Emprego da citometria de fluxo na avaliação do perfil imunofenotípico de pacientes com leucemia linfocítica crônica

Chronic lymphocytic leukemia (B-CLL) is a clonal proliferation of mature B lymphocytes characterized by indolent clinical course. Biologically this clonallity is characterized by low expression of surface immunoglobulin (sIg) with restriction to a single immunoglobulin light chain associated with high expression of CD5 antigen and positivity to B cell antigens lymphocytes such as CD19, CD20 and CD23 and negativity to FMC7. The immunological profile and morphological analysis of lymphoid cells are the main means for the differential diagnosis of B-CLL from other chronic lymphoproliferative diseases. The aim of this study was to evaluate the expression pattern of a variety of membrane antigens in leukemic cells originating from patients with B-CLL. In this study, peripheral blood samples from 80 patients with B-CLL were analyzed by multiparametric flow cytometry in addition to routine hematologic exams, using a panel of monoclonal antibodies (MoAb): CD45/CD14, CD3/CD19/CD45, CD4/CD8 / CD3, CD20/CD5/CD3, CD3/CD16-56/CD45, CD2/CD7, FMC7/CD23, CD103/CD22/CD20, HLADR/CD38, CD10/CD19, CD1a, CD11b and also IgM/gD, kappa and lambda immunoglobulin light chains for the detection of surface immunoglobulin and clonal restriction for immunoglobulin light chain. The Hematological data were obtained from the hematological analyzer and cytomorphological analysis in blood film stained by Leishmann. The study samples consisted of 45 men and 35 women, ages ranging from 55 to 84 years (mean 65 years). Complete white blood count showed count ranging from 10.0 to 42.0 x 109/l. (mean 50.0 x 109/l) and lymphocytes count greater than 5.0 x 109/l in all cases. The neoplastic cells displayed B-CLL phenotype (CD5+/CD19+/CD20+/HLADR+/CD23+) in the vast majority of the cases, associated to failed to stain for T cell markers (CD1a, CD2, CD4, CD3, CD7, CD8), CD103, CD14 and FMC7. Leukemic cells of most patients also expressed low intensity of IgM and IgD with restricted kappa light chain, in most cases (59,7%). This observation highlights the importance of immunophenotyping for correct diagnosis of chronic lymphoproliferative syndromes and the panel of MoAb used was sufficient for diagnostic confirmation of B-CLL

Quantificação de subpopulações linfocitárias no sangue do cordão umbilical de eqüinos

Este estudo visou a determinar os valores eritroleucométricos e quantificar as subpopulações linfocitárias no sangue do cordão umbilical (SCU) e no sangue jugular de eqüinos neonatos. Foi realizada a colheita de SCU e do sangue jugular de 20 potros ao nascimento. As amostras foram submetidas às determinações dos valores eritroleucométricos e à quantificação de subpopulações de linfócitos-T, pela técnica citofluorométrica. Não foram verificadas diferenças significativas (P<0,05) entre os valores médios de tais parâmetros, entre o sangue jugular de neonatos e o SCU eqüino. O valor total para neutrófilos segmentados, no SCU e na jugular dos neonatos, foi inferior ao reportado para eqüinos ao nascimento. As contagens de linfócitos CD5+ e CD4+ no SCU e jugular de neonatos eqüinos foram inferiores àquelas admitidas para o sangue periférico de eqüinos adultos, indicando um componente imunológico imaturo. No entanto, a contagem de linfócitos CD8+ foi semelhante à descrita em sangue periférico de eqüinos adultos. A proporção CD4:CD8 obtida nesse ensaio, tanto para o SCU (2,64±0,91), como no sangue jugular de eqüinos neonatos (2,41±0,81), demonstrou uma dominância das células T CD4+ sobre os linfócitos T CD8+.

T lymphocyte subsets of the umbilical cord blood of dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hemograma e hemogasometria de eqüinos submetidos à obstrução experimental de jejuno

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Características físico-químicas e citológicas do líquido sinovial da bainha tendínea digital de eqüinos

Foram estudadas as características físico-químicas e citológicas do líquido sinovial da bainha tendínea digital de nove eqüinos hígidos. Verificou-se que o líquido é viscoso, amarelo claro, límpido, livre de partículas e que não coagula à temperatura ambiente. Sua concentração média de ácido hialurônico foi 60,20mg/dl, a taxa de glicose, similar à plasmática e sua concentração protéica não ultrapassou 1,74g/dl, com relação média albumina:globulina de 0,94. O número médio de células nucleadas foi de 313 células/µl, com predominância de grandes células mononucleares e linfócitos. Houve correlação significativa (r = - 0,649, P<0,01) entre o aumento da concentração de ácido hialurônico e a diminuição percentual de linfócitos. As mensurações das características pesquisadas no líquido sinovial da bainha tendínea digital de eqüinos são de execução simples e passíveis de implantação na rotina de atendimentos clínico-cirúrgicos.

Old beagle dogs have lower faecal concentrations of some fermentation products and lower peripheral lymphocyte counts than young adult beagles

The effects of age on microbiota composition, gut fermentation end-product formation and peripheral lymphocyte numbers were compared between old and young adult Beagle dogs fed four kibble diets differing in yeast cell wall contents. The experiment had a double 4 x 4 Latin square design, one with four mature dogs (4 years old) and the other with four old dogs (10 years old), with four replicates (diets) per dog. In each period a 15d adaptation period preceded a 5d total collection of faeces for the digestibility trial. on day 21, fresh faecal samples were collected for the determination of bacterial enumeration, pH, biogenic amine and short-chain fatty acid. Flow cytometry was used for immunophenotypic evaluation. Dogs were fed four kibble diets with similar composition with 0, 0.15, 0.30 and 0.45% of yeast cell wall (as-fed), respectively. Data were evaluated using general linear models of Statistical Analysis Systems statistical software (P<0.05). No evidence of a difference in faecal bacteria counts between ages was found (total aerobes, total anaerobes, Bifidobacterium, Lactobacillus, Clostridium and Escherichia coli: P. 0.15). Faecal concentrations of butyrate, histamine, agmatine and spermine were lower (P <= 0.05) and faecal pH was higher (P=0.03) in older dogs than in mature adult dogs, suggesting an alteration in bacterial metabolic activity, or in the rate of intestinal absorption of these compounds. Concentrations of T-lymphocytes, T-cytotoxic lymphocytes and B-lymphocytes were also lower (P <= 0.01) in older dogs than in mature adult dogs. The study confirmed alterations in peripheral lymphocytes and revealed a reduced concentration of some fermentation end products in the colon of old dogs.

Hematological values and body, heart and liver weights of male and female broiler embryos of young and old breeder eggs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aspectos morfométricos da cicatrização do cólon descendente de eqüinos submetidos a enterorrafias aposicionais com poliglactina 910 e com cianoacrilato

Foi realizado estudo morfométrico do cólon descendente de 15 eqüinos hígidos, nove fêmeas e seis machos, submetidos a enterorrafias aposicionais com fio de poliglactina 910 ou com cianoacrilato. Os animais foram distribuídos aleatoriamente em cinco grupos de três animais cada e sacrificados aos três, sete, 14, 35 e 70 dias de pós-operatório para colheita dos segmentos intestinais operados e posterior processamento histológico de rotina. A análise morfométrica foi feita em microscópio óptico adaptado a um sistema de análise de imagens, onde se quantificou o infiltrado inflamatório, mediante contagens de neutrófilos, eosinófilos, macrófagos, linfócitos e plasmócitos, não sendo observadas diferenças (P>0,05) entre as enterorrafias. Houve diferenças significativas entre os dias de pós-operatório para as contagens de neutrófilos, eosinófilos, macrófagos e linfócitos. Também se mensuraram as espessuras das regiões operadas (cicatriz) e das áreas contíguas não operadas (controle), registrando-se diferenças entre os dias de pós-operatório e as regiões (P<0,05), mas não entre as enterorrafias. O fio de poliglactina 910 e o cianoacrilato apresentam a mesma eficácia para serem usados em enterorrafias aposicionais do cólon descendente de eqüinos.

Kinetics of cellular component in inflammatory response induced by different stimuli in the swim bladder of pacu, Piaractus mesopotamicus Holmberg 1887 (Characidae)

The present study assessed the kinetics of cell accumulation at the site of inflammation induced by thioglycolate, Escherichia coli lipopolysaccharide (LPS) and heat-inactivated Aeromonas hydrophila, in the pacu, Piaractus mesopotamicus (Characidae), swim bladder. A quantitative, as well as qualitative, assessment was done of all the cells present in the exudate at 6, 24, and 48 h (n = 8) after inoculation of inflammatory agents. The results show that the thioglycolate was the irritant to induce higher total inflammatory cell accumulation when compared to the control group, 6 h after insult (P < 0.05). Inoculation of heat-inactivated Aeromonas hydrophila induced progressive accumulation of total inflammatory cells, with cell number peaking after 24 h and being significantly higher than observed in the other groups (P < 0.05). Injection of LPS also induced greater cell accumulation when compared to the control group (P < 0.05), although in lower numbers than those induced by the other two irritants. All irritants injected induced significantly greater accumulation of lymphocytes and thrombocytes when compared to the control group (P < 0.05).

Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological findings

Clinical signs, humoral and cellular immune responses, and microscopic and gross tissue alterations resulting from acute experimental Ehrlichia canis infection in dogs were studied. Four dogs were inoculated with E canis and four were used as uninfected controls. After a 10-14-day incubation period, infected dogs developed pyrexia up to 41 degreesC for 6-8 days. Antibody titers to E. canis antigen were demonstrable in all inoculated dogs at 30 days post-infection. Necropsy of infected animals revealed pale mucous membranes, generalized lymphadenopathy, splenomegaly, edema and ascites. Microcopically, the main lesions were: lymphoreticular hyperplasia in cortical areas of lymph nodes and spleenic white pulp, periportal accumulation of mononuclear cells and centrolobular fatty degeneration of the liver. Kidneys presented with glomerulonephritis characterized by interstitial, mononuclear infiltration. Immunophenotyping of lymphocytes from lymph nodes and spleen sections displayed alterations in IgG, IgM, CD3+ and CD8+ cells population in infected dogs. (C) 2003 Elsevier B.V. All rights reserved.

Haematological and gill responses in parasitized tilapia from valley of Tijucas River, SC, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Antigen-presenting cells in draining lymph nodes of goats repeatedly infested by the Cayenne tick Amblyomma cajennense nymphs

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Leukocyte entry into the CNS of Leishmania chagasi naturally infected dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Inflammatory responses of Nile tilapia Oreochromis niloticus to Streptococcus agalactiae: effects of vaccination and yeast diet supplement

This study evaluated the effects of dietary supplementation with 0.3% Saccharomyces cerevisiae yeast cell wall and of vaccination against Streptococcus agalactiae on the cellular component of acute inflammation induced in the coelomic cavity of Nile tilapia Oreochromis niloticus and on survival of the fish after challenge. A total of 84 tilapia of mean (+/- SD) weight 125.0 +/- 1.5 g were distributed among twelve 310 l fiberglass tanks according to a 2 x 2 x 3 factorial design in the following manner: with and without supplementation; 2 stimulations (oily solution without S. agalactiae vaccine and vaccination); 15 d later all fish were intracoelomically challenged with 10(8) CFU ml(-1) of a homologous strain of S. agalactiae, and evaluated after 6, 24 and 48 h, with 7 replicates. The fish received the non-supplemented or supplemented diet for a total of 77 d. The vaccination was performed on the 60th day, intracoelomically, as a single injection of 0.5 ml of the vaccine containing 10(8) CFU ml(-1). Fifteen days later, all the fish were challenged with S. agalactiae by means of an intracoelomic inoculation of 10(8) CFU ml(-1). No mortality was observed among the supplemented fish. The fish that were fed the non-supplemented diet and immunized with the bacterium presented a mortality rate of 28.5%. Among the non-supplemented and non-immunized fish, the mortality rate was 38.09%. Supplementation, in both vaccinated and non-vaccinated fish, induced larger accumulations of thrombocytes, lymphocytes and macrophages at the inflammatory focus. The results suggest that supplementation with 0.3% yeast cell wall, in both vaccinated and non-vaccinated fish, improved the inflammatory response of the fish and protected against the challenge. Vaccination increased the defense response, but the effect was stronger when associated with supplementation with S. cerevisiae.

Immune response pattern of the popliteal lymph nodes of dogs with visceral leishmaniasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)