Biomedical subjects

J Kirkley

Publications and source records attributed to J Kirkley.

15 recordsLinked to original sources

PubMed1997

The predominance of beta (CC) chemokine transcripts in idiopathic inflammatory muscle diseases.

Cytokines and chemokines that upregulate major histocompatibility complex class I antigens, recruit lymphocytes, and enhance T-cell-mediated myotoxicity may be important in the pathogenesis of dermatomyositis and polymyositis. We searched for cytokine and chemokine transcripts in inflammatory muscle specimens from 14 newly diagnosed or treated patients. Control specimens from six patients without inflammatory muscle disease were analyzed for transcripts of interleukins-1 beta, -2, -4, -6, -10, and -15, and interferon-gamma, tumor necrosis factor-alpha, transforming growth factor-beta 1, macrophage inflammatory proteins-1 alpha and -1 beta (MIP-1 alpha, MIP-1 beta), and the chemokine "regulated on activation, normally T expressed and secreted" (RANTES). Surprisingly, the proinflammatory and lymphocyte cytokines were detected only sporadically in myositis muscle specimens, and their presence did not correlate with disease activity or treatment status of the patient. In contrast, MIP-1 alpha and MIP-1 beta were detected in 13 and 6 myositis biopsies, respectively, and RANTES, another beta (CC) chemokine, was detected in eight myositis biopsies. This study and other reports of low levels of acute-phase cytokines in myositis patients suggest that the proinflammatory cytokines do not play a major role in ongoing muscle damage. The CC chemokines studied here, in particular MIP-1 alpha, might contribute to ongoing muscle inflammation, and the pathogenesis of inflammation in myositis may follow a previously unrecognized pathway.

Cells, Cultured↗

PubMed1987

Monoclonal antibody definition of multiple polymorphic epitopes on human leukocyte antigen-DRw52.

Monoclonal antibodies have revealed complexity within the human leukocyte antigen class II antigens. We have studied epitopes present on a DR3 homozygous B-lymphoblastoid cell line using five polymorphic monoclonal antibodies produced and characterized in our laboratory. Serological analysis on a panel of B-cell lines revealed that the antibodies have different, but related, specificities (NDS9-anti-DR3, NDS10-anti-DR5, less than 3, less than w6, NDS11-anti-DR3, 5, w6, NDS12-anti-DR3, 5, w6, w8, NDS13-anti-DR3, 5, w6, w8+). Competitive radioimmunoassays and two-dimensional gel analyses demonstrated that whereas the epitopes recognized by the broadly reactive antibodies NDS10, 11, 12, and 13 reside on the same molecule, the epitope detected by NDS9 is present on a molecule with different electrophoretic mobility. Thus, using polymorphic monoclonal antibodies, we have defined multiple epitopes associated with DR3, which have different distributions at the population level.

Antibodies, Monoclonal↗

PubMed1983

A monoclonal antibody recognizing HLA-DR2 on malignant and activated cells.

A monoclonal antibody, designated NDS15.38, which recognizes a polymorphic determinant of HLA-DR, was produced from a fusion in which mice were immunized with the human B lymphoblastoid cell line GIR2 (HLA type A1, B8, 27, Cw2, DR2,7). NDS15.38 functions efficiently as an affinity column and purifies a two-chain complex of molecular weight 33 000 and 30 000 under reducing conditions. The monoclonal antibody reacts with HLA-DR2-positive B lymphoblastoid cell lines and B lymphocytes from patients with chronic lymphatic leukemia in an indirect radioactive binding assay. However, NDS15.38 does not appear to react with peripheral blood B lymphocytes from normal individuals. Using a peroxidase staining technique, NDS15.38 was shown to react with phytohemagglutin (PHA)-stimulated lymphocytes and with apparently activated B cells in the germinal centers of lymph nodes from individuals who were tissue typed as HLA-DR2. Thus it appears that NDS15.38 recognizes a polymorphic determinant of HLA-DR on malignant and stimulated cells, but not on resting cells.

Animals↗

PubMed1980

Monoclonal antibody to a human leukocyte-specific membrane glycoprotein probably homologous to the leukocyte-common (L-C) antigen of the rat.

The monoclonal antibody (F 10-89-4) described in this study recognizes an antigen which by quantitative absorption analysis is absent from human brain, kidney, liver, heart, erythrocytes, platelets and normal serum, but is present on spleen, lymph node, chronic lymphatic leukemia cells, bone marrow, thymus and granulocytes at a ratio of 1:1:0.8:0.3:0.1, respectively. Analysis with the fluorescence-activated cell sorter showed that 100% of thymocytes, lymph node lymphocytes, blood mononuclear cells and granulocytes carry the antigen, while 83% of bone marrow cells are positive. There was marked heterogeneity in the amount of labeling of thymocytes, with 3 major peaks. There was also heterogeneity of labeling of blood mononuclear cells and lymph node lymphocytes, with a weakly staining hump containing approximately 20% of the cells in the case of lymph node lymphocytes. Double labeling experiments demonstrated that the weakly staining cells of blood and lymph node were B lymphocytes, while frozen sections of thymus showed that the antigen was expressed most weakly in subcapsular cortical thymocytes, and most strongly on medullary thymocytes. Biochemical studies established that the antigen bound to lentil lectin columns, and sodium dodecyl sulfate polyacrylamide gel electrophoresis studies using NaB3H4-labeled blood mononuclear cells established that the antigen was a major glycoprotein of lymphocytes, and that its molecular weight was in the region of 190000 to 215000.

Absorption↗

PubMed1980

Monoclonal antibody to a human brain-granulocyte-T lymphocyte antigen probably homologous to the W 3/13 antigen of the rat.

The monoclonal antibody (F 10-44-2) described in this report recognizes an antigen which by quantitative absorption analysis is found predominantly on spleen, lymph node, bone marrow, thymus, granulocytes and brain, the amount of antigen on these tissues being approximately the same within a factor of 2 or 3. Analysis with the fluorescence-activated cell sorter showed that 29% of thymus cells, 61% of bone marrow cells, 95% of blood mononuclear cells, 98% of lymph node lymphocytes and 100% of granulocytes carried the antigen. With blood mononuclear cells and lymph node lymphocytes, there were two distinct peaks, with one peak labeling very weakly. Double labeling experiments established that the weakly labeled peak contained the B lymphocytes. Studies on frozen sections of thymus established that positive thymocytes were found only in the medulla indicating that the antigen appears late in T lymphocyte maturation. The lymphatic nodules (B lymphocyte areas) of spleen and lymph node appeared virtually negative on frozen sections showing that there was too little antigen on the B lymphocyte surface for confident detection by fluorescence microscopy. Sodium dodecyl sulfate polyacrylamide gel eletrophoresis of NaB3H4-labeled blood mononuclear cells established that the antigen was a major glycoprotein of the leukocyte membrane and that its mol. wt. was 105000. This antigen shows a striking similarity in biochemistry and tissue distribution to the W 3/13 antigen of the rat and is likely to be the human homologue of this antigen.

Absorption↗

PubMed1980

125I anti-immunoglobulin binding assay for the detection and characterization of anti-platelet antibodies.

The binding assay as described in this paper is a very versatile system, and in this study it has been evaluated specifically for the detection of allo- or autoantibodies to platelets in man. The basic assay involves the incubation of a standard number of platelets with dilutions of test sera and the detection of platelet bound immunoglobulin by a second incubation with 125I labeled, immunoadsorbent purified rabbit F(ab') anti-human F(ab')2 (RAH). Of most importance, by varying the number of target platelets in the titrations and looking for binding plateaus, one can readily define conditions of optimum sensitivity for particular serum/platelet combinations. In addition, the assay can be used in conjunction with quantitative absorptions to subdivide complex sera into subspecificities and to give an estimate of the relative amounts of particular antigens on different platelets or other tissue or cell suspensions. One can also use saturating concentrations of RAH in the second incubation, in which case the amount of platelet bound radioactivity is directly related to the amount of first antibody bound to the platelets, and this can be manipulated to give information about serum antibody concentrations and amounts of antigen on the target tissue. The problem of ABO antibodies in this system, optimal conditions for platelet storage for the assay, and techniques for reducing assay backgrounds resulting from immunoglobulin adsorbed to the platelet surface are all evaluated.

ABO Blood-Group System↗

PubMed1977

Regulation of Leishmania populations within the host. I. the variable course of Leishmania donovani infections in mice.

The course of infection with Leishmania donovani was followed in seven strains of laboratory mice with measurement of the liver parasite burdens over 20 weeks. The acute parasite population growth rate varied greatly between, but not within, strains. Four strains were relatively resistant with less than an eight-fold increase while the three acutely susceptible strains showed over an eight-fold increase in the first month. Thereafter, one initially susceptible strain showed a dramatic fall in parasite numbers with histological liver damage while another strain maintained an immense parasite load for up to 2 years involving mononuclear phagocytes throughout the body. The system provides a model for studying genetic control of resistance to intracellular infection and the range of responses is compared with human leprosy and cutaneous leishmaniasis.

Animals↗

PubMed1972