Tumor necrosis factor, macrophage colony-stimulating factor, and interleukin 1 production within sponge matrix allografts.

Neither the presence nor the specific role of secretory cytokines in in vivo allograft rejection has been extensively studied. We quantitated the levels of colony-stimulating factors, tumor necrosis factor, and interleukin 1 within the rejecting allograft. BALB/c (H-2d) mice were implanted with polyurethane sponges containing either allogeneic C57BL/6 (H-2b) or syngeneic splenocytes, or splenocyte-free media. At various days postgrafting, the sponges were harvested, and the cells infiltrating the grafts were analyzed for specific antidonor cytolytic activity, while IL-1, TNF, and CSF levels were measured in the graft exudate fluid. Allogeneic grafts had significantly higher concentrations of CSF, TNF, and IL-1 than syngeneic of splenocyte-free grafts. A specific radioimmunoassay revealed that macrophage colony-stimulating factor (M-CSF) is the primary CSF produced in the grafts. Peak TNF levels preceded peak M-CSF and IL-1 levels, which coincided with the initial appearance of allospecific cytotoxic T lymphocytes. Maximal CTL activity was seen on day 13, when the levels of these cytokines had already begun to fall. Specific bioassays for multi-CSF (IL-3), granulocyte CSF, granulocyte-macrophage CSF, IL-2, and IL-4 failed to detect these cytokines in the sponge fluid at any time. We hypothesize that TNF, M-CSF, and IL-1 probably play regulatory roles in the immunologic events at the site of allograft challenge.     

Interleukin 4 enhances osteoblast macrophage colony-stimulating factor,but not interleukin 6, production

To determine if interleukin 4's (IL-4) recently discovered skeletal effects could be explained by its effects on osteoblasts, we have examined IL-4's impact on macrophage colony stimulating factor (M-CSF) and interleukin 6 (IL-6) secretion by the murine osteoblastic cell line MC3T3-E1. Interleukin-4 increased colony-forming activity in MC3T3 supernatants two-threefold with colony cytomorphology, cytohistochemistry, and blockade of the effect by anti-M-CSF antibody, indicating that the IL-4-induced activity was M-CSF. MC3T3 M-CSF supernatant activity increased in a time-dependent manner with positive IL-4 effects seen after a 24-hour exposure. The maximal IL-4 effective dose was 100 U/ml where conditioned media from IL-4-treated cells contained twofold more M-CSF than control cells (400 U/ml versus 200 U/ml M-CSF) as detected by a sandwich M-CSF ELISA. Northern blots showed that IL-4 (200 U/ml) rapidly increased steady-state M-CSF mRNA levels with maximal induction observed by 2 hours followed by a decline to near basal levels by 24 hours. IL-4 also dose dependently increased M-CSF mRNA levels with maximal induction (fourfold) seen at 100 U/ml IL-4. In contrast to its impact on MC3T3 M-CSF production, IL-4 (200 U/ml) did not stimulate MC3T3 IL-6 secretion whereas IL-1 (1 pM) stimulated a 500-fold increase in MC3T3 IL-6 release. When utilized to treat newborn calvarial osteoblast-enriched cultures, IL-4 dose dependently augmented M-CSF production, with the maximal effect seen at 200 U/ml where IL-4-treated, osteoblast-conditioned media contained almost 500 U/ml M-CSF, compared with 200 U/ml M-CSF in controlconditioned media. These observations indicate that the range of IL-4 cellular targets in skeletal tissues include osteoblastic cells, and that this cytokine increases osteoblast expression of M-CSF, a hematopoietic cytokine pivotal for monocyte/macrophage differentiation. Furthermore, IL-4's impact on osteoblast-produced M-CSF levels is selective because IL-6 levels were unaltered by IL-4 treatment.     

Interleukin-4 regulates macrophage interleukin-12 protein synthesis through a c-fos mediated mechanism

BACKGROUND: Interleukin-4 (IL-4) treatment after lipopolysaccharide (LPS) induction inhibits macrophage (Mphi) IL-12 synthesis; however, IL-4 pretreatment (PreTx) primes the Mphi for increased LPS-induced IL-12 production. In this study we study the role of c-fos in the IL-4 priming of Mphi IL-12 synthesis. METHODS: With a murine in vitro peritoneal M phi model, we studied the effect of either c-fos deficiency (wild type, WT; homozygous c-fos knockout, Homo KO) or c-fos overexpression to study the role of c-fos in IL-4 priming of LPS-induced M phi IL-12 synthesis. RESULTS: (1) We first show that IL-4 PreTx results in a 72% decrease in Mphi c-fos mRNA compared with vehicle PreTx. (2) With respect to IL-12 p70 protein, IL-4 PreTx in the WT group increased LPS-induced Mphi IL-12 p70 2.2-fold compared with vehicle PreTx. Compared with vehicle PreTx in the WT group, vehicle PreTx in the Homo KO group followed by LPS stimulation resulted in a 2.8-fold increase in IL-12 p70 in the Homo KO group. IL-4 PreTx did not significantly increase IL-12 p70 over vehicle PreTx in the Homo KO group. (3) We studied the effect of c-fos overexpression on LPS-induced Mphi IL-12 production when primed with IL-4. Overexpression of c-fos completely inhibited IL-4 primed LPS-induced IL-12 p70 protein synthesis. CONCLUSIONS: These data demonstrated that down-regulation of c-fos is an integral part of the IL-4 priming process for Mphi IL-12 production.     

Glomerular interleukin 1 production is dependent on macrophage infiltration in anti-GBM glomerulonephritis

Both macrophages and glomerular mesangial cells have the potential to synthesize interleukin 1 (IL-1), however, their respective contributions to IL-1 production in anti-GBM glomerulonephritis (GN) are unknown. To address this problem, IL-1 production by glomeruli from rabbits with macrophage-associated anti-GBM GN (passive autologous anti-GBM GN [PAGBMGN]) and macrophage independent (heterologous phase) anti-GBM GN was studied. Macrophage-infiltrated nephritic glomeruli produced IL-1 bioactivity which was inhibitable by an anti-IL-1 antibody, and had a molecular weight consistent with rabbit IL-1. Glomerular IL-1 production in PAGBMGN was markedly augmented (1.43 +/- 0.79 U/10(3) glomeruli [gloms]/24 hr) compared to normal glomeruli (0.13 +/- 0.06 U/10(3) gloms/24 hr, P less than 0.05) or glomeruli from rabbits with macrophage independent GN (0.11 +/- 0.07 U/10(3) gloms/24 hr, P less than 0.05). IL-1 production by glomeruli from leukocyte depleted rabbits with PAGBMGN (0.16 +/- 0.07 U/10(3) gloms/24 hr) was not significantly elevated compared to normal glomeruli. Glomerular macrophages from rabbits with PAGBMGN produced more IL-1 (3.62 +/- 1.63 U/10(3) cells/24 hr) than blood monocytes (0.51 +/- 0.30 U/10(3) cells/24 hr) or alveolar macrophages (0.24 +/- 0.12 U/10(3) cells/24 hr) from the same animals. These results show that in experimental anti-GBM GN where injury is macrophage dependent, IL-1 production is also macrophage dependent and infiltrating glomerular macrophages are the major source of IL-1. Further, as glomerular IL-1 production was not significantly augmented in GN in the absence of macrophages, glomerular deposition of immunoglobulin and complement alone do not stimulate significant IL-1 production by intrinsic glomerular cells in experimental anti-GBM GN.     

转录因子c-fos mRNA及其蛋白在大鼠烧伤创面的表达

目的 研究烧伤后创面组织转录因子c-fos mRNA及其蛋白在伤后不同时相点表达的规律，探讨c—fos在启动烧伤创面损伤与修复中的作用。方法 在Wistar大鼠背部制作30％TBSA深Ⅱ度烧伤模型，取正常皮肤组织与烧伤后不同时相点创面皮肤组织，分别用原位杂交和SP免疫组化方法检测烧伤后c—fos mRNA及其蛋白表达的变化规律及特征。结果 烧伤后c—fos mRNA及蛋白表达明显增多，fos蛋白在伤后3h即到达峰值，而其mRNA则在伤后6h到达峰值。结论 烧伤可以诱导c—fos mRNA和蛋白的表达，但其基因表达增加延后于其蛋白表达增加，提示首先出现的fos蛋白是通过对已存在的fos分子进行翻译后修饰而合成的。     

Polymeric IgA increases the synthesis of macrophage migration inhibitory factor by human mesangial cells in IgA nephropathy.

BACKGROUND: It has been suggested that polymeric IgA (pIgA) or IgA immune complexes play a significant pathogenic role in IgA nephropathy (IgAN). Macrophage migration inhibitory factor (MIF) shares many activities with other pro-inflammatory cytokines. In human glomerulonephritis, including IgAN, glomerular expression of MIF is found to correlate with progressive renal injury. We hypothesized that deposition of pIgA within the kidney may lead to enhanced synthesis of MIF by mesangial cells. METHODS: In this study we examined the effect of pIgA and monomeric IgA (mIgA) from randomly selected patients with IgAN in clinical quiescence on the gene expression and protein synthesis of MIF in cultured human mesangial cells (HMC). RESULTS: Both pIgA and mIgA from IgAN patients or matched healthy controls increased MIF gene expression and protein synthesis in a dose-dependent fashion. The magnitude of MIF protein induction by pIgA (100 microg/ml) was similar to that of tumour necrosis factor-alpha (TNF-alpha) at 10 pg/ml. In all subjects, the induction of MIF was higher for pIgA when compared with mIgA (P < 0.01). Furthermore, the up-regulation of MIF synthesis by either pIgA or mIgA was significantly higher in IgAN patients than in healthy controls (P < 0.05). Similarly, pIgA and mIgA were able to induce TNF-alpha gene expression and protein synthesis in mesangial cells. Incubation of mesangial cells with neutralizing antibody to TNF-alpha reduced the MIF synthesis induced by pIgA. CONCLUSION: We demonstrate that pIgA is capable of inducing MIF and TNF-alpha production in HMC, which may play a major pathogenic role in IgAN. Induction of MIF can be partially blocked by neutralizing antibody to TNF-alpha, suggesting the possibility that up-regulation of MIF synthesis in HMC is mediated via an amplifying proinflammatory loop involving TNF-alpha.     

Blockade of prostaglandin production increases cachectin synthesis and prevents depression of macrophage functions after hemorrhagic shock.

Although hemorrhage severely depresses macrophage functions, it is not known whether the increased TNF-alpha or PGE2 production is responsible for it. To study this C3H/HeN mice were bled to mean blood pressure of 35 mmHg for 60 minutes, resuscitated, and treated with either ibuprofen (1.0 mg/kg body weight) or vehicle (saline). Hemorrhage increased plasma prostaglandin E2 (PGE2) levels by 151.7% +/- 40.0% (p less than 0.05) and significantly decreased peritoneal macrophage (pM phi) antigen presentation (AP) by 60.5% +/- 7.3%, Ia expression by 52.3% +/- 7.6%, and interleukin-1 (IL-1) synthesis by 60.5% +/- 12.3% compared to shams. However ibuprofen treatment reduced PGE2 plasma levels by 61.3% +/- 12.1% and significantly increased AP (+237.0% +/- 95.3%), Ia expression (+72.8% +/- 27.5%), IL-1 synthesis (+235.7% +/- 134.7%), and cachectin synthesis (+485.8% +/- 209.0%) compared to vehicle-treated animals. These results indicate that prostaglandins but not cachectin are involved in the suppression of pM phi functions following hemorrhage because blockade of prostaglandin synthesis improved depressed macrophage functions despite enhanced cachectin synthesis.     

Administration of high-dose macrophage colony-stimulating factor increases bone turnover and trabecular volume fraction

Macrophage colony-stimulating factor (M-CSF) is a hematopoietic growth factor that plays a critical role in early osteoclastogenesis. To characterize the skeletal effects of M-CSF, we administered soluble M-CSF to mice. It was hypothesized that M-CSF would stimulate bone formation through coupled activity of osteoclasts and osteoblasts. Twenty-four male C57BL/6 J mice (n = 12/group, aged 7 weeks) received subcutaneous injections of human M-CSF [5 mg/(kg day)] or inert vehicle (VEH) for 21 days. M-CSF increased serum bone turnover markers (+57% TRAP-5b and +44% osteocalcin). Microcomputed tomography revealed an anabolic effect on tibial trabecular bone, with higher bone volume fraction (+35%), connectivity density (+79%), and number (+18%), as well as lower trabecular separation (−18%). M-CSF had no significant effect on cortical bone mineral content, geometry, or strength. There was no change in quantitative histomorphometry parameters of femoral cortical bone. These results reveal the complex, site-specific effects of M-CSF. In particular, we have demonstrated an anabolic effect of M-CSF on trabecular bone achieved through coupled activation of osteoblasts. However, in contrast to previous studies, M-CSF was found to have no effect on cortical bone. M-CSF was demonstrated to significantly influence both bone modeling and remodeling in relatively young animals.     

Priming interleukin 8 production: role of platelet-activating factor and p38

HYPOTHESIS: Platelet-activating factor (PAF) activates p38, an important intracellular signal transduction kinase, and primes human mononuclear cells for the production of interleukin 8 (IL-8), a potent chemoattractant and activator of neutrophils. METHODS: Human mononuclear cells were isolated from healthy adults by Ficoll-paque density-gradient centrifugation. Interleukin-8 in the supernatant was measured by enzyme-linked immunosorbent assay. Dual phospho-specific p38 antibody was used to detect activated p38 by Western blotting. RESULTS: Lipopolysaccharide (LPS) and PAF activated p38. There was a shorter latency to peak p38 activation with PAF vs LPS stimulation, 5 vs 30 minutes. Platelet-activating factor-induced p38 activation was calcium dependent because it was inhibited by ethyleneglycoltetracetic acid. Lipopolysaccharide, 0.01 to 1.00 ng/mL, induced significant IL-8 production. Although PAF did not induce significant IL-8 production, it potentiated LPS-induced IL-8 production. Production of IL-8, in response to LPS alone or in combination with PAF, was inhibited by SB202190, a specific p38 inhibitor. CONCLUSIONS: Although LPS and PAF activated p38, only LPS induced IL-8 production; PAF acted as a priming agent. It seems that p38 activation is necessary but not sufficient for IL-8 production by human mononuclear cells. Identifying and evaluating the activation state of inflammatory signal transduction pathways might lead to methods for controlling and preventing neutrophil-induced tissue injury without interfering with the normal host immune response.     

转录因子12对前列腺癌进展及预后的影响

目的:探讨转录因子12(TCF12)对前列腺癌进展及预后的影响。方法:利用在线生信分析网站UALCAN、基因表达谱动态分析(GEPIA)、基因、蛋白质相互作用关系检索工具(STRING)及METASCAPE分析TCF12在前列腺组织中表达(高表达组和低表达组)及其与预后的关系,预测TCF12相关基因发挥作用的可能信号通...     

Increased production of macrophage migration inhibitory factor by T cells in patients with IgA nephropathy.

BACKGROUND/AIM: Several studies have demonstrated an upregulation of macrophage migration inhibitory factor (MIF) synthesis in experimental glomerulonephritis. To our best knowledge, no investigation of MIF production by T cells from patients with IgA nephropathy (IgAN) has been reported so far. MIF is one of the immunoregulatory cytokines involved in T-cell activation and delayed-type hypersensitivity. In this study, we examined MIF production of T cells from patients with IgAN to investigate the contribution of the cells to elevated serum MIF content and to its pathologic characteristics. METHODS: We measured MIF production by T cells from the peripheral blood of 20 healthy controls and 20 patients with IgAN before and approximately 10 days after the beginning of steroid therapy. The disease controls included 20 patients with minimal-change nephrotic syndrome (MCNS) before therapy. MIF concentrations in the supernatants of T-cell cultures were measured with a specific enzyme-linked immunosorbent assay (ELISA). We also investigated the relationship between MIF levels and disease activity. RESULTS: The mean level for MIF in patients with IgAN was significantly elevated compared with that of healthy controls and also higher than that of patients with MCNS. When T cells were stimulated by concanavalin A, MIF production by T cells of patients with IgAN was more enhanced than in control subjects or patients with MCNS. We also investigated the relationship between MIF levels and pathological features in IgAN patients. Our findings reveal that MIF levels correlated with the grade of glomerular crescent formation and immunofluorescent C3 deposits in the glomeruli. Moreover, MIF overproduction was significantly related to the acute exacerbation stage of IgAN patients. Elevated MIF levels during the acute phase or exacerbations were found to be decreased during spontaneous or steroid therapy-induced remission in all IgAN patients examined. CONCLUSIONS: We provide data indicating that a T-cell population isolated by negative selection from peripheral blood mononuclear cells shows increased in vitro production of MIF in IgAN, and a reduction in that production when patients are treated with corticosteroids. In this paper, we describe the hypothetical role of MIF in the pathophysiology of IgAN.     

Decreased interleukin generation in patients with cancer of the digestive system. A correlation between interleukin 1 and interleukin 2 production

The generating capacity of interleukin 1 (IL-1) and interleukin 2 (IL-2) by peripheral blood mononuclear cells (PBMNC) was measured in 40 patients with digestive cancer (20 localized and 20 disseminated) and 20 age- and sex-matched control subjects. The localized carcinoma patients showed normal IL-1 production and a significantly depressed IL-2 production (p<0.05) when compared to the healthy individuals. The disseminated carcinoma patients exhibited a significant impairment of both IL-1 and IL-2 production in comparison with the healthy controls (IL-1: p<0.001, IL-2: p<0.001) and the localized carcinoma patients (IL-1: p<0.001, IL-2: p<0.001). A significant correlation was observed between IL-1 and IL-2 generation in all the cancer patients (r=0.458, p<0.01). These results suggest that progressive tumor growth may result in decreased interleukin production by the host PBMNC, and that related mechanisms, which are more susceptable to lymphocytes than monocytes, may be involved in the impairment of both IL-1 and IL-2 production.     

Decreased interleukin generation in patients with cancer of the digestive system. A correlation between interleukin 1 and interleukin 2 production

The generating capacity of interleukin 1 (IL-1) and interleukin 2 (IL-2) by peripheral blood mononuclear cells (PBMNC) was measured in 40 patients with digestive cancer (20 localized and 20 disseminated) and 20 age- and sex-matched control subjects. The localized carcinoma patients showed normal IL-1 production and a significantly depressed IL-2 production (p less than 0.05) when compared to the healthy individuals. The disseminated carcinoma patients exhibited a significant impairment of both IL-1 and IL-2 production in comparison with the healthy controls (IL-1: p less than 0.001, IL-2: p less than 0.001) and the localized carcinoma patients (IL-1: p less than 0.001, IL-2: p less than 0.001). A significant correlation was observed between IL-1 and IL-2 generation in all the cancer patients (r = 0.458, p less than 0.01). These results suggest that progressive tumor growth may result in decreased interleukin production by the host PBMNC, and that related mechanisms, which are more susceptible to lymphocytes than monocytes, may be involved in the impairment of both IL-1 and IL-2 production.     

mRNA and protein expression of transcription factor c—fos in burned rats and their effects on wound healing

Objective:To explore the expression of mRNA and its protein in burned rats and their effects of burn wound healing.Methods:A partial-thickness burn of 30% total body surface area was created on the back of 40 Wistar rats.In situ hybridization and immunohistochemical methods were used to exaluate the location and the amount of the c-fos mRNA and its protein in normal skin and the burned skin,respectively,at 3h,6h,1d,3d,7d and 14d after burn.Results:Under a light microscope,both the expression of c-for mRNA and its protein could be found in the normal skin,but their induction levels were much higher in the burned skin.The level of for protein expression reached peak at 3h after burn while that of c-for mRNA reached peak at 6h after burn.Conclusions:The expression of c-fos can be induced by burns.And the peak level expression of c-for mRNA comes later than that of c-fos protein.It indicates that the action of fos protein is induced by post-translational modification of pre-existing fos molecules.