RA synovial tissues [23]. However, the role of Th22cells in systemic lupus erythematosus (SLE) has not been reported yet.In present study, we measured the frequency of Th22cells and their secretion of IL-22in peripheral blood (PB) of patients with SLE and discussed the underlying mechanism of their roles in SLE.Method65patients diagnosed of SLE were recuited.30sex and age matched health Volunteers were accepted as healthy controls(HC).Artti-coagula-ted peripheral blood was collected and peripheral blood mononuclear cells(PBMC)were isolated.PBMC was cultured in RPMI1640complete culture medium with PMA and Ionomycin to stimulate the expression of intracellular cytokine in the presence of protein transport inhibitor-Golgiplug. Th1, Th17and Th22cells were identifiedby the expression of surface CD4and intracellular cytokine INF-r. IL-17and IL-22respectively.Serum concentrations of different cytokines were quantified by enzyme-linked immunosorbent assay (ELISA) kits for IL-22and IL-17Clinical data was collected, organ involvement was asessed and SLEDAI score was evaluated according to SLEDAI2000system.Results1. Th17cells was significantly increased in SLE patients compared to healthy controls (P=0.003), and significant difference between active and inactive SLE patients (P=0.000) could be seen. However, Th22cells or Thl cells were unchanged compared to healthy controls.2. Th22cells were significantly elevated in patients with sole lupus skin disease, while they decreased in patients with sole lupus nephritis [(2.03±1.02%) and (0.60±0.19%), respectively versus (1.28±0.40%);(P=0.000,0.020respectively)]3. Thl7cells was significantly increased in both patients with sole lupus skin disease and patients with sole lupus nephritis compared with healthy controls [(2.02±0.82%) and (1.98±0.62%) respectively, versus (1.13±0.65%);(P=0.000,0.000respectively)]. For Thl cells, there was no obvious difference between two groups of patients and healthy controls4. Serum IL-22was unchanged in SLE [median,196.26pg/ml (range36.80-531.70)], compared with healthy controls [median,187.61pg/ml (range56.00-429.00); P=0.725], and no significant difference in serum IL-22between active and inactive SLE patients could be seen, the serum level of IL-22was significantly increased in patients with sole lupus skin disease [median,289.34pg/ml (range89.70-531.70); P=0.001], while it decreased in patients only with sole lupus nephritis [median,82.44pg/ml (range,36.80-178.50); P=0.024]5. Increased level of IL-17was seen in SLE [median,73.08pg/ml (range,17.00.-176.00)], compared to healthy controls [median,46.76pg/ml (range,16.05-173.20); P=0.001]. the level of IL-17in active SLE patients was significantly higher than that in inactive SLE patients P=0.002], and compared to healthy controls, the level of IL-17was increased in both SLE patients with sole lupus skin disease [median,77.78pg/ml (range,35.00-172.00); P=0.004] and SLE patients with sole lupus nephritis [median,85.20pg/ml (range,23.10-160.40); P=0.000]6. Positive correlation both between Th22cells and serum level of IL-22and between Th17cells and serum level of IL-17in patients with SLE(r=0.855, p=0.000; r=0.771,p=0.000).7. Th17cells and serum level of IL-17positively correlated with SLEDAI in SLE patients (r=0.279, p=0.012; r=0.211, p=0.046). Th22, Thl or IL-22failed to show a statistical correlation with SLEDAI (p=0.120, p=0.070and p=0.280respectively). ConclusionAbnomality of T helper subgroups and their cytokines exists in SLE.Patients with active disease had higher expression of Th17cells and increased level of IL-17correlating with the disease activity of SLE. Th17cells could act as biomarker as acticve disease. Expression of Th22cells and IL-22was higher in patients with sole lupus skin disease, and lower in patients with sole lupus nephritis. Therefore, Th22or IL-22might be a more important index to predict the tissue involvement of SLE.To investigate their proprty in SLE,more studies about the expression of Th22or IL-22in tissue are needed. Part2Up-Regulation of Renal IL-22/IL-22R Expression in MRL/lpr MiceBackgroundSystemic lupus erythematosus is a systemic autoimmune disease, characterized by a multitude of autoantibody production, complementactivation, and immune-complex deposition, which causes tissue and organ damage. Many different factors contribute to the pathogenesis of SLE, and cytokine is one of the most important factors. IL-22belongs to the IL-10family of cytokine. Special immune cell populations including Th cells secreated IL-22. The main biological role of IL-22includes the increase of innate immunity, protection from damage, and enhancement of regeneration. Recent studies have showed in the serum of patients with SLE, IL-22levels were significantly decreased compared with normal controls. However, our results in part I have demonstrated unchanged serum IL-22levels in patients with SLE. In the study, to invesitage the role of IL-22in SLE, we detect the expression of IL-22/IL-22R in MRL/lpr mice.Materials and MethodsSerum samples and kidney tissue were obtained from MRL/1pr mice and age-matched C57BL/6J(control) mice at age6weeks,12weeks and18weeks. The following were carried out:1. Anti-dsDNA antibodies levels and IL-221evels in serum were determined by enzyme linked immunosorbent Assay(ELISA).2. Renal and lymph node morphologic features were examined by light microscopy3. The mRN A expression of IL-22/IL-22R were invesitaged by RT-PCR. 4. Immunohistologic analyses were employed to examine the localization of IL-22/IL-22R in kidneys and lymph tissues.Reslut1. MRL/lpr mice showed characteristic alterations of serum immune parameters, with progressive increases in the levels of antibodies with age, compared with age-matched C57BL/6J(control) mice. The levels of anti-dsDNA antibodies in MRL/lpr mice were significantly higher than what in C57BL/6J mice at ages12weeks and18weeks2. Conventional histologic staining of kidneys and lymph node demonstrated minimal abnormalities in MRL/lpr mice at ages6weeks. MRL/lpr mice showed progressive development of renal damage, and obvious follicle proliferation and germinal center of lymph node, which is noticeable at ages12weeks and reaches significance at ages18weeks. The observed lesions included the presence of enlarged hypercellular glomeruli, with increased numbers of both resident cells and infiltrating leukocytes. A pareallel increase in mesangial matrix was also noted. Prominent interstitial mononuclear cell infiltrating was also observed, with predominantly perivascular localization in the cortex and medulla of the kindey. Crescents were identified in the later phases. In contrast, no histopathologic abnormalities can be observed in C57BL/6J mice.3. Serum IL-22levels in MRL/lpr mice were unchanged, in compared with what in C57BL/6J mice.4. Higher expression of IL-22, IL-22RmRNA was found in MRL/lpr mice than what in C57BL/6J mice.5. Strong expression of IL-22, IL-22R protein was found mainly located in glomeruli of MRL/lpr mice, especially in the enlarged hypercellular glomeruli, whereas weak and negative expression was found in glomeruli of C57BL/6J mice. 6. No expression of IL-22, IL-22R protein was found in lymph node in MRL/lpr mice and C57BL/6J mice.ConculationSerum IL-22levels did not differ between MRL/1pr mice and C57BL/6J mice.Whereas higher expression of IL-22, IL-22RmRNA were prominent in the kindey of MRL/lpr mice. IL-22, IL-22R proteins were more distinguished in the enlarged hypercellular glomeruli. The results the present study demonstrated that IL-22/IL-22R axis may be over-activated in murine lupus nephritis, suggesting that IL-22/IL-22R may contribute to the pathogenesis of lupus nephritis... |
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