Propolis anti-inflammatory effects on MAGE-1 and retinoic acid-treated dendritic cells and on Th1 and T regulatory cells

Background: Propolis exhibits huge potential in the pharmaceutical industry. In the present study, its effects were investigated on dendritic cells (DCs) stimulated with a tumor antigen (MAGE-1) and retinoic acid (RA) and on T lymphocytes to observe a possible differential activation of T lymphocytes, driving preferentially to Th1 or Treg cells. Methods: Cell viability, lymphocyte proliferation, gene expression (T-bet and FoxP3), and cytokine production by DCs (TNF-α, IL-10, IL-6 and IL-1ß) and lymphocytes (IFN-γ and TGF-ß) were analyzed. Results: MAGE-1 and RA alone or in combination with propolis inhibited TNF-α production and induced a higher lymphoproliferation compared to control, while MAGE1 + propolis induced IL-6 production. Propolis in combination with RA induced FoxP3 expression. MAGE-1 induced IFN-γ production while propolis inhibited it, returning to basal levels. RA inhibited TGF-ß production, what was counteracted by propolis. Conclusion: Propolis affected immunological parameters inhibiting pro-inflammatory cytokines and favoring the regulatory profile, opening perspectives for the control of inflammatory conditions.(AU)

Nodular lymphocytic conjunctivitis in a horse - case report / Conjuntivite linfocítica nodular em equino - relato de caso

The case of an equine with nodular lymphocytic conjunctivitis is described. A 14-year-old crossbred mare was seen presenting with development of a mass in the nasal angle of the right eye, behind the third eyelid. The tutor reported slow growth over 4 years, always accompanied by epiphora, and that no treatment had been performed prior to consultation. The histopathological and immunohistochemical results found a nodular, subepithelial structure, composed predominantly of densely packed small lymphocytes. Through the exams, associated with studies with monoclonal anti B lymphocyte antibodies and polyclonal anti T lymphocyte antibodies, the diagnosis of nodular lymphocytic conjunctivitis was reached. Only clinical pharmacological treatment was chosen, based on the use of topical and intralesional hydrocortisone acetate. After one month of treatment the mass completely disappeared without sequelae.

Descreve-se o caso de um equino com conjuntivite linfocítica nodular. Uma égua, mestiça, de 14 anos, foi atendida apresentando desenvolvimento de uma massa no ângulo nasal do olho direito, atrás da terceira pálpebra. O tutor relatou um crescimento lento durante 4 anos, sempre acompanhado de epífora, para o qual não foi realizado tratamento prévio à consulta. Os resultados histopatológico e imuno-histoquímico constataram uma estrutura nodular e subepitelial, composta predominantemente por linfocitos pequenos densamente agrupados. Por meio dos exames, associados a estudos com anticorpos monoclonais antilinfocitos B e anticorpos policlonais antilinfocitos T, chegou-se ao diagnóstico de conjuntivite linfocítica nodular. Optou-se apenas pelo tratamento clínico farmacológico, baseando-se na utilização de acetato de hidrocortisona tópica e intralesional. Após um mês de tratamento, a massa involuiu por completo, sem deixar sequelas.

CD4+ and CD8+ T lymphocyte counts and ratio in cats with chronic gingivostomatitis and naturally infected with feline immunodeficiency virus: a preliminary study / Razão e contagem de linfócitos T CD4+ e linfócitos T CD8+ em gatos com gengivoestomatite crônica e infectados naturalmente pelo vírus da imunodeficiência felina: estudo preliminar

This study aimed to evaluate the CD4+ and CD8+ T lymphocytes counts and CD4+: CD8+ ratio in a colony of cats with chronic gingivostomatitis (CGS). We used forty domestic short-haired cats inhabiting the same colony. Ten cats with CGS were immunodeficiency virus-positive (group IV), and ten with CGS were immunodeficiency virus-negative (group III). As a control, twenty cats without CGS were used: ten cats were immunodeficiency virus-positive (group II) and ten cats were immunodeficiency virus-negative (group I). We employed flow cytometry to count CD4+ and CD8+ T lymphocytes. In cats infected with the immunodeficiency virus, the presence of CD4+ lymphocytes were lower both for animals with and without CGS. Conversely, not immunodeficiency virus-infected cats with CGS had a higher amount of CD4+ when compared to seronegative animals without CGS. The counts of CD8+ T lymphocytes showed no significant difference among cats with CGS, whether infected with immunodeficiency virus or not. The CD4+: CD8+ ratio was only different for group III, which was higher than any other group. No difference was observed for total lymphocyte number and CD8+ among groups. By contrast, mean CD4+ levels were different, with cats from groups III and IV showing higher levels than those from groups I and II. The flow cytometry could be a useful tool for the diagnosis and prognosis of cats with CGS infected by the immunodeficiency virus.

Este estudo teve como objetivo avaliar a contagem e a razão de linfócitos T CD4+ e CD8+ em uma colônia de gatos com gengivoestomatite crônica (CGS). Foram analisados quarenta gatos domésticos que habitavam a mesma colônia. Dez gatos com CGS foram positivos para o vírus da imunodeficiência (grupo IV), e dez com CGS foram negativos para o vírus da imunodeficiência (grupo III). Como controle, vinte gatos sem CGS foram usados: dez gatos foram positivos para o vírus da imunodeficiência (grupo II) e dez gatos foram negativos para o vírus da imunodeficiência (grupo I). Empregou-se a citometria de fluxo para contagem de linfócitos T CD4+ e CD8+. Nos gatos infectados pelo vírus da imunodeficiência, a presença de linfócitos CD4+ foi menor tanto para os animais com e sem CGS. Por outro lado, gatos não infectados e com CGS apresentaram maior quantidade de linfócitos CD4+ quando comparados a animais soronegativos sem CGS. A contagem de linfócito T CD8+ não mostrou diferença significativa entre gatos com CGS, infectados ou não com o vírus da imunodeficiência. A razão CD4+:CD8+ foi diferente apenas para o grupo III, que foi maior do que qualquer outro grupo. Não foi observada diferença para o número total de linfócitos e CD8+ entre os grupos. Em contraste, os níveis médios de CD4+ foram diferentes, com os gatos dos grupos III e IV apresentando níveis mais elevados do que os dos grupos I e II. A citometria de fluxo pode ser uma ferramenta útil para o diagnóstico e prognóstico de gatos com CGS infectados pelo vírus da imunodeficiência.

Tanshinone IIA ameliorates myocardial ischemia/reperfusion injury in rats by regulation of NLRP3 inflammasome activation and Th17 cells differentiation

Purpose: Tanshinone IIA is a well-known lipophilic active constituent refined from traditional Chinese medicines, danshen. It has been previously demonstrated to possess various biological properties, including anti-inflammatory, antioxidant, promoting angiogenesis effect and so on. However, the mechanism of tanshinone IIA on myocardial ischemia-reperfusion injury (MI/RI) remains unclear. In this study, we investigated the effect of tanshinone IIA on MI/RI. Methods: MI/RI rat models were set up. Echocardiographic evaluation and hematoxylin and eosin staining were performed to analyze the cardiac function and morphology of MI/RI. Western blot was conducted for the detection of protein expression of pyrin domain containing 3 (NLRP3) and caspase-1 in heart tissues. Flow cytometry and real-time polymerase chain reaction were used for the detection of proinflammatory cytokines and Th17 cells differentiation. Results: We found that tanshinone IIA alleviated the myocardial damage of MI/RI, ameliorated the overall and local inflammatory reaction, and produced a cardioprotective effect by inhibiting of NLRP3 inflammasome activation and Th17/Treg cells differentiation. Conclusions: Our results highlighted the cardio-protective effect of tanshinone IIA on MI/RI and uncovered its underlying mechanism related to the NLRP3 inflammasome inhibition and the modulation of Th17/Treg cells differentiation.

The ethanol extract of Periplaneta Americana L. improves ulcerative colitis induced by a combination of chronic stress and TNBS in rats

Purpose: To investigate the effects of Periplaneta americana L. on ulcerative colitis (UC) induced by a combination of chronic stress (CS) and 2,4,6-trinitrobenzene sulfonic acid enema (TNBS) in rats. Methods: The experiment UC model with CS was established in rats by a combination of chronic restraint stress, excess failure, improper, and TNBS. The body weight, disease activity index (DAI), colonic mucosal injury index (CMDI), histopathological score (HS) and pro-inflammatory mediators were measured. The content of corticotropin-releasing hormone (CRH) in hypothalamus or adrenocorticotropic hormone (ACTH) and corticosteroids (CORT) in plasma were evaluated by enzyme-linked immunosorbent assay. The proportion of T lymphocyte subsets was detected by flow cytometry, and gut microbiota was detected by 16S rDNA amplicon sequencing. Results: Weight loss, DAI, CMDI, HS and proinflammatory mediators were reversed in rats by P. americana L. treatment after UC with CS. Increased epidermal growth factor (EGF) was observed in P. americana L. groups. In addition, P. americana L. could reduce the content of CRH and ACTH and regulate the ratio of CD3+, CD3+CD8+ and CD3+CD4+CD25+/CD4+ in spleen. Comparably, P. americana L. changes composition of gut microbiota. Conclusions: The ethanol extract of Periplaneta Americana L. improves UC induced by a combination of CS and TNBS in rats.

Linfoma de células T na cavidade nasal de caprino / T-cell lymphoma in the nasal cavity in goat

Background: Lymphomas are considered uncommon in goats, being the multicentric form with the highest number of cases for the species. Primary intranasal lymphomas are often diagnosed in dogs, cats, and humans. In the literature, there is only a description of a multicentric case involving the frontal sinuses and mucosa of the nasal cavity in a goat; therefore, it is important to describe unusual cases of this disease for the inclusion of new clinical and pathological characteristics in the ruminant clinic medicine. The objective of this work is to describe a case of T-cell lymphoma in the nasal cavity of a young goat. Case: The animal had dyspnea and respiratory noise for 15 days. Clinical examination showed nodulation in the right nasal cavity associated with serosanguinous secretion. Tracheostomy was performed; however, after 30 days the animal was euthanized. A sagittal plane of the head showed a pinkish-gray mass in the right and left nasal cavity, with a smooth, multilobulated surface, smooth adhering to the rostral portion of the dorsal concha and occluding the dorsal nasal meatus. Submandibular lymph nodes were slightly enlarged. Histopathological examination of the nasal cavity revealed a non-encapsulated, poorly delimited and ulcerated tumor composed of round cells arranged in a mantle supported by a discrete fibrovascular stroma extending the mucosa and lamina propria. Cells were round with sparse, eosinophilic and poorly delimited cytoplasm. Nuclei varied from round to elongated with condensed chromatin and evident nucleoli. Occasionally, aberrant nuclei, reniform shape and multinucleated cells were seen. Pleomorphism was moderate characterized by anisocytosis and anisocariosis. Typical and atypical mitosis were frequent (0-4 per field of highest magnification [400x]). Amidst the neoplasm, there were multifocal areas of necrosis and hemorrhage associated with a mild lymphocytic inflammatory infiltrate. Immunohistochemistry showed positive immunostaining for Vimentin antibodies and CD3, and negative for pan CK and CD20. Discussion: The lymphomas immunophenotyping is little used when it comes to farm animals, and there are few studies that use this technique for the definitive diagnosis of these neoplasms for small ruminants. The use of this technique must be considered in each case, in order to determine the pathogenesis, the accurate diagnosis and the origin of the neoplastic lymphocytes. In goats, T-cell lymphomas are the most diagnosed, although cases of multicentric B-cell lymphomas with ocular involvement have been diagnosed. In view of the clinical picture of the case described, infectious rhinitis already described in goats, such as aspergillosis and protothecosis, should be included as differential diagnoses. However, the anatomopathological findings facilitate the direction of the diagnosis, since infectious rhinitis presents as nodules / ulcerated masses or focal areas of necrosis associated with purulent secretion and in the histopathological examination it is possible to observe the intralesional etiological agents. In addition, the enzootic ethmoidal tumor must be included, as it has similar clinical signs and affects young animals, but they are adenomas/adenocarcinomas that affect the ethmoidal nasal shells induced by a retrovirus. Lymphomas in the caprine species are rare in the Northeastern semi-arid, but that in the present diagnostic routine occasionally occurs, being important the first description of its nasal shape for its inclusion in the differential diagnoses of diseases that present with clinical obstruction and dyspnea for the species.

Clonality test by PCR - PARR in real time of canine lymphomas

Lymphoma is a neoplasm of hematopoietic origin that affects canines. The proper establishment of prognosis and rapid institution of treatment are essential for a better quality of life, and immunophenotyping is one of the tools used for this purpose. The objective of this study was to perform a clonality test for immunophenotypic characterization of canine lymphomas using the polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) technique in real-time from samples fixed in formalin and embedded in paraffin. The 23 analyzed samples were fixed in formalin and embedded in paraffin canine lymphoma from the collection Laboratory of Histopathology of the Animal Pathology Area of the Departament of Veterinary Medicine - Federal Rural University of Pernambuco (UFRPE). Samples were processed, their DNA was extracted, quantified, diluted, and standardized at a concentration of 50 ng/µL. After extraction, all samples were subjected to conventional PCR for endogenous control (detection of the IgM target region), in which the extracted DNA was amplified in a final volume of 25 µL. The 128 bp amplified product was detected by 1.5% agarose gel electrophoresis. Of the 23 samples analyzed for the detection of the conserved region referring to the endogenous gene, 91.30% (21/23) amplified the conserved region Cµ by conventional PCR, and two samples 8.70% (2/23) were negative. Endogenous control positive samples were subjected to real-time PCR-PARR for detection of IgH Major and IgH Minor for B lymphocytes (LB), and TCRy for lymphocytes T (LT) target regions. All reactions were performed in duplicate to reduce the risk of false-positive or false-negative results due to technical errors. Samples previously confirmed by immunohistochemistry were used as positive controls for T cell and B cell lymphoma, and MilliQ water was used as a negative reaction control. After amplification, the melting curve gradually increased the temperature by 1o C/5 s to 95o C during continuous fluorescence monitoring. Of the 21 samples analyzed, 100.00% (21/21) demonstrated clonal amplification. Of these, 57.15% (12/21) were positive for phenotype B, and 42.85% (9/21) were positive for phenotype T. Due to the importance of researching and confirming samples from files fixed and embedded in paraffin samples in laboratories, PCR-PARR is a good tool for this purpose. In the present study, real-time PCR analysis demonstrated greater sensitivity in the characterization of the immunophenotype of lymphomas from old samples fixed in formalin and embedded in paraffin. The temperature of melting curve analysis may vary depending on the amount of DNA and its quality. In the present study, it was found that the average melting temperature in the samples varied between ± 3o C when compared to that in the control sample for LB and LT, 83.5o C and 80o C, respectively: in the literature, there is a relative difference in this temperature, which may vary up to 4o C. Real-time PCR-PARR was satisfactory in the characterization of the immunophenotype of canine lymphomas from formalin-fixed and paraffin-embedded samples; therefore, its use is recommended for both retrospective studies. The use of PCR-PARR associated with histopathological and/or cytopathological examination in cases of canine lymphomas strongly helps pathologists, provide a safe establishment of the immunophenotype, minimize errors, and optimize the diagnosis, thus directly contributing to the establishment of the prognosis.(AU)

Effects of Glutamine on Lymphocyte Proliferation and Intestinal Mucosal Immune Response in Heat-Stressed Broilers

To investigate the protective effect of glutamine (Gln) on lymphocyte proliferation and the intestinal mucosal immune response in heat-stressed broilers, 360 21-day-old Arbor Acres (AA) broilers were assigned to 4 groups in a completely randomized design, each of which included 6 replicates with 15 birds per replicate for 21 days. The chickens were fed a basal diet under no stress (NS group), a basal diet under heat stress (HT group), or a basal diet under heat stress with the addition of either 0.5 % or 1.0 % Gln. The results showed that the broilers in the HT group exhibited fewer proliferating peripheral lymphocytes, a lower growth performance, phagocytic rate and index of neutrophils, fewer goblet cells in whole intestine and intraepithelial lymphocyte (IEL) cells in the ileum, a lower sIgA content in the duodenum and the jejunum, a lower immunoglobulin content of serum and intestinal mucosa, than those of the NS group (p<0.05). Diets supplemented with Gln increased growth performance, the number of proliferating peripheral lymphocytes, the phagocytic rate and phagocytic index of neutrophils, the number of whole intestine goblet cells and ileum IEL cells, the sIgA contents of the duodenum and the jejunum, and the immunoglobulin contents of serum and intestinal mucosa (p<0.05) in broilers exposed to HT. In conclusion, Gln can enhance intestinal immune function in broiler chickens by stimulating T and B lymphocyte proliferation, increasing the number of goblet cells and IEL cells, as well as increasing the content of sIgA and immunoglobulin secretion.(AU)

Effects of Glutamine on Lymphocyte Proliferation and Intestinal Mucosal Immune Response in Heat-Stressed Broilers

To investigate the protective effect of glutamine (Gln) on lymphocyte proliferation and the intestinal mucosal immune response in heat-stressed broilers, 360 21-day-old Arbor Acres (AA) broilers were assigned to 4 groups in a completely randomized design, each of which included 6 replicates with 15 birds per replicate for 21 days. The chickens were fed a basal diet under no stress (NS group), a basal diet under heat stress (HT group), or a basal diet under heat stress with the addition of either 0.5 % or 1.0 % Gln. The results showed that the broilers in the HT group exhibited fewer proliferating peripheral lymphocytes, a lower growth performance, phagocytic rate and index of neutrophils, fewer goblet cells in whole intestine and intraepithelial lymphocyte (IEL) cells in the ileum, a lower sIgA content in the duodenum and the jejunum, a lower immunoglobulin content of serum and intestinal mucosa, than those of the NS group (p<0.05). Diets supplemented with Gln increased growth performance, the number of proliferating peripheral lymphocytes, the phagocytic rate and phagocytic index of neutrophils, the number of whole intestine goblet cells and ileum IEL cells, the sIgA contents of the duodenum and the jejunum, and the immunoglobulin contents of serum and intestinal mucosa (p<0.05) in broilers exposed to HT. In conclusion, Gln can enhance intestinal immune function in broiler chickens by stimulating T and B lymphocyte proliferation, increasing the number of goblet cells and IEL cells, as well as increasing the content of sIgA and immunoglobulin secretion.

Immune response in Santa Inês sheep during Fasciola hepatica infection / Resposta imune em ovinos da raça Santa Inês durante a infecção por Fasciola hepatica

In the present study, we describe the proliferative response and the IFN-γ, IL-10, IL-4 cytokine expression profiles in peripheral blood, and IgG production, after the experimental infection of Santa Inês sheep with 250 Fasciola hepatica metacercariae that were evaluated at 60 and 210 days post-infection(dpi). The cytokine expression profile at 60 dpi was characterized by the production of IFN-γ and IL-10, which is indicative of an initial mixed Th1/Th2 response. The modulation of the response occurred at 210 dpi with a predominance of IL-10 and IL-4 over IFN-γ. Mononuclear cells from peripheral blood stimulated with F. hepatica antigen exhibited proliferative capacity at 60 dpi, whereas the response was consistent with modulation from Th1 to Th2 in the chronic phase. The IgG antibody response was more marked at 60 days than at 210 dpi, confirming the mixed-response profile. The late modulation of the T lymphocyte response in association with the predominance of IL-10 and IL-4 at 210 dpi suggests the involvement of these cytokines after the establishment of the parasites in the bile ducts. The transition from a mixed to a regulatory response in the chronic phase was also accompanied by a reduced number of eggs per gram of feces.(AU)

No presente estudo, observou-se a resposta proliferativa e os perfis de expressão de citocinas IFN-γ, IL10, IL-4 no sangue periférico, e a produção de IgG, aos 60 e 210 dias após infecção (dpi) experimental de ovinos Santa Inês com 250 metacercárias de Fasciola hepatica. O perfil de expressão de citocinas aos 60 dpi foi caracterizado pela produção de IFN-γ e IL-10, o que é indicativo de uma resposta mista inicial Th1/ Th2. A modulação da resposta ocorreu aos 210 dpi, com predominância de IL-10 e IL-4 sobre IFN-γ. As células mononucleares do sangue periférico estimuladas com antígeno total de F. hepatica adulta exibiram capacidade proliferativa aos 60 dpi, enquanto a resposta foi consistente com uma modulação de Th1 para Th2 na fase crônica. A resposta do anticorpo IgG foi maior aos 60 dpi do que aos 210 dpi. A modulação tardia da resposta dos linfócitos T em associação com a predominância de IL-10 e IL-4 aos 210 dpi sugere o envolvimento dessas citocinas após o estabelecimento dos parasitos nos ductos biliares. A transição de uma resposta mista para uma resposta com pefil regulador na fase crônica também foi acompanhada por redução dos ovos por grama de fezes.(AU)

Central nervous system infiltration in acute lymphoblastic leukemia in a dog

Background: Acute lymphoblastic leukemia (ALL) is a malignant neoplasia in which there is proliferation of lymphoid progenitor cells in the bone marrow, blood, and extramedullary sites. This disorder has a fast and progressive development; in dogs, cases of infiltration of ALL cells in the central nervous system (CNS) are uncommon and rare. Diagnosis can be achieved with the help of the clinical history and physical, radiographic, hematological, myelographic, and cerebrospinal fluid (CSF) tests in patients with or without neurological clinical signs. The present report aims to describe a case of ALL and the presence of lymphoblasts in the CSF of a dog with neurological clinical signs. Case: An 8-year-old Lhasa Apso dog was examined at the Veterinary Hospital of Universidade Federal do Paraná, Curitiba campus. At the physical examination, the animal exhibited apathy and paralysis of pelvic limbs, which progressed to tetraplegia. Abdominal palpation revealed presence of hepatosplenomegaly and absence of lymphadenomegaly. No alterations were observed in radiographs of the cervical, thoracic or lumbar spine. A complete blood count revealed presence of non-regenerative anemia (hematocrit = 22%), extreme lymphocytosis (185,229 cells/µL), lymphoblasts at a level of 72% (133,364 cells/µL), and thrombocytopenia (66,000 platelets/µL). The biochemical tests revealed increased alkaline phosphatase (859 IU/L). The levels of alanine aminotransferase, creatinine, urea, total protein, albumin, and globulin were normal. The diagnosis of ALL was achieved with the help of a myelogram. The myelogram findings included 39% of mature lymphocytes and 59% of lymphoblasts exhibiting large size, spherical shape, poorly delimited borders, with a high nucleus/cytoplasm ratio, marked cytoplasmic basophilia, and 2 to 3 evident nucleoli; metarubricytes (1%) and promyelocytes (0.6%) were also observed. The CSF contained an increased number of nucleated cells (27 cells/µL) comprising lymphocytes (43%), macrophages (33%), and segmented neutrophils (24%). Of the 11.6 lymphocytes per µL of CSF, 8.1 were lymphoblasts, which indicates infiltration of ALL cells in the CNS. The animal died one day after collection of bone barrow and CSF. Discussion: Relevant alterations observed in this case included the neurological signs caused by the infiltration of neoplastic cells in the CNS, severe leukocytosis and lymphocytosis, with large amounts of lymphoblasts in the blood and predominance of lymphoblasts in the bone marrow, which are alterations typically found in ALL. The animal also exhibited non-regenerative anemia and thrombocytopenia, which were secondary to infiltration of leukemic cells in the bone marrow. The CSF exhibited pleocytosis (27 cells/ µL), and 30% of the cells observed were lymphoblasts. Lymphoblast infiltration in the CNS of leukemic dogs is rare, and other studies have reported absence of neurological signs or neurological signs different from those observed in the present study. CSF analysis in indicated in cases of leukemia to assess leukemic cell infiltration in the CNS. In the case reported here, the plasma level of alkaline phosphatase was increased (859 IU/L) as a consequence of hepatomegaly and hepatic cholestasis. ALL is a very aggressive, proliferative neoplasia, and the resulting lymphoblasts infiltrated the CNS of the animal. In cases of ALL, performing complete blood count, myelogram, and CSF analysis is indicated whether the patients exhibit neurological signs or not.(AU)

Histopathological and immunophenotypical characterization of canine multicentric lymphoma in Brazil: a study of 203 cases / Caracterização histopatológica e imunofenotípica do linfoma multicêntrico canino no Brasil: um estudo de 203 casos

The immunophenotype is regarded as an independent prognostic factor in high-grade lymphomas, seeing that lymphomas of T-cell origin are associated with shorter survival time. Although a number of studies have evaluated the immunophenotypical profile of lymphoma in the USA and Europe, Brazilian research on the matter remains scarce. Exact characterization of the histopathological type is crucial to establish proper treatment and prognosis. This study evaluated the database of immunohistochemistry laboratories that perform immunophenotyping of canine lymphoma in Brazil. A total of 203 cases of multicentric lymphoma were classified according to the WHO classification. Immunophenotyping was able to identify 71.4% lymphomas of B-cell line, 27.1% of T-cell line and 1.5% of non-B cells and non-T cell lines. Diffuse large B-cell lymphoma was the most common with 59.1% of the cases. Among T-cell lymphomas, lymphoblastic was the most common (11.33% of the cases). Even though canine lymphomas tend to be high-grade, indolent lymphomas comprised 11.82% of the cases and T-zone lymphoma was the most prevalent (8.86%). The immunophenotype of multicentric lymphoma in Brazil is similar to those in other parts of the world, which suggests similar etiologic factors to the development of this disease.(AU)

O imunofenótipo é considerado um fator prognóstico independente em linfomas de alto grau, visto que os linfomas de origem de células T estão associados a menor tempo de sobrevida. Apesar de vários estudos terem avaliado o perfil imunofenotípico do linfoma nos EUA e na Europa, a pesquisa brasileira sobre o assunto ainda é escassa. A caracterização exata do tipo histopatológico é crucial para estabelecer o tratamento e o prognóstico adequados. Este estudo avaliou a base de dados de laboratórios de imuno-histoquímica que realizam imunofenotipagem do linfoma canino no Brasil. Um total de 203 casos de linfoma multicêntrico foi classificado de acordo com a classificação da OMS. A imunofenotipagem foi capaz de identificar 71,4% dos linfomas da linhagem de células B, 27,1% da linhagem de células T e 1,5% das linhagens de células não B e não T. O linfoma difuso de grandes células B foi o mais comum em 59,1% dos casos. Entre os linfomas de células T, o linfoblástico foi o mais comum (11, 33% dos casos). Embora os linfomas caninos tendam a ser de alto grau, os linfomas indolentes representaram 11,82% dos casos e o linfoma da zona T foi o mais prevalente (8,86%). O imunofenótipo do linfoma multicêntrico no Brasil é semelhante ao de outras partes do mundo, o que sugere fatores etiológicos semelhantes ao desenvolvimento dessa doença.(AU)

Histopathological and immunophenotypical characterization of canine multicentric lymphoma in Brazil: a study of 203 cases / Caracterização histopatológica e imunofenotípica do linfoma multicêntrico canino no Brasil: um estudo de 203 casos

The immunophenotype is regarded as an independent prognostic factor in high-grade lymphomas, seeing that lymphomas of T-cell origin are associated with shorter survival time. Although a number of studies have evaluated the immunophenotypical profile of lymphoma in the USA and Europe, Brazilian research on the matter remains scarce. Exact characterization of the histopathological type is crucial to establish proper treatment and prognosis. This study evaluated the database of immunohistochemistry laboratories that perform immunophenotyping of canine lymphoma in Brazil. A total of 203 cases of multicentric lymphoma were classified according to the WHO classification. Immunophenotyping was able to identify 71.4% lymphomas of B-cell line, 27.1% of T-cell line and 1.5% of non-B cells and non-T cell lines. Diffuse large B-cell lymphoma was the most common with 59.1% of the cases. Among T-cell lymphomas, lymphoblastic was the most common (11.33% of the cases). Even though canine lymphomas tend to be high-grade, indolent lymphomas comprised 11.82% of the cases and T-zone lymphoma was the most prevalent (8.86%). The immunophenotype of multicentric lymphoma in Brazil is similar to those in other parts of the world, which suggests similar etiologic factors to the development of this disease.(AU)

O imunofenótipo é considerado um fator prognóstico independente em linfomas de alto grau, visto que os linfomas de origem de células T estão associados a menor tempo de sobrevida. Apesar de vários estudos terem avaliado o perfil imunofenotípico do linfoma nos EUA e na Europa, a pesquisa brasileira sobre o assunto ainda é escassa. A caracterização exata do tipo histopatológico é crucial para estabelecer o tratamento e o prognóstico adequados. Este estudo avaliou a base de dados de laboratórios de imuno-histoquímica que realizam imunofenotipagem do linfoma canino no Brasil. Um total de 203 casos de linfoma multicêntrico foi classificado de acordo com a classificação da OMS. A imunofenotipagem foi capaz de identificar 71,4% dos linfomas da linhagem de células B, 27,1% da linhagem de células T e 1,5% das linhagens de células não B e não T. O linfoma difuso de grandes células B foi o mais comum em 59,1% dos casos. Entre os linfomas de células T, o linfoblástico foi o mais comum (11, 33% dos casos). Embora os linfomas caninos tendam a ser de alto grau, os linfomas indolentes representaram 11,82% dos casos e o linfoma da zona T foi o mais prevalente (8,86%). O imunofenótipo do linfoma multicêntrico no Brasil é semelhante ao de outras partes do mundo, o que sugere fatores etiológicos semelhantes ao desenvolvimento dessa doença.(AU)

Multicentric T-cell lymphoma in a creole filly / Linfoma multicêntrico de células T em uma potra crioula

A 2-year-old creole filly was referred to us for evaluation because of the clinical suspicion of infection by Streptococcus equi. It presented with progressive weight loss and increased volume of the submandibular, retropharyngeal, and precrural lymph nodes. General clinical examinations and laboratory tests revealed dehydration, anemia, leukopenia, hyperfibrinogenemia, and thrombocytopenia. The initial treatment for equine adenitis did not achieve significant results, and new hematological and biochemical tests and lymph node cytology by puncture were performed. Cytology revealed cells compatible with neoplastic lymphocytes, resulting in the suspicion of lymphoma. The animal died from general weakness and was sent for autopsy. Macroscopically, generalized lymphadenomegaly and splenomegaly were observed, with multiple nodules and tumor lesions in the splenic parenchyma. There was an irregular nodule in the medullary layer of the right kidney. The liver and lungs were slightly enlarged, with petechiae and multifocal suffusions. Histopathological evaluation of different organ specimens revealed intense proliferation of the neoplastic lymphoid cells, invading the adjacent tissues, with moderate cellular pleomorphism. Immunohistochemistry of the lymph node sections with neoplastic infiltration revealed multicentric T-cell lymphoma. In horses, cases of lymphomas are rare and should be differentiated from other causes that induce lymphadenomegaly in this species.(AU)

Uma potra da raça crioula, com dois anos de idade foi encaminhada para avaliação clínica com suspeita de infecção por Streptococcus equi. A paciente apresentava emagrecimento progressivo e aumento dos linfonodos submandibulares, retrofaríngeos e pré-crurais. A potra foi submetida a exames clínicos gerais e de laboratório, apresentando desidratação, anemia, leucopenia, hiperfibrinogenemia e trombocitopenia. O tratamento inicial para adenite equina não obteve resultados significativos, e novos exames hematológicos, bioquímicos e citologia da punção de linfonodos foram realizados. O resultado da citologia revelou células compatíveis com linfócitos neoplásicos, o que levou a suspeita de linfoma. O animal morreu em consequência da debilidade de seu estado geral e foi encaminhado para necropsia. Macroscopicamente, observou-se linfadenomegalia generalizada, esplenomegalia com múltiplos nódulos no parênquima do baço, bem como lesões tumorais caracterizadas por rim direito apresentando nódulo irregular na camada medular, fígado levemente aumentado e pulmões com petéquias e sufusões multifocais. Amostras de variados órgãos foram submetidos a avaliação histopatológica, a qual revelou intensa proliferação de células linfoides neoplásicas com invasão de tecidos adjacentes com moderado pleomorfismo celular. Secções de linfonodos com infiltrado neoplásico foram submetidas à avaliação imuno-histoquímica (IHQ) a qual determinou tratar-se de linfoma de células T multicêntrico. Em equinos, esse tipo de neoplasma é pouco frequente e deve ser diferenciado de outras causas que induzem linfadenomegalia nessa espécie.(AU)

Cutaneous non-epitheliotropic large T-cell lymphoma in an English Bulldog

Cutaneous lymphoma is histologically classified in epitheliotropic and non-epitheliotropic, the first showing higher incidence in dogs, and the second, in cats. Non-epitheliotropic lymphoma presents lymphocyte aggregates in the dermis and subcutaneous tissue, however cutaneous annexes are not infiltrated. It is usually more aggressive than epitheliotropic lymphomas. The aim of this study was to report a case of non-epitheliotropic lymphoma in a 9-year-old, female, English Bulldog presented with non-ulcerated skin nodules adhered to deep tissues. Microscopic and immunophenotypic features supported the diagnosis of non-epitheliotropic large T-cell lymphoma. Treatment was initiated with modification of the LOPP protocol, replacing procarbazine by dacarbazine (600 mg/m²) for up to six cycles, with a three-month survival. In the 11th week of treatment, after recurrent episodes of vomiting and diarrhea, abdominal ultrasound was performed and revealed an infiltrative mass in the stomach’s greater curvature topography, showing an expansive and accentuated increase in one week, when euthanasia was elected.

Cutaneous non-epitheliotropic large T-cell lymphoma in an English Bulldog

Cutaneous lymphoma is histologically classified in epitheliotropic and non-epitheliotropic, the first showing higher incidence in dogs, and the second, in cats. Non-epitheliotropic lymphoma presents lymphocyte aggregates in the dermis and subcutaneous tissue, however cutaneous annexes are not infiltrated. It is usually more aggressive than epitheliotropic lymphomas. The aim of this study was to report a case of non-epitheliotropic lymphoma in a 9-year-old, female, English Bulldog presented with non-ulcerated skin nodules adhered to deep tissues. Microscopic and immunophenotypic features supported the diagnosis of non-epitheliotropic large T-cell lymphoma. Treatment was initiated with modification of the LOPP protocol, replacing procarbazine by dacarbazine (600 mg/m²) for up to six cycles, with a three-month survival. In the 11th week of treatment, after recurrent episodes of vomiting and diarrhea, abdominal ultrasound was performed and revealed an infiltrative mass in the stomachs greater curvature topography, showing an expansive and accentuated increase in one week, when euthanasia was elected.(AU)

Clinical, cytological, histopathological and immunohistochemical features of a T-Zone lymphoma in a mixed-breed dog

Background: Non-Hodgkin lymphoma are a group of lymphoid neoplasms originating from the proliferation of precursors or mature, T, B and/or NK lymphocytes. T-Zone lymphoma (TZL) is characterized as an indolent lymphoma due to its slow progression and poor chemotherapy’s response. Dogs affected by this neoplasm may live for many years without clinical signs and are often underdiagnosed. The aim of this study was to report a TZL case in a 9-year-old male mixed breed dog, submitted to clinical follow-up and chemotherapy. Case: A 9-year-old male mixed-breed dog was presented due to the observation of an increased left mandibular lymph node. The previous cytological examination was suggestive of reactive hyperplasia and histopathological examination, by incisional biopsy, compatible with lymphocytic low-grade lymphoma. Physical examination revealed enlarged and firm left mandibular lymph node and adequate physical condition. A cytological examination was performed on the mandibular and both popliteal lymph nodes and revealed many small lymphocytes with hyperchromatic chromatin, rarely evident nucleolus and whose cytoplasm often projected in the form of a “hand-mirror” or “comet tail”, compatible with lymphocytic lymphoma (low grade) and suggestive of TZL, in the three assessed lymph nodes. The histopathological and immunohistochemical examination, of the mandibular lymph node were chosen to confirm the diagnosis. In histopathology it was observed that 40% of the sample contained a monotonous cell population, composed by small lymphocytes, with some presenting “hand-mirror” morphology. Two mitotic figures were evidenced per field of high magnification (40x), inferring a low-grade disease. Immunohistochemical analysis revealed neoplastic proliferation with immunolabeling of CD3 lymphocytes and positivity for Ki-67 in 48% of neoplastic cells, but negative for CD20, CD79a, CD45, MUM-1 and...

Clinical, cytological, histopathological and immunohistochemical features of a T-Zone lymphoma in a mixed-breed dog

Background: Non-Hodgkin lymphoma are a group of lymphoid neoplasms originating from the proliferation of precursors or mature, T, B and/or NK lymphocytes. T-Zone lymphoma (TZL) is characterized as an indolent lymphoma due to its slow progression and poor chemotherapys response. Dogs affected by this neoplasm may live for many years without clinical signs and are often underdiagnosed. The aim of this study was to report a TZL case in a 9-year-old male mixed breed dog, submitted to clinical follow-up and chemotherapy. Case: A 9-year-old male mixed-breed dog was presented due to the observation of an increased left mandibular lymph node. The previous cytological examination was suggestive of reactive hyperplasia and histopathological examination, by incisional biopsy, compatible with lymphocytic low-grade lymphoma. Physical examination revealed enlarged and firm left mandibular lymph node and adequate physical condition. A cytological examination was performed on the mandibular and both popliteal lymph nodes and revealed many small lymphocytes with hyperchromatic chromatin, rarely evident nucleolus and whose cytoplasm often projected in the form of a “hand-mirror” or “comet tail”, compatible with lymphocytic lymphoma (low grade) and suggestive of TZL, in the three assessed lymph nodes. The histopathological and immunohistochemical examination, of the mandibular lymph node were chosen to confirm the diagnosis. In histopathology it was observed that 40% of the sample contained a monotonous cell population, composed by small lymphocytes, with some presenting “hand-mirror” morphology. Two mitotic figures were evidenced per field of high magnification (40x), inferring a low-grade disease. Immunohistochemical analysis revealed neoplastic proliferation with immunolabeling of CD3 lymphocytes and positivity for Ki-67 in 48% of neoplastic cells, but negative for CD20, CD79a, CD45, MUM-1 and...(AU)

Concentrations of matrix metalloproteinase-2 and interleukin-1 beta in the aqueous humor of dogs with normal and cataractous eyes / Concentração de metaloprotease-2 e de interleucina-1 beta no humor aquoso de cães com olhos normais e cataratosos

We aimed to determine the concentration of MMP-2 and IL-1ß in the aqueous humor of dogs with healthy eyes (n=8) and in those with mature (n=8) and hyper mature cataracts (n=8). Correlations between cytokines, cytokines, and intraocular pressure (IOP), as well as cytokines with ages of patients of each group, were also assessed. In patients with cataract, aqueous humor was collected at the end of the electroretinographic procedure. In healthy dogs, aqueous humor was collected before elective surgeries. Cytokine levels were determined using ELISA. IOP was assessed by applanation tonometry. IOP of patients with mature and hyper mature cataracts were lower than the ones measured in normal eyes (P=0.158). MMP-2 aqueous humor concentration was higher in patients with hyper mature cataracts, in comparisons with healthy patients (P=0.04). Average IL-1 ß aqueous concentration was higher in patients with cataracts (P<0.0001). Significant higher values of IL-1 ß were observed in patients with hyper mature, than in the ones with mature cataracts (P=0.0085). Correlations between MMP-2 and IL-1 ß (r=-0.38, P=0.06), MMP-2 and IOP (r=-0.149, P=0.484), and IL-1 ß and the ages of patients were not observed (P>0.05). IL-1 ß and IOP correlated negatively (r=-0.42, P=0.04). MMP-2 and the ages of patients correlated only in dogs with mature cataracts (r=0.772, P=0.02). It can be concluded that the increment in the aqueous humor concentration of IL-1 ß found in dogs with mature and hyper mature cataracts, in addition to the negative relationship of this cytokine with IOP, suggests that IL-1 ß is involved in the pathogenesis of LIU. Despite dogs with hypermature cataracts presented significant higher concentrations of MMP-2 in the aqueous humor, such cytokine did not correlate with IL-1 ß and IOP. In our study, a relationship between aqueous humor cytokines and the ages of patients was only confirmed between MMP-2 and the ages of dogs with mature cataracts.(AU)

Objetivou-se determinar as concentrações da metalloprotease-2 (MMP-2) e de interleucina-1 ß (IL-1 ß) em cães com olhos saudáveis (n=8) e naqueles com catarata madura (n=8) e hipermatura (n=8). Correlações entre ambas as citocinas, entre as citocinas e a pressão intraocular (PIO), assim como entre as citocinas e a idade dos pacientes dentro de cada grupo foram averiguadas. Nos pacientes com catarata, o humor aquoso foi colhido ao final da eletrorretinografia. Nos cães saudáveis, o humor aquoso foi colhido antes do início de cirurgias eletivas. Os níveis das citocinas foram determinados por ELISA e a PIO por tonometria de aplanação. A PIO dos pacientes com catarata madura e hipermadura foram mais baixas que aquelas dos pacientes controle (P=0.158). A concentração de MMP-2 no humor aquoso foi maior nos pacientes com catarata hipermtura, comparativamente aos pacientes saudáveis (P=0.04). A concentração de IL-1 ß no humor aquoso foi mais elevada nos cães com catarata (P<0.0001). Nos pacientes com catarata hipermatura, os valores de IL-1 ß foram significativamente mais altos que aqueles dosados nos pacientes com catarata madura (P=0.0085). Correlações entre MMP-2 e IL-1 ß (r=-0.38, P=0.06), MMP-2 e PIO (r=-0.149, P=0.484) e IL-1 ß e as idades dos pacientes não foram observadas (P>0.05). A IL-1 ß se correlacionou negativamente com a PIO (r=-0.42, P=0.04). Correlação entre MMP-2 e a idades dos pacientes foi observada apenas nos cães com catarata madura (r=0.772, P=0.02). Conclui-se que o aumento na concentração de IL-1 ß no humor aquoso de cães com catarata madura e hipermatura, associado à correlação negativa entre essa citocina e a PIO, sugerem que a mesma está envolvida na patogênese da uveíte induzida pela lente. Apesar dos cães com catarata hipermadura apresentarem concentrações significativamente maiores de MMP-2 no humor aquoso, essa citocina não se correlacionou com a IL-1 ß e a PIO. Em nosso estudo, correlação entre as citocinas dosadas no humor aquoso e a idade dos pacientes foi confirmada apenas entre MMP-2 e a idade dos cães com catarata madura.(AU)

Concentrations of matrix metalloproteinase-2 and interleukin-1 beta in the aqueous humor of dogs with normal and cataractous eyes / Concentração de metaloprotease-2 e de interleucina-1 beta no humor aquoso de cães com olhos normais e cataratosos

We aimed to determine the concentration of MMP-2 and IL-1ß in the aqueous humor of dogs with healthy eyes (n=8) and in those with mature (n=8) and hyper mature cataracts (n=8). Correlations between cytokines, cytokines, and intraocular pressure (IOP), as well as cytokines with ages of patients of each group, were also assessed. In patients with cataract, aqueous humor was collected at the end of the electroretinographic procedure. In healthy dogs, aqueous humor was collected before elective surgeries. Cytokine levels were determined using ELISA. IOP was assessed by applanation tonometry. IOP of patients with mature and hyper mature cataracts were lower than the ones measured in normal eyes (P=0.158). MMP-2 aqueous humor concentration was higher in patients with hyper mature cataracts, in comparisons with healthy patients (P=0.04). Average IL-1 ß aqueous concentration was higher in patients with cataracts (P<0.0001). Significant higher values of IL-1 ß were observed in patients with hyper mature, than in the ones with mature cataracts (P=0.0085). Correlations between MMP-2 and IL-1 ß (r=-0.38, P=0.06), MMP-2 and IOP (r=-0.149, P=0.484), and IL-1 ß and the ages of patients were not observed (P>0.05). IL-1 ß and IOP correlated negatively (r=-0.42, P=0.04). MMP-2 and the ages of patients correlated only in dogs with mature cataracts (r=0.772, P=0.02). It can be concluded that the increment in the aqueous humor concentration of IL-1 ß found in dogs with mature and hyper mature cataracts, in addition to the negative relationship of this cytokine with IOP, suggests that IL-1 ß is involved in the pathogenesis of LIU. Despite dogs with hypermature cataracts presented significant higher concentrations of MMP-2 in the aqueous humor, such cytokine did not correlate with IL-1 ß and IOP. In our study, a relationship between aqueous humor cytokines and the ages of patients was only confirmed between MMP-2 and the ages of dogs with mature cataracts.(AU)

Objetivou-se determinar as concentrações da metalloprotease-2 (MMP-2) e de interleucina-1 ß (IL-1 ß) em cães com olhos saudáveis (n=8) e naqueles com catarata madura (n=8) e hipermatura (n=8). Correlações entre ambas as citocinas, entre as citocinas e a pressão intraocular (PIO), assim como entre as citocinas e a idade dos pacientes dentro de cada grupo foram averiguadas. Nos pacientes com catarata, o humor aquoso foi colhido ao final da eletrorretinografia. Nos cães saudáveis, o humor aquoso foi colhido antes do início de cirurgias eletivas. Os níveis das citocinas foram determinados por ELISA e a PIO por tonometria de aplanação. A PIO dos pacientes com catarata madura e hipermadura foram mais baixas que aquelas dos pacientes controle (P=0.158). A concentração de MMP-2 no humor aquoso foi maior nos pacientes com catarata hipermtura, comparativamente aos pacientes saudáveis (P=0.04). A concentração de IL-1 ß no humor aquoso foi mais elevada nos cães com catarata (P<0.0001). Nos pacientes com catarata hipermatura, os valores de IL-1 ß foram significativamente mais altos que aqueles dosados nos pacientes com catarata madura (P=0.0085). Correlações entre MMP-2 e IL-1 ß (r=-0.38, P=0.06), MMP-2 e PIO (r=-0.149, P=0.484) e IL-1 ß e as idades dos pacientes não foram observadas (P>0.05). A IL-1 ß se correlacionou negativamente com a PIO (r=-0.42, P=0.04). Correlação entre MMP-2 e a idades dos pacientes foi observada apenas nos cães com catarata madura (r=0.772, P=0.02). Conclui-se que o aumento na concentração de IL-1 ß no humor aquoso de cães com catarata madura e hipermatura, associado à correlação negativa entre essa citocina e a PIO, sugerem que a mesma está envolvida na patogênese da uveíte induzida pela lente. Apesar dos cães com catarata hipermadura apresentarem concentrações significativamente maiores de MMP-2 no humor aquoso, essa citocina não se correlacionou com a IL-1 ß e a PIO. Em nosso estudo, correlação entre as citocinas dosadas no humor aquoso e a idade dos pacientes foi confirmada apenas entre MMP-2 e a idade dos cães com catarata madura.(AU)