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Biomedical subjects

S Serke

Publications and source records attributed to S Serke.

83 records · Page 5Linked to original sources
PubMed1989

Lymphocytosis of large granular lymphocytes associated with anemia and neutropenia: proof of monoclonality of the LGL-population, but benign clinical course.

A 37-year-old man with severe transfusion-requiring anemia and neutropenia associated with lymphocytosis of large granular lymphocytes (LGLs) has been followed over a period of 3.5 years. Detailed phenotyping of the LGL has been performed, revealing a phenotype of CD3+, CD8+, CD16+, HLA-DR+, and Leu-7+. Southern-blot analysis of the T-cell receptor beta-chain locus detected a gene rearrangement, thus providing proof of monoclonality of the peripheral blood LGLs.

Adult↗
PubMed1989

A flow cytometric assay for the determination of cell proliferation with a monoclonal antibody directed against DNA-methyltransferase.

The enzyme DNA-methyltransferase is responsible for the methylation of a newly synthesized DNA-strand. A monoclonal antibody directed against DNA-methyltransferase was used to determine cell proliferation by means of flow cytometry. The reactivity of DNA-methyltransferase antibody was compared with the known proliferation markers transferrin-receptor and Ki67. All three methods showed comparable reactivity with the erythroblastic cell line K562 (86%, 81%, 76% respectively). In a second set of experiments peripheral blood mononuclear cells were stimulated with phytohaemagglutinin; in three experiments, a mean of 63% of the cells reacted with DNA-methyltransferase antibody after 72 h of culture as compared to a mean of 6% in the case of unstimulated control cells. HL60 cells were incubated with DMSO and harvested on day 5 of culture. The results obtained show that in differentiated cells the fraction positive with DNA-methyltransferase antibody decreased to levels below 10%. It is concluded that the technique described is a fast and easy method for the flow cytometric determination of cellular proliferation.

Antibodies, Monoclonal↗
PubMed1988

In situ immunocytochemical staining method for haemopoietic cells grown in vitro.

Characterization and enumeration of haemopoietic cells grown in vitro is routinely performed either on unstained cultures or on cultures stained by cytochemical agents. This report describes a novel method for the immunocytochemical analysis of cells grown in plasma clots. The stain is performed in situ by subjecting the whole plasma clot in the culture dish to the staining procedure. The growth of early haemopoietic progenitor cells was monitored in cultures from normal human peripheral blood and proliferating progenitor cells were identified with an anti-human transferrin receptor monoclonal antibody followed by a two-layer immunoperoxidase stain. The number of transferrin receptor positive clusters demonstrable after 5 days of culture was similar to the number of haemopoietic colonies detectable after 12 days of culture.

Biomarkers↗
PubMed1988

An immunocytochemical method for the detection of fluorochrome-labelled DNA probes hybridized in situ with cellular RNA.

Various detection systems for in situ hybridization of nucleic acids are currently used. We report here an immunocytochemical detection system which is based on the detection of FITC-labelled DNA/mRNA hybrids and takes advantage of FITC molecules attached covalently to the DNA probes prior to hybridization. In situ hybridization on cytocentrifuge spots is followed by the application of an anti-fluorescein antibody thus permitting detection of mRNA/DNA-FITC hybrids. The anti-FITC antibody reaction is demonstrated by an indirect immunocytochemical peroxidase-staining method. The T lymphoblast cell line Jurkat and the cDNA for the TCR-beta chain were chosen to establish the technique.

Cell Line↗
PubMed1987

Lymphocyte activation by phytohaemagglutinin and pokeweed mitogen. Identification of proliferating cells by monoclonal antibodies.

Using a two-colour immunofluorescence technique, we have investigated the mitogenic effects of phytohaemagglutinin-M (PHA-M) and of pokeweed nitrogen (PWM) on human lymphocyte subsets. These were identified by CD1, CD3, CD4, CD8, and CD16 monoclonal antibodies, and proliferation was demonstrated by a polyclonal anti-transferrin antibody. Evidence has been obtained for the generation of a population expressing both the CD4 and CD8 antigens simultaneously, in short-term cultures of peripheral blood mononuclear cells in the presence of PWM and of PHA-M.

Antibodies, Monoclonal↗
PubMed1986

Malignant fibrous histiocytoma associated with peripheral blood eosinophilia. In vitro studies demonstrating tumor-derived eosinophilopoietic activity.

Peripheral blood eosinophilia is a well-recognized paraneoplasia in many kinds of hematological and nonhematological malignancies. We report the case of a patient with a malignant fibrous histiocytoma. With increasing tumor burden, the patient developed a marked peripheral blood eosinophilia. Using an in vitro assay for growth of eosinophilic colonies, both the serum and the tumor of the patient proved to contain an eosinophilopoietic activity.

Cell Division↗
PubMedDate not supplied

Analysis of CD34-positive hemopoietic progenitor cells from normal human adult peripheral blood: flow-cytometrical studies and in-vitro colony (CFU-GM, BFU-E) assays.

Hemopoietic progenitor cells are present in minute numbers in the peripheral blood of healthy adults. By in vitro colony-assays evaluating BFU-E- and CFU-GM-growth, their numbers have been estimated to be about 1,000 per 1.0 ml of whole blood. Employing a CD34-moAb, detecting an antigen present on virtually all hemopoietic progenitor cells, and by using multiparameter flow-cytometry, we have designed a flow-cytometric method for the quantitation of CD34(+)-cells in blood. Comparative studies on bone marrow and blood have shown that CD34(+)-cells from both sources display almost identical light-scatter characteristics. They differ, however, with regard to the coexpression of the CD33- and the CD19-antigens. In vitro colony-assays for BFU-E and CFU-GM in single cultures have shown that about 25% of the CD34(+)-cells from blood were clonogenic in vitro. Our data indicate that the CD34(+)-cells from peripheral blood differ substantially from bone marrow CD34(+)-cells.

Adult↗