Detection Of Perforin And CTLA-4 In The Study Of Immune State In Tuberculous And Malignant Pleural Effusion Microenvironment

Backgroud pleural effusion is a frequent clinical sign, including tuberculous and malignant pleural effusion, these two effusion have higher lymphocyte proportion.Many studies have proved that immune state of malignant effusion microenviroment is suppressive. Cytotoxic T Lymphocyte, as the major effective cell in cell mediated immunity, express Perforin which has an important role of cytotoxicity and initiating cell apoptosis. cytotoxic T lymphocyte associated antigen-4 (CTLA-4) is costimulatory molecules which has an important role of down regulation of T lymphocyte, blockade of CTLA-4's binding with B7 can enhance cell immunity. What is the relationship between immunosuppresive microenviroment and CTL's expressions of Perforin and CTLA-4, it deserves to explorate.Objective:(1).To study the lymphocyte classification of pleural effusions.(2).To detect the expression of perforin and ctla-4 by CTL of pleural effusion and the concentration of perforin and ctla-4.(3).To explorate the relationship between immune state of microenviroment and expression of Perforin and CTLA-4.Methods:1. sample collection and managmentSixty inpatients' pleural effusions had been collected from September 2007 to June 2008, these inpatients were from Renmin Hospital of Wuhan University, among these samples thirty were malignant,thirty were tuberculous. All samples were gotten by thoracentesis before treatment, each above 50 ml, anticoagulation with heparin and then filtering with 200 screen mesh nylon web.2.sample's detection(1)Detection of Perforin concentration of pleural effusion by ELISA;(2) detection of CTLA-4 of pleural effusion by ELISA;(3)Isolating cell and labelling and determination by flow cytometry.Results: (1)Ther are no difference between patient age and man female ratio; (2)there are no difference between the number of nucleated cell and the lymphocyte subset;(3)the concetration of perforin in tuberculous effusion is 9816±673.0pg/ml, the concetration of perforin in malignant pleural effusion is 4755±790.9pg/ml, there is significant difference between these two(p=0.0001);(4)the concetration of CTLA-4 in tuberculous effusion is 1.885±0.3937ng/ml, he concetration of CTLA-4 in malignant effusion is 5.508±1.343ng/ml, there is significant difference between these two(p=0.0185);(5)the perforin expression rate of CTL in tuberculous effusion is 15.62±1.541%, while the perforin expression rate of CTL in malignant effusion is 8.010±1.051%, there is significant difference between these two(P=0.0007);(6)the CTLA-4 expression rate of CTL in tuberculous effusion is 13.70±1.663%%, while the CTLA-4 expression rate of CTL in malignant effusion is 6.400±0.9508%, there is significant difference between these two(P=0.0052).Conclusion(1)there is a lower perforin concetration in malignant pleural effusion;(2)the perforin expression rate of CTL in malignant effusion is lower than the rate of CTL in tuberculous effusion;(3)there is a higher sCTLA-4 concetration in malignant pleural effusion;(4)the CTLA-4 exprssion rate of CTL in malignant effusion is lower than the rate of CTL in tuberculous effusion;(5)Perforin and CTLA-4 may play an important role in the formation of malignant effusion microenvirment's immune state, the determination of Perforin and CTLA-4 may have a very important clinical significance in the differential diagnosis of pleural effusions;(6)It is practicable to use Cytotoxic T Lymphocyte of Effusion-Associated Lymphocytes for studying tumor-induced immunosuppression.