immunology of infectious and parasitic diseases - XXXVII Congress ...

INCREASE TLR2 RECEPTOR, ROS AND NO PRODUCTION IN
MONONUCLEAR CELLS FROM DOGS AFTER STIMULATION WITH P-
MAPA.
LARISSA MARTINS MELO(PG) 1 ; LETICIA DA CRUZ SANCHES(PG) 1 ;
KATHLENN LIEZBETH DE OLIVEIRA DA SILVA(PG) 1 ; JULIANA PEROSSO
BORGES(PG) 1 ; BRUNA BRITTO DE OLIVEIRA(IC) 1 ; MARCOS DE ARRUDA
SOMENZARI (PG) 1 ; VALÉRIA MARÇAL FÉLIX DE LIMA(PD) 1
(1) Laboratório de Imunologia, Depto. Clínica, Cirurgia e Reprodução Animal –
Faculdade de Medicina Veterinária -UNESP - Araçatuba -SP
Introduction: Leishmania (L.) chagasi is the etiologic agent of visceral
leishmaniasis (VL), which can be transmitted to humans, and the dogs are main
domestic reservoirs. VL in Brazil represents a serious public health problem.
The infection in dogs suppress cellular immune response and the therapeutic
arsenal against CVL is limited. It is therefore important studying new
alternatives for the treatment of infected dogs, which may reduce the incidence
of the disease in endemic areas. The new immunomodulator P-MAPA improve
the immunocompetence when the immune system was impaired.
Methods and Results: This investigation was performed in Araçatuba, the city
is located in the São Paulo state. It is a region known to be endemic for canine
VL. A total of 15 adult dogs were selected based on positive serological tests for
visceral leishmaniasis by ELISA and the presence of at least three clinical
signs. A group of 6 healthy dogs, from a non-endemic area were included in the
study as negative controls. These animals showed negative serological tests for
visceral leishmaniasis by ELISA.The peripheral blood mononuclear cells were
isolated from both group and cultured in RPMI-1640 with different
concentrations of P-MAPA 20, 100 and 200 µg/ml in a humid environment at
37°C with 5% CO2 for 24h. Dyed for determination of TLR2, TLR4 using the
monoclonal antibodies conjugated to fluorochromes. The ROS production was
measured using carboxy-H2DFFDA (Molecular Probe ®) according to
manufacturer's instructions. After data acquisition in EasyCyte mini ® (Guava,
Hayward, CA), the analysis of the data was held in the Software Express Plus ®
Guava. For determination NO were using the culture supernatant of
mononuclear cells stimulated with P-MAPA by Griess reaction. The data were
analyzed with non parametric test. Results were considered as significant when
p