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1.
New leucocyte common antigens expressed on all the normal chicken leucocytes have been characterized using two monoclonal antibodies designated as K-11 and K-55. These monoclonal antibodies stained virtually 100% of the leucocytes derived from various lymphoid organs including the spleen, thymus, bursa of Fabricius, caecal tonsil and peripheral blood, as well as a monocytic cell line (MC29), a B cell line (LSCC-RP9), and a T cell line (CU12). However, they did not stain mature erythrocytes, intestinal epithelial cells, or chicken embryonic fibroblasts. The two monoclonal antibodies showed different staining patterns and detected non-overlapping epitopes on MC29 cells in two color immunofluorescence analysis. Western blot analysis under non-reducing conditions showed that the monoclonal antibody K-11 recognized three splenic leucocyte proteins with molecular weights of 92, 42 and 41 kDa, whereas the monoclonal antibody K-55 recognized two proteins with molecular weights of 97 and 42 kDa. The data indicate that the monoclonal antibodies K-11 and K-55 recognize novel leucocyte-common antigens which have lower molecular weights than the previously reported leucocyte-common antigen family. 相似文献
2.
M A Qureshi L Miller H S Lillehoj M D Ficken 《Veterinary immunology and immunopathology》1990,26(3):237-250
A new chicken mononuclear cell line (MQ-NCSU) has been established. The starting material used to initiate this cell line was a transformed spleen from a female Dekalb XL chicken which had been experimentally challenged with the JM/102W strain of the Marek's disease virus. After homogenization, a single cell suspension of splenic cells was cultured using L.M. Hahn medium supplemented with 10 microM 2-mercaptoethanol. Under these culture conditions, a rapidly proliferating cell was observed and then expanded after performing limiting dilution cultures. These cells were moderately adherent and phagocytic for sheep red blood cells and Salmonella typhimurium. When tested against a panel of monoclonal antibodies (mAb) using the flow cytometry, MQ-NCSU cells stained readily with anti-chicken monocyte specific (K-1) mAb but did not stain with mAb detecting T-helper, T-cytotoxic/suppressor, and NK cells. MQ-NCSU cells expressed very high levels of Ia antigens and transferrin receptors. In addition, cell-free supernatant obtained from MQ-NCSU culture contained a factor which exhibited cytolytic activity against tumor cell targets. Based on their cultural, morphological, and functional characteristics and mAb reactivity profile, we conclude that MQ-NCSU cell line represents a malignantly-transformed cell which shares features characteristic of cells of the mononuclear phagocyte lineage. 相似文献
3.
H Salmon B Charley C Labonnardiere M Olivier K Kelley A Paraf 《Veterinary immunology and immunopathology》1989,23(1-2):113-128
After carbonyl iron treatment and gradient isolation, spleen and blood pig lymphocytes exhibited NK activity and produced IFN after viral induction. Removal of plastic-adherent cells, including the majority of B cells, did not change these activities. The plastic-non-adherent cells were further separated into two subsets of roughly similar size by panning using a monoclonal, anti-T, and anti-null cell antibodies (81 + cells). NK activity and IFN production were found in the 81 - cell fraction. A significantly higher proportion of null lymphocytes from blood and of splenic Fc-gamma receptor-bearing lymphocytes was also found among the 81 - cell fraction as compared to the 81 + fraction, without any change among other subsets. Similar proportions of helper (PT4+), cytotoxic (PT8+) and total T cells (MSA4+) were found among lymphocytes bound to target K562 cells and among the whole lymphocyte population. In contrast, lymphocytes that bound K562 cells demonstrated a striking increase in the proportion of Fc-gamma receptor-positive cells of high affinity. These results show that NK cells and IFN-producing cells are mainly included in the same blood and spleen fraction, and suggest that among 81 - cells only those expressing an Fc-gamma receptor of high affinity are active. 相似文献
4.
The morphologic and biological properties of porcine cells mediating natural killer (NK) activity were determined. In a previous study, we demonstrated that lymphocytes from the peripheral blood of pigs greater than 1 week of age possessed NK activity to K562 tumor cells and that lymphocytes from the blood and spleen of pigs greater than 1 day of age were able to mediate natural cytotoxicity against parainfluenza-3 (PI3) virus-infected Vero cells (Yang and Schultz, 1986a). Discontinuous density gradients were used to enrich NK cells. NK cytotoxicity was mainly present in high-density Percoll fractions (50 to 55% and 55 to 60%); little or no NK activity was present in lower density fractions. The NK cell enriched lymphocytes responded to the mitogens PHA, ConA and PWM. NK cells were sensitive to the suppressive effect of corticosteroid, but Protein A did not affect NK activity. The amount of cytotoxicity directly corresponded to the degree of binding that occurred between the NK enriched lymphocyte population and the target cells. Cytochemical and morphological studies demonstrated that these bond cells which are believed to be responsible for the NK activities, were mainly small to medium lymphocytes lacking azurophilic cytoplasmic granules. These findings were confirmed by ultrastructural studies of effector and "target-binding" cells. The results of the present study suggested that the cells mediating NK activity in pigs have the morphological and density characteristics of small and medium sized lymphocytes; findings that differ from those described for NK cells in human and other animal species. 相似文献
5.
Natural killer (NK) cell lysis of target cells by an Fc receptor‐mediated mechanism has not been conclusively demonstrated in cattle (Campos and Rossi, Vet. Immunol. Immunopathol. 8, 351–362, 1985), although it is well recognized in other species (Sulica et al., Nat. Immun. 14, 123–133, 1995). To resolve this problem, bovine peripheral blood mononuclear cells were completely depleted of adherent monocyte/macrophage type cells. The resulting enriched population of lymphocytes, was totally devoid of adherent monocytes, but contained up to 2 % NK‐like cells. On their own, this population had very low background levels of cytotoxicity for virus‐infected target cells in 51chromium release assays, but following the addition of virus‐specific antibodies, high levels of lysis were observed. This enhanced level of antibody‐dependent cytotoxicity demonstrated that bovine NK‐like cells can mediate killing of targets by an Fc receptor‐mediated mechanism as has been demonstrated for NK cells from other species. 相似文献
6.
Further phenotypic characterization of target cells for bovine leukemia virus experimental infection in sheep 总被引：3，自引：0，他引：3
Y Aida M Miyasaka K Okada M Onuma S Kogure M Suzuki P Minoprio D Levy Y Ikawa 《American journal of veterinary research》1989,50(11):1946-1951
To determine the phenotype of target cells for bovine leukemia virus (BLV) infection in sheep, we analyzed blood lymphocytes from BLV-infected clinically healthy and leukemic sheep by use of monoclonal antibodies. In clinically healthy and leukemic sheep that were BLV-infected, the blood concentration of T lymphocytes was within normal values, but the number of B lymphocytes was increased in several cases. In addition, the number of blood lymphocytes expressing the BLV antigen correlated well with that of B lymphocytes. Double immunofluorescence staining demonstrated that lymphocytes expressing BLV antigens bore B-cell but not T-cell surface markers. Moreover, neoplastic cells in the lymph nodes of leukemic sheep were stained immunohistochemically with an anti-B monoclonal antibody but not with any of anti-T monoclonal antibody tested, indicating that tumor cells are of B-lymphocyte origin. Collectively, these results show that BLV antigen-positive cells obtained from BLV-infected sheep that have no clinical signs and BLV-induced lymphosarcoma cells belong to the B-lymphocyte lineage. 相似文献
7.
8.
The present study examined the properties of NK activity in Yorkshire swine. The results support other porcine studies which indicate the swine NK system has both similarities and differences to this system in other species. Profiles of NK activity indicated swine NK cells are highly reactive against the YAC-1 lymphoma, the K-562 myeloid leukemia, the P-815 mastocytoma, and the TU-5 virally transformed fibroblast. In contrast, the MOLT-4 and SB leukemias are NK resistant. Kinetic studies indicated that in Yorkshire swine, NK lysis begins 6 h after mixing effectors and targets. The kinetics of the lytic reaction differ both from other breeds of swine and from other species, where cytotoxicity is readily measured in 4-h assays. The delayed lysis was not due to delayed target cell recognition, because Yorkshire swine NK cells are rapidly bound to tumor targets. The delayed lysis seems to be due to a refractoriness in the NK lytic mechanism. This delay may relate to the morphologic finding that the target-binding cell in Yorkshire swine appeared quite different from the large granular lymphocyte (LGL) reported as the NK effector in humans and rodents. Indeed, in light microscopic studies the typical tumor-binding cell in Yorkshire swine is a small, apparently nongranular, lymphocyte. Analysis of NK activity at the single cell level was performed with single effector-tumor conjugates immobilized in agarose. Generally, lysis by target binders paralleled sensitivity to lysis in 51Cr release tests, indicating lysis in agarose may be used as an NK index in swine. Like other species, swine NK cells were found to be nonadherent lymphocytes with a characteristic tissue distribution. Peripheral blood and spleen had the highest levels of NK activity. Lymph node cells displayed a small amount of NK activity which was limited to the YAC-1 target, while thymocytes showed no appreciable NK activity against any of the cell lines tested. 相似文献
9.
Allogeneic PM/86 melanoma cells of Munich Troll miniature swine have been used for the demonstration of porcine peripheral blood NK cell activity. Compared with the specific lysis of xenogeneic K562-, U937- and Vero-target cells, NK cell-mediated cytotoxicity (NK-CMC) against PM/86 melanoma tumor cells was significantly lower in a 16 h chromium release assay. The target cell susceptibility to peripheral blood NK-CMC of both adult Troll miniature swine and German Landrace sows was very similar. Cold target inhibition assays revealed the allogeneic PM/86 melanoma cells to be the most powerful inhibitors of NK-CMC. Nylon wool non-adherent lymphocytes produced interferon (IFN)-alpha in different quantities upon contact with NK susceptible target cells. The NK effector cells could be stimulated to a higher lytic activity against all susceptible targets by a moderate dose of natural human interleukin-2 (nhuIL-2). The role of NK-CMC in melanoma tumor rejection and/or prevention of metastases is yet unknown in swine although porcine melanoma serves as a good model for the disease in man. 相似文献
10.
Characterization of bovine haemopoietic progenitor cells using monoclonal antibodies and fluorocytometry 总被引：1，自引：0，他引：1
G Fritsch R T Nelson P Muiya J Naessens S J Black 《Veterinary immunology and immunopathology》1991,27(4):277-292
Monoclonal antibodies against bovine leucocyte cell surface differentiation antigens were used in combination with a fluorescence activated cell sorter to enrich bovine haemopoietic progenitor cells present in bone marrow cell populations prior to in vitro culture. After two sequential centrifugations of the bone marrow cell suspension through Ficoll-Paque, the interface fraction was stained with a cocktail of monoclonal antibodies directed against mature monocytes/macrophages, granulocytes and lymphocytes. Using appropriate electronic window settings on a FACStar Plus, cells with a high 90 degrees light scattering property (granular cells), a low forward light scattering property (erythrocytes and reticulocytes) and cells positive for monoclonal antibodies specific for lineage-restricted leucocyte markers were removed and the negative cell fraction collected. These negatively-selected cells were stained with monoclonal antibodies specific for a pan-leucocyte or a MHC class II marker and the positive cell population was collected in a second sort and subsequently submitted to culture. All erythroid and granulocyte/macrophage colony forming cells expressed MHC class II antigens, as well as the pan-leucocyte antigen. These same progenitors did not bind any of a variety of monoclonal antibodies directed against lineage-specific antigens on lymphocytes, granulocytes or monocytes/macrophages, although they did bind monoclonal antibodies recognizing MHC class I antigens. Between 85% and 91% of the isolated cells seeded were capable of forming erythroid or granulocyte/macrophage colonies within 5 to 10 days, thus increasing the plating efficiency of these cell types in bone marrow populations by at least 60 fold. 相似文献
11.
A 4-MDa component, recovered from uterine luminal secretions of gilts on d 15 of pregnancy, was assessed for suppression of the lytic responses from natural killer (NK) and lymphokine-activated killer (LAK) effector cells. Each cell type originated from preparations of peripheral blood lymphocytes (PBL), and the LAK cells were generated from the incubation of PBL with interleukin-2. The PBL and LAK cells were cultured for 5 d with and without the 4-MDa component. Following culture, the cells were incubated (22 h) with NK-sensitive K-562 target cells at varying effector:target cell ratios (25:1 to 200:1). Lytic activity was assessed with the chromium-51 release assay. Additional experiments were conducted in order to determine whether suppressor activity of the 4-MDa component was time-dependent and associated with transforming growth factor-beta2 (TGF-beta2). For effector:target cell ratios combined, the 4-MDa component suppressed the lytic activity of PBL but failed to affect the LAK cells. Suppression of NK-mediated lysis occurred by d 3 of the 5-d culture period. In addition, suppressor activity of the 4-MDa component was reversed by a neutralization antibody to TGF-beta2. In conclusion, the 4-MDa component with TGF-beta2 activity suppressed the lytic responses of porcine NK cells. 相似文献
12.
Guimarães MC Guillermo LV Matta MF Soares SG DaMatta RA 《Veterinary immunology and immunopathology》2011,140(3-4):317-322
Macrophages are fundamental cells of the innate immune system, which, through phagocytosis and nitric oxide production, eliminate pathogens. The aim of the present study was to determine if macrophages from chicken families divergently selected to high and low antibodies response differ in nitric oxide production and phagocytic capacity. Blood monocytes derived macrophages were activated with lipopolysaccharide and supernatant from chicken spleen lymphocytes cultured with Concanavalin A (containing chicken interferon). Nitric oxide production was evaluated in culture supernatants. Phagocytic capacity of activated and non-activated macrophages was assayed using yeasts and IgY opsonized sheep red blood cells. Activated and non-activated macrophages from the high antibodies response family produced higher nitric oxide levels, internalized more yeast and significantly more opsonized sheep red blood cells than macrophages from the low antibodies response family. Moreover, activated macrophages became more elongated and widely spread. These findings indicate that macrophages from the high antibodies response family were more active suggesting that the differences in antibody response also depend on macrophage function. 相似文献
13.
P Quere 《Veterinary immunology and immunopathology》1992,32(1-2):149-164
Marek's disease virus (MDV)-transformed lymphoblastoid cells (MDCC-MSB1, -PA9 and -RP1) added to chicken splenic lymphocytes after treatment with mitomycin, suppress the lymphoproliferative response to T-cell mitogens (concanavalin A or phytohemagglutinin) by 40-70%. This suppressive activity was observed in syngeneic as well as in allogeneic combinations of cell lines and responder lymphocytes. The suppressive effect disappeared when the addition of MD-transformed cell lines to the responder cultures was delayed for 24 h. Treatment with glutaraldehyde, instead of mitomycin, greatly weakened the suppressive activity of the MD lymphoblastoid cells. A reduction of interleukin 2 (IL-2)-like activity produced by responder lymphocytes was observed after mixing with mitomycin-treated lymphoblastoid cells, but also, although slightly less, with the same glutaraldehyde-treated cells. Nevertheless no membrane fluorescence was observed, using INN-CH16 monoclonal antibody on MDV-induced lymphoblastoid cell lines to check up on the presence of IL-2 receptor-like structure. All the three lines exhibited a CD4+, CD8- phenotype. 相似文献
14.
P G Greenlee S E Calvano F W Quimby A I Hurvitz 《Veterinary immunology and immunopathology》1987,15(4):285-296
Using automated flow cytometry, 23 commercially available antibodies (all but one of them monoclonal) raised against surface antigens of specific populations of human, rat, and mouse lymphocytes were tested for cross-reactivity to peripheral blood lymphocytes from five clinically healthy adult dogs. Of all the antibodies tested, only the polyclonal anti-asialo GM1 directed against mouse NK cells, and the monoclonal antibodies anti-HLA-DR directed against the human class II antigen and anti-B1, a human pan B cell marker, consistently labeled subpopulations of canine lymphocytes. 相似文献
15.
The monoclonal antibody (MAb), C5B6, recognizes the CD11c/CD18 molecule on the surface of bovine peripheral blood monocytes. C5B6 was reactive with 69-83% monocytes, all granulocytes, and less than 5% of lymphocytes from cattle. Of the lymphocyte series, the antibody had specificity for large lymphocytes and two lines expressing T cell markers, but was not reactive with small lymphocytes, thymocytes, a tumor cell line of B-cell lineage, an interleukin 2 (IL2)-dependent T cell line, fibroblasts, or human, sheep, goat or pig peripheral blood mononuclear cells. No dual fluorescence was seen using C5B6 and antibodies to bovine IgM, CD2, CD4 or CD8. Immunoprecipitation of 125I labeled peripheral blood mononuclear cells with C5B6 antibody defined two bands: 150,000 and 95,000 Da. Antibody to the beta chain (CD18) of the leukocyte adhesion receptor family precipitates the 95 kDa beta subunit and the three associated alpha subunits (180, 165 and 150). The bands obtained using MAb C5B6 correlated with the p150/95 bands observed using an antibody that precipitated the alpha and beta chains of the leukocyte adhesion receptor family. Functionally, the primary but not the secondary proliferative response to alloantigens was inhibited by C5B6 MAb. No effect was seen using C5B6 MAb in cytotoxicity assays or in the secondary proliferative response to Brucella abortus or bovine herpes virus type 1. 相似文献
16.
Weiss DJ 《American journal of veterinary research》2001,62(8):1229-1233
OBJECTIVE: To evaluate monoclonal antibodies that may be useful for immunophenotyping myeloid cells in bone marrow of dogs. SAMPLE POPULATION: Bone marrow specimens obtained from 5 dogs. DESIGN: Specimens were labeled with monoclonal antibodies that detected CD18, major histocompatability antigen class-II (MHC class-II), CD14, and Thy-1. Cells labeled with each of the antibodies were isolated by use of a fluorescence-activated cell sorter. Differential cell counts of sorted cells were used to determine cells that were labeled by each of the various antibodies. RESULTS: Myeloid cells labeled with anti-CD18 antibody included granulocytes, lymphocytes, and monocytes-macrophages. Immature and mature granulocytes were labeled. Lymphocytes, monocytes-macrophages, and eosinophils were labeled with anti-Thy-1 antibody. Cells labeled with anti-MHC-class II antibody included approximately 9% of bone marrow cells, which consisted almost exclusively of lymphocytes and monocytes-macrophages. Approximately 4% of bone marrow cells were labeled with anti-CD14 antibody, with > 90% of sorted cells being monocytes-macrophages. CONCLUSIONS AND CLINICAL RELEVANCE: Four monoclonal antibodies for use in detecting subpopulations of canine bone marrow cells were evaluated. These antibodies should be useful in differentiating the origin of leukemic cells in dogs. 相似文献
17.
Sheep antibodies against a pig E-rosette-forming lymphoblastic T lymphoma raised by two intravenous injections of 10(10) cells showed little lymphocytotoxic activity which could be absorbed with red cells, alveolar macrophages or kidney or liver cell homogenates. Bone marrow absorption yielded subpopulation specific antibody which binds to E rosette-forming cells (E.RFC) using either complement-mediated cytotoxicity or indirect antiglobulin rosette formation. In 30 blood lymphocyte preparations from 20 pigs with a range of approximately 20-85% E rosettes the mean E% 43.5 +/- 2.7 agreed with the % antigen+ cells by cytotoxicity mean = 42.6 +/- 2.7 and in each individual sample these figures also agreed closely. In samples of blood lymphocytes enriched and depleted for E rosettes, results of %E+ also agreed closely with % antigen+ cells. This relationship also held for thymocytes and the specific antibodies could be completely absorbed with thymocytes. These data show that the antibody identified peripheral and thymic E.RFC. Bound to lymphocytes the antibody inhibited E rosette-formation with sheep red blood cells (SRBC) in saline (S) and dextran (DS) and with pig RBC in dextran and in Ficoll, but did not affect B cells shown by immunofluorescence, direct antiglobulin rosette formation or Fc rosette-formation, either in saline or dextran, (which include T gamma cells). E rosette inhibition was dependent on antibody concentration, showing single and double sigmoid curves for S and DS rosettes respectively, consistent with differing ease of inhibition of the strong and weak rosette formation. The same spectrum of inhibition of rosette formation by antibody binding followed subsequent incubation with C'6-deficient rabbit serum, but with C'-sufficient serum resulted in loss of cells which require dextran for Fc rosette-formation (T gamma). Thus the serum reveals E rosette-forming T cells and their subpopulations, perhaps by binding to the SRBC receptor. 相似文献
18.
Min W Lillehoj HS Li G Sohn EJ Miyamoto T 《Veterinary immunology and immunopathology》2002,88(1-2):49-56
The chicken IL-15 gene was recently cloned and shown to encode a polypeptide with T cell growth factor activity similar to IL-2. To further characterize the chemical and biological properties of chicken IL-15, we generated a panel of monoclonal antibodies against bacterially expressed protein and characterized their binding specificities. All antibodies were reactive by ELISA with recombinant IL-15, but not IL-2, and identified a 15kDa recombinant chicken IL-15 by Western blot analysis. Two antibodies inhibited IL-15-induced proliferation of splenic lymphoblast cells. These monoclonal antibodies will be useful for further structural and immunological studies of chicken IL-15. 相似文献
19.
Identification of monoclonal antibodies for immunohistochemical staining of feline B lymphocytes in frozen and formalin-fixed paraffin-embedded tissues. 
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下载免费PDF全文 C E Monteith B J Chelack W C Davis D M Haines 《Canadian journal of veterinary research》1996,60(3):193-198
Commercially-available monoclonal antibodies to B lymphocytes were evaluated for immunohistochemical staining of feline B lymphocytes in frozen and formalin-fixed, paraffin-embedded tissues using an avidin biotin complex immunoperoxidase immunohistochemical technique. Three monoclonal antibodies: F46A and F72A raised to "carnivore" B lymphocytes and RA3.6B2 raised to murine B lymphocytes, stained B lymphocyte-dependent areas of frozen feline lymphoid tissue. In addition, antibody RA3.6B2 stained B lymphocyte dependent areas in formalin-fixed, paraffin-embedded feline tissues. There was no staining of T lymphocyte-dependent areas in either frozen or formalin-fixed tissues. Dual parameter flow cytometry, using an anti-pan-T lymphocyte antibody, revealed that greater than 99% of the cells stained by RA3.6B2 are a population distinct from T lymphocytes. F46A was shown to stain a sub-population of those cells stained with RA3.6B2. These antibodies may be useful in the identification of feline B lymphocytes using immunohistochemistry and flow cytometry and thereby provide additional tools to study B lymphocyte ontogeny and the significance of lymphocyte phenotype in lymphoid neoplasia in cats. 相似文献
20.
Natural cell-mediated cytotoxicity to cells infected with infectious bovine rhinotracheitis virus 总被引：1，自引：1，他引：0
Cell-mediated cytotoxicity against viral-infected cells was demonstrated in a 6-hr 51Cr release assay. Peripheral blood mononuclear leukocytes from both infectious bovine rhinotracheitis virus (IBRV)-infected and noninfected cattle exhibited preferential lysis against IBRV-infected primary bovine embryonic kidney (BEK) cells compared to cells infected with pseudorabies virus and noninfected BEK cells. Addition of specific antibody to the assay did not enhance cytotoxicity. The effector cell was a nonadherent cell which was either spontaneously enriched or generated during in vitro cultivation. Maximal cytotoxic activity was detected in peripheral blood mononuclear cells cultured for 3 to 5 days. Several factors affected the magnitude of cytotoxicity during the assay: target cell type, concentration of viral inoculum, duration of effector and target cell contact. It is suggested that target cell lysis was a form of natural cell-mediated cytotoxicity mediated by a cell which has different characteristics from the typical human and murine NK cell. 相似文献