首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献，搜索用时 140 毫秒
1.
Background— Morphology and cytochemistry are the foundation for classification of leukemias in dogs and cats. Advances in automated hematology instrumentation, immunophenotyping, cytogenetics, and molecular biology are significantly improving our ability to recognize and classify spontaneous myeloproliferative and lymphoproliferative disorders. Objective— The purpose of this study was to assess the utility of flow cytometry‐based light scatter patterns provided by the Cell‐Dyn 3500 (CD3500) automated hematology analyzer to predict the lineage of leukemic cells in peripheral blood of dogs and cats. Methods— Leukemic cells from 15 dogs and 6 cats were provisionally classified using an algorithm based on the CD3500 CBC output data and were subsequently phenotyped by enzyme cytochemistry, immunocytochemistry, indirect flow cytometry, and analysis of antigen receptor gene rearrangement. Results— The algorithm led to correct predictions regarding the ontogeny of the leukemic cells (erythroid/megakaryocytic potential, myeloid leukemia, monocytic leukemia, chronic granulocytic leukemia, lymphoid leukemia) in 19/21 animals. Mismatches in the WBC impedance count and the WBC optical count in conjunction with microscopic assessment of blasts in the blood were useful for predicting myeloproliferative disorders with erythroid or megakaryocytic potential. The leukocyte light scatter patterns enabled distinction among myeloid leukemias (represented by acute myelomonocytic leukemia, acute monocytic leukemia, chronic granulocytic leukemia) and lymphocytic leukemias (including acute and chronic lymphocytic leukemias). One case of acute lymphocytic leukemia was misidentified as chronic lymphocytic leukemia. Conclusions— Algorithmic analyses can be applied to data generated by the CD3500 to predict the ontogeny of leukemic cells in the peripheral blood of dogs and cats. This rapid and quantitative technique may be used to improve diagnostic decisions, expand therapeutic choices, and increase prognostic accuracy.  相似文献   

2.
Cytochemical staining for leukocyte alkaline phosphatase(LAP), nonspecific esterase (NSE), nonspecific esterase with fluoride inhibition (NSE-F), periodic acid Schiff (PAS) reactivity, and peroxidase (PO) was valuable in identification of the neoplastic cell type in 10 leukemic cats. Staining both blood and bone marrow smears was often necessary for making the correct diagnosis. Cytochemical staining resulted in changing the morphologic diagnosis of leukemia in two of the 10 cats. Also, increased LAP activity, probably a marker for myelocytic leukemia in the cat, was observed in bone marrow cells from three nonleukemic, FeLV-positive cats.  相似文献   

3.
Induction of arginase activity in mouse myeloid leukemic M1 cells by treatment with recombinant human tumor necrosis factor (rH-TNF) or TNF-elicited mouse serum (TNS) were examined in vitro. M1 cells differentiated into macrophage-like cells by addition of rH-TNF or TNS. The differentiated cells expressed phagocytic function and did not grow anymore. Cytolytic effect of rH-TNF or TNS was not observed. The differentiation of M1 cells into phagocytic cells by the TNS treatment was more rapidly than that by the rH-TNF treatment though TNS contained 25-time less amounts of TNF indicating species specificity of TNF action on myeloid leukemic cells or TNS containing other differentiation factor(s). 3H-ornithine formation from 3H-arginine is catalyzed by arginase (EC. 3.5.3.1). The enzyme product increased in the M1 cell culture medium by the treatment with rH-TNF or TNS. The arginase activity statistically correlated with the appearance percentage of differentiated cells with phagocytic function. These results suggest that in vitro differentiation of M1 cells is accompanied by induction of arginase activity.  相似文献   

4.
Flow cytometric evaluation of hemophagocytic disorders in canine   总被引：1，自引：0，他引：1  
Background — Hemophagocytic macrophages in canine bone marrow are observed in malignant histiocytosis as well as benign hemophagocytic histiocytosis. Cytomorphologic evaluation alone may be inadequate to consistently differentiate between benign and malignant forms of hemophagocytic disorders. Objective — The purpose of this study was to evaluate the ability of flow cytometry and immunophenotyping to differentiate between benign and malignant types of hemophagocytic disorders in dogs. Methods — Blood smears and bone marrow differential cell counts were evaluated for 10 dogs with hemophagocytic disorders. Bone marrow samples were labeled with monoclonal antibodies to CD18, MCH class‐II, Thy‐1, CD14, CD3, and CD21. Using flow cytometry, forward‐angle versus side‐angle light scatter plots were analyzed and immunophenotypes were determined. Results — Scatter plots from 3 dogs with a necropsy diagnosis of malignant histiocytosis revealed 2 atypical cell clusters. One cluster contained cells of similar size or larger than immature myeloid cells and metamyelocytes. Cells in the other cluster were highly granular, with granularity similar to or greater than that of metamyelocytes. In bone marrow from dogs with malignant histiocytosis that was labeled with anti‐CD14 antibody, macrophages represented 29–48% of nucleated cells. Seven dogs had a clinical or histopathologic diagnosis of benign hemophagocytic syndrome. Three of the dogs had normal cell distribution in scatter plots. Two dogs had 2 abnormal cell clusters: 1 within the immature myeloid and metamyelocyte gates and the other with granularity similar to or greater than that of metamyelocytes. The remaining 2 dogs had an atypical cell population, mostly within the immature myeloid gate. For dogs with benign hemophagocytic syndromes, 6–17% of cells in the bone marrow were CD14 positive. Conclusions — The cellular distribution in scatter plots and the total number of macrophages in bone marrow may be useful in differentiating malignant histiocytosis from benign hemophagocytic syndromes in dogs.  相似文献   

5.
Leukemic cells from 17 dogs with spontaneous leukemia were stained with leukocyte alkaline phosphatase, alpha naphthyl acetate esterase with and without fluoride, peroxidase, and periodic acid-Schiff. Cytochemistry was necessary for identification or confirmation of leukemic cell type in most dogs and resulted in changing the light microscopic morphologic diagnosis in eight of 17 dogs. Leukemic cell types diagnosed were myelomonocytic leukemia in seven dogs, monocytic leukemia in five dogs, lymphocytic leukemia in four dogs, and myelocytic leukemia in one dog.  相似文献   

6.
B-lymphocytes from 7 normal cattle and from 3 persistent lymphocytotic, 6 lymphosarcomatosus, and 3 bovine leukemia virus-infected animals with diverse defects in humoral immunity were examined for their capacity to undergo terminal differentiation in vitro. Pokeweek mitogen-stimulated cells from normal and leukemic cattle were stimulated to synthesize and secrete immunoglobulins, and these were detected by radial immunodiffusion. Lymphocytes from normal cattle synthesized more IgM than those from persistent lymphocytotic-, lymphosarcomatous-, and bovine leukemia virus-infected cattle. Leukemic cattle have a higher percentage of cells containing intracytoplasmic immunoglobulins; most cells are believed to contain IgG. The serum IgM in normal cattle is significantly (P less than 0.05) higher than that in leukemic cattle. The results indicate that there is an abnormality either in synthesis or secretion of IgM or in abnormal catabolism of Ig from leukemic B-lymphocytes.  相似文献   

7.
诱导鸡胚精原干细胞向成骨细胞分化的研究   总被引：1，自引：0，他引：1  
为了探讨体外培养的鸡胚精原干细胞(SSCs)的多向分化潜能,取孵化18~20 d的鸡胚睾丸,无菌获取SSCs,体外培养传代,通过地塞米松、β-甘油磷酸钠、维生素C诱导鸡胚SSCs向成骨细胞分化,钙结节Von Kossa s法、改良的钙钴法及免疫组化法进行成骨细胞鉴定。结果显示SSCs被诱导15~21 d后分化为成骨细胞,诱导形成率为75%~80%;诱导后的细胞Von Kossa s染色可见细胞间布满黑色颗粒,提示有矿化基质沉积;改良钙钴法碱性磷酸酶染色胞浆呈深棕色或深黑色,免疫组化法染色细胞呈阳性。因此可以得出在不同的诱导条件下,SSCs体外可被定向诱导分化为成骨细胞,证明其具有多向分化潜能。  相似文献   

8.
Monocytic leukemia in a horse   总被引：2，自引：0，他引：2  
On clinical examination, a six-year-old Hassian gray gelding with a history of impaired performance, slight cough, colic, and edema of the ventral abdomen, prepuce and the legs had reduced skin turgor, pale mucous membranes, forced costoabdominal breathing, reduced venous return, enlarged lymph nodes, and splenomegaly. Hematologic findings revealed anemia, leukocytosis and a high percentage of monocytoid leukemic cells. Generalized lymphadenopathy, splenomegaly, ascites, hydrothorax, and a diffusely thickened gut wall were found at necropsy. Massive infiltration with monocytoid leukemic cells was detected in lymph nodes, spleen, bone marrow, liver, gut wall, kidneys, and choroid plexus. Incubation of living cells obtained from a leukocyte concentrate with latex particles revealed phagocytosis in the leukemic cells on light and electron microscopy. The leukemic cells also had a marked alpha-naphthyl-acetate and naphthol-AS-acetate esterase activity, but were only weakly positive to naphthol-AS-D-chloroacetate esterase. A very weak alkaline phosphatase activity only was demonstrated in a few leukemic cells. On scanning electron microscopy, the leukemic cells had prominent ruffles and ridge-like profiles. These features of the leukemic cells excluded lymphocytic and granulocytic leukemia, and monocytic leukemia was diagnosed.  相似文献   

9.
Macrophages play a pivotal role in the pathophysiology of many diseases by mediating the host immune response to infections and intoxications. The species-specific activation of macrophages and the differential response in cytokine production impedes the extrapolation of results between species. Therefore, the aim of this study was to isolate and immortalise macrophages from equine bone marrow (BM) cells in order to study equine-specific signalling pathways. The isolated BM-derived macrophages (referred to as e-CAS cells) showed proliferation kinetics similar to that of standardised cell lines and were maintained in culture for >76 passages. To characterise the cells, a number of typical parameters of macrophages were tested. Morphological evaluation (May-Grünwald Giemsa staining) and non-specific esterase activity indicated the e-CAS cells to be macrophages. The presence of CD14 and their ability to phagocytose Escherichia coli bioparticles further confirmed their identity, as did their ability to produce cytokines, reactive oxygen and nitrogen intermediates in response to LPS. These data show that the established cell line (e-CAS) shows the characteristics of equine macrophages and may, therefore, prove to be a unique in vitro model for studying the cellular biology of equine inflammation.  相似文献   

10.
Leukoproliferative disorders reported in horses include lymphoma, lymphocytic leukemia, plasma cell myeloma, granulocytic leukemia, monocytic leukemia, myelomonocytic leukemia, and eosinophilic leukemia. Lymphoma affects horses of all ages, whereas leukemias often occur in younger horses. Clinical signs are often nonspecific including depression, anorexia, fever, and weight loss. Specialized diagnostic techniques such as cytochemistry and immunophenotyping better define the cellular origin of leukoproliferative disorders, which is essential for developing appropriate therapeutic protocols and rendering an accurate prognosis.  相似文献   

11.
A transplantable mononuclear cell leukemia (MCL) was established from spontaneous MCL in an F344 rat. In this work, we paid special attention to a nodular tumor, named MCL-YSK, developed at the subcutaneous transplant site. MCL-YSK was serially passaged in subcutaneous tissue of syngeneic rats up to the 19th generation. Transplants from MCL-YSK grew into nodules 3 cm in diameter and 11.3 g in weight 9 weeks after subcutaneous implantation. Neoplastic cells forming the nodules had azurophilic cytoplasmic granules, which ultrastructurally appeared to be lysosomes. The cells reacted positively for acid phosphatase and nonspecific esterase, but not for alkaline phosphatase, alpha-1 antitrypsin and lysozyme, nor reacted with anti-rat monocyte/macrophage monoclonal antibody and anti-rat CD-8 monoclonal antibody. They possessed Fc-receptor. Leukemic cells first appeared in the peripheral blood 6 weeks after transplantation when subcutaneous nodules reached an average diameter of one cm. Subsequently, leukemic changes progressed in recipients as MCL-YSK grew larger. The recipients died during the period from 8 to 12 weeks after transplantation, showing anemia, depression, splenohepatomegaly and lymph node enlargement. Diffuse or focal proliferation of sprinkled tumor cells was present in many organs. Hematologic changes suggestive of hemolytic anemia, elevated plasma enzymes and decreased drug-metabolizing enzymes, indicative of hepatic malfunction, were seen in transplant recipients. MCL-YSK was easily transplanted into athymic nude mice. The transplanted mice showed leukemic changes similar to those observed in rats with transplanted MCL-YSK.  相似文献   

12.
The influence of interferon (IFN)-alpha on the in vitro differentiation of myeloid porcine dendritic cells (DC) was evaluated as the ability of the DC to stimulate to cell proliferation in a mixed leukocyte reaction (MLR), and as their ability to produce cytokines at exposure to bacterial and viral preparations. Porcine monocytes were enriched from purified peripheral blood mononuclear cells (PBMC) by plastic adherence and cultured in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 or in GM-CSF, IL-4 and IFN-alpha. After 5 days of culture, the cells developed a dendritic morphology and the proportion of cells expressing MHC class II and B7 molecules was increased as determined by flow cytometry. Dendritic cells, differentiated for 5 days in GM-CSF, IL-4 and IFN-alpha, were able to stimulate both allogeneic and syngeneic PBMC to proliferation in an MLR. The DC produced the Th1 associated cytokines IFN-alpha at Sendai virus stimulation, and IL-12 at stimulation with plasmid DNA (pre-incubated in the presence of lipofectin), heat-inactivated Actinobacillus pleuropneumoniae, UV-inactivated Aujeszky's disease virus and live Sendai virus. The heat-inactivated bacteria and Sendai virus also induced production of the Th2 associated cytokines IL-10 and IL-6. The addition of IFN-alpha during differentiation of DC in GM-CSF and IL-4 enhanced their ability to stimulate allogeneic and syngeneic MLR, but did not alter their ability to produce cytokines.  相似文献   

13.
The aims of the present study were to establish a culture system for goat skeletal muscle stem cells and to examine their myogenic and adipogenic properties in vitro. Cells were isolated from the skeletal muscle of the Shiba goat and cultured in vitro. Most of the cells were positive for myogenic markers, such as Pax7, MyoD, and desmin, and immunocytochemistry revealed they differentiated to form myotubes expressing myosin heavy chain, indicating they were highly myogenic. Myogenic differentiation was strongly suppressed by the addition of basic fibroblast growth factor, while proliferation was unaffected. When the cells were cultured in adipogenic differentiation medium, some of the cells differentiated into mature adipocytes that stained with Oil Red-O. These cells were immunocytochemically positive for adipogenic markers, including peroxisome proliferator-activated receptor-gamma (PPAR gamma) and CCAAT/enhancer-binding protein-alpha (C/EBP alpha). These results clearly demonstrate the presence of both myogenic and adipogenic stem cells in goat skeletal muscle.  相似文献   

14.
Cell-mediated immune protection in chickens against Pasteurella multocida   总被引：2，自引：0，他引：2  
Immune protection by cellular immunity in chickens against Pasteurella multocida was investigated by in vivo and in vitro experiments using spleen cells and culture supernatants of immunised chickens. Intraperitoneal or intravenous transfer of immune splenic cells into normal chickens induced transmission of an as effective protection as that exhibited in immunised chickens. Immune protection was also obtained by intravenous treatment of chickens with culture supernatant fluid from immune splenic cells of hormonally bursectomised chickens. The in vitro experiment showed that intracellular bacterial proliferation was inhibited in peritoneal macrophages from immunised chickens, or from normal chickens sensitised with culture supernatant fluid of immune splenic cells, and the macrophages were protected from disruption by infection. Peritoneal macrophages sensitised with culture supernatant fluid from unimmunised splenic cells, or peritoneal macrophages from unimmunised chickens, allowed considerable intracellular proliferation of bacteria with almost complete breakdown of the macrophages within 24 hours after bacterial challenge. These data suggest that the protective immunity of chickens against P multocida was dependent on cell-mediated immunity by mediators such as the macrophage activating factor from T lymphocytes.  相似文献   

15.
BACKGROUND: Flow cytometry may be used to determine immunophenotype or lineage of leukemic cells, but few antibodies are available that are specific for cells of monocytic and granulocytic lineage. OBJECTIVE: The purpose of this study was to evaluate the flow cytometric staining patterns of 3 commercial monoclonal antibodies for monocytes and granulocytes in clinically healthy dogs and in dogs with acute myeloid leukemia (AML). METHODS: Mouse antihuman macrophage antibody (MAC387), mouse anti-human myeloperoxidase (MPO), and a canine neutrophil-specific antibody (NSA) were evaluated using flow cytometry on blood from 6 clinically healthy control dogs, and on blood (n = 7) and/or bone marrow (n = 2) from 8 dogs with AML. A diagnosis of acute leukemia was confirmed by >30% blasts in bone marrow or >30% blasts in peripheral blood, together with bi- or pancytopenia, circulating CD34-positive blast cells, and clinical signs of disease. Leukemic samples also were evaluated using a wide panel of monoclonal antibodies. RESULTS: MAC387 stained neutrophils and monocytes from control dogs, although the staining profiles for the 2 cell types differed. MPO and NSA resulted in strong positive staining of neutrophils; MPO also stained monocytes weakly. Lymphocytes did not stain with any of the antibodies. One case was classified as AML of granulocytic lineage (AML-M1), 6 cases were classified as acute monocytic leukemia (AML-M5), and 1 case was classified as acute myelomonocytic leukemia (AML-M4). Neoplastic myeloblasts in the dog with granulocytic AML were positive for MPO, NSA, MAC387, and CD4. All monoblasts from the dogs with AML-M5 were positive for CD14, 5 of 6 were positive for MAC387, and 2 were positive for MPO. NSA staining was negative in the 2 dogs with AML-M5 in which it was evaluated. In the dog with AML-M4 variable percentages of blast cells were positive for CD14, MPO, MAC387, CD4, and NSA. CONCLUSIONS: Antigens identified by antibodies to MAC387, MPO, and NSA were expressed not just by normal mature neutrophils and monocytes, but also by neoplastic myeloblasts and monoblasts. These 3 antibodies may be useful as part of a wider panel for immunophenotyping AML in dogs.  相似文献   

16.
This is one of the first characterizations of channel catfish (Ictalurus punctatus) leukocytes by enzyme cytochemistry. Leukocytes demonstrated cytoplasmic staining patterns very similar to mammalian leukocytes when stained with acid phosphatase, alpha-naphthyl butyrate esterase, beta-glucuronidase, alpha-naphthyl acetate esterase, Sudan Black B and anti-immunoglobulin specific immunohistochemistry. Lymphocytes, monocytes, macrophages, neutrophils, and surface immunoglobulin positive (surface Ig+) cells were present in channel catfish renal hematopoietic tissue and spleen and demonstrated distinctive cytoplasmic foci staining patterns, cytoplasmic blushing or cell membrane staining. Monocytes, macrophages, lymphocytes and surface Ig+ cells were present in the thymus. Thymic and splenic cellular organization appeared very similar to these same mammalian tissues. In the thymus, acid phosphatase positive cells were distributed throughout the parenchyma, while alpha-naphthyl butyrate esterase and beta-glucuronidase positive cells were concentrated in the cortex and the medulla, respectively. Surface immunoglobulin positive cells occurred in the cortex. In the spleen, acid phosphatase positive cells were scattered throughout the parenchyma, while alpha-naphthyl butyrate esterase positive cells were scattered throughout the parenchyma and adjacent to splenic arterioles. Beta-glucuronidase and surface immunoglobulin positive cells were restricted to immediately adjacent to splenic arterioles. Sudan Black B positive cells were scattered throughout the parenchyma, while alpha-naphthyl acetate esterase positive cells occurred adjacent to peri-arteriole lymphoid sheaths and appear very similar to mammalian metallophils.  相似文献   

17.
The combination of flow cytometric scatterplot analysis and specific monoclonal antibodies was used to evaluate the lineage of cells from six dogs with proliferative disorders of bone marrow. Scatterplot analysis was used to identify mature and immature myeloid and erythroid cells. The immunophenotype of cells in the immature myeloid gate was determined by labeling cells with four monoclonal antibodies. These results were compared to results of cytologic and cytochemical evaluation. The immunophenotype of a dog with a diagnosis of myelogenous leukemia was a cluster of differentiation-18 (CD-18) positive, CD-14 negative, Thy-1 negative, and a major histocompatibility complex (MHC) class II negative. The immunophenotype of a dog with a diagnosis of myelomonocytic leukemia was CD-18 positive, CD-14 positive, Thy-1 positive, and MHC class II positive. Although this phenotype clearly differentiated myelomonocytic leukemia from myelogenous leukemia, it was similar to the immunophenotype of dogs with a diagnosis of malignant histiocytosis or hemophagocytic syndrome. The immunophenotype of two dogs with myelodysplastic syndrome was CD-18 positive and CD-14 negative. Results for Thy-1 and MHC class II were variable. As additional lineage-specific monoclonal antibodies become available, immunophenotyping should become a valuable tool for determination of the lineage of cells in canine myeloproliferative disorders.  相似文献   

18.
Cytochemical characterization of feline leukemic cells   总被引：1，自引：0，他引：1  
Hematopoietic cells in blood and/or bone marrow from 23 leukemic cats and ten control cats were characterized using a battery of cytochemical enzyme stains. The results of cytochemical staining led to modification of diagnosis based on clinical, hematologic and histopathologic findings in four (17%) of the leukemias. Sudan black-B and chloroacetate esterase served as granulocytic markers in both the control and leukemic groups. Peroxidase activity was seen in the granulocytes and monocytes of control animals but not in the blasts of leukemic cats. Diffuse alpha naphthyl butyrate esterase staining marked monocytes in both control and leukemic animals. Cytochemical staining was found to be a valuable aid in the classification of leukemias in the cat.  相似文献   

19.
胚胎干细胞是存在于早期胚胎或原始生殖嵴,具有发育全能性和无限增殖能力的一类细胞。作为实验材料,该类细胞已广泛应用于基因的表达与调控、细胞分化的机制、动物克隆及转基因动物生产等研究领域。与其他哺乳动物胚胎干细胞的研究相比,虽然鱼类胚胎干细胞的研究起步较晚,但近年来该领域的研究进展迅速。本文综述了鱼类胚胎干细胞的分离培养、生物学特性、体外分化能力及嵌合体形成等方面的研究进展。  相似文献   

20.
Histological and to some extent additional enzyme-histochemical studies were conducted into 28 cases of Marek's disease with minor, moderate, and intensive lymphoma formation. The investigations were undertaken with a view to differentiating T-lymphocytes and macrophages by differentiated activity on acid unspecific alpha naphthylacetate esterase. Lymphoma specimens were cytologically evaluated under semi-quantitative or quantitative aspects. Evidence was provided to the existence of correlations between intensity of Marek's disease lymphomas and their relative contribution to lymphocytes, lymphoblasts, and macrophages. The amount of lymphoblasts was found to go up along with advancing lymphoma proliferation, while the amounts of lymphocytes and macrophages went down. Moderately pronounced lymphomas contained 16 percent macrophages on average, whereas only about 8 percent were recordable from highly pronounced lymphomas. The high presence of macrophages in less developed lymphomas is considered to reflect unspecific defence reaction against neoplastic lymphatic cells in the phase of low tumour malignancy.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号