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

EXPRESSION OF MMP-2 AND 9 IN SKIN OF DOGS WITH VISCERAL
LEISHMANIASIS
ANA PAULA PRUDENTE JACINTHO 1 ; MAYARA CAROLINE ROSOLEM 1 ;
MARCIO DE BARROS BANDARRA 1 ; PAMELA RODRIGUES REINA
MOREIRA 1 ; GISELE FABRINO MACHADO 2 ; GUILHERME DIAS DE MELO 2 ,
DANISIO PRADO MUNARI 3 ; ROSEMERI DE OLIVEIRA VASCONCELOS 3* .
(1). Programa de Pós-graduação em Medicina Veterinária, FCAV/UNESP,
Jaboticabal, SP; (2).FMVA/UNESP, Araçatuba, SP; (3). FCAV/UNESP,
Faculdade de Ciências Agrárias e Veterinárias, Jaboticabal, SP;
*rosevasc@fcav.unesp.br
Introduction: Dogs infected by Leishmania (L.) chagasi may have clinical signs
for this zoonosis and they are the main source of infection for man. The skin is
the first organ infected by the parasite. The aim of this study was to evaluate the
expression of MMP-2 and MMP-9 by immuno-histochemistry and zymography
in the skin of dogs naturally affected by protozoa and compare with
immunodetection of TNF-, TGF- and Leishmania sp. and with the alterations
of dermal extracellular matrix.
Methods and Results: This study included 36 dogs, without preference for
age, breed or sex, from the Zoonosis Control Center of Araçatuba (SP), where
they were euthanized due to the positive diagnosis of visceral leishmaniasis
(VL). One control group was formed with dogs from non-endemic area for VL
(CEUA nº. 020373/09). In immunohistochemical analysis we used the
antibodies MMP 2 and 9 (Abcam), TNF-α and TGF-β (Santa Cruz Biotecnology)
and for the parasite load was used canine hyperimmune serum (J. Immunol.
Meth. 292:17-23, 2004). The gelatinolytic activity was assessed by zymography
(Vet. Immunol. Immunopathol. 136:340-345, 2010).The Picrosirius red staining
was used to differentiate collagen types I and III, in dermis of the muzzle, ear
and abdomen, in asymptomatic (GA), oligosymptomatic (GO) and symptomatic
dogs (GS). The parasite load (P=0.0001) and intensity of inflammation were
higher in regions of the ear (P=0.1709) and muzzle (P=0.0235) mainly in dogs
GS. In the dogs GA and GO the skin inflammation was discrete or absent. The
active form of MMP-9 predominated in the GS group and same skin areas. The
macrophages were significantly different from lymphocytes in the cutaneous
inflammatory infiltrate (P =0.01) in ear and muzzle of GS dogs. In this case
there were granulomas associated with degradation of mature collagen (type I)
and discrete deposition of the young collagen (type III), mainly in dogs with high