Production of Rantes/CCL5 in human gingival fibroblasts challenged with tumor necrosis factor alpha

Chemokines are small-secreted proteins that stimulate the directional migration of leukocytes and thereby mediate the inflammatory process. The present study investigates the capacity of human gingival fibroblasts to produce the beta chemokine Rantes/CCL5. In situ hybridization, immunohistochemistry and ELISA were used to measure the induction of Rantes/CCL5 at the mRNA and protein levels, both in unstimulated gingival fibroblasts as well as in fibroblasts treated with the proinflammatory cytokines tumor necrosis factor (TNF)alpha or interleukin (IL)-1beta. TNFalpha in different concentrations (0.1-10 ng/ml) induced Rantes/CCL5 mRNA expression and protein production in 24-h cultures of human gingival fibroblasts. The expression of Rantes/CCL5-mRNA and protein production, induced by TNFalpha, was evident at 6 h and thereafter increased continuously during the study period (24 h). IL-1beta (3-300 pg/ml) also enhanced the production of Rantes/CCL5 in gingival fibroblasts. The amount of Rantes/CCL5 induced by IL-1beta (300 pg/ml), however, was less than that induced by TNFalpha (10 ng/ml). The study suggests that human gingival fibroblasts, by producing the chemokine Rantes/CCL5, participate in the regulation of the host response during the inflammatory process in the periodontal tissue.     

Upregulation of the chemokines Rantes, MCP-1, MIP-1a and MIP-2 in early infection with Trypanosoma brucei brucei and inhibition by sympathetic denervation of the spleen

We examined the induction of 4 chemokines during early experimental African trypanosomiasis using in situ hybridization and immunocytochemistry. mRNA expression and protein production of Rantes, MCP-1, MIP-1a and MIP-2 were studied in splenocytes obtained at 0 h, 4 h and 12 h post-infection. Splenic denervation was performed to study the role of the central nervous system in early infection. The mRNA for Rantes increased at 4 h and declined at 12 h, but the protein level was high at both time-points. MCP-1 and MIP-la had elevated mRNA and protein levels at 12 h post-infection. MIP-2 mRNA was high at both 4 h and 12 h, but the protein level was only increased at 12 h. Splenic denervation, but not sham operation, suppressed these responses. The upregulation of these chemokines during very early infection suggests a chemokine role in the developing immunopathology The sympathetic nervous system may, however, participate in modulation of such early immune responses.     

Differential in situ distribution of interleukin-8, monocyte chemoattractant protein-1 and Rantes in human chronic periapical granuloma

In situ distribution of three prototype chemokines interleukin (IL)-8, monocyte chemoattractant protein (MCP)-1 and Rantes was determined in chronic human periapical granulomas by immunohistochemistry using monoclonal antibodies. IL-8 was found primarily in the cytoplasm of the Malassez epithelial cells. MCP-1 immunoreactivity was confined to the endothelial cells that lined small venules. Each of the three investigated chemokines, including Rantes, exhibited a characteristic binding pattern to the extracellular matrix of the lesion. The observed chemokines may play a role in establishing the cellular composition of chronic apical periodontitis, thus augmenting the intensity of local inflammation and tissue damage.     

一种新型重组免疫毒素DT390-Rantes真核表达质粒的构建及功能

目的 构建一种新型重组免疫毒素DT390-Rantes的真核表达质粒,并对这种免疫毒素的功能进行初步研究。方法 通过RT-PCR的方法,从小鼠肝脏中获得Rantes基因;将Rantes基因插入到含有DT390片段的真核质粒SRα中,构建成重组质粒;重组质粒转入大肠杆菌JM109,筛选出含有正确插入片段的克隆;限制性内切酶酶切图谱分析重组质粒,并进行DNA序列分析;用脂质体介导法转染NIH3T3细胞,免疫荧光检测重组质粒的表达情况;MTT法测定DT390-Rantes的表达活性。结果 经过酶切分析及DNA测序证实,Rantes基因正确插入到真核表达质粒SRα中,并在NIH3T3细胞中表达;MTT法证实,重组免疫毒素DT390-Rantes能在体外杀伤活化的T细胞。结论 成功地构建了一种新型重组免疫毒素真核表达质粒DT390-Rantes-SRα,该质粒可在真核细胞中瞬时表达,其转染上清对活化的T细胞具有杀伤作用。     

In flammatory and physiological roles of chemokines

Chemokines, members of the family of chemotactic peptides, have a well documented function in different inflammatory diseases where they induce leukocyte emigration into lesions. Several recent observations indicate that, in addition to pathological states, chemokines are also produced and secreted under physiological conditions by various exocrine glands in amounts sufficient for their full biological effect. The glands involved in chemokine production and secretion include eccrine sweat glands, lactating mammary glands, lacrimal and salivary glands. It is suggested that analogous to their role in inflammatory diseases, chemokines produced by the exocrine glands are responsible for the induction of homeostatic leukocyte migration into mucosal epithelia and skin and also, mammary glands and milk. In addition, the mechanism by which chemokines induce leukocyte homing under physiological circumstances is discussed.     

一种新型重组免疫毒素DT390-Rantes真核表达质粒的构建及功能

目的 构建一种新型重组免疫毒素DT390-Rantes的真核表达质粒，并对这种免疫毒素的功能进行初步研究。方法通过RT-PCR的方法．从小鼠肝脏中获得Rantes基因；将Rantes基因插入到含有DT390片段的真核质粒SIh中．构建成重组质粒；重组质粒转入大肠杆菌JM109，筛选出含有正确插入片段的克隆；限制性内切酶酶切图谱分析重组质粒，并进行DNA序列分析；用脂质体介导法转染NIH3T3细胞，免疫荧光检测重组质粒的表达情况；MTT法测定DT390-Rantes的表达活性。结果经过酶切分析及DNA测序证实，Rantes基因正确插入到真核表达质粒SR仅中，并在NIH3T3细胞中表达；MTT法证实，重组免疫毒素DT390-Rantes能在体外杀伤活化的T细胞。结论成功地构建了一种新型重组免疫毒素真核表达质粒DT390-Rantes-SRa，该质粒可在真核细胞中瞬时表达．其转染上清对活化的T细胞具有杀伤作用。     

Differential in situ distribution of interleukin‐8, monocyte chemoattractant protein‐1 and Rantes in human chronic periapical granuloma

In situ distribution of three prototype chemokines interleukin (IL)‐8, monocyte chemoattractant protein (MCP)‐1 and Rantes was determined in chronic human periapical granulomas by immunohistochemistry using monoclonal antibodies. IL‐8 was found primarily in the cytoplasm of the Malassez epithelial cells. MCP‐1 immunoreactivity was confined to the endothelial cells that lined small venules. Each of the three investigated chemokines, including Rantes, exhibited a characteristic binding pattern to the extracellular matrix of the lesion. The observed chemokines may play a role in establishing the cellular composition of chronic apical periodontitis, thus augmenting the intensity of local inflammation and tissue damage.     

Relationship Between Eosinophilia and Levels of Chemokines (CCL5 and CCL11) and IL-5 in Bronchoalveolar Lavage Fluid of Patients With Mustard Gas-Induced Pulmonary Fibrosis

Objective Therefore, this study was designed to analyze the bronchoalveolar lavage (BAL) fluid concentrations of IL-5, RANTES (CCL5) and eotaxin (CCL11) and also to examine the relationship between the percentage and absolute number of the BAL eosinophils and these measured chemokines in patients with sulfur mustard (SM) gas-induced pulmonary fibrosis (PF). Patients Fifteen veterans with mustard gas-induced PF and 14 normal veterans as control group. Intervention Pulmonary function tests, tests for D_LCO, computed tomography scans of the chest, analyses of BAL fluids for RANTES (CCL5), eotaxin (CCL11), and IL-5 were performed in all cases. Results Eosinophilic alveolitis was the predominant feature (p < 0.0001). There were significant differences in CCL5, CCL11, and IL-5 levels of BAL fluid between patients with PF and controls (p < 0.0001, p < 0.0001, and p = 0.001, respectively). The concentrations of CCL5 and CCL11 showed positive correlations with percentage (r = 0.57 and p = 0.03; r = 0.52 and p = 0.04, respectively) and absolute counts (r = 0.54 and p = 0.04, r = 0.53 and p = 0.04, respectively) of BAL eosinophils. There were significant positive correlations between the concentrations of IL-5 and the proportion and total cell number of eosinophils in BAL (r = 0.67 and p = 0.01; r = 0.59 and p = 0.02, respectively) too. Conclusion A significant correlation between BAL CCL5, CCL11, and IL-5 levels and eosinophils in patients with pulmonary fibrosis due to SM gas inhalation has been demonstrated, suggesting that these C–C chemokines and IL-5 contribute to the recruitment of eosinophils cells in the lung in these victims.