Quantitative PCR and unconventional serological methods to evaluate clomipramine treatment effectiveness in experimental Trypanosoma cruzi infection

• Published in: Experimental and molecular pathology 2016-10, Vol.101 (2), p.274-280
• Main Authors: Bazán, Paola Carolina, Lo Presti, María Silvina, Strauss, Mariana, Báez, Alejandra Lidia, Miler, Noemí, Paglini, Patricia Adriana, Rivarola, Héctor Walter
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Protozoan - blood; Chagas disease; Chagas Disease - blood; Chagas Disease - diagnostic imaging; Chagas Disease - drug therapy; Chagas Disease - parasitology; Chronic phase; Clomipramine - therapeutic use; Clomipramine treatment; DNA, Protozoan - blood; Electrocardiography; Female; Male; Mice; Parasitemia - drug therapy; qPCR; Real-Time Polymerase Chain Reaction - methods; Reference Standards; Serology; Serology - methods; Survival Analysis; Treatment Outcome; Trypanosoma cruzi; Trypanosoma cruzi - immunology; Trypanosoma cruzi - physiology
• ISSN: 0014-4800; 1096-0945
• DOI: 10.1016/j.yexmp.2016.09.006

Clomipramine (CLO), a tricyclic antidepressant drug, has been used for the treatment of mice infected with Trypanosoma cruzi. In this work we evaluated the effectiveness of CLO treatment upon T. cruzi-infected mice in the chronic phase of the experimental infection using Quantitative polymerase chain reaction (qPCR) and recombinant ELISA. Sixty Swiss albino mice were inoculated with 50 trypomastigote forms of T. cruzi (Tulahuen strain). CLO treatment consisted of 5mg/kg/day during 60days by intraperitoneal injection, beginning on day 90 post infection (p.i) when the mice presented electrocardiographic (ECG) alterations compatible with the chronic phase of the disease. The evolution of experimental infection and the treatment efficacy were studied through survival, electrocardiography, serology using a mixture and individual (1, 2, 13, 30, 36 and SAPA) recombinant proteins from epimastigotes and trypomastigotes of T. cruzi; and qPCR on days 180 and 270 p.i. CLO treatment in the chronic phase decreased the parasite load, reduced the levels of antibodies against antigen 13 throughout 270days p.i and reversed the ECG abnormalities in the treated animals, from 100% of the mice with alterations at the beginning of the treatment to only 20% of the mice with alterations by day 270 p.i. This study shows that qPCR and the use of recombinant antigens are more sensitive to evaluate the effectiveness of the treatment and proves that clomipramine may be considered as a new chemotherapy for the chronic phase of the disease. [Display omitted] •qPCR and recombinant antigens are effective to evaluate trypanocidal treatments.•Conventional PCR and qPCR detect T. cruzi in the chronic phase of infection.•Clomipramine may be considered for the treatment of the chronic phase of the disease.