A standardized cytological and immunochemical method for the analysis of fine-needle spleen aspirates: assessment of leukocyte population changes in canine visceral leishmaniosis.
A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirative biopsy was standardized in this work. The procedure includes erythrocyte lysing, preparation of cytospin films and staining by histochemical and immunocytochemical techniques. The cellular profiles of spleen preparations were compared with those observed in peripheral blood samples subjected to the same procedure. Two groups were compared, one consisting of 14 healthy uninfected and the other of 15 polysymptomatic Leishmania chagasi/infantum-infected dogs, from an endemic area for visceral leishmaniosis. Cell populations were identified by conventional hematoxilin-eosin and Wright' stainings, and by immunocytochemistry using monoclonal antibodies against canine CD45RA and CD45RB, phagocytes and a pan-leukocyte antigen. Larger neutrophil (P < 0.0001) and monocyte/macrophage (P = 0.0036) relative counts and lower lymphocyte relative counts (P < 0.0001) were found in the spleen, and not in the blood, of the animals with leishmaniosis than in those of the healthy animals. The proportions of CD45RB+ cells were higher, and of CD45RA+ cells were lower, both in the spleen and in the blood of animals with leishmaniosis than in those of healthy dogs (P < 0.05). Additionally, hematoxilin-eosin-stained cytospins of spleen aspirates from Leishmania-infected animals permitted the easy visualization of amastigote forms inside phagocytes, under light microscopy.
Barrouin-Melo
Stella Maria
SM
Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Rua Valdemar Falcão 121, Salvador 40295-001, Brazil. barrouin@ufba.br
Larangeira
Daniela Farias
DF
Santos
Silvana Ornelas
SO
Chagas-Júnior
Adenizar Delgado
AD
Paixão
Mariza
M
Aguiar
Paulo Henrique Palis
PH
dos-Santos
Washington Luís Conrado
WL
Pontes-de-Carvalho
Lain
L
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D044963:Biopsy, Fine-Needle / Q000379:methods / Q000662:veterinary*; D004283:Dog Diseases / Q000097:blood / Q000276:immunology* / Q000469:parasitology*; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D007124:Immunoenzyme Techniques / Q000662:veterinary; D018314:Leishmania infantum / Q000276:immunology*; D007898:Leishmaniasis, Visceral / Q000097:blood / Q000276:immunology* / Q000469:parasitology / Q000662:veterinary*; D017493:Leukocyte Common Antigens / Q000032:analysis; D007958:Leukocyte Count / Q000662:veterinary; D008297:Male; D013154:Spleen / Q000276:immunology / Q000469:parasitology* / Q000473:pathology
D000913:Antibodies, Protozoan; D017493:Leukocyte Common Antigens