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R J Marder

Publications and source records attributed to R J Marder.

32 records · Page 2Linked to original sources
PubMed1987

Recent progress in clinical quantitative cytology.

The last several years have seen changes in quantitative cytologic procedures that have significantly expanded the clinical research and diagnostic applications of quantitative cytologic analysis. Intact nuclei can be isolated from formalin-fixed paraffin-embedded tissue, and the DNA distribution of these nuclei can be analyzed either with flow cytometry or with slide-based image-processing techniques. In addition, more informative DNA analysis has been achieved using antibodies to bromodeoxyuridine-modified DNA, proliferation-associated nuclear antigens, or tissue-type-specific cytoplasmic antigens as second stains for two-parameter analysis. Improvements in both staining and instrumentation have also increased the usefulness of flow cytometry in analysis of "rare events," ie, cell types comprising less than 0.1% to 0.5% of a total cell suspension. Commercially available slide-based image-processing systems have been developed that provide sophisticated cell and tissue analysis functions using inexpensive personal-computer-based systems.

Antibodies↗
PubMed1986

Histiocytosis X. Flow cytometric DNA-content and immunohistochemical and ultrastructural analysis.

A 76-year-old man developed a generalized orange-red nodular eruption associated with constitutional symptoms. A biopsy specimen of a nodule revealed an extensive infiltration of histiocytes with relatively abundant cytoplasm and folded nuclei. Electron microscopy showed Langerhans' cell granules, which confirmed the diagnosis of histiocytosis X. Results of immunohistochemical studies revealed a pattern of antigen expression usually found in histiocytosis X, including Ia, T6, and S100. Analysis of the DNA content of the cells with flow cytometry revealed an aneuploid peak. The patient responded partially to topical mechlorethamine hydrochloride therapy.

Adult↗
PubMed1986

Phenotypic analysis in diffuse, large cell lymphoma. Clinical and histologic associations.

To investigate the possible relationships between immunologic phenotype, histologic subtype, and clinical features in diffuse, large cell lymphoma (DLCL), a computerized registry has been established for the prospective collection of immunologic, histologic, and clinical data. A combination of immunofluorescence and immunoperoxidase technics on single-cell suspensions, frozen tissues, and B5-fixed, paraffin-embedded specimens was used to study the first 33 biopsies. A definitive phenotype was established in all but two cases. Monoclonal antibody reagents reactive in B5-fixed, paraffin-embedded tissue sections helped assign a B-cell lineage in four cases lacking surface or cytoplasmic immunoglobulin, monoclonal light chains, and T-cell markers. There was no statistically significant association between the immunologic phenotype (whether mature B or not) and any clinical or histologic parameter, including response to therapy and survival. Bone marrow involvement was found to be associated significantly with both vague nodularity and a cleaved cell subtype. Through the use of a multifaceted approach to the immunophenotypic analysis of the DLCLs, a distinct lineage and stage of differentiation could be assigned to most biopsy specimens. That such analysis has significant clinical implications for patients with DLCL could not be demonstrated in this series.

Adult↗
PubMed1986

Prognostic implications of ploidy and proliferative activity in diffuse large cell lymphomas.

Paraffin-embedded surgical biopsies from 50 patients with newly diagnosed diffuse large cell lymphoma (DLCL) were examined for proliferative activity and DNA aneuploidy by flow cytometry. These results were correlated with the clinical characteristics of these patients and the course of their disease. High proliferative activity, defined as less than 80% of cells in G0 or G1, was found to be the single most important pretreatment adverse prognostic factor in these patients. This relationship remained significant after correcting for poor performance status and advanced Ann Arbor stage, the other factors found to be associated with a shortened survival. DLCLs with high proliferative activity were more probable to present with extranodal involvement than those with lower proliferative activity. The mitotic count as determined by light microscopy did not correlate with flow cytometry-defined proliferative activity and may be a less accurate method for assessing this important biological characteristic in DLCL. DNA aneuploidy was detected in 62% of cases but did not appear to have any prognostic significance. Biopsies from patients who presented with lymphomatous bone marrow involvement, however, invariably demonstrated an aneuploid stemline. These results suggest that differences in proliferative activity may be an important biological basis for the variable prognosis seen in DLCL.

Adolescent↗
PubMed1985

Immunohistochemical analysis of human lymphomas with monoclonal antibodies to B cell and Ia antigens reactive in paraffin sections.

Monoclonal antibodies (MoAbs) to B cell- and T cell-specific antigens uniformly have been restricted in their use to cell suspension or frozen section techniques because the antigens that they identify are either masked or lost in the fixation or paraffin-embedding processes. Antiimmunoglobulin antisera, although readily identifying cytoplasmic immunoglobulin in paraffin sections, has not been as useful as originally hoped since the majority of B cell lymphomas express surface immunoglobulins, which requires cell suspensions or frozen sections for detection. Because cell suspension procedures disrupt tissue architecture and frozen section techniques grossly distort morphology, neither method allows the combination of optimal morphologic and immunologic classification of lymphomas. We recently reported two MoAbs, LN-1 and LN-2, that react with B cells in paraffin sections. LN-1 reacts with the surface membrane and cytoplasm of germinal center B cells. LN-2 reacts uniquely with the nuclear membrane and cytoplasm of mantle zone and germinal center B cells and interdigitating histiocytes. We have also identified a new MoAb, LN-3, that reacts with the HLA-DR antigen in paraffin sections. We now report the use of LN-1, LN-2, and LN-3 in the analysis of paraffin sections from 58 non-Hodgkin's lymphomas and 15 cases of Hodgkin's disease. The types of cells reactive with these MoAbs in neoplastic lymphoid proliferations largely recapitulate their benign morphologic and immunologic counterparts. As a panel, LN-1, LN-2, and LN-3 were reactive with 98% of B cell lymphomas, and LN-1 and LN-2 were negative on all T cell lymphomas. In addition to identifying the cell of origin of these malignant proliferations, these MoAbs were also useful for identifying architectural features in neoplastic lymph nodes. Thus, these reagents provide the ability to assess the immunologic phenotype of neoplastic lymphocytes in conjunction with the critical morphologic criteria requiring paraffin embedding.

Antibodies, Monoclonal↗
PubMed1984

Immunohistochemical localization of the immunodominant differentiation antigen lacto-N-fucopentaose III in normal adult and fetal tissues.

Monoclonal antibodies were produced by immunizing rats with human small cell lung carcinoma (SCLC) cell lines. Monoclonal antibodies 600D11 and 624A12 were found to be directed against the ceramide pentasaccharide that contains the lacto-N-fucopentaose III (LNFP III) sequence of sugars, an isomer of the Lewis A blood group antigen. LNFP III is an immunodominant antigen whose reactivity is maintained in formalin-fixed paraffin-embedded sections (PS). LNFP III has been recognized in a number of human tumors including: SCLC; adenocarcinomas of the breast, gastrointestinal tract, genitourinary tract, and lung; renal cell carcinoma; neuroblastoma; and myelogenous leukemia. We now report the normal adult and fetal tissue distribution of the LNFP III antigen by immunoperoxidase staining on PS utilizing 600D11 and 624A12. Binding was demonstrated in bronchial epithelium and bronchial glands; squamous epithelium of the esophagus; gastric crypts, duodenal enterocytes and Brunners glands; argentaffin cells; jejunal and colonic goblet cells; pancreatic acinar cells; salivary glands; endocervical and exocervical cells; skin epidermis; myelinated motor fibers; cells of the adrenal medulla and anterior pituitary gland; polymorphonuclear leukocytes (PMNs); tissue macrophages and renal proximal tubules and loops of Henle. Staining was localized to cell membranes and within the cytoplasm, with greatest intensity at the apical and basal portions of the cells. These staining patterns were noted in adult and neonatal tissues, and initial expression could be traced to approximately the second trimester of fetal development. Knowledge of the normal tissue distribution of this immunodominant antigenic determinant may offer insight into its structural and functional role in benign and malignant tissues.

Adolescent↗
PubMed1984

Assay, purification and further characterization of 7S C1q-precipitins (C1q-p) in hypocomplementemic vasculitis urticaria syndrome and systemic lupus erythematosus.

C1q-precipitins (C1q-p) are comprised of 7S IgG with C1q-binding activity found in sera of patients with hypocomplementemic vasculitis urticaria syndrome ( HVUS ) and systemic lupus erythematosis (SLE). We have utilized C1q-coated polystyrene beads to selectively isolate C1q-p and to establish a sensitive and quantitative assay of C1q-p and immune complex activity. Purified C1q-p was comprised of polyclonal IgG which retained 7S sedimentation and solid phase C1q-binding activity at physiological ionic strength both in the presence and absence of normal human sera. No precipitation interaction was observed between C1q-p and fluid-phase C1q or C1 under the conditions tested. Purified C1q-p had no activity in the Raji cell immune complex activity. C1q-p activity also was observed in and purified from SLE serum; this activity was distinguishable from 7S immune complex activity detected by Raji cells which was also present in SLE serum. These studies indicate that C1q-p is a 7S IgG molecule found in HVUS as well as some SLE sera and has activity in C1q-binding but not in Raji cell-binding immune complex assays. These data also suggest that C1q-p is a monomeric, polyclonal IgG with preferential affinity for bound C1q. In addition to its potential role in immune complex disease, C1q-p may also provide an important tool for studying the interaction of immunoglobulin and C1q, and should contribute important information to understanding the pathobiology of immune complex disease.

Antigen-Antibody Complex↗
PubMed1984

Two new monoclonal antibodies (LN-1, LN-2) reactive in B5 formalin-fixed, paraffin-embedded tissues with follicular center and mantle zone human B lymphocytes and derived tumors.

Two monoclonal antibodies (LN-1, LN-2) reactive with B lymphocytes in B5 formalin-fixed, paraffin-embedded tissue sections have been produced by utilizing cell extracts from pokeweed mitogen-stimulated peripheral blood lymphocytes and diffuse histiocytic lymphoma SU-DHL-4 cells, respectively. Both monoclonal antibodies were initially identified by indirect immunofluorescence screening techniques on paraformaldehyde-acetone-fixed cell preparations. Specificity screens with 36 well-characterized human lymphoma and leukemia cell lines showed that both LN-1 and LN-2 stained cell lines of B cell lineage but were unreactive with those of T cell or, with one exception, myeloid derivation. Null cell acute lymphoblastic leukemia cell lines were found to be LN-2+ but LN-1-. The B cell specificity of these reagents was confirmed on 15 lymphoma and 17 leukemia biopsy specimens by using indirect immunofluorescence techniques. Immunoperoxidase staining of sections from B5-fixed, paraffin-embedded human lymphoid tissues showed that LN-1 bound to the cell membrane and cytoplasm of germinal center cells whereas LN-2 stained the nuclear membrane and cytoplasm of germinal center and mantle zone B lymphocytes as well as interfollicular histiocytes and thymic medullary dendritic cells. Both monoclonal antibodies failed to stain cortical thymocytes, lymph node T cells, and peripheral blood T and myeloid cells. Immunoperoxidase staining of 20 nonlymphoid human organs and tissues revealed that LN-1 reacted positively with red blood cell precursors of the bone marrow, ciliated epithelial cells of the bronchus, distal tubular cells of the kidney, and ductal cells from several organs including the breast and prostate. In contrast, LN-2 was unreactive with all human nonlymphoid organs and tissues including the bone marrow. Indirect immunofluorescence staining of a panel of 26 solid tumor cells lines showed that LN-1 was reactive with the majority of epithelium-derived cell lines, glioblastomas, and astrocytomas but was unreactive with neuroblastomas, small cell carcinoma of the lung, and sarcomas. LN-2 was unreactive with 25 of 26 of the solid tumor cell lines by these techniques. Immunobiochemical studies have shown that LN-1 recognizes a cell surface sialoantigen whereas LN-2 is directed against a 35,000 dalton nuclear membrane protein. Because of their high specificity for B cell tumors and their ability to stain B5-fixed, paraffin-embedded tissues, LN-1 and LN-2 are useful reagents for the diagnosis and classification of the human lymphomas and leukemias.

Antibodies, Monoclonal↗
PubMed1980

A hypocomplementemic vasculitic urticarial syndrome. Report of four new cases and definition of the disease.

We describe four new patients with a unique syndrome of persistent urticaria, with leukoclastic angiitis, severe angioedema, occasional life-threatening laryngeal edema, arthritis, arthralgia, neurologic abnormalities and pronounced persistent hypocomplementemia. The complement abnormalities involved markedly reduced levels of the Clq subunit of the first component of complement (Cl) in the presence of near normal levels of Clr and Cls subunits of Cl; modest to marked depletion of the fourth component of complement (C4), the second component of complement (C2) and the third component of complement (C3); and normal levels of the fifth through ninth components of complement (C5 through C9) and properdin factors B and D. A striking serologic abnormality found in all patients was the presence of low molecular weight (7S) proteins which precipitated with Clq in agarose gels; these previously were shown to be comprised at least in part of immunoglobulin G. The present experience is offered to help to define the clinical, histopathologic and serologic characteristics of this entity, designated hypocomplementemic vasculitic urticarial syndrome, and to emphasize its distinctiveness and prevalence.

Adult↗
PubMed1978

Low molecular weight C1q-precipitins in hypocomplementemic vasculitis-urticaria syndrome: partial purification and characterization as immunoglobulin.

A lupus-like syndrome involving chronic urticaria with cutaneous vasculitis, systemic symptoms, hypocomplementemia with preferential depletion of C1q, and low m.w. (7S) C1q-precipitins has recently been defined. The C1q-precipitin activity (C1q-p) seems to represent a diagnostic marker of the disease, but its chemical nature is not yet clear. We have partially purified and characterized C1q-p from the serum of two patients with this syndrome and compared its activity with the C1q-precipitating activity of aggregated human gamma-globulin (AHGG) anti-C1q antibodies, and several polynucleotides including DNA and polyinosinic acid. C1q-p was found to partition with IgG during precipitation by ammonium sulfate and low ionic strength buffer as well as during column chromatography on DEAE-cellulose and G-200 Sephadex. Like AHGG, but in complete contrast to the polynucleotides, the C1q-precipitating activity of C1q-p was sensitive to pepsin, trypsin, and acidic conditions, but unaffected by DNAse or RNAse; the C1q-precipitating activity of anti-C1q antibody was not diminished by any of these procedures. Thus, C1q-p consists of gamma-migrating protein of low m.w., and its C1q-precipitating activity is indistinguishable from that of AHGG. These results are consistent with the concept that C1q-p is comprised, at least in part, of IgG that binds C1q via the Fc portion of the molecule.

Binding, Competitive↗
PubMed1976

C1q deficiency associated with urticarial-like lesions and cutaneous vasculitis.

We describe a 46 year old women with a seven year history of urticarial-like symptoms and cutaneous vasculitis with marked deficiency of C1q in the presence of normal levels of C1r and C1s and high titers of low molecular weight (7S) C1q precipitins. Hemolytic C1 activity, which was greatly reduced, was restored upon the addition of purified C1q. The other complement components were present in moderately reduced or normal levels. This patient bears resemblance to several other persons previously described with urticarial-like lesions in association with selective deficiency of C1q. The similarity of the clinical features, pathologic diagnosis, C1q levels disproportionately deficient in relation to other complement components and low molecular weight C1q precipitins support the conclusion that these are causally related in a symptom complex. The underlying basis is yet to be defined.

Complement C1↗
PubMedDate not supplied

The clinical evaluation of plasma fibronectin as a marker for nutritional depletion and repletion and as a measure of nitrogen balance.

Plasma fibronectin has been suggested as a possible marker for nutritional repletion or depletion. This study was undertaken to evaluate the usefulness of plasma fibronectin in patients who received intense nutritional support. Twenty-seven patients referred to our Nutritional Support Services were followed for 3 to 5 wk; 22 received parenteral hyperalimentation alone, two received enteral alone, and three received a combination of both. Plasma fibronectin, serum albumin, serum transferrin, total lymphocyte counts, and 24-hr urine nitrogen balance studies were performed weekly; anthropometric measurements were performed every other week. Plasma fibronectin concentration, measured by laser nephelometry, showed a significant rise (p less than 0.005) in all patients after 1 wk of nutritional therapy; however, there was no significant difference among the subsequent weeks. Plasma fibronectin did not correlate with nitrogen balance studies, serum albumin, or total lymphocyte counts. A correlation between serum transferrin and plasma fibronectin was found not to be clinically useful. Thus, plasma fibronectin is sensitive to nutritional repletion after 1 wk of therapy, but is not useful thereafter. The relationship among nutritional status, immunologic function, plasma fibronectin, and other serum proteins are discussed.

Adult↗