Detecção de Leishmania (Viannia) em polimorfonucleares e mononucleares no sangue de pacientes com leishmaniose tegumentar

Abstract

The leishmaniases are parasitic diseases, occurring in 98 countries, including Brazil. In Brazil, American cutaneous leishmaniasis (ACL) has been reported in all states and is endemic in Paraná. Most cases of leishmaniasis in Paraná are caused by Leishmania (Viannia) braziliensis that has the potential to develop mucosal form. The appearance of mucosal injury usually occurs years after the healing of the primary lesion and can occur because of inadequate treatment of primary lesions or a failure in cellular immunity, affecting the persistence of the parasite in the body. Thus, the objective was to evaluate the presence of Leishmania (Viannia) in polymorphonuclear and mononuclear cells in the blood of patients with LTA, using the technique of Polymerase Chain Reaction (PCR) and specific primers in order to better understand the mechanisms of persistence of the parasite and therefore the pathogenesis of the disease. To obtain cells was used the reagent Mono-Poly Resolving Medium. After polymorphonuclear and mononuclear separation and washing, DNA was extracted by guanidine isothiocyanate and phenol. It was used MP3H and MP1L primers for amplification. It were studied 94 patients, 63.83% were men and, 36.17% women. From 94 patients, 71 (75.53%) had primary lesion and no history of CL and, 23 (24.47%) reported previous infection. From 71 patients with primary lesions, 30 (31.91%) were diagnosed according to tests recommended by the Health Ministry (Brazil). From 23 patients who reported previous infection, 7 (30.43%) presented reactivation of the primary lesion, 2 (8.69%) presented new skin lesion and, 14 (60.86%) returned for treatment control. PCR was positive in mononuclear cells of two patients who presented active primary lesion with 3 to 6 months of time evolution and these patients did not present history of CL. The presence of Leishmania (Viannia) DNA in mononuclear cells indicates that, in some patients, the parasite is able to escape to the macrophage elimination mechanisms and remain in the body and, hematogenic spread can occur with development of mucosal lesion.