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Biomedical subjects

I McConnell

Publications and source records attributed to I McConnell.

At least 109 records · Page 6Linked to original sources

Monoclonal antibodies to sheep lymphocytes. I. Identification of MHC class II molecules on lymphoid tissue and changes in the level of class II expression on lymph-borne cells following antigen stimulation in vivo.

A rat monoclonal antibody is described that recognizes sheep MHC Class II antigen and appears to recognize a determinant that is non-polymorphic. This antibody precipitates polypeptides of apparent molecular weight 33,000 and 28,000/29,000 (under non-reducing conditions). The binding of this antibody was inhibited by an anti-human DP, DQ, DR beta-chain monoclonal (DA6.231) and its reactivity on murine cells suggests specificity for the I-E gene product. Using this antibody we have studied the distribution of Class II molecules in sheep and also the variations in Class II expression by cells in both peripheral and efferent lymph, produced as a result of in vivo antigen inoculation.

Animals↗

Marginal zone B cells express CR1 and CR2 receptors.

Opsonized yeast is known to bind strongly to the marginal zones in frozen sections of rat spleen (Kumararatne, D. S. et al., Eur. J. Immunol. 1981. 11: 858). This study reports an analysis of the cells involved in this binding. Sheep red cells coated respectively with C3b, C3bi or C3d were used as indicator cells. These showed homogeneous binding of both C3b and C3bi to marginal zones and germinal centers. C3d-coated red cells bound in a uniform speckled pattern to marginal zones. They also bound to germinal centers and the small lymphocyte zones of the follicles. Selective depletion experiments were undertaken to show that binding to marginal zones was a property of the IgM+ and IgD- B cells characteristic of this area. Binding to germinal centers was attributable to follicular dendritic cells. The C3d receptors in follicles were shown to be on IgM+ and IgD+ small B lymphocytes.

Animals↗

Distinct populations of class II molecules identified by monoclonal antibodies.

Binding inhibition and sequential immunoadsorption studies with two rat monoclonal antibodies have been used to demonstrate the existence of distinct populations (isotypes) of HLA-DR like (class II) molecules in human B lymphoma cell lysates. The structures of the molecules isolated by these two reagents have been compared by peptide 'mapping' and appear to be closely similar.

Antibodies, Monoclonal↗

Specific selection of antigen-reactive lymphocytes into antigenically stimulated lymph nodes in sheep.

Sheep were primed to a variety of antigens and the efferent lymphatic from a popliteal lymph node was cannulated. The cannulated node was challenged repeatedly with PPD and all the lymph and cells removed from the animal. During this time the PBL were monitored for reactivity to all antigens (purified protein derivative of tuberculin [PPD], johnin, and keyhole limpet hemocyanin [KLH]) by the vitro transformation assay. The response of these cells to PPD was found to be gradually eliminated after repeated challenge of the cannulated node with that antigen. The response to the other antigens was unimpaired. No depletion of this response to PPD occurred in cannulated sheep when the antigen was given into a noncannulated node. In vivo delayed-type hypersensitivity skin test and helper T cell assays confirmed that there is a specific selection of antigen-reactive cells from the recirculating lymphocyte pool into antigen-stimulated lymph nodes.

Animals↗

Activation of the alternative complement pathway by normal and transformed cells in homologous and heterologous sera.

A variety of both normal and viral transformed or infected cells have been tested for their ability to activate the alternative pathway of complement in either homologous or heterologous sera. Normal cell lines fail to produce alternative pathway activation in homologous sera but readily activate the alternative pathway in heterologous sera. In contrast, human cells, persistently infected with measles virus or chicken cells transformed with oncogenic avian viruses, produce a marked activation in homologous sera. It is suggested that in certain circumstances, the alternative pathway of complement can discriminate between normal and abnormal "self" cells of foreign cell surfaces.

Animals↗

Malignant cells isolated from Burkitt's lymphoma but not other forms of leukemia activate the alternative complement pathway in human serum.

Cells isolated directly from primary biopsies of Burkitt's lymphoma were found to activate the alternative complement pathway in hypogammaglobulinemic human serum. In contrast, cells isolated from patients with either acute lymphoblastic leukemia, chronic lymphocytic leukemia or chronic myeloid leukemia did not activate. No defects in the ability of Burkitt's lymphoma sera to support alternative pathway activation were detected.

Burkitt Lymphoma↗

Reactivity of monoclonal antibodies with human myeloid precursor cells.

Five monoclonal antibodies have been tested for their ability to bind to myeloid precursor cells in normal human bone marrow. Indirect immunofluorescence and the fluorescence activated cell sorter was used to separate cells according to their reactivity for trial culture in vitro in order to grow granulocyte-macrophage colony forming cells (CFUc). Two antibodies (OKT3 and OKT11) which react strongly with bone marrow T lymphocytes were found to be unreactive with CFUc. YD1/23 reacts very strongly with both T and B lymphocytes but is only weakly reactive with CFUc. In contrast, OKT10 and YE2/36 did react with CFUc. The consequences of these findings and the potential clinical use of these antibodies in bone marrow transplantation are discussed.

Adult↗

Increased numbers of cells with suppressor T cell phenotype in the peripheral blood of patients with infectious mononucleosis.

Peripheral blood cells from six patients with acute infectious mononucleosis were studied by two-colour immunofluorescence using antibodies to human T cells, inducer and suppressor-cytotoxic T cell subsets and Ia-like antigens. The absolute number of T cells with the suppressor-cytotoxic phenotype was substantially increased in each case; many of these cells also expressed Ia-like antigens and had the morphology of the large atypical cells characteristic of infectious mononucleosis. These activated suppressor T cells of infectious mononucleosis may therefore represent a control mechanism to prevent viral-induced proliferation of B cells.

Cytotoxicity Tests, Immunologic↗

Lymphocyte traffic through antigen-stimulated lymph nodes. II. Role of Prostaglandin E2 as a mediator of cell shutdown.

Prostaglandin E2 (PGE2) can mediate changes in lymphocyte traffic through lymph nodes. By measuring PGE2 levels in efferent lymph from cannulated sheep lymph nodes it was shown that increased synthesis of PGE2 always correlated with cell shutdown; infusion of PGE2 alone induced cell shutdown and inhibition of PGE2 synthesis by the node abolished cell shutdown induced by antigen.

Animals↗

Lymphocyte traffic through lymph nodes during cell shutdown.

Antigenic challenge of lymph nodes in sheep has marked effects on lymphocyte traffic through lymph nodes. The non-specific effects include a marked reduction in lymphocyte output in efferent lymph without a corresponding decrease in lymph flow--a phenomenon known as cell shutdown. With certain antigens there is a total disappearance of B lymphocytes during cell shutdown. The phenomenon can be reproduced in unprimed lymph nodes whenever localized complement activation occurs within the node. This also induces the release of prostaglandins, particularly PGE2. These results suggest that cell shutdown might be a two-step process involving both complement and prostaglandins. Repeated stimulation of nodes with antigen also has considerable effects on the traffic of antigen-specific lymphocytes. Antigen localized within the node can promote the selective entry into the node of T lymphocytes specific for the challenge antigen. Consequently there is a net loss from the whole animal of T cells reactive to the challenge antigen. These results are discussed in relation to lymphocyte recirculation through antigen-stimulated lymph nodes.

Animals↗

Immunological clearance of 75Se-labelled Trypanosoma brucei in mice. II. Mechanisms in immune animals.

Using trypanosomes labelled with [75Se]-methionine a series of experiments was conducted to investigate the respective roles of antibody, macrophage activtion and complement in the removal of trypanosomes from the circulation of immune mice. It was found that clearance in such animals is largely accomplished by antibody-mediated hepatic phagocytosis, which, at least in passively immunized animals, is dependent on opsonization involving C3. No evidence was found to suggest that intravascular lysis or activated macrophages are involved in immune clearance.

Animals↗

Activation of the alternative complement pathway by human B cell lymphoma lines is associated with Epstein-Barr virus transformation of the cells.

Activation of the alternative complement pathway by human B cell lymphoma lines is correlated with the presence of Epstein Barr virus (EBV) in the cell genome. EBV-negative B cell lymphoma lines produce little activation of the alternative pathway as measured either by C3 deposition on the cell surface or C3 conversion and consumption of alternative pathway activity in the supernatant serum. By contrast, EBV-positive sublines derived by in vitro EBV conversion of EBV-negative parental lines produce considerable activation of the alternative pathway. This membrane-associated complement-activating mechanism reflects an EBV-induced membrane change in these cells and may provide a mechanism whereby EBV-transformed cells are controlled in vivo.

B-Lymphocytes↗

Purification of antigen-dependent macrophage migration inhibition factor (MIF) from lymph draining a tuberculin reaction.

An antigen-dependent factor showing migration inhibition (MIF) and gold uptake stimulation (GUS) activities which has been previously described (Lowe & Lachmann, 1974) has been further purified from efferent lymph collected from cannulated nodes of BCG-sensitized sheep undergoing a delayed hypersensitivity response to PPD. During purification, fractions containing MIF activity also exhibited GUS activity. Initial purification by salt precipitation showed that antigen-dependent MIF activity was in the 40-90% ammonium sulphate precipitate. Non-specific activity and contaminating immunoglobulin were found in the 0--20% and 20--40% precipitates. Gel filtration on Sephadex G-200 and affinity chromatography on Concanvalin A-Sepharose have shown that antigen-dependent MIF is a glycoprotein of approximately 70,000 molecular weight (Lowe & Lachmann, 1974). Traces of contaminating antibody in the glycoprotein fraction were removed by immuno-adsorption on monospecific anti-sheep IgG-Sepharose. Antigen-dependent MIF was also purified by affinity chromatography on PPD-Sepharose. The eluted fractions with all the antigen-specific activity, contained less than 1% of the applied material. Analysis by polyacrylamide gel electrophoresis showed that the major protein component in the purified MIF preparation has a molecular weight and electrophoretic mobility identical with that of sheep albumin. Although this represents a high degree of purification of antigen-dependent MIF it seems that albumin is still present as a contaminant and that the protein associated with MIF activity is present in trace quantities.

Ammonium Sulfate↗