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Biomedical subjects

C S Xu

Publications and source records attributed to C S Xu.

6 recordsLinked to original sources

Increase in plasma thrombomodulin and decrease in plasma von Willebrand factor after regular radiotherapy of patients with cancer.

In this study, we investigated plasma levels of thrombomodulin (TM) and von Willebrand factor (vWF) in 51 patients suffering from cancer or tumor undergoing 60 cobalt radiotherapy. Plasma TM and vWF antigen were measured by immunoradiometric assay and ELISA, respectively. During radiotherapy, an increase in plasma TM in patients was observed, which was radiation-dose dependent and there was a positive correlation between plasma TM level and radiation doses. However, the level of plasma vWF in the patients was decreased during radiotherapy and there was an inverse correlation between the amount of plasma vWF and radiation doses. Our data indicate that plasma TM is an useful molecular marker for early detection of radiation injury to endothelial cells in patients undergoing radiotherapy.

Adolescent

Detection of a Thy-1 precursor in human T lymphoma cell lines.

In an attempt to gain insight into the biosynthesis and processing of human Thy-1, rabbit antisera (R alpha TP) were raised against a synthetic peptide (TP) of 13 amino acid residues of identical amino acid sequence to that of residues 110-122 of the putative Thy-1 precursor deduced from the cDNA sequencing. Immunoblotting assay showed that the IgG fraction of R alpha TP (R alpha TP-IgG) recognized the synthetic peptide without demonstrable cross-reactivity with isolated human brain Thy-1 and detergent-solubilized membrane proteins of human brain cells. Immunohistochemical studies showed that both R alpha TP-IgG and HB-2S-1 (a monoclonal antibody which reacts with both membrane-bound Thy-1 on viable cells and detergent-solubilized Thy-1) stained cells of two human T lymphoma cell lines (HUT-78 and HUT-102) but they did not stain cells of a human B lymphoma cell line (Raji). In contrast, in cell surface indirect immunofluorescence assay, HB-2S-1 reacted with HUT-78 and HUT-102 cells while R alpha TP-IgG did not. Taken together, these data indicate the existence of a precursor form of human Thy-1 which is processed prior to anchoring to the cell surface.

Antigens, Surface

Detection of Thy-1 on cell surface of human T lymphoid cell lines by a monoclonal antibody.

A mouse IgG2b(kappa) monoclonal antibody (MAb) HB-2S-1 against human brain Thy-1 was secreted by a hybridoma clone after fusion of mouse myeloma cells with spleen cells from a mouse that went through a prolonged immunization procedure before fusion. When tested against isolated human Thy-1 by the enzyme-linked immunosorbent assay (ELISA), MAb HB-2S-1 in culture supernatant showed a titer of over 100,000, and a titer of over 10 million in ascites of a mouse injected with the hybrid clone. By immunoblotting, this antibody was found to bind a doublet of protein bands of approximately 25,000 daltons among all proteins solubilized by deoxycholate (DOC) from membrane of human brain cells. When tested on human lymphoid cell lines by immunofluorescence, MAb HB-2S-1 strongly stained four T lymphoma cell lines, C91-Pl, HUT-102, HUT-78, and C5-MJ; and weakly two leukemia cell lines, MOLT-3 and Jurkat(clone E6-1). It did not stain a third T leukemia cell line, CCRF-CEM; a human B cell line, Raji; a plasmacytoma cell line, HMy2; or a myelomonocytic cell line, HL-60. Peripheral blood lymphocytes from ten normal human adults and the viable T cells isolated from another normal individual were also negative.

Animals

Identification and characterization of Thy-1 homologues from bovine thymocytes.

Two forms of Thy-1 homologues of apparent mol. wt of 25,000 (designated BTp25) and 45,000 (designated BTp45) were isolated from bovine thymocyte membrane by solubilization, affinity chromatography with Con A, and preparative SDS-PAGE. Both forms reacted with a rabbit antiserum to murine Thy-1 in an enzyme-linked immunosorbent assay (ELISA). BTp45 is most likely a dimer of BTp25, since the two are indistinguishable in their amino acid compositions. Comparison of amino acid compositions of BTp25 and BTp45 to that of rodent and human Thy-1 by the S delta Q index revealed significant relatedness among these molecules. BTp25 and BTp45 demonstrate more structural homology to rodent Thy-1 than to human Thy-1. Detailed chemical analyses indicate that bovine Thy-1 homologues contain neutral sugars and fatty acids covalently bound to the polypeptide chain; therefore, they are lipoglycoproteins.

Amino Acids

Release of Thy-1 from human and murine T-cell lines by a specific phospholipase.

Proteins on the outer surface of cultured human and murine lymphoblastoid T cells were labelled with 125I. The labelled cells were incubated with the enzyme phosphatidylinositol-specific phospholipase C (PI-PLC). Proteins cleaved from the cell membrane by the enzyme were immunoprecipitated with anti-Thy-1 antibodies, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and identified by autoradiography. A doublet of Thy-1 bands of approximately 16,000 daltons were detected. The result suggests that: Thy-1 is present on the human and murine T cells which we tested, and Thy-1 is attached to the cell membrane via a phosphatidylinositol domain.

Animals