Pristane-induced Lupus Mice Research Articles

Effect of active immunization with IL-17A on B cell functionand infection risk in pristane-induced lupus model.

The aim of this study was to determine the effect of active immunization of interleukin (IL)-17A to inhibit B cell functions and monitor the risk of infection in a pristane-induced lupus mice model. Female Balb/c mice were given a single intraperitoneal injection of 0.5mL pristane. IL-17A was coupled to keyhole limpet hemocyanin (KLH) and given to mice in three different doses: D0 (0μg/mL), D1 (1μg/mL), and D2 (10μg/mL). The vaccine was given three times with 3-week intervals. At day 42, mice were injected intraperitoneally with methicillin-resistant Staphylococcus aureus (MRSA) and monitored for 3weeks. Plasma cells proliferation, Th17 and plasma cell percentages were measured by flow cytometry; anti-IL-17A antibody titers, IL-17A, and anti-double-stranded DNA (anti-dsDNA) levels were measured by enzyme-linked immunosorbent assay; and MRSA colonization was measured by bacterial counter. Anti-IL-17A antibody titers were significantly higher in D2 compared to D0 (P=0.012). Serum IL-17A levels were also significantly lower in D2 compared to D0 (P=0.000) while Th17 percentages were not significantly different between groups. D2 was also had significantly lower anti-dsDNA (P=0.021), lower plasma cell percentages (P=0.000) and lower B cell proliferation rate (P=0.001) compared to D0. Analysis for the risk of infection also revealed that D2 did not increase the risk of infection compared to D0 (P=0.504). Active immunization with IL-17A coupled to KLH was able to induce a high titer of neutralizing antibodies against IL-17A and inhibit B cell functions without increasing the risk of infection in a pristane-induced lupus mice model.

Regulatory T Cells Compensation Failure Cause the Dysregulation of Immune Response in Pristane Induced Lupus Mice Model.

IntroductionRegulatory T cells’ (Tregs’) role remains unclear in the pathogenesis of systemic lupus erythematosus (SLE). This study was aimed at monitoring the percentage of Tregs within 32 weeks and monitoring its relationship with the percentage of other T helper (Th) cell subsets and the levels of autoantibodies and pro-inflammatory cytokines in a murine SLE model induced by pristane.MethodsForty-eight female BALB/c mice were divided into a healthy control (HC) and a pristine-induced (PI) group. SLE was induced by a single 0.5 cc pristane intraperitoneal injection. Six from each group were sacrificed every eight weeks until 32 weeks post-pristane injection. Treg, Th1, Th2 and Th17 percentages from the spleen were measured using flowcytometry. ANA, IL-6 and IFN-α levels were measured from serum using ELISA.ResultsThe Treg percentage from the PI group increased significantly at 16 weeks compared to the HC group, while Th1, Th2 and Th17 percentages decreased. Tregs in the PI group began to reduce from the 24th to 32nd weeks, followed by an elevation of the Th1, Th2 and Th17 percentages. Tregs were negatively correlated with Th1 and Th2. Tregs in the PI group had a negative correlation with ANA and IFN-α levels from serum, whereas Tregs had a positive correlation with IL-6 levels.ConclusionThe compensation of Tregs observed at 16 weeks after pristane injection failed, marked by a decreasing number of Tregs, followed by an increase of Th subsets, pro-inflammatory cytokines and autoantibodies. This compensatory failure of Tregs could be affected by pro-inflammatory cytokines, such as IFN-α and IL-6.

Oral supplementation of melatonin protects against lupus nephritis renal injury in a pristane-induced lupus mouse model

AimsSince lupus nephritis (LN) etiopathogenesis is not fully understood, herein we investigated the morphological basis of LN in mice induced with pristane. Main methodsTo evaluate the melatonin effects in these animals, we studied the renal cytoarchitecture by means of morphological analyses, immunofluorescence expression of specific markers related to fibrosis, oxidative stress, inflammation and apoptosis. Key findingsWe observed that pristane-LN mice have serious alterations in the kidney cytoarchitecture, i.e. tubular degeneration, glomerular hypercellularity, matrix mesangial expansion and interstitial inflammation. The pristane-induced LN mice treated with melatonin exhibited a well preserved cytoarchitecture. SignificanceOur results document that LN etiopathogenesis is related to both tubular damage and glomerular lesions. We suggest that it is essential to take in consideration both these lesions for LN diagnosis and classification. Clearly, we show that the use of melatonin may be a possible therapeutic strategy for improvement the renal injury in this disorder.

Immune modulation effects of curcumin in pristane-induced lupus mice

Background: Curcumin, a polyphenolic compound derived from food spice turmeric has been widely used in Asian traditional medicine for its medicinal properties as antitumor, antioxidant, and anti-inflammatory properties. Meanwhile, intraperitoneal (i.p.) injection of the hydrocarbon oil pristane into normal mice leads to a lupus-like autoimmune syndrome. We aimed to investigate the effects of curcumin on systemic lupus erythematosus (SLE) clinical manifestation, adaptive immune system components, proinflammatory cytokines, and autoantibody production in pristane-induced lupus mice. Methods: Fifty female BALB/c mice, 6–8 weeks old were divided into 2 groups: Forty mice received a single i.p. injection of 0.5 cc pristane for lupus induction and ten mice as healthy controls. Starting at 16 weeks after injection, forty pristane-induced lupus mice were divided into four groups based on doses of curcumin received intragastrically: 0, 12.5, 50, and 200 mg/kg bw/day daily for 16 weeks. At 32 weeks after injection, all of mice were assessed for arthritis score, proteinuria level, body weights, adaptive immune system components (Th1, Th2, Th17, and Treg percentages) from spleen using flow cytometry; proinflammatory cytokines and autoantibody production, including interleukin-6 (IL-6), interferon-alpha (IFN-α), and antinuclear antibody (ANA) from serum using enzyme-linked immunosorbent assay. Results: Arthritis score and proteinuria level were decreased in curcumin-treated mice. However, body weights were not significantly different between the groups. The decreased of Th1, Th2, and Th17 percentages were seen after treatment with 200 mg/kg bw/day of curcumin (P = 0.031,P = 0.017, andP = 0.005, respectively). However, only slight increase of Treg percentages was seen after curcumin treatment. Treatment with 200 mg/kg bw/day of curcumin decreased serum IL-6 and IFN-α levels (P = 0.007 andP = 0.003). Furthermore, ANA levels were also decreased significantly after treatment with 200 mg/kg bw/day of curcumin (P = 0.013). Conclusion: Our findings suggested that curcumin could prove useful as a therapeutic intervention in SLE.

Immunomodulation Effects of Bryophyllum Pinnatum on Pregnant Pristane-Induced Lupus Mice Model

Objective: To determine the effect of Bryophyllum pinnatum treatment in modulating immune response and the pregnancy outcomes of pregnant pristane-induced lupus mice model. Methods: Sixteen Balb/c mice were intraperitoneally injected with single 0.5 cc pristane to induce lupus manifestations. After 12 weeks of injection, mice were mated and considered as gestational day 0 (GD0). Mice were divided into 4 groups based on the dosages of Bryophyllum pinnatum : control (no treatment), B1 (10.5 mg/kg), B2 (21 mg/kg), and B3 (42 mg/kg). The treatment was given orally every day started from GD9 until 9 days. At the end of the study, blood pressure and fetal size were measured. Serum anti-dsDNA and urine albumin levels were measured by ELISA. Spleen T helper (Th) and mature B cells percentages were measured by flow cytometry. Results: Administration of Bryophyllum pinnatum reduced the percentages of Th1 (p=0.006), Th2 (p=0.005), Th17 (p=0.000), and mature B cells (p=0.007) in dose-dependent manner. B1 and B2 had significantly lower of systolic blood pressure compared to control (p=0.026 and p=0.022 respectively). Significantly lower of anti-dsDNA levels were found in B1 group compared to control (p=0.014). However, no significantly different of urine albumin levels were found between groups. Bryophyllum pinnatum also significantly increased the fetus body weight in dose-dependent manner (p=0.000). Conclusion: Treatment of Bryophyllum pinnatum could improve the pregnancy outcome and modulate the immune response in pregnant pristane-induced lupus mice. Therefore, Bryophyllum pinnatum is a potential herb which can be developed as an immunosuppressive agent in the future.

Effect of Baicalin on the Ex vivo Production of Cytokines in Pristane-Induced Lupus Mice

Effect of Baicalin on the Ex vivo Production of Cytokines in Pristane-Induced Lupus Mice

Immunosuppressive CD11b+Ly6Chi monocytes in pristane-induced lupus mouse model.

Myeloid-derived suppressor cells with immunosuppressive functions have been described to be associated with one of the mechanisms by which malignant tumors escape immune surveillance. However, little is known about the role of myeloid-derived suppressor cells in autoimmunity. In the current study, when we attempted to characterize the peritoneal cells in pristane-induced lupus model, as reported previously, we observed that there were markedly increased CD11b(+)Ly6C(hi) monocytes. Surprisingly, this type of monocytes was almost phenotypically identical to the reported monocytic myeloid-derived suppressor cells. Further analysis on how these CD11b(+)Ly6C(hi) cells affected T cell response showed that they strongly suppressed T cell proliferation in vitro in a manner dependent on cell-cell contact, NO, and PGE2. In addition, we found that CD11b(+)Ly6C(hi) monocytes inhibited Th1 differentiation but enhanced development of forkhead box p3(+)CD4(+) regulatory T cells. Consistent with the in vitro experimental results, the in vivo adoptive cell transfer study showed that infusion of pristane-treated syngeneic CD11b(+)Ly6C(hi) monocytes significantly suppressed the production of anti-keyhole limpet hemocyanin antibodies induced by keyhole limpet hemocyanin immunization. In addition, we found that CD11b(+)Ly6C(hi) monocytes were also increased significantly in spleen and peripheral blood and showed immunosuppressive characteristics similar to their peritoneal counterparts. Our findings indicate that CD11b(+)Ly6C(hi) monocytes in a pristane-induced lupus mouse model are monocytic myeloid-derived suppressor cells instead of inflammatory monocytes, as demonstrated previously. To our knowledge, this is the first to describe myeloid-derived suppressor cells in a pristane-induced lupus mouse model, which may lead to a better understanding of the role of CD11b(+)Ly6C(hi) monocytes in this specific pristane-induced lupus model.

Low Birth Weight and Maternal and Neonatal Deaths are Complications of Systemic Lupus Erythematosus in Pregnant Pristane Induced Lupus Mice

Objectives: This study aims to investigate the relationship between systemic lupus erythematosus and pregnancy in pristane induced lupus mice model, including the pregnancy outcome for mothers and neonates. Materials and methods: Sixteen female Balb/c mice 6-8 week-old were separated into two groups: pristane induced lupus (PIL) mice group that received a single intraperitoneal injection of 0.5 ml of pristane and healthy control (HC) group that received phosphate-buffered saline injection. At one month after injection, all mice were mated with 8-10 week-old male Balb/c mice. All mice from both groups were observed for body weight and survival. On the day of births; number of neonates born, birth weight, and length of neonates were calculated. Neonates were also monitored for survival rate and their ages along the study. Results: There was no statistically significant difference in body weight before and after delivery, survival rate during pregnancy, number of neonates born, and gestational ages of mothers from both groups. Mothers began to die in the postpartum period. On third day after delivery, 0% of mothers from PIL group survived whereas 100% mothers from HC group survived (p=0.000). Of neonates born from PIL group, 15% survived at the end of fifth day after birth whereas 100% of neonates from HC group still survived (p=0.000). Neonates from PIL group also had significantly lower body weight and body length at birth compared to HC group (p=0.000 and p=0.000, respectively). Conclusion: Maternal and neonatal death, low birth weight, and low birth length are complications of systemic lupus erythematosus at pregnancy in pristane induced lupus mice model.

Resveratrol possesses protective effects in a pristane-induced lupus mouse model.

BackgroundSystemic lupus erythematosus (SLE) is a multisystemic autoimmune disease characterized by the production of autoantibodies. To date, no therapy has been found to satisfactorily treat SLE. SIRT1 deficiency results in the development of an autoimmune syndrome in mice, including a high titer of anti-nuclear antibody in serum, immunoglobulin deposition in the kidney, and immune complex glomerulonephritis. Resveratrol is an activator of SIRT1 and possesses anti-inflammation and immune-regulatory properties.ObjectiveTo evaluate the preventative effects of resveratrol on a pristane-induced lupus animal model and assess its putative immune modulation effects.MethodsBALB/c mice received a single intraperitoneal injection of 0.5 ml of pristane on day 1 and then various doses of resveratrol were given to the mice daily starting on day 2 and continuing for seven months. The autoantibodies in serum and supernatants were measured. Single cells isolated from spleen, isolated CD4+ T cells, and CD19+ B cells were cultured with or without resveratrol in vitro and assessed by flow cytometry.ResultsResveratrol attenuated proteinuria, immunoglobuin depositon in kidney, and glomerulonephritis as well as IgG1 and IgG2a in serum in pristane-induced lupus mice. Resveratrol also suppressed CD69 and CD71 expression on CD4+ T cells as well as CD4+ T cell proliferation, induced CD4+ T cell apoptosis, and decreased CD4 IFNγ+ Th1 cells and the ratio of Th1/Th2 cells in vitro. In vitro antibody production and proliferation of B cells were also inhibited.ConclusionResveratrol possesses protective effects in pristane-induced lupus mice and may represent a novel approach for the management of SLE.

OP0103 Decreased Lupus Severity in Mir155 Deficient Mice

BackgroundMicroRNAs (miRs) recently came into view as an important class of regulators of gene expression that are associated with a variety of biological functions. Deregulation of endogenous miR155 was observed...

Pristane-induced lupus as a model of human lupus arthritis: evolvement of autoantibodies, internal organ and joint inflammation

Arthritis is frequently seen in human lupus, but rarely in lupus models. Pristane-induced lupus (PIL) can be induced in various mouse strains such as BALB/c and C57BL/6. We herein characterize clinical and histological features of arthritis in the context of systemic lupus and provide a prudent comparison with models of rheumatoid arthritis (RA). A total of 57 BALB/c mice received pristane (PIL group) and were analyzed for serum autoantibodies (anti-chromatin-, -histone, -Sm, -dsDNA), as well as for clinical features and histopathology of joints, lungs and kidneys. Joint pathology was quantified by image analysis and tissue cytometry. Ten C57BL/6 mice (Bl/6-PIL) and historical groups of two different RA models were analyzed accordingly. In BALB/c PIL, clinical arthritis started at three months, occurred finally in 79% of PIL (but not in controls, p<0.001) and correlated with areas of inflammation, erosion, cartilage damage, osteoclast numbers and total severity score (for all: r>0.7, p<0.001). After eight months, 58% of PIL (but no controls, p<0.001) had mild-erosive arthritis. In contrast to RA, the most frequent inflammatory cell type of the pannus was granulocytes (17.7%), PIL had lower numbers of osteoclasts, erosions rarely affected both layers of the cortical bone and there was no progression to complete joint destruction (even after one year of observation). Serum autoantibodies (auto-abs) preceded arthritis and became significantly elevated in all PIL; affected joints showed increased deposits of IgG (and IgM) within the inflammatory tissue, indicative of an ab-mediated process. PIL mice with arthritis also showed signs of pulmonary (100%) and renal (46%) lupus. In contrast to BALB/c, Bl/6-PIL mice did not develop any signs of arthritis. PIL in BALB/c mice is characterized by severe organ involvement, typical autoabs and by a mild-erosive arthritis with similarities to, but also with distinct differences from, RA. PIL may help to study arthritis along with other key features of systemic lupus erythematosus after therapeutic interventions or in knock-out models based on a BALB/c but not on a C57BL/6 background.

AB0127 Arthritis in a model for systemic lupus: involvement of joints, inner organs and course of autoantibodies in pristane-induced lupus.

ObjectivesArthritis is frequently seen in human lupus, but rarely in lupus models. Pristane-induced lupus (PIL) can be induced in various mouse strains such as BALB/c and C57Bl/6. We herein characterize...

SAT0164 Pristane-induced lupus in BALB/C mice: A model for lupus arthritis

BackgroundSystemic lupus erythematosus (SLE) is frequently featured by musculoskeletal issues as arthralgia and arthritis, which account for the most important burdens in patient-based questionnaires. In most cases, lupus arthritis appears...

Regulatory Effect of Melatonin on Cytokine Disturbances in the Pristane-Induced Lupus Mice

Systemic lupus erythematosus (SLE) develops in relation to many environmental factors. In our opinion, it is more important to investigate the effect of melatonin on the environmental- related SLE. In the present study, 0.5 ml pristane were used to induce SLE in female BALB/c mice. Melatonin (0.01, 0.1, 1.0 mg/kg) was orally administered immediately after pristane-injection for 24 weeks. IgM anti ssDNA and histone antibodies were detected after 0, 1, 2, 4, 8 weeks pristane injection. The levels of IL-2, IL-6 and IL-13 were detected after 24 weeks. Renal lesions were also observed. The results showed that melatonin antagonized the increasing levels of IgM anti ssDNA and histone autoantibodies. Melatonin could also decrease the IL-6 and IL-13 production and increase the IL-2 production. Besides, melatonin could lessen the renal lesions caused by pristane. These results suggested that melatonin has a beneficial effect on pristane-induced lupus through regulating the cytokines disturbances.

Prostaglandin E2-mediated dysregulation of proinflammatory cytokine production in pristane-induced lupus mice

Systemic lupus erythematosus (SLE) is characterized by inflammatory and dysregulatory immune responses including overactive B cells, overproduction of proinflammatory cytokines, and T cell hyperactivity. PGE(2) modulates a variety of immune processes at sites of inflammation, including production of inflammatory cytokines. However, the role of PGE(2) in dysregulatory inflammatory and immune responses in lupus remains unclear. We investigated whether PGE(2) mediates production of inflammatory cytokines in pristane-induced lupus BALB/c mice. Our results showed that levels of serum and BAL PGE(2) and LPS-stimulated production of PGE(2) by peritoneal macrophages were remarkably increased in pristane-induced lupus mice compared to healthy controls. Exogenous PGE(2) enhanced production of IL-6, IL-10, and NO but decreased TNF-alpha by macrophages and augmented IFN-gamma, IL-6, and IL-10 by splenocytes from pristane-induced lupus mice compared to healthy controls. Exogenous PGE(2) also enhanced production of IFN-gamma, IL-6, and IL-10 by thymocytes from pristane-induced lupus mice. Indomethacin (Indo), a PGE(2) synthesis inhibitor, greatly inhibited LPS-induced production of IL-6 and IL-10 by macrophages from pristane-induced lupus mice, while enhanced TNF-alpha. Indo remarkably inhibited Con A-increased production of IFN-gamma, IL-6, and IL-10 by splenocytes and thymocytes from pristane-induced lupus mice. Therefore, our findings suggest that endogenous PGE(2) may mediate dysregulation of production of proinflammatory cytokines, such as IL-6, IL-10, and IFN-gamma, and NO in pristane-induced lupus mice.

Immunoregulatory abnormalities of T cells and hyperreactivity of B cells in theIn Vitro immune response in pristane-induced lupus mice

Systemic lupus erythematosus (SLE) is characterized by overactive B cells that differentiate into autoantibody-forming cells, aberrant T cell function that provides helping B cells produce autoantibodies, and overproduction of proinflammatory cytokines. However, immunodysregulation in lupus pathogenensis remains incomplete. We examined mitogen-stimulated production of proinflammatory cytokines, cell proliferation, T cell activation, and T cell apoptosis in vitro in pristane-induced lupus BALB/c mice compared to normal mice. LPS-stimulated production of IL-6 and IL-10 by splenocytes and macrophages from pristane-induced lupus mice were remarkably up-regulated compared to normal mice, whereas production of macrophage TNF-alpha was significantly down-regulated. Moreover, in vitro production of IL-2, IL-6, IL-10 and IFN-gamma by Con A-stimulated splenocytes, cell proliferation in LPS- or Con A-stimulated- thymocytes and splenocytes, and expression of CD69+CD4+ T cells in Con A-stimulated splenocytes were greatly increased in cells derived from pristane-induced lupus mice compared to normal mice. In addition, splenic T cells and CD4+ T cells in thymocytes from pristane-induced lupus mice were more resistant than nonautoimmune normal cells to Con A-induced apoptosis. Our findings indicate that immunoregulatory abnormalities of T cells and hyperreactivity of B cells in the in vitro immune responses in pristane-induced lupus mice may explain some of lupus pathogenesis.