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Biomedical subjects

M R Brandon

Publications and source records attributed to M R Brandon.

At least 91 records · Page 5Linked to original sources
PubMed1989

Subpopulations of lymphocytes in the mammary gland of sheep.

The subpopulations of lymphocytes in the pregnant and non-pregnant mammary glands of the sheep were delineated by a panel of monoclonal antibodies. The most striking feature observed was that in the mammary gland of both pregnant and non-pregnant sheep the great majority of the lymphocytes in the ductal and alveolar epithelium were agranulated CD8+ CD5- cells. A small subpopulation of granulated lymphocytes in the epithelium expressed the CD45R antigen but not the major histocompatibility complex (MHC) class II molecules. Other subpopulations, especially B lymphocytes, were present in much lower concentrations and were located mainly in the periductal and intralobular connective tissues. Patches of lymphocytes clustering around venules were observed and the majority of them were shown to be CD5+ CD4+, while some were CD5+ CD8+ but none were CD45R+ (B cell). It is suggested that selective traffic of T cells occurs at these sites.

Animals↗
PubMed1989

Lymphocyte subpopulations in the blood of sheep persistently infected with border disease virus.

The surface phenotypes of peripheral blood lymphocytes in groups of lambs and adult sheep persistently infected with Border disease virus (P-I BD) were compared with those of healthy controls. The proportion and number of lymphocytes bearing surface immunoglobulin (sIg+) and expressing class II MHC antigen (B cells) were significantly increased. A significant increase in CD1+ lymphocytes was also evident. Conversely, the proportion of T lymphocytes in P-I BD lambs was reduced. A marked reduction in the proportion of circulating lymphocytes expressing class I MHC antigen was also observed. These findings were not affected by differences in the strain of the virus responsible for the persistent infection.

Aging↗
PubMed1989

Site-directed differences in the immune response to the fetus.

Major differences in the maternal immune response to the fetus were observed in the placentomes and in the interplacentomal regions of the pregnant sheep uterus. Firstly, fewer lymphocytes were detected in the placentomes compared to the interplacentomal regions and to non-pregnant uterine tissue (Lee, Gogolin-Ewens & Brandon, 1988). Secondly, a large population of CD45R+ granulated lymphocytes was uniformly distributed in the interplacentomal uterine epithelium throughout pregnancy but never in the syncytial layer of the placentomes. Thirdly, monoclonal antibodies specific for the CD5 antigen consistently stained the endothelium of blood vessels within the placentomes but never blood vessels in the interplacentomal areas. Finally, OLA class I antigens were present on the interplacentomal uterine epithelial cells and on the maternal stromal cells, but no staining of the trophoblast or syncytium was observed. These observations suggest that different mechanisms to prevent immune rejection of the fetus may operate in the placentomes where trophoblast invasion of the maternal tissue occurs compared to the interplacentomal regions.

Animals↗
PubMed1988

Lymphocyte subsets show marked differences in their distribution between blood and the afferent and efferent lymph of peripheral lymph nodes.

The surface phenotypes (CD1, CD4, CD5, CD8, SBU-T19, MHC class I, MHC class II, and sIg) of cells in blood, lymph nodes, and lymph were determined to examine simultaneously the distribution of lymphocyte subsets circulating in blood, afferent lymph, and efferent lymph of a peripheral lymph node. Marked differences in the percentage of certain lymphocyte subsets were apparent within the compartments examined, suggesting that lymphocyte subsets leave the blood with differing efficiencies. Lymphocyte subsets also appeared to be extracted from the blood at different rates by lymph node as opposed to subcutaneous vascular endothelium. Endothelial cells in different vascular beds may express different numbers of molecules complementary to a set of migration-related cell surface molecules specific for each lymphocyte subset. Accordingly, the vascular endothelium would be the key factor in regulating nonrandom cell migration.

Animals↗
PubMed1988

Lymphocyte phenotypes in the abomasal mucosa of sheep infected with Haemonchus contortus.

Lymphocyte subpopulations in the abomasal mucosa of worm-free and parasitized sheep were assessed in situ. A preponderance of T-lymphocytes, with approximately equal numbers of cells expressing CD5, CD4 and CD8 antigens, was found. Most of the lymphocytes expressing CD8 lacked CD5. Using a panel of 15 monoclonal antibodies to ovine leucocyte antigens, abomasal lymphoid follicles in the mucosa were shown to resemble lymph node follicles phenotypically. Abomasal epithelial cells contained major histocompatibility complex (MHC) class II antigen. Infection or hyperimmunization of pasture-reared sheep with the gastric nematode Haemonchus contortus increased the numbers of mucosal mast cells and eosinophils but did not alter the phenotypic composition or number of mucosal lymphocytes or the pattern of expression of MHC class II antigens.

Abomasum↗
PubMed1988

Identification of a unique lymphocyte subpopulation in the sheep uterus.

A panel of monoclonal antibodies was used to define the lymphocyte subpopulations in the sheep uterus at various stages of the oestrous cycle. A striking finding was that the majority of lymphocytes in the uterine and endometrial glandular epithelia belonged to a unique lymphocyte subpopulation that expressed the CD45R antigen but was negative for major histocompatibility complex (MHC) class II molecules and expressed low or undetectable levels of the CD5 antigen. When examined under the electron microscope using the immunogold technique, the CD45R+ lymphocytes were found to have one to three membrane-bound granules in their cytoplasm. Other lymphocyte subpopulations found in the uterus at various stages of the oestrous cycle were localized mainly in the caruncular and intercaruncular stroma. The unique CD45R+ granular lymphocyte subpopulation may be equivalent to the 'natural killer' cells reported in mouse and man, and may have an important role in local immunity of the female reproductive tract.

Animals↗
PubMed1988

Presence of a distinct CD8+ and CD5- leucocyte subpopulation in the sheep liver.

The relative proportions of CD5, CD4, CD8, SBU-T19 and surface immunoglobulin (sIg)-positive lymphocytes in normal sheep livers were determined by flow cytometry analysis of isolated liver leucocytes and compared to lymphocyte subpopulations found in peripheral blood and hepatic lymph nodes. In contrast to the blood, very low numbers of SBU-T19+ cells were found in both the liver and hepatic lymph nodes. The ratio of helper versus suppressor/cytotoxic T cells in the liver was less than 1, which was much lower than in blood or hepatic lymph nodes. A large proportion of liver lymphocytes were characterized by a low intensity staining for the CD8 antigen (CD8L). Two-colour flow cytometry analysis revealed that the CD8L cells were negative for the CD5 pan T-cell marker. It is suggested that these CD8LCD5- cells are natural killer (NK) cells similar to those found in the gut. An influx of CD8LCD5- cells was observed in leucocyte-infiltrated liver lesions at later stages of a rejection response to the cestode Taenia hydatigena.

Animals↗
PubMed1988

Lymphocyte phenotypes in the intestinal mucosa of sheep infected with Trichostrongylus colubriformis.

Monoclonal antibodies to ovine lymphocyte surface antigens were used in an immunohistochemical study of the intestine of sheep. In the epithelium CD8+ cells predominated whereas the majority of lamina propria T lymphocytes were CD4+. Infection of sheep with the parasitic nematode Trichostrongylus colubriformis including sufficiently large numbers of parasites to induce protective immunity did not alter the number of CD4+ or CD8+ lymphocytes in the intestinal mucosa. In contrast, exposure of naive sheep to a single large infection of T. colubriformis resulted in a substantial decrease in number of CD8+ cells and moderate decreases in number of CD4+ cells in the duodenal but not the jejunal mucosa. MHC class II antigens were not detected either in or on epithelial cells of the sheep small intestine.

Aging↗
PubMed1987

N-terminal amino acid sequence analyses of the alpha and beta polypeptides from four distinct subsets of sheep major histocompatibility complex class II molecules.

Four monoclonal antibodies, SBU.II 28-1, 37-68, 38-27, and 42-20, each recognizing a distinct, non-overlapping subset of sheep class II molecules, were used to purify class II molecules from a single sheep. Four class II alpha subunits designated 28-1 alpha, 37-68 alpha, 42-20 alpha, and 38-27 alpha and five class II beta subunits designated 28-1 beta, 37-68 beta 1, 37-68 beta 2, 42-20 beta, and 38-27 beta were compared by N-terminal sequence analyses. Two distinct alpha subunits were identified; the 28-1 alpha, 37-68 alpha, and 42-20 alpha subunits all had identical N-terminal amino acids sequences, which exhibited about 75% homology with HLA-DR alpha and mouse E alpha polypeptides. In contrast, the 38-27 alpha sequence exhibited about 80% sequence homology with HLA-DQ alpha and mouse A alpha polypeptides. In general, sheep beta subunits displayed insufficient sequence homology to enable correlation with human beta-chain sequences; however, the 38-27 beta-chain sequence showed homology with the HLA-DQ beta sequence. The conserved sequence surrounding the site for N-linked glycosylation within human/mouse beta polypeptides (residues 19 to 21) was not present in sheep beta sequences and in contrast with the beta-chains of mouse and man, sheep beta polypeptides contained between 1 and 3 positionally variable cysteine residues (residues 13 to 15 inclusive). Individual sheep beta subunits exhibited extensive sequence heterogeneity and each consisted of a unique population of beta polypeptide species. At least 16 different beta polypeptide sequences were identified from a single sheep and the existence of no fewer than nine non-allelic beta genes was inferred from the sequence data. We have previously provided evidence suggesting that the sheep has multiple major histocompatibility complex class II alpha and beta genes related to those of all three HLA-D subregions. The present results suggest that a number of these genes encode HLA-DQ-like heterodimers and that a sheep DR-like alpha gene product is shared with the products of a large and heterogeneous sheep beta gene family.

Amino Acid Sequence↗
PubMed1987

Genetic organization of the ovine MHC class II region.

To study the class II genes of the major histocompatibility region of the sheep genome, human HLA class II genes corresponding to the known subregions in man (DR, DQ, DP, DO, and DZ) were used for Southern hybridization analysis of sheep DNA and to probe a sheep genomic library. Hybridizing bands were noted for all probes except DP alpha. DQ alpha and beta and DR beta appear to be present as multicopy genes, while DR alpha-, DZ alpha-, and DO beta- like genes appear to be single copy. All bands detected with the DP beta probe were also detectable with other beta chain probes. From eight lambda-bacteriophage clones of a sheep genomic library nine distinct class II genes were identified. These genes were characterized by differential hybridization analysis and restriction mapping. Two genes were DR beta-like, three DQ alpha-like and four DQ beta-like. The extensive cross-hybridization observed with beta chain probes was not seen with alpha chain probes. The results of this study suggest that the major histocompatibility complex class II region of the sheep has a similar genetic organization to that of man, with the provisional exception of the DP subregion.

Animals↗
PubMed1987

A monoclonal antibody to the p220 component of sheep LCA identifies B cells and a unique lymphocyte subset.

The leukocyte common antigen (LCA, CD45) of humans and rodents is expressed exclusively by leukocytes, and has been implicated in a number of immune functions (1-4), although its precise function is still unknown. Three monoclonal antibodies (mAbs) were produced which identified different epitopes on the LCA of sheep. mAbs 1-11-32 and 38-42 reacted with determinants of LCA expressed on all leukocytes, but showed differential reactivity with thymocytes. Another antibody, 20-96, identified an epitope of LCA expressed mainly on B cells, but also on a unique lymphocyte subset contained mostly in peripheral blood, which was 20-96high, sIg-, CD4-, CD8-, SBU-T19-, but CD5+. These cells constituted only 5-6% of PBL. The cellular lineage of this latter subset is uncertain since these cells appeared to be unrelated to B cells, and were absent from the thymus. Unlike the other two mAbs to LCA, 20-96 was not reactive with macrophages and granulocytes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of material immunoprecipitated by the "pan" LCA-specific mAbs revealed lymphocyte forms with molecular weights (MW) of 220, 210, and 190K, whereas 20-96 immunoprecipitated only a 220K MW form. The expression and MW of LCA on thymocytes or ileal Peyer's patch (IPP) cells differed from those on peripheral lymphocytes. B cells in IPP, which constitute 98% of cells, expressed the 20-96 determinant at low density, in contrast to its high expression on peripheral B cells. LCA from IPP existed in two forms of 220 and 190K MW, whereas LCA from peripheral B cells was entirely 220K MW, and thymus 210 and 190K MW.

Animals↗
PubMed1987

Antigens of activated rat T lymphocytes including a molecule of 50,000 Mr detected only on CD4 positive T blasts.

Mouse monoclonal antibodies (MAbs) have been prepared against rat T cell blasts. One MAb called MRC OX-40 recognized an antigen that differed from any previously described in that its expression was detected only on T blasts that also expressed the CD4 antigen. The OX-40 MAb did not detect an activation determinant of CD2 or CD4 molecules but recognized a distinct chain of mol. wt 50,000. The OX-40 MAb augmented T cell proliferation at late stages on in vitro responses. Other MAbs without obvious counterparts in other species were MRC OX-48 and MRC OX-49,50 which recognized cell surface molecules of mol. wts of about 95,000 and 90,000, respectively. The OX-48 antigen was not expressed on resting lymphocytes but was found on a subset of T and B blasts and also on other leucocytes. The OX-49,50 antigen was found on most haemopoietic cells but was expressed at greatly increased levels after lymphocyte activation and this was also the case for MRC OX-47 antigen which is of unknown Mr. The MRC OX-39 MAb was found to bind the rat IL-2 receptor; expression of this antigen was detected on thymic dendritic cells as well as on T blasts. The phenotype of rat T blasts compared to resting cells was also examined and changes in expression of L-CA, Thy-1, OX-2 and CD8 antigens were seen in addition to the changes found with the above MAbs.

Animals↗
PubMed1987

A functional and biochemical analysis of bovine class II MHC antigens using monoclonal antibodies.

Monoclonal antibodies (MAbs) reacting with bovine (2) ovine (3), murine (1) or human (1) Class II MHC antigens were examined for reactivity with bovine peripheral blood leucocytes (PBL) and lymph node cells (LNC) by immunofluorescence, immunoprecipitation and the capacity to inhibit mixed lymphocyte responses (MLR), lectin- and antigen-induced blastogenesis. The 6 MAbs identified comparable percentages of Class II positive lymphocytes in PBL (40.8 to 54.2%) and LNC (6 to 11.5%) regardless of BoLA-A phenotype. Immunohistological staining of Class II MAb was localized principally to the lymphoid follicles in lymph nodes and to isolated epithelial reticular cells in the thymus. The anti-Class II MAb immunoprecipitated alpha- and beta- chains of 26-29K and 32-34K, respectively. These MAb inhibited proliferative responses in the MLR by between 25 and 74%, and diminished blastogenesis induced by specific antigens (purified protein derivative + PPD and ovalbumin) and B-lymphocyte mitogens (PPD, lipopolysaccharide and dextran sulphate) by between 45 and 75%, regardless of BoLA-A phenotype. In contrast, proliferation in response to concanavalin A and phytohaemagglutinin were unaffected by the anti- Class II MAb. Similarly these MAb did not affect lysis by cytotoxic T-lymphocytes, the activity of which was depressed by anti-Class I MAbs and monospecific alloantisera.

Animals↗
PubMed1987

Monoclonal antibodies to sheep MHC class I and class II molecules: biochemical characterization of three class I gene products and four distinct subpopulations of class II molecules.

The availability of a panel of monoclonal antibodies to sheep MHC class I and class II gene products has allowed for the first time an assessment of the relative complexity of the sheep MHC. By using four monoclonal antibodies to MHC class I, and seven monoclonal antibodies to MHC class II molecules together with one-dimensional SDS-PAGE, sequential immunoprecipitation and 2-dimensional gel analysis, three class I gene products and four distinct subsets of class II molecules have been identified. Sheep class I molecules showed heterogeneity on 2-dimensional gels and as in mouse and man, represented the products of at least three different non-allelic class I genes. Interestingly, the sheep beta 2 microglobulin molecule also displayed heterogeneity, consistent with either two primary gene products or allelic variation. Four sheep class II monoclonal antibodies identified distinct, non-overlapping subsets of sheep class II molecules of Mr 32-36 K (alpha chain) and 25-28 K (beta chain). These class II molecules were co-expressed on sheep B lymphocytes and represented the primary products of different sheep MHC class II genes. The class II molecules within three of these subsets displayed allelic polymorphism essentially restricted to their beta polypeptides, while the fourth subset of class II molecules showed allelic variation in both their alpha and beta polypeptides. The results of this study represent the first evidence for gene duplication and heterogeneity within the sheep MHC. The identification of three primary class I gene products and four distinct subsets of class II molecules suggests three class I loci and up to four distinct class II subregions within the sheep MHC. Potentially large numbers of allelic variants of these different gene products may be expressed in normal sheep.

Animals↗
PubMed1987

Lymphocyte antigens of sheep: identification and characterization using a panel of monoclonal antibodies.

A panel of monoclonal antibodies has been developed and used to identify and characterize the lymphocyte antigens of sheep. These studies have shown that sheep lymphocyte antigens display similar, if not identical, tissue distributions to their analogues in other species. Some of the major sheep antigens, including CD5, CD4, CD8, SBU-T19, Pgp-1, LCA and the MHC antigens, are described in detail.

Animals↗
PubMed1987

Biochemical and molecular analysis of sheep MHC class II molecules.

A panel of monoclonal antibodies was used for structural and immunodepletion analysis of sheep MHC class II molecules. The results indicate the antibodies recognize molecules of molecular weight 32-34,000 (alpha chain) and 26-28,000 (beta chain). Immunodepletion analysis indicates that the antibodies may recognize up to four distinct class II molecules some of which are structurally distinguishable using SDS-PAGE. Southern blot analysis using HLA-D region DR, DQ, DP, DO and DZ cDNA probes showed that a number of the cDNA probes hybridized specifically to sheep DNA indicating the presence of closely related genes in sheep. Together the results suggest that the sheep MHC class II region contains distinct MHC class II genes similar to those found in man.

Animals↗
PubMed1987

Monoclonal antibodies to sheep MHC class II molecules recognize all HLA-D or subsets of HLA-D region products.

Six murine monoclonal antibodies raised against sheep MHC class II molecules were analyzed for reactivity with HLA-D subregion products. All the antibodies reacted with human peripheral blood lymphocytes, monocytes, and B-lymphoblastoid cell lines homozygous for various HLA-DR specificities, suggesting that the antibodies recognized nonpolymorphic determinants on HLA class II molecules. SDS-PAGE and two-dimensional NEPHGE/SDS-PAGE analyses of molecules immunoprecipitated from 35S-methionine-labeled, DR-homozygous B-lymphoblastoid cell lines showed that the monoclonal antibodies precipitated typical class II molecules (Mr 32-34K and 25-29K). From comparison with antibodies of known HLA-D subregion specificity, two of the sheep antibodies appeared to react with the products of single HLA-D subregions, while another showed balanced reactivity with all HLA-D molecules. Antibody SBU.II 38-27 reacted exclusively with HLA-DQ molecules, antibody SBU.II 28-1 with HLA-DP molecules, and antibody SBU.II 49-1 with HLA-DR, -DQ, and -DP molecules. However, analysis of immunoprecipitates from surface-iodinated WT-49 cells (DR3 homozygous) using SBU.II 28-1 and the DP-specific monoclonal antibody B7/21, suggested that the two antibodies immunoprecipitated different alpha polypeptides. The two antibodies SBU.II 38-27 and 28-1 appear to be at least as specific as existing reagents, if not more so. As such, they are of value in their potential contribution to our understanding of the molecular characteristics and ultimately the functions of the HLA-DQ and -DP subregion products, as well as the identification/characterization of HLA-D equivalents in other species.

Adult↗