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Biomedical subjects

R C Atkins

Publications and source records attributed to R C Atkins.

At least 199 records · Page 11Linked to original sources
PubMed1984

PHM 14: a novel monoclonal antibody that reacts with both normal and neoplastic human B cells but not B-CLL.

The reactivity of PHM 14, a monoclonal antibody to B cells produced in our laboratory was compared with Leu 10, a commercially produced antibody. Flow microfluorometric analysis showed that both PHM 14 and Leu 10 reacted with normal B cells, a variety of B cell lines and with B cell leukaemias and lymphomas, except in the case of B-CLL where PHM 14 reacted with a minority of cases, whereas Leu 10 reacted with a majority. PHM 14 and Leu 10 did not react with normal monocytes, granulocytes, T cells, T cell lines or with T cell leukaemias or lymphomas. Specificity of PHM 14 for B cells was further supported by the demonstration that PHM 14-positive blood cells, sorted from PHM 14-negative blood cells, reacted with polyclonal anti-immunoglobulins and with a panel of B cell markers but did not react with markers for T cells, monocytes, granulocytes or NK cells. Our results suggest that both PHM 14 and Leu 10 should be useful diagnostic reagents in the investigation of B cell-derived leukaemias and lymphomas.

Antibodies, Monoclonal↗
PubMed1984

Cellular composition of crescents in human rapidly progressive glomerulonephritis identified using monoclonal antibodies.

The relative contributions of glomerular epithelial cells, macrophages, and other cell types to the formation of cellular crescents characteristic of human rapidly progressive glomerulonephritis remain controversial. To identify and quantitate the cell types present during different stages of glomerular crescent formation, immunoperoxidase labelling of cryostat sections from renal biopsies with cellular (n = 9) or sclerosed (n = 3) crescents was performed using monoclonal antibodies to cell-specific antigens of leucocytes, epithelial cells, and other glomerular cell types. Fresh cellular crescents consisted of macrophages (34.5 +/- 7.0%; mean +/- SEM) plus lesser proportions of polymorphs (12.8 +/- 4.7%) and epithelial cells (10.4 +/- 1.5%). Sclerosed crescents contained fewer macrophages (5.1 +/- 1.0%), but similar proportions of polymorphs (11.1 +/- 2.9%) and epithelial cells (11.5 +/- 2.1%). Lymphocytes were not detected within crescents. Many of the remaining unlabelled cells morphologically resembled fibroblasts and expressed surface fibronectin, though fibroblast-specific cell markers were not available. These results show that macrophages and not epithelial cells constitute the major cell type within cellular crescents. Therapeutic manoeuvres directed against macrophages may, therefore, be of clinical value in the management of human crescentic rapidly progressive glomerulonephritis.

Adult↗
PubMed1984

Production of monoclonal antibodies to fibronectin, type IV collagen and other antigens of the human glomerulus.

A series of monoclonal antibodies to human glomerular antigens was prepared by immunisation of a mouse with isolated whole glomeruli, followed by boosting with particulate glomerular basement membrane and fusion of murine spleen cells with the NSI-myeloma line. Hybridoma supernatants were screened jointly by a radioimmunoassay involving binding to isolated glomeruli, and by a 4-layer immunoperoxidase technique applied to polyester wax-embedded sections. Seven monoclonal antibodies with different specificities (PHM7-PHM13) were established and repeatedly cloned. Each antibody displayed a distinctive distribution within the glomerulus, including different patterns of staining of mesangial cells, mesangial matrix and glomerular basement membrane, in addition to extra-glomerular basement membranes and extracellular matrix. All antibodies also stained cellular outgrowths of isolated glomeruli cultured in vitro, and showed additive binding to cultured cells by radioimmunoassay. Physical characterization using absorptions with purified substrates, plus specific chemical and enzymatic digestions, indicated that PHM12 is directed against type IV collagen. PHM13 is directed against fibronectin as shown by absorption with purified fibronectin and immunoprecipitation of a 220 000 MW glycoprotein. The remaining 5 monoclonal antibodies, which react with carbohydrate (PHM7) or protein (PHM8-PHM11) determinants, were shown to be nonreactive with type IV collagen, fibronectin or other known glomerular components including sialic acid, laminin, amyloid P-component or various glycosaminoglycans. These monoclonal antibodies therefore appear to define a new series of human glomerular antigens, or possibly closely related antigenic determinants, which are synthesized by glomerular cells and incorporated into the mesangium and glomerular basement membrane. These antibodies, by providing markers for at least 2 antigens known to be important in glomerular cell-matrix interactions, should prove useful in research into the mechanisms involved in renal pathology.

Antibodies, Monoclonal↗
PubMed1984

Monoclonal anti-T-cell antibodies react with circulating myeloid leukemia cells and normal tissue macrophages.

Rabbit and monoclonal antibodies to human myeloid leukemia cells, monocytic leukemia cells and human thymocytes have shown the existence of common T-cell/myeloid/monocyte antigens. For this reason, the specificity of a series of monoclonal antibodies to human T-cells (OKT 1, 3, 4, 5, 6, 8, 9, 10; and NA1/34) was tested by immunofluorescence (cytofluorograph) and complement-mediated cytotoxicity against human myeloid leukemia and normal blood cells and leukemic cell lines. In addition, an immunohistological analysis of the specificity of OKT4, 9.3, Leu 3a, OKT3 and NA1/34 antibodies was performed using normal lymphoid tissues and a sensitive immunoperoxidase technique. Normal human peripheral blood mononuclear cells reacted with OKT3 ("pan T-cell", mean 54%), OKT4 ("helper T-cell", mean 35%) and OKT 5/8 ("suppressor T-cell", mean 18%) as previously reported. However, OKT3 reacted with the cell lines K562 (myeloid), RC2a and THP-1 (monocytoid) and U937 (macrophage) as well as with cells from 9/65 myeloid leukemia patients. OKT4 reacted with the cell lines HL60 (promyelocyte), RC2a and U937 and also with cells from 6/60 myeloid leukemia patients. OKT5 reacted with the cell lines K562 and THP-1. OKT1 ("pan T-cell") reacted with THP-1 and with myeloid and monocytic leukemia samples (5/32) as did OKT6 ("cortical thymocyte") (3/32). OKT10 ("common thymocyte") reacted with a range of leukemia cell lines (B-cell, pre- B-cell and macrophage) as well as 7/21 myeloid leukemia samples. In tissue sections Leu 3a, (9.3 and OKT4 to a lesser extent), stained paracortical lymphocytes, plus subcapsular and medullary macrophages, and dendritic cells present within the paracortex.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1984

Monoclonal antibody analysis of glomerular hypercellularity in human glomerulonephritis.

To determine the contribution of infiltrating circulating leucocytes to glomerular hypercellularity, and to further investigate the immune and inflammatory mechanisms involved in human glomerulonephritis, a series of renal biopsies were evaluated using cell-specific monoclonal antibodies. In ninety-three renal biopsies from patients with glomerulonephritis, intraglomerular leucocytes were identified by immunoperoxidase localization of monoclonal antibodies to the leucocyte-common antigen, and antigens characteristic of T-cell and T-cell subsets, B-cells, monocytes and granulocytes. Normal glomeruli contained a mean of 2 leucocytes, predominantly monocytes, per glomerular cross-section. No significant increase in leucocytes was found in 41 biopsies with non-proliferative types of glomerulonephritis. However, in renal biopsies from 22 of the 46 patients with proliferative forms of glomerulonephritis, there was a significant increase in glomerular leucocytes. These biopsies were from 5 patients with post-infectious glomerulonephritis (mean of 30 leucocytes per glomerulus), 11 patients with crescentic glomerulonephritis (mean of 16 leucocytes per glomerulus) and 6 patients with mesangial proliferative glomerulonephritis due to systemic lupus erythematosus (mean of 5 leucocytes per glomerulus). The increased intraglomerular leucocytes consisted of macrophages and granulocytes. T and B-cells were generally not found within glomeruli. Thus, glomerular hypercellularity in proliferative glomerulonephritis is in part due to infiltration by inflammatory cells. No evidence was found to directly incriminate cellular immune mechanisms in the pathogenesis of the glomerular lesions of glomerulonephritis since T-cells were not identified within glomeruli.

Antibodies, Monoclonal↗
PubMed1983

Composition of interstitial cellular infiltrate identified by monoclonal antibodies in renal biopsies of rejecting human renal allografts.

Monoclonal antibodies and a four-layer immunoperoxidase technique were used to analyze and quantitate the various infiltrating cell types found in percutaneous renal biopsies of 25 patients undergoing acute renal allograft rejection. Cell markers included monoclonal antibodies to the human leukocyte-common antigen (PHM 1), mononuclear phagocytes (PHM 2, FMC 32), T cells and T cell subsets (OKT 3, OKT 4, and OKT 8), B cells (7.2), polymorphs (FMC 10, FMC 13), and plasma cells (OKT 10). The proportion of labelled interstitial cells was expressed as a percentage of the total number of infiltrating leukocytes identified with PHM 1, and correlated with the histologically graded intensity of rejection. In mild rejection 32% of the infiltrating cells were T lymphocytes, of which 90% were OKT-8-positive cytotoxic-suppressor cells, and 52% were macrophages. Similarly, in moderate rejection T cells composed 42% of the infiltrate (with 67% of T cells expressing OKT 8 antigen), and macrophages formed 38% of the total cells. By contrast, in severe rejection, the T cell component was decreased to 15% of the cells, of which 78% were OKT-8-positive; these were preponderantly macrophages (60%) and polymorphs (22%). These studies demonstrate that cytotoxic-suppressor T cells and macrophages are the major cells mediating acute interstitial graft rejection.

Antibodies, Monoclonal↗
PubMed1983

Continuous ambulatory peritoneal dialysis (CAPD): an established treatment for endstage renal failure.

This paper is a study of 117 patients with endstage renal failure, treated by continuous ambulatory peritoneal dialysis (CAPD) over periods of 1-56 months. The study has shown CAPD to be an effective form of dialysis with a number of advantages over intermittent peritoneal dialysis and hemodialysis (better control of salt and water status, hypertension and anemia, steady state biochemistry and greater ease of self-dialysis). Peritoneal clearance and ultrafiltration have remained adequate in all but a few patients. Hypoproteinemia, poor nutrition, obesity and abdominal herniae have been problems in a small percentage of patients. Hyperlipidemia has developed in half the patients but improved with diet. Peritonitis remains the major barrier to the more widespread use of CAPD, although its incidence can be considerably reduced by use of better connectors, bacterial filters and choice of patients.

Adolescent↗
PubMed1983

Monoclonal antibodies to human glomerular cells: a marker for glomerular epithelial cells.

A monoclonal antibody, PHM 5, directed against human glomerular epithelial cells, was prepared after immunisation of a mouse with isolated human glomeruli and fusion of spleen cells with a mouse myeloma. Binding of antibody to isolated glomeruli was detected by radioimmune indirect binding assay, and specificity for glomerular epithelial cells was shown using a four-layer peroxidase-antiperoxidase technique applied to epoxy resin sections of the human kidney cortex. PHM 5 appears to detect a human-specific cell surface carbohydrate antigen not previously described, and can be used to identify epithelial cells in renal biopsy sections and in culture outgrowths of isolated glomeruli.

Animals↗
PubMed1983

Membrane antigens of human cells of the monocyte/macrophage lineage studied with monoclonal antibodies.

Three monoclonal antibodies, FMC17, FMC32, and FMC33 directed against human cells of the monocyte-macrophage lineage are described. The antibodies react strongly with blood monocytes and weakly, if at all, with granulocytes. Lymphoid cells are not stained. In tissue sections macrophages and interdigitating reticulum cells are stained. Lymphoid leukemia cells generally do not react with the antibodies, while myeloid leukemia cells give a variable pattern, with relatively differentiated cells more likely to react than undifferentiated cells. Differences between the 3 antibodies in their reactivity with leukemic cells and tissue macrophages indicate that they are directed against distinct antigens, which may serve as differentiation markers in the monocyte/macrophage lineage.

Antibodies, Monoclonal↗
PubMed1983

Antigenic heterogeneity of human mononuclear phagocytes: immunohistologic analysis using monoclonal antibodies.

Eight monoclonal antibodies to cell surface antigens of human monocytes were evaluated as immunologic markers for recognition of macrophages in sections of normal and diseased tissues, using immunoperoxidase and enzyme histochemical techniques. Monoclonal antibodies assessed were PHM2, PHM3, FMC17, FMC32, FMC33, FMC34, OKM1, and 63D3. Sites studied were human bone marrow, blood, lymph node, spleen, thymus, liver, kidney, lung, and peritoneal lavages, rejecting renal allografts containing inflammatory macrophages, and granulomata showing epithelioid and multinucleate giant cell formation. All antibodies bound to at least some tissue macrophages and, except for FMC32 and FMC33 antibodies, which were identically distributed, each antibody had a distinctive tissue distribution. Some antigens were shared by other bone-marrow-derived cells (megakaryocytes and cortical thymocytes), endothelium, epithelium, and dendritic cells. Antigenic differences were also detected between mononuclear phagocytes present at different sites, different stages of differentiation, and likely different states of activation. These studies provide evidence of major antigenic differences between various populations of human mononuclear phagocytes. They therefore indicate the need for careful evaluation of experiments involving the recognition of macrophages in tissue sections and smears based solely on the use of antimonocyte monoclonal antibodies.

Animals↗
PubMed1982

A comparison of fixatives and immunohistochemical technics for use with monoclonal antibodies to cell surface antigens.

A variety of fixatives and four immunohistochemical technics were compared for demonstration of binding of monoclonal antibodies to cell surface antigens in tissue sections. Three recently described monoclonal antibodies, directed against the human leukocyte-common antigen (PHM 1) and antigens present on mononuclear phagocytes (PHM 2, PHM 3), were used. Antigen stability to fixation and heat were first assessed by quantitating the resultant effect on binding of the monoclonal antibodies to a macrophage cell line in vitro, using a radiolabeled second antibody. All fixatives affected their binding, but least effect was seen with formaldehyde-based fixatives and ethanol. Sections of formaldehyde- and ethanol-fixed lymph nodes were then labeled with monoclonal antibodies, and four developing methods were compared: indirect immunofluorescence, indirect immunoperoxidase, 3-layer heterologous peroxidase-antiperoxidase (PAP), and a 4-layer PAP technic. The 4-layer PAP proved to be the most sensitive technic since all three antibodies were localized successfully while both indirect technics only detected PHM 1, and the heterologous PAP method was not successful with any monoclonal antibody. The best combination of specific labeling and adequate morphology was seen with periodate-lysine-paraformaldehyde (PLP) fixation and the 4-layer PAP method.

Antibodies, Monoclonal↗
PubMed1982

Involvement of the macrophage in experimental chronic immune complex glomerulonephritis.

Serial renal biopsies for glomerular culture, histochemical staining for beta-glucuronidase, electron microscopy (EM) and light microscopy, were used to study macrophage involvement in experimental chronic immune complex (IC) glomerulonephritis (GN) induced in rabbits by daily intravenous injections of bovine serum albumin (BSA). In the 26 animals studied, proliferative GN of variable severity was induced, with mild disease in 5 animals, moderate proliferation in 15 and crescentic GN in 6. Macrophages first appeared in glomerular culture outgrowths during the 2nd and 3rd weeks, coincident with the onset of proteinuria and rising serum creatinine concentration. Large numbers of macrophages (in excess of 20 per glomerulus) were seen by the 5th weeks and persisted to the 9th week. The number of macrophages in outgrowths was not significantly greater in animals with crescentic disease. EM demonstrated macrophages in capillary loops, and in glomeruli with crescents, macrophages could be seen in the urinary space. Histochemical staining for beta-glucuronidase also demonstrated macrophages in the glomerular tuft and in crescents when present. These results indicate that macrophages constitute a considerable proportion of the glomerular hypercellularity seen in chronic IC glomerulonephritis.

Animals↗
PubMed1982

The immunohistochemical demonstration of major histocompatibility antigens in the human kidney using monoclonal antibodies.

The distribution of HLA-ABC and DR antigens in human kidneys was studied using monoclonal antibodies to monomorphic determinants and a 4-layer immunoperoxidase technique applied to frozen and paraffin sections. Enzyme digested paraffin sections provided the best localization. HLA-ABC antigens were located on all renal endothelium, Bowman's capsule and in proximal tubules. HLA-DR antigen was restricted to capillary and venous endothelium plus, at low density, in proximal tubules. Minor differences in distribution were found with different monoclonal antibodies, but dendritic cells were not detected using monoclonal antibodies to HLA-ABC, HLA-DR or leukocyte-common antigens.

Antibodies, Monoclonal↗
PubMed1981

Monoclonal antibodies to human macrophage and leucocyte common antigens.

Three monoclonal antibodies have been made to identify cells of the human mononuclear phagocyte system in fluids and tissues. The first, PHM 1, recognises a surface antigen common to all leucocytes. The other 2 antibodies, PHM 2 and PHM 3, bind to monocytes and macrophages but not to polymorphonuclear cells (PMN). PHM 2 labels all monocytes, macrophages and a small population of T-cells. PHM 3 labels most monocytes and macrophages but no other blood cells. The application of these monoclonal antibodies to the identification of mononuclear phagocytes in blood and liver using an unlabelled antibody immunoperoxidase (PAP) technique is demonstrated.

Antibodies, Monoclonal↗