ABSTRACT
We propose to analyze the relation between the cellular immune response of Chagas' disease patients after in vitro stimulation of peripheral blood mononuclear cells (PBMC) with recombinant antigens cytoplasmatic repetitive antigen (CRA) or flagellar repetitive antigen (FRA) of T. cruzi and the chronic clinical forms of disease. Cells were stimulated using phytohemagglutinin, CRA, FRA, or a soluble antigen of Epimastigota (Ag-Epi) for 24 hr, 72 hr, or 6 days. The proliferation of cells was evaluated after 6 days of culture by quantification of incorporated 3H-thymidine. Cytokines were measured in the supernatants obtained after 24 hr (tumor necrosis factor [TNF]-alpha and interleukin [IL]-4), 72 hr (IL-10), and 6 days (interferon [IFN]-gamma) using enzyme-linked immunosorbent assay (ELISA). Cells of the Chagas patients stimulated with the recombinant antigens exhibited higher proliferation responses compared with that of non-Chagas (NC) individuals. However, when proliferation was compared between patients with the cardiac form (CF) or indeterminate form (IF), it was not possible to establish a difference in the response. So far as the cytokines secreted in the culture supernatants after stimulation in vitro with T. cruzi antigens were concerned, the results showed that CRA, as well as Epi-Ag, were able to stimulate the production of TNF-alpha and IFN-gamma in Chagas patients as compared with NC individuals. However, the cytokine levels after stimulation with the T. cruzi antigens were not different between the patients with CF and IF. CRA was capable of inducing a T helper type 1 (Th1) immune response, with elevated production of TNF-alpha and IFN-gamma in Chagas patients that are carriers of CF and IF clinical forms.
Subject(s)
Antigens, Protozoan/immunology , Chagas Disease/immunology , Chagas Disease/parasitology , Immunity, Cellular/immunology , Recombinant Proteins/immunology , Trypanosoma cruzi/immunology , Adult , Aged , Aged, 80 and over , Animals , Carrier State/parasitology , Cells, Cultured , Female , Humans , Immunity, Cellular/drug effects , Interferon-gamma/immunology , Interleukin-10/immunology , Lymphocytes/drug effects , Lymphocytes/immunology , Lymphocytes/parasitology , Male , Middle Aged , Mitogens/pharmacology , Subcellular Fractions/drug effects , Subcellular Fractions/parasitology , Trypanosoma cruzi/drug effects , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
The Brazilian Ministry of Health has made tests for HIV1 and HIV2, HTLV I and HTLV II, HCV, HBV, T. cruzi, T. pallidum and Plasmodium in endemic areas, mandatory for all blood collection bags used in the country. However, blood-borne infectious diseases are not investigated in blood recipients before transfusion. For this study, a serological evaluation of recipients before transfusion was carried out. Prior to transfusion, serum samples from 159 blood recipients were analyzed using the same tests used in the serological screening of blood donors. The blood recipients were divided into three groups: Group 1 (G1), patients who had never received blood, Group 2 (G2), patients who had received multiple transfusions and Group 3 (G3) one-off recipients. SPSS v.8 was used for statistical analysis. Values of p<0.05 were taken to be significant. The results showed that 62 blood recipients tested positively for one or more blood-borne infectious diseases. In addition, several recipients were unaware of their serological status before the transfusion. The identification of blood-borne infectious diseases in recipients before transfusion could avoid the State being held responsible by recipients who were unaware that they were carriers of such diseases and only found out about their contamination after transfusion.
O Ministério da Saúde brasileiro determina a realização de testes sorológicos para HIV 1 e 2, HTLV I e II, HCV, HBV, T. cruzi, T. pallidum e Plasmodium nas áreas endêmicas, em todas as bolsas de coleta de sangue utilizadas no País. Entretanto, as doenças infecciosas transmissíveis através do sangue não são investigadas nos receptores de sangue (RS) antes da transfusão. Neste estudo, realizamos uma avaliação sorológica dos RS anterior à transfusão. Amostras de soro de 159 RS foram analisadas aplicando-se os mesmos testes utilizados na triagem sorológica dos doadores de sangue. Os RS foram divididos em três grupos: Grupo 1 (G1), pacientes que nunca receberam sangue, Grupo 2 (G2), pacientes politransfundidos e Grupo 3 (G3) receptores eventuais. Para a análise estatística utilizou-se o programa SPSS v.8. Valores de p<0,05 foram considerados significantes. Os resultados mostraram que 62 RS apresentaram positividade para uma ou mais doenças infecciosas transmissíveis pelo sangue. Além disso, vários RS desconheciam seu estado sorológico anterior à transfusão. A identificação de doenças infecciosas transmissíveis pelo sangue em RS anterior à transfusão poderia evitar a responsabilidade do Estado pelos RS que desconheciam ser portadores de tais doenças e apenas tiveram conhecimento de sua contaminação após a transfusão.
