Canine stem cell factor augments expression of matrix metalloproteinase-9 by CD34 cells.

BACKGROUND: Canine models have proved to be predictive of clinical findings in human bone marrow (BM) transplantation; consequently, the utilization of dogs is an excellent tool for supporting therapeutic purposes. Considering the role of growth factors in homing and mobilization of hematopoietic progenitors, the aim of this work was to evaluate whether canine stem cell factor (cSCF) contributes to matrix metalloproteinase (MMP)-9 secretion by CD34 cells. METHODS: The study was carried out in a cell population selected by immunomagnetic techniques using the anti-canine CD34 monoclonal antibody (MAb) 3B4 produced by us. Secretion of MMP-9 was evaluated by zymography. RESULTS: Analyzes of canine CD34(+) cells guaranteed that the MAb 3B4 was optimum for selecting a subset population with defined characteristics of primitive hematopoietic cells. The isolated cells were able to proliferate onto irradiated pre-established stroma, giving rise to mature neutrophils. There was also a 20-fold enrichment in the long-term culture-initiating cell content when the isolated population was added to irradiated cultures, with respect to the starting mononuclear cell population. DISCUSSION: We have provided the first evidence that canine BM CD34(+) cells constitutively express MMP-9 and the role of cSCF in up-regulating the secretion of this enzyme. The fact that cSCF augments expression of MMP-9 together with the ability of the isolated CD34(+)cells to proliferate onto irradiated pre-established stroma enables further investigations to determine whether the secretion of MMP-9 mediated by cSCF is one of the factors that enhance migration, homing and repopulation of primitive hemopoietic cells.

Canine long-term bone marrow culture neutrophil production and functionality.

This in vitro study has been conducted to determine the optimal experimental conditions under which to produce canine neutrophils in long-term bone marrow cultures (LTBMC), establish functional parameters of neutrophils obtained from LTBMC and peripheral blood and to ascertain whether these cells display physiological similarities. Our aim is to provide an experimental model, enabling a correlation between hemopoietic injury and neutrophil functionality. The authors demonstrate for the first time that canine neutrophils grown in cultures are able to produce oxyradicals capable of killing bacterial products. Moreover, culture-grown neutrophils contain gelatinase granules, a marker of terminal neutrophil differentiation, and express a specific surface antigen. The results described in this article illustrate the development of a dynamic system that mimics physiological hemopoiesis.

Antibody response in dogs experimentally infected with Leishmania infantum: infection course antigen markers.

Five-month-old beagle dogs were experimentally infected with amastigotes of Leishmania infantum and kept for 14 months. Infection course was monitored by clinical examination, serum protein variations, and levels of specific antibodies against Leishmania estimated by indirect immunofluorescence test and Western blotting (WB). Infected animals developed notable changes in serum protein levels reaching maximum protein concentrations 2-3 months postinfection (p.i.) related to the gamma-globulin fraction. Specific antibody titers were in good agreement with the serum protein rise, reaching immunofluorescence values of over 1:800 3 months p.i. Serial Western blotting analysis with L. infantum promastigotes protein showed a strong response against immunodominant antigens of 50-57, 42, and 29 kDa during most of the studied period with immunofluorescence titers of over 1:100 and in addition the response was remarkably homogeneous among the infected dogs. Immunoreactivity patterns displayed time-related variations; the response against 29 and 50-57 kDa was seen very early, followed by the reaction around 42, 76, and 86 kDa. In addition the recognition of peptides around 34-35.4 and 26 kDa was restricted to the acute phase of the experimental infection. Preliminary results obtained in naturally infected dogs seem to support the predictive value of the WB.

Erosive colitis in experimental canine Leishmaniasis.

Six 4 month-old beagles were inoculated with Leishmania infantum, three of them intraperitoneally (Group A) and the other three intravenously (Group B). The animals from Group A were killed at 109, 433 and 592 days after inoculation and animals from Group B 109, 171 and 334 days after inoculation. At 350 days (Group A) and 275 days (Group B) after inoculation, dogs started to develop a chronic large bowel diarrhoea. At necropsy a diffuse colonic and rectal wall thickening of gradually increasing severity was observed. Histological examination of the colon showed a diffuse inflammatory infiltration of the mucosa and submucosa by macrophages with amastigotes, lymphocytes, plasma cells and some neutrophils and eosinophils. The surface epithelium developed increasingly extensive degeneration, which caused the development of erosions on the mucosal surface. The crypts of Lieberkühn decreased in number and showed degeneration of the crypt epithelium.