No abstract available.
Monokines* ; Neutrophil Activation* ; Neutrophils*

BACKGROUND: Monocytes and T helper cells play major roles in the immunologic dysfunction of atopic dermatitis (AD). Many studies have been done on the cytokine pattern to evaluate abnormalities or differences of immune cells in AD, but the results were conflicting among studies and most of these previous reports were performed with various kinds of mitogen-stimulation. OBJECTIVE: The purpose of this study was to investigate spontaneous cytokine pattern in peripheral blood mononuclear cells (PBMC) from patients with AD. We focused on the expression of monokines that had effects on monocytes and T cells. METHODS: We measured mRNA expression of IL-10, GM-CSF and TNF-alpha in freshly isolated PBMC with semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The intensity of cytokine cDNA was normalized to that of beta-actin product as a standard marker. RESULTS: IL-10 mRNA expression was significantly enhanced in AD compared:with control subjects (p<0.05). Spontaneous mRNA expression of TNF-alpha was significantly lower in AD patients (p <0.01). The level of GM-CSF mRNA expression was heterogeneous and spontaneous mRNA expression was slightly increased in AD although the difference did not reach the level of statistical significance. CONCLUSION: Our data was able to represent in vivo cytokine expression state of PBMC in atopic dermatitis. Increased expression of IL-10 and GM-CSF may have been associated with monocyte dysfunction in AD although increase in the expression of GM-CSF mRNA was not statistically significant. TNF-alpha mRNA expression was decreased in AD and increased IL-10 was suggested to exert an inhibitory effect on the expression of TNF-alpha mRNA.
Actins ; Dermatitis, Atopic* ; DNA, Complementary ; Granulocyte-Macrophage Colony-Stimulating Factor ; Humans ; Interleukin-10 ; Monocytes ; Monokines ; RNA, Messenger* ; T-Lymphocytes ; T-Lymphocytes, Helper-Inducer ; Tumor Necrosis Factor-alpha

Silicone gel breast implants may induce local(fibrous capsular contracture) or systemic(rheumatoid arthritis, systemic sclerosis, etc) complications. The exact mechanism of fibrous capsular contracture has not been fully understood. In the present study, we tried to find out the effect of silicone gel on the fibroblast proliferation which has been known as a major contributing factor in fibrous capsular contracture formation. In vitro, activated macrophages are known to secrete monokines which affect fibroblast proliferation and collagen synthesis. And tumour necrosis factor-alpha(TNF-alpha) and interleukin-6(IL-6), which were released by macrophages, were reported as potent stimulator of fibroblast proliferation. The goal of this study is to investigate the role of macrophages and tumour necrosis factor-alphaor interleukin-6 in the interaction of fibroblasts and silicone gel. We designed four groups, two experimental and two control, using Institute for Cancer Research(ICR) mouse peritioneal macrophage and silicone gel. For the preparation of the conditioned medium of macrophages, peritoneal macrophages were prepared and cultured for 24 hours on the silicone gel-coated and naked (not coated) surface [silicone gel-macrophage conditioned medium(SCM; experimental group) and normal polystyrene-macrophage conditioned medium(NCM; control group) respectively]. To correct the effect of 10% fetal bovine serum which was included in Rapid Prototyping and Manufacturing Institute (RPMI) 1640 medium and draw the effect only by macrophages, the RPMI 1640 medium with 10% fetal bovine serum was cultured by the same method on the silicone gel-coated and naked surface (silicone gel-macrophage free conditioned medium; SFM and normal polystyrene-macrophage free conditioned medium; NFM respectively). Each conditioned medium was added onto NIH 3T3 fibroblasts culture at a final 25% concentration of total culture medium and followed by the cultivation for 24 hours. For antibody neutralizing experiments, each conditioned medium was preincubated with polyclonal rabbit anti-mouse TNF-alpha antibody or polyclonal rat anti-mouse IL-6 antibody for 1 hour and then, conditioned medium with antibody was added to the culture medium of NIH 3T3 fibroblasts by the same method. After 24 hours cultivation, total number of viable fibroblast(cell growth), DNA synthesis and collagen synthesis of fibroblasts with each medium were measured by sulforhodamine B(SRB) assay, 3H-thymidine and 3H-proline incorporation respectively. The results were as follows: 1. In the experiment about the effect of the conditioned medium on the fibroblast activity, the experimental group(SCM), compared with the control group(NCM), showed a significant increase of the cell growth (p<0.01), a significant decrease of DNA synthesis(p<0.001), but no significant difference in the collagen synthesis. 2. In the experiment about the effect of polyclonal rabbit anti-mouse TNF-alpha antibody on the fibroblast activity, after the addition of antibody the experimental group, compared with the control group, showed a significant decrease of the cell growth(p<0.001), a significant increase of DNA synthesis(p<0.01), but no significant difference in the collagen syn thesis. 3. In the experiment about the effect of polyclonal rat anti-mouse IL-6 antibody on the fibroblast activity, after the addition of antibody the experimental group, compared with the control group, showed a significant decrease of the cell growth(p<0.001), a significant increase of DNA synthesis(p<0.0001), but no significant difference in the collagen synthesis. In conclusion, culture supernatants (conditioned medium) of peritoneal macrophages, activated by silicone gel, stimulate the NIH 3T3 fibroblast proliferation. TNF-alpha and IL-6, products of macrophage, are involved in the stimulation of NIH 3T3 fibroblast proliferation in an in vitro condition.
Animals ; Arthritis ; Breast Implants ; Collagen ; Contracture ; Culture Media, Conditioned ; DNA ; Fibroblasts* ; Interleukin-6* ; Macrophages ; Macrophages, Peritoneal* ; Mice ; Monokines ; Necrosis ; Rats ; Scleroderma, Systemic ; Silicone Gels* ; Tumor Necrosis Factor-alpha*

BACKGROUND: The pathogenesis of silicosis has been focused on the interaction between alveolar macrophages and silica particle. Although fibrosis in silicosis has been studied extensively, the mechanism is still not fully understood. There is increasing evidence that monokines and arachidonic acid metabolites produced by macrophage are involved in pathogenesis of silicosis. Recently, it was reported that prostaglandin E2 produced from macrophage counteracts the stimulatory effects of other monokines on fibroblast proliferation or collagen production. Until now, it was remained uncertain by which mechanism silica particle may activate alveolar macrophage to an enhanced release of prostaglandin E2. METHODS: In order to investigate the relationship between the activity of alveolar macrophage and the production of PGE2 from activated alveolar macrophage, the authors measured hydrogen peroxide and PGE2 from alveolar macrophages activated by silica in vitro and from alveolar macrophages in the silicotic nodules from rat. Experimental silicosis was induced by intratracheal infusion of silica(SiO2) suspended in saline (50 mg/ml) in Sprague-Dawley rats. RESULTS: 1) The silicotic nodules with fibrosis were seen from the sections of rat lung at 60 days after intratracheal injection with 50 mg aqueous suspension of silica. 2) In vitro, silica caused the dose dependent increase of hydrogen peroxide(p<0.05) and PGE2(p>0.05) release from alveolar macrophages. Alveolar macrophages from rats with silicotic nodules released more hydrogen peroxide and PGE2 than those of control group(p<0.05). CONCLUSION: These results suggest that silica particle could activate macrophage directly and enhanced the release of PGE2 and hydrogen peroxide from the alveolar macrophage.
Animals ; Arachidonic Acid ; Collagen ; Dinoprostone* ; Fibroblasts ; Fibrosis ; Hydrogen ; Hydrogen Peroxide ; Lung ; Macrophages ; Macrophages, Alveolar* ; Monokines ; Rats ; Rats, Sprague-Dawley ; Silicon Dioxide* ; Silicosis

Collagen induced arthritis (CIA)is an animal model that in many ways resembles rheumatoid arthritis (RA). CIA can be induced in susceptible animals by immunization with type II collagen (CII). Like RA,CIA is characterized by intense joint inflammation and destruction.On histological examination,there is synovitis accompanied by erosion of cartilage and subchondral bone. Autoanti-bodies to CII initiate joint inflammation by binding to articular cartilage,forming antigen-antibody complexes locally and activating hemolytic complement. Susceptibility to CIA in mice is linked to the expression of specific class II MHC Molecules,which dictate the T cell determinants on CII,and therefore,the subsets of T cells that can be activated by CII.In addition to activation of B cells reactive to CII,the T cells stimulate monocytes/macrophages.These cells amplify the inflammatory cascade by secretion of proinflammatory monokines, including TNF-alpha and IL-1,leading to the production of other proinflammatory proteins,including matrix metalloproteinases (MMPs).The importance of CIA lies in its possible relationship to arthritis in humans.Progress in understanding CIA has contributed to the development of new therapies for RA.In addition,it has been found that mice with human HLA-DR1,DR4 and HLA-DQ8 transgenes,which have been demonstrated to be the susceptibility markers for RA, confer susceptibility to CIA.These observations coupled with the finding of T cells and B cells reactive with CII in the inflamed joints of RA patients establish the potential role of CII autoimmunity in the pathogenesis of RA.
Animals ; Antigen-Antibody Complex ; Arthritis ; Arthritis, Experimental* ; Arthritis, Rheumatoid* ; Autoimmunity* ; B-Lymphocytes ; Cartilage ; Collagen ; Collagen Type II* ; Complement System Proteins ; Humans ; Immunization ; Inflammation ; Joints ; Matrix Metalloproteinases ; Mice ; Models, Animal ; Monokines ; Synovitis ; T-Lymphocytes ; Tumor Necrosis Factor-alpha