Levels of parasitemia in Chagas Disease patients under organ transplantation, immunosuppressive drug therapy and co-infection with human immunodeficiency virus disease

• Grant number: 14/03966-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2014
• End date: December 31, 2014
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Maria Aparecida Shikanai Yasuda
• Grantee: Caroline Medeji Ricardo dos Santos
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Chagas Disease is now an active infection in urban areas of endemic and non-endemic countries. The migration of infected people from endemic areas to big cities generated new sources of infections, congenital transmission as well as blood derivatives. In parallel, the reactivation of chronic trypanosomiasis was reported in patients with chronic disease and comorbidities as HIV infection/aids (acquired immunodeficiency syndrome), or immunosupression under organ transplants or chemotherapy for neoplasia and autoimmune diseases. Considering the high morbi-mortality of Chagas disease reactivation in immunosuppressed patients, this report aims to analyze the levels of parasitemia in immunocompromised patients (coinfection HIV/Trypanosoma cruzi, organ transplants, and other forms of immunosuppression) in comparison with nonimmunocompromised patients (Chronic Chagas disease). Patients will be selected from the Division of Clinics for Infectious and Parasitic Diseases, Transplant Infirmaries, and other units from the Hospital das Clínicas da Faculdade de Medicina da USP. About 120 patients will be included, on the basis of at least two, positive serological tests (ELISA, IFI, or indirect hemagglutination tests) for Trypanosoma cruzi epimastigote antigens. Patients with reactivation or acute disease with positive direct microscopy exam on peripheral blood or leucocytes buffy coat or liquor or other biological samples and/or presence of the parasite on biopsy sample could be included in the immunosuppressed or nonimmunosuppressed group. A sample of 10 mL of total blood will be collected on 10 mL of Guanidine-HCl-EDTA pH 8,0. The DNA extraction will be performed by QIAmp DNA Mini system (Quiagen) and both qualitative PCR, with primers that amplify kDNA and quantitative DNA with primers from the nuclear DNA TCZ3/TCZ4 with the SYBR Green Advantage qPCR Premix will be performed. It is a challenge to find out the magnitude of parasitemia observed in patients under organ transplants or drug immunosupression for the treatment of autoimmune diseases. It is also possible to compare these findings with the levels of parasitemia observed in patients with HIV/T. cruzi coinfection, whose risk for reactivation has been partially established in prospective studies in clinical practice. Therefore, on the basis of parasitemia detected by Real-Time PCR, it is also expected to estimate if other immunosuppressed non-HIV patients are at risk of reactivation. (AU)