Development of a sandwich ELISA for evaluating soluble OX40L (CD252) in human sera of different ages or with Graves' disease.
ABSTRACT OX40 ligand (CD252), a molecule originally identified as human gp34, is an important costimulatory molecule during immune response. Here, we describe a sandwich ELISA for the detection and quantification of soluble OX40L using anti-OX40L monoclonal antibodies 1G1 and 4C12 as capture and detecting antibody, respectively. With this ELISA, the existence and concentration of soluble forms of OX40L (sOX40L) was demonstrated for the first time. It was found that soluble OX40L is detectable in the sera of elderly persons (above 60 years old) and patients with Graves' disease which has the highest mean serum concentration of sOX40L, suggesting the potential diagnostic significance of sOX40L in the disease. Surprisingly, the quantitation of sOX40L was correlated with the age and among these subjects, those of 70s and 80s have much higher sOX40L concentration than those of 60s.
- Cytokine 01/1996; DOI:10.1109/CLEOE.1996.562516 · 2.87 Impact Factor
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ABSTRACT: We have previously demonstrated that normal human T cells either long-term repeatedly stimulated or freshly activated in vitro in the presence of TGF-beta express the cell surface T-cell costimulating molecule OX40 ligand (OX40L). To further elucidate the kinetics of OX40L expression by human T cells, we have examined whether cell proliferation was required for the expression of OX40L. Thus, normal fresh peripheral blood mononuclear cells were stimulated with immobilized anti-CD3 antibody in the presence of the DNA synthesis-blocking agents such as mitomycin C, 5-fluorouracil, or X-ray irradiation. Flow cytometric analyses demonstrated that a significant frequency of these DNA-damaged activated primary CD4+ and CD8+ T cells became OX40L+ as early as 1 hour after treatment. The OX40L induction on the DNA-damaged activated T cells was inhibited by treatment with either RNA or protein synthesis inhibitors, actinomycin D, or cycloheximide, respectively. Induced OX40L on T cells was functional because it bound recombinant OX40. These data indicate that human primary T cells are programmed to rapidly express functional OX40L molecules after stimulation under DNA-damaging conditions, demonstrating that the induction of OX40L by T cells is independent of cell proliferation. The clinical implications of these new findings are discussed.Human Immunology 09/2008; 69(9):533-42. DOI:10.1016/j.humimm.2008.07.001 · 2.28 Impact Factor
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ABSTRACT: Golgi protein-73 (GP73) is a newly identified candidate serum marker for liver diseases. The utility of this biomarker remains limited, largely due to the lack of quantitative information. The aims of this study were to quantify serum GP73 (sGP73) in healthy individuals and in patients with liver diseases, and to validate sGP73 as a biomarker for early diagnosis of liver disease. Recombinant GP73 was used to generate monoclonal (mAb) and polyclonal antibodies (pAb). Using these antibodies in a quantitative enzyme-linked immunosorbent assay, GP73 was measured in serum from 263 patients with various forms of liver and other diseases. The median sGP73 in patients with liver disease was significantly higher (P < 0.001) than in healthy individuals and in patients with other diseases. When sGP73 was used to detect liver disease, it had a sensitivity of 82% and a specificity of 80% at the optimal cut-off value of 85.5 microg/L. The area under the receiver-operating characteristic curve was 0.9. sGP73 concentration in patients with liver disease was three-fold higher than in healthy individuals. However, sGP73 concentrations did not differ significantly between patients from each liver disease group. Furthermore, sGP73 was not significantly elevated in patients with diseases other than liver disease compared with healthy individuals. These results suggest that sGP73 may be used as a serum marker for the diagnosis of liver disease.Annals of Clinical Biochemistry 12/2008; 46(Pt 1):38-43. DOI:10.1258/acb.2008.008088 · 2.08 Impact Factor