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The Role of Dendritic Cells and CC Chemokine Ligand-20 in Rat with Chronic Obstructive Pulmonary Disease

Author: GuYanHui
Tutor: OuYangYao
School: Zunyi Medical College,
Course: Department of Respiratory Medicine
Keywords: Chronic Obstructive Pulmonary Disease dendritic cells CD80 CD86 OX-62 MHC-Ⅱ chronic obstructive pulmonary disease dendritic cell chemokine CCL20
CLC: R563.9
Type: Master's thesis
Year: 2011
Downloads: 33
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Abstract

Part I:The expression of CD80, CD86, OX-62 and MHC-II in the broncho-alveolar lavage fluid and peripheral blood of COPDObjective:To study the expression of Dendritic Cells (DCs) in broncho-alveolar lavage fluid (BALF) and peripheral blood of rats with Chronic Obstructive Pulmonary Disease (COPD).Methods:Wistar rats were randomly divided into three groups:control group(n=24), COPD model group(n=40) and smoking cessation group(n=10). The rats of COPD models group were established via cigarette inhalation about four weeks and intratracheal 200ug/200ul of lipopolysaccharide (LPS) solution twice totally; after 28 days, the rats of COPD model group which were stopped to smoking and injection of LPS were quitting smoking group. Then pathomorphology of rats’lung and bronchiole were investigated by HE staining at 7-day,14-day,21-day,28-day and 42-day respectively. The cell count determined by CD80, CD86, OX-62 and MHC-Ⅱwere analyzed by FCM in the BALF and peripheral blood collected from COPD rat groups and control groups at 28 days.Results:The HE staining result suggest that there were inflammation in the airway at 7 days,14 days,21 days and 28 days, which were severe gradually. The changes of pathology of the COPD model at 28-day were the same as those of COPD patients, and there were no significant difference between the groups of 28 days and 42 days. In BALF, the percentages of CD80 and CD86 in COPD models groups were significantly increased compared with the control group, there was a difference statistically significant (p<0.05), but there were no significant difference of OX-62 and MHC-Ⅱbetween the two groups. In peripheral blood, the percentages of CD80, CD86 and OX-62 in COPD model group were significantly increased compared with the control group, there was a difference statistically significant (p<0.05), the percentages of MHC-Ⅱwere significantly decreased compared with the control group, there was a difference statistically significant (p<0.01). The percentages of CD80, CD86, OX-62 and MHC-Ⅱin the BALF and peripheral blood were no correlation between the two groups.Conclusion:The COPD rat models were established successfully by this combined methods, and the model were still stable relatively after ending the combination two weeks later. The significantly upregulation of CD80 and CD86 in the COPD rat model groups, suggesting that DCs may be involved in COPD about inflammatory reaction. Part II:The role of DCs and CCL20 in the pathogenesis of COPDObjective:To study the changes of CCL20 and DCs in the BALF and peripheral blood of COPD rat model group and CCL20 inhibited rat model group, and further to make clear of the pathogenesis of COPD about DCs and CCL20.Methods:Wistar rats were randomly divided into three groups:control group(n=6), COPD model group(n=10) and CCL20 inhibited model group(n=10). The rats of COPD groups and CCL20 inhibited models groups were established via cigarette inhalation about four weeks and intratracheal 200ug/200ul of LPS solution twice totally, and there were intraperitoneal injection of CCL20 in the CCL20 inhibited models groups. Then pathomorphology of rats’ lung and bronchiole were investigated by HE staining. The contents of CCL20 were analyzed with ELISA in the BALF and peripheral blood collected from CCL20 inhibited models group and COPD rat group.The cell count determined by CD80, CD86, OX-62 and MHC-II were analyzed with FCM in the BALF and peripheral blood collected from CCL20 inhibited models group and COPD rat group.Results:The contents of CCL20 in BALF about CCL20 inhibited model group were significantly increased compared with the control group, there was a difference statistically significant (p<0.001), but there was no difference compared with COPD group; in peripheral blood, the contents of CCL20 about CCL20 inhibited model group were significantly increased compared with the control group, there was a difference statistically significant (p<0.05), and were significantly decreased compared with the control group, there was a difference statistically significant (p<0.01). In peripheral blood, the percentages of CD80 about CCL20 inhibited model group were significantly increased compared with the control group, there was a difference statistically significant (p<0.05), but there was no difference compared with COPD model group. The percentages of CD86 and OX-62 were significantly decreased compared with COPD model group, there was a difference statistically significant (p<0.05), but there was no difference compared with the control group. The percentages of MHC-Ⅱwere significantly decreased compared with the control group, there was a difference statistically significant (p<0.05), but there was no difference compared with COPD model group. In BALF, the percentages of CD80 about CCL20 inhibited model group were significantly increased compared with the control group, there was a difference statistically significant (p<0.05), were significantly decreased compared with the control group, there was a difference statistically significant (p<0.01). The percentages of CD86 were significantly increased compared with the control group, there was a difference statistically significant (p<0.05), but there was no difference compared with COPD model group. There were no difference about OX-62 and MHC-Ⅱamong the three groups in BALF. The percentages of CD80, CD86, OX-62 and MHC-Ⅱin the BALF and peripheral blood were no correlation among the three groups. There was positive correlation about CCL20 in the BALF and peripheral blood of CCL20 inhibited model group.Conclusion:In CCL20 inhibited model group, the contents of CCL20 were increased in peripheral blood and BALF compared with control groups,and were decreased in BLAF compared with COPD groups, suggesting that CCL20 may be involved in the pathogenesis of COPD,CCL20 monoclonal antibody may play a inhibit role in this pathogenesis; the percentages of CD86 and OX-62 were decreased in peripheral blood compared with COPD groups, the percentages of CD80 were decreased in BALF compared with COPD groups,but were decreased compared with control groups, suggesting that CCL20 monoclonal antibody can decreased the number of DCs in the peripheral blood and in the airway of COPD, so it will help to reduce the systemic inflammatory response of COPD.

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