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Biomedical subjects

B S Wilson

Publications and source records attributed to B S Wilson.

At least 91 records · Page 5Linked to original sources
PubMed1985

A rapid immunostaining method utilizing preformed antibody-avidin-biotin-peroxidase complexes.

A rapid immunostaining method utilizing a preformed antibody-avidin-biotin-peroxidase complex that produces staining in paraffin sections and in frozen sections of tissues in 15 minutes is described. Although the complex did not produce staining with the common leukocyte monoclonal antibody, consistent staining was seen with most antibodies used. The complex gave consistent and reproducible staining for at least three months when maintained in storage at 4 degrees C. The results show that many available antibodies can be adapted to a rapid one-step staining procedure employing the avidin-biotin-peroxidase complex system.

Adrenal Glands↗
PubMed1985

Monoclonal antibodies to the 140,000 mol wt glycoprotein of B lymphocyte membranes (CR2 receptor) initiates proliferation of B cells in vitro.

Several mouse monoclonal IgG antibodies (AB1, AB2, AB3, and AB5) were developed that reacted with a 140,000 mol wt glycoprotein on the surface of cultured RAJI B lymphoid cells. The antibodies reacted with purified normal human peripheral blood B cells and CLL Ig+ B cells and showed specific germinal center and mantle zone staining in tissue sections of secondary lymphoid organs. Immunodepletion studies using 125I surface-labeled Raji cell membrane antigens demonstrated that the antigen identified by AB5 is the same 140,000 mol wt glycoprotein detected by anti-B2 that has recently been shown to react with the C3d fragment or CR2 receptor. (Iida et al: J Exp Med 158:1021, 1983). Addition of the AB series and anti-B2 monoclonal antibodies to cultures of purified human peripheral blood B cells resulted in the uptake of 3H-thymidine at two to six times background control levels provided that irradiated autologous T cells were added to the culture. Stimulation was not evoked by other monoclonal antibodies to B cell surface molecules (ie, B1, BA-1, BA-2, and HLA-DR). Pepsin-generated F(ab')2 fragments of anti-CR2 antibodies were essentially as effective as the intact IgG molecule in stimulating B cells. Induction of B cell proliferation by antibody binding to CR2 suggests that the C3d receptor may have an integral role in regulation of humoral immune response.

Animals↗
PubMed1985

Immunohistochemical localization of chromogranin in human hypophyses and pituitary adenomas.

Chromogranin was demonstrated by immunohistochemistry in the cytoplasm of human beta-thyrotropin, human beta-follicle-stimulating hormone-, human beta-luteinizing hormone-, and human alpha-subunit-containing cells of the non tumorous human adenohypophyses. Some surgically removed human beta-thyrotropin-, human beta-follicle-stimulating hormone-, human beta-luteinzing hormone-, and human alpha-subunit-producing pituitary adenomas, as well as some null cell-adenomas, exhibited chromogranin immunoreactivity, whereas adenomas storing human growth hormone, human prolactin, or corticotropin were negative. Chromogranin immunopositivity was variable in extent and intensity; not every glycoprotein-producing cell could be immunostained in the nontumorous adenohypophysis and the majority of chromogranin-containing adenomas showed only focal positivity. No explanation can be offered for this variability. The demonstration of chromogranin by the avidin-biotin-peroxidase technique may be helpful in the immunohistochemical characterization of some glycoprotein hormone-producing pituitary adenomas, as well as null-cell adenomas of the human pituitary.

Adenoma↗
PubMed1985

Distribution of chromogranin and S100 protein in normal and abnormal adrenal medullary tissues.

The distribution of chromogranin and S100 protein was studied in 30 adrenal pheochromocytomas and 19 normal adrenal medullary tissues. Immunostaining in the tumors was compared with staining in sections of histologically normal medullae. Chromogranin showed diffuse cytoplasmic staining in all chromaffin cells. Chromogranin staining was consistently more intense in normal medullae, while less intense staining was present in most tumors from all four groups. S100 protein was present in the cytoplasm and nuclei of sustentacular cells surrounding chromaffin cells and in nerve branches. Many S100 protein-positive cells were present in normal medullae, in the two hyperplastic medullae, and in pheochromocytomas from patients with multiple endocrine neoplasia, type 2. Very few sustentacular cells were present in the other pheochromocytomas. These results indicate that S100 staining may be helpful in separating pheochromocytomas in patients with multiple endocrine neoplasia, type 2 disease from benign and malignant sporadic tumors.

Adrenal Medulla↗
PubMed1985

Detection of HLA-DR antigens in paraffin-embedded thyroid epithelial cells with a monoclonal antibody.

The human Class II major histocompatibility (MHC) antigens, or Ia antigens, which are thought to regulate immune cell interaction, can be detected in paraffin-embedded tissues by immunoperoxidase staining with a recently developed monoclonal antibody (LK8D3). HLA-DR antigens were observed in lymphoid tissues, Langerhans cells of the skin, some epithelial cells, and pulmonary alveolar macrophages. The expression of HLA-DR antigens was analyzed in formalin-paraffin sections by immunoperoxidase in 86 normal and abnormal thyroid epithelial tissues. All patients with Hashimoto's disease (8/8) and most patients with Graves' disease (6/8) expressed HLA/DR antigens in the thyroid epithelial cells and in adjacent inflammatory cells. Most papillary carcinomas (12/18), including 3 of 5 follicular variant of papillary thyroid carcinomas, had HLA-DR antigens detected in epithelial cells; whereas medullary thyroid carcinomas (0/5), follicular carcinomas (0/5), and multinodular goiters (0/4) did not have detectable HLA-DR immunoreactivity. A few other thyroid lesions had HLA-DR antigens detected in epithelial cells, including anaplastic carcinomas (2/5), Hurthle-cell tumors (1/16), and thyroid lymphomas (2/2). Monoclonal antibody LK8D3 and two other commercially available monoclonal antibodies against HLA-DR-stained tissues equally well in cryostat sections, but only antibody LK8D3 was effective in formalin-fixed paraffin-embedded tissue sections. These results indicate that epithelial cells from thyroids of patients with autoimmune diseases commonly express HLA-DR antigens. The presence of HLA-DR antigens in most papillary thyroid carcinomas may be helpful diagnostically in cases of follicular variants of papillary carcinomas. The role of HLA-DR expression in autoimmune thyroid disease and in papillary thyroid carcinoma remains to be determined.

Antibodies, Monoclonal↗
PubMed1985

Immunocytochemical characterization of a monoclonal antibody that recognizes mitosing cells.

The isolation and characterization of a monoclonal antibody (C5F10) which identifies dividing cells in normal and neoplastic tissues (carcinomas, sarcomas, and lymphoreticular malignancies) in formalin-fixed, paraffin-embedded sections is described. The antibody also recognizes rapidly dividing cells in normal and transformed cells in culture. A combination of autoradiography with 3H-thymidine and immunochemical localization of C5F10 showed that cells in S-phase of the cell cycle were weakly stained for C5F10, while dividing cells stained intensely with this antibody. The target structure of C5F10 appears to be different from the commonly recognized microtubule, intermediate filament and microfilament proteins and from the proliferating cell nuclear antigen (cyclin). This antibody may be a useful tool for readily detecting dividing cells in cell cultures and in tissue sections and may prove useful in studies to analyze the molecular basis of cell growth.

Animals↗
PubMed1984

Immunohistochemical detection of chromogranin and neuron-specific enolase in pancreatic endocrine neoplasms.

Twenty pancreatic endocrine tumors and one duodenal somatostatinoma were stained with polyclonal antisera against eight hormones, neuron-specific (NSE), and with a monoclonal antibody against the endocrine granule protein chromogranin. Silver staining was performed on all tumors with the Grimelius method. All the tumors and all cells in the normal pancreatic islet showed NSE immunoreactivity. Chromogranin immunoreactivity was found predominantly in the glucagon cells of normal islets, in one glucagonoma, four gastrinomas and in 10 tumors associated with multiple hormonal production, but not in five insulinomas or in one somatostatinoma. Grimelius reactivity was identical to chromogranin immunoreactivity in all cases except that more cells in the tumors and in the normal islets were positive for chromogranin. Both chromogranin and NSE were detected in a metastasized pancreatic endocrine tumor which had metastasized to a peripancreatic lymph node. The metastatic tumor failed to stain with antisera to all eight hormones. These results indicate that chromogranin and NSE are excellent general markers for pancreatic endocrine tumors and that the Grimelius stain and other related argyrophilic reactions may involve binding to chromogranin.

Antibodies, Monoclonal↗
PubMed1984

Expression of Ia-like (DR) antigen on human alveolar macrophages isolated by bronchoalveolar lavage.

Alveolar macrophages isolated by bronchoalveolar lavage from nonsmoking patients with sarcoidosis, interstitial pulmonary fibrosis (DIF), and normal volunteers were examined for surface expression of Ia-like (DR) antigen using a monoclonal antibody technique. We found increased percentages of alveolar macrophages expressing DR antigen in the 8 patients with sarcoidosis (77 +/- 3%) and the 7 patients with DIF (78 +/- 3%) over that in the 9 healthy, nonsmoking volunteers (21 +/- 5%, p less than 0.05). Four patients, 2 with DIF and 2 with sarcoidosis, were studied before and 3 months after initiation of therapy. A decrease in DR antigen expression was found to be associated with clinical improvement. Alveolar macrophages from 3 patients were studied serially in culture, with a steady decrease in DR antigen expression with time. We conclude (1) that alveolar macrophages from patients with sarcoidosis and DIF are in a state of heightened activity, and (2) that DR antigen expression may be an indicator of disease activity.

Adult↗
PubMed1984

Acute lung injury in rat caused by immunoglobulin A immune complexes.

Mouse IgG and IgA, with reactivity to dinitrophenol conjugated to carrier protein, have been isolated from myeloma proteins by means of a variety of affinity techniques. The IgA was predominantly in the dimeric form. The in vitro and in vivo biological activities of IgA-containing immune complexes were assessed in the rat. IgA-containing immune complexes were demonstrated, in a dose-dependent manner in vitro, to activate neutrophils and to generate O.-2. In addition, these immune complexes showed evidence of complement activation in vitro, by the use of immunofixation techniques. When IgA was instilled into the airways of rats and antigen was injected intravenously, acute lung injury occurred, as reflected by increases in lung permeability and morphological changes consisting of blebbing of endothelial cells, intra-alveolar hemorrhage, and fibrin deposition. The lung changes were directly proportional to the amount of IgA instilled into the airways and failed to occur if intravenous injection of antigen was omitted. Lung injury did not occur in animals that received an intravenous injection of antigen in the absence of an airway injection of IgA. Lung injury related to IgA-containing immune complexes was complement dependent but neutrophil independent. In companion studies with mouse IgG-containing immune complexes, acute lung injury also occurred and had morphological features similar to those associated with IgA-induced lung injury except that, in the case of IgG immune complex-induced damage, neutrophils were more evident. Acute lung injury induced by IgG-containing immune complexes, whether of mouse or rabbit origin, was complement and neutrophil dependent. The similarities and differences between IgG- and IgA-associated acute immune complex-induced injury of rat lung were reinforced by the use of morphometry techniques. Studies with another monoclonal IgA antibody-containing antigen-binding activity to phosphorylcholine also demonstrated the ability of IgA antibody to cause acute lung injury in the rat. Neither antibody alone nor antigen (phosphorylcholine linked to bovine serum albumin) alone produced evidence of lung injury. These studies indicate for the first time that immune complexes containing IgA have lung-damaging properties and that the pathogenic mechanisms are different from those associated with IgG-associated immune complex-induced acute lung injury.

Animals↗
PubMed1984

Pituitary adenomas. An immunohistochemical study of hormone production and chromogranin localization.

Tumors from 42 surgically resected pituitaries and from 13 autopsy cases were studied immunohistochemically with polyclonal antisera to 7 anterior pituitary hormones and with a newly developed monoclonal antibody directed against human chromogranin for evaluation of the distribution of chromogranin in normal and neoplastic pituitaries. In addition, a prospective study was done for assessment of the prevalence, morphology, and endocrine cell types of pituitary tumors in 100 autopsy subjects. When these 55 pituitary adenomas were examined with monoclonal antibody (LK2H10) directed against human chromogranin, selective staining of normal adenohypophyseal cell types and pituitary tumors was observed. Most null-cell adenomas (12/14) were positive for chromogranin, whereas all prolactin (PRL)-producing adenomas (19/19) were negative. Growth hormone (GH) adenomas were focally positive (9/9). All oncocytomas (2/2), 1 thyrotropin (TSH) adenoma, and a follicle-stimulating hormone/luteinizing hormone adenoma were positive for chromogranin. One or more adenomas were present in 14% of the autopsy cases. The tumors occurred most frequently in patients in the fifth through the seventh decades of life. Immunohistochemical staining of 13 adenomas revealed 1 TSH, 1 ACTH, and 4 PRL-producing tumors, whereas 7 other tumors, which were null-cell or undifferentiated adenomas, failed to stain for any of the seven principle pituitary hormones. These results indicate that antibody LK2H10 to human chromogranin is useful in the immunohistochemical characterization of pituitary adenomas. Incidental pituitary microadenomas from autopsy-derived pituitaries most commonly produce PRL, or they belong to the null-cell or undifferentiated tumor group.

Adenoma↗
PubMed1984

Immunoperoxidase staining for Ia-like antigens in paraffin-embedded tissues from human melanoma and lung carcinoma.

The human Ia-like antigens that are predominantly expressed by cells associated with immunologic function has been considered as a diagnostic marker of malignant transformation of some nonlymphoid tissues. Immunoperoxidase staining of formalin-fixed and paraffin-embedded tissue sections with a monoclonal antibody to Ia-like antigens was chosen for assessment of the value of this marker for diagnosis in surgical pathology. Monoclonal antibody LK8D3 developed against a human melanoma cell line bearing Ia-like antigens was found to react in serologic and immunochemical studies with an antigenic determinant of Ia-like antigens that was relatively stable to formalin fixation and paraffin embedding. Avidin-biotin complex peroxidase staining of formalin-paraffin sections with LK8D3 showed focal expression of Ia-like antigens in 3 of 12 melanomas, whereas all 8 cases of intradermal nevi were negative. Immunoperoxidase staining of formalin-paraffin sections of lung carcinomas with antibody LK8D3 was related to the histologic subtype of tumors. Thus, squamous cell carcinomas showed only very focal staining for Ia-like antigens in 5/9 cases, while widespread and intense Ia-like immunoreactivity was seen in 3/5 cases of lung adenocarcinomas, including two bronchioalveolar carcinomas. The presence of Ia-like antigens in lung adenocarcinoma may not be entirely associated with malignant transformation, because normal alveolar lining cells were stained with the antibody.

Adenocarcinoma↗
PubMed1984

Detection of chromogranin in neuroendocrine cells with a monoclonal antibody.

A monoclonal antibody ( LK2H10 ) produced against a human pheochromocytoma reacted immunohistochemically with 126 normal and neoplastic endocrine tissues with secretory granules which were formalin-fixed and paraffin-embedded. Antibody LK2H10 did not react with 46 other endocrine tissues or tumors without secretory granules nor with 113 normal and neoplastic nonendocrine cells and tumors. Tumors with abundant secretory granules showed intense and diffuse staining, and tumors with few granules, such as Merkel cell carcinomas, neuroblastomas, and small cell carcinomas of lung, showed focal staining. Antibody LK2H10 did not react with melanomas, nevi, posterior pituitary, peripheral nerve tissues, or neurons. The target structure of LK2H10 was identified as human chromogranin, of which the major fraction was chromogranin A (mol wt 68,000 daltons). Preabsorption with purified chromogranin A blocked immunoperoxidase staining by LK2H10 in normal adrenal medulla, in the anterior pituitary, and in a pheochromocytoma. Ultrastructural immunohistochemistry with LK2H10 showed that chromogranin was present in cytoplasmic secretory granules. These results indicate that chromogranin is widely distributed in the secretory granules of most polypeptide-producing endocrine tissues, and it is readily detected with the use of monoclonal antibody LK2H10 . The detection of this marker can be very helpful as a diagnostic aid for neuroendocrine cells and tumors.

Adrenal Gland Neoplasms↗
PubMed1983

Specific endocrine tissue marker defined by a monoclonal antibody.

One of two mouse monoclonal antibodies (LK2H10) produced by hybridoma technology against a human endocrine tumor (pheochromocytoma) demonstrated specific immunoreactivity for 69 normal and neoplastic endocrine and tissues known to contain secretory granules. This immunoreactivity was specific, since other normal tissues, tumors from endocrine cells without granules, and tumors from other nonendocrine tissues were negative when tested with antibody LK2H10. The antibody reacted with human fetal adrenal medulla and human pancreatic endocrine cells and with adrenal medullary cells from monkeys and pigs. The antigen detected by antibody LK2H10 is associated with cytoplasmic secretory granules, has an estimated molecular weight of 68,000, and may be related to human chromogranin.

Antibodies, Monoclonal↗
PubMed1983

Immunochemical characterization of a human high molecular weight--melanoma associated antigen identified with monoclonal antibodies.

Sodium dodecyl sulfate polyacrylamide gel analysis of a high molecular weight (HMW) human melanoma associated antigen (MAA) defined by murine monoclonal antibodies revealed a number of distinct polypeptides ranging from 80,000 up to 280,000 daltons, in addition to an extremely heterogeneous group of components distributed over a wide range in apparent molecular weight (300,000-700,000 daltons). The 280,000 dalton and the larger heterogeneous molecular weight material are glycosylated since they are labeled with 3H-sugars. The HMW-MAA is readily solubilized in the absence of detergents and the entire series of polypeptides fractionates together in the void volume of a Sephadex G200 column. Peptide maps of the various polypeptides of the HMW-MAA, generated by Staphylococcus aureus V-8 protease, are essentially the same except that some of the proteolytic fragments derived from the lower molecular weight polypeptides (80,000 daltons) are present in greater amounts than are similar fragments derived from the larger molecular weight polypeptides; the latter finding suggests that the complexity in molecular weight of the MAA may reflect combinations of several base subunits. Proteolytic cleavage of the HMW-MAA generates a number of peptides ranging in molecular weight from 77,000 daltons to less than 12,000 daltons, which still react with monoclonal antibodies and can distinguish monoclonal antibodies specific for different antigenic determinants of this MAA.

Antibodies, Monoclonal↗
PubMed1982

Heterogeneity of human melanoma-associated antigens defined by monoclonal antibodies and conventional xenoantisera.

Immunochemical analysis of cultured human melanoma cell detergent extracts and spent culture medium with conventional xenoantisera and monoclonal antibodies identified four types of 94,000 (94K) dalton molecules and two types of high-molecular-weight melanoma-associated antigens by the following characteristics: (1) association with other components, (2) mobility in SDS-PAGE under reducing and nonreducing conditions, (3) antigenicity, and (4) presence in spent culture medium. Conventional xenoantisera were found to contain antibody populations to antigenically distinct structures, some of which have similar apparent molecular weights. Immunodepletion studies showed that the antigenic determinant detected by the monoclonal antibody 225.28S to a high-molecular-weight melanoma-associated antigen was expressed on a subpopulation of the antigens defined by the conventional xenoantiserum #8995. These data prove that antibodies reactive with antigens of similar molecular weight cannot be assumed to identify the same structures, and indicate that tumor-associated antigens may be heterogeneous in the expression of antigenic determinants defined by monoclonal antibodies.

Animals↗
PubMed1982

Tissue distribution, molecular profile, and shedding of a cytoplasmic antigen identified by the monoclonal antibody 465.12S to human melanoma cells.

The mouse immunoglobulin G2 monoclonal antibody (MoAb) 465.12S reacts with a cytoplasmic antigen present in human melanoma cells but not detectable in melanocytes. Indirect immunofluorescent staining of a large number of surgically removed normal adult and fetal tissues with the MoAb 465.12S detected the cytoplasmic antigen in epithelial cells from several organs. The intensity of staining was greater in adult tissues than in the corresponding fetal tissues. Furthermore, the MoAb 465.12S stained nearly all of the surgically removed tumors tested but did not stain many of the normal tissues from which they originated. In almost all cases, the intensity and frequency of staining wa greater for tumor cells than for corresponding normal tissues. From cultured carcinoma and melanoma cells, the MoAb 465.12S immunoprecipitated four glycopolypeptides with molecular weights of 94,000, 75,000, 70,000, and 25,000. Incorporation of 3H-labeled sugars into the various components of the cytoplasmic antigen revealed that the M.W. 75,000 component was unusual in that it contained only glucosamine and mannose. The antigenic determinant defined by the MoAb 465.12S appears to be protein rather than carbohydrate in nature since it is heat sensitive and is expressed on the antigens synthesized by cells in presence of tunicamycin. Analysis of the spent culture medium of carcinoma and melanoma cell lines revealed that the cytoplasmic antigen is readily shed by these cells and consists of a major M.W. 94,000 and a minor M.W. 72,000 component. Treatment of cultured melanoma cells with the antibiotic tunicamycin showed that glycosylation of the cytoplasmic antigen is required for its shedding and/or stability in the spent culture medium.

Antibodies, Monoclonal↗