[Screening and analysis of genes encoding hepatocellular carcinoma associated tumor antigens].

Objectives: To screen and clone the genes encoding hepatocellular carcinoma associated tumor antigens.

Methods: A hepatocellular carcinoma cDNA express library was constructed with ZAP vector and analyzed by serological analysis of recombinant cDNA expression library (SEREX) with sera from autologous and allogenous patients. Monoclonalized positive phage clones were converted into pBK-CMV phagemid forms by in vivo excision.

[Effect of CD28 costimulator on T cell receptor(TCR)-induced apoptosis of thymocytes].

Aim: To analyze apoptosis and its degree of thymocytes at early and intermediate phases induced by anti-TCRalphabeta mAb or anti-TCRalphabeta mAb+anti-CD28 mAb stimuli, and to analyze the influence of CD28 costimulator on TCR-induced apoptosis of thymocyte subsets.

Methods: Thymocytes were freshly prepared and were cultured for 20 h in the presence of anti-TCRalphabeta mAb+anti-CD28 mAb or anti-TCRalphabeta mAb along, The cultured cells were stained with fluorescein labelled Annexin V, PI, anti-CD4 mAb and anti-CD8 mAb, then color reagents. The apoptotic cells were analyzed by FACS.

Multiple variants and a differential splicing pattern of kinectin in human hepatocellular carcinoma.

To extend the search for hepatocellular carcinoma (HCC) associated antigens with immunogenicity for clinical applications, we constructed a cDNA expression library using resected human HCC tissue sample and screened it by serological analysis of recombinant cDNA expression library (SEREX) with autologous and allogeneic sera. A total of 24 distinct antigens were isolated and kinectin was the antigen most frequently identified. We found that kinectin was alternatively spliced at four sites and obtained all eight theoretical forms of variant, six by SEREX and two by RT-PCR, from the different splicing combinations of the last three sites.

Production and secretion of calcitonin gene-related peptide from human lymphocytes.

Calcitonin gene-related peptide (CGRP) is a 37-amino acid neuropeptide, which is mainly present in primary sensory nerves. Although our previous study has shown that rat lymphocytes can synthesize beta-CGRP, there is no evidence demonstrating whether CGRP can be synthesized by human lymphocytes. In this study, the production of CGRP from human lymphocytes from spleen and blood were investigated by using CGRP-specific radioimmunoassay (RIA), and RNase protection assay (RPA).