[Expression and antigen binding activity of anti-human CD86 diabody].

Aim: To construct a recombinant eukaryotic expression vector of anti-human CD86 diabody gene and express anti-CD86 diabody through Chinese hamster ovary (CHO) cells, then analyze the capability of the diabody to recognize the tumor cells expressing CD86 and its biological effect.

Methods: The antibody heavy and light chain viable region gene (V(H); and V(L);) were cloned from hybridoma cell 1D1 which secreted anti-human CD86 monoclonal antibody. Anti-human CD86 diabody gene V(H);-(GGGGS)-V(L); was constructed by SOE-PCR.

[Effect of CD80-scFv on recognition of CD80 molecule and proliferation of different tumor cells in vitro].

Aim: To prepare CD80-scFv in CHO cells and investigate its effect on the recognition and proliferation of different tumor cells.

Methods: The CD80-scFv was purified from culture supernatant without fetal calf serum (FCS) by IMAC affinity chromatography, and then bound to CD80 molecule on the Daudi, U251, A375 and 8266 cells detected by flow cytometry. Different concentrations of CD80-scFv were added in the training system of different tumor cells, and we analyzed the influence on the proliferation of these cells by MTT assay.

[Changes in the expression of T lymphocyte subsets in ageing mice induced by D-galactose].

Aim: To explore the changes and significance of spleen T cell subsets in ageing mice induced by D-galactose.

Methods: Ageing mice model was successfully established by 100 g/L D-galactose. The content of IFN-γ and IL-4 in serum was measured by ELISA.

[The changes of spleen B cells of D-galactose-induced aging mice].

Aim: This paper is to analyze the changes of important membrane type molecules of the spleen B cells and their significance on the basis of biology identification by establishing subacute aging mice model with D-galactose.

Methods: Health Kunming mice are injected with 100 ng/L D-galactose, 0.25 mL/20 g, once per day, for 42 days consecutively, into their back necks.

[Changes and effects of significant membrane molecules on thymic T cell of aging mice induced by D-galactose].

Aim: To establish subacute aging mice model by D-galactose and to explore the changes and effects of significant membrane molecules on thymic T cell.

Methods: Female Kunming mice of 8 weeks old were injected with D-galactose of 12.5 mL/(kg.

[Construction of human CXCR3B gene transfected cell line, identification and study of its biological function].

Aim: To construct recombinant human CXCR3B gene expression vector and obtain L929-CXCR3B gene transfected cell line for stably expressing human CXCR3B.

Methods: Human CXCR3B gene of full length was amplified by PCR from the plasmid pMD19-T/huCXCR3A. Then, it was inserted into eukaryotic expression vector pIRES2-EGFP to construct recombinant vector pIRES2-EGFP/huCXCR3B.

The negative co-signaling molecule b7-h4 is expressed by human bone marrow-derived mesenchymal stem cells and mediates its T-cell modulatory activity.

Though experimental evidence shows that human bone marrow-derived mesenchymal stem cells (hBMSCs) are able to suppress T-cell activation and proliferation, the precise mechanisms are still not completely understood. Here, we investigated the role of the negative costimulatory molecule B7-H4 in the immunosuppressive effect of hBMSCs on T-cell activation. We showed that B7-H4 expresses abundantly on hBMSCs assessed by reverse transcription, immunofluorescence staining, and flow cytometric analysis.

[Inhibitory and lethal effects of human-mouse chimeric antibody against CD80 on the growth of B lymphoma cell lines Raji and Daudi].

Aim: To study the inhibitory and lethal effects of human-mouse chimeric antibody against CD80 (named ch-4E5) on the growth of B lymphoma cell lines Daudi and Raji.

Methods: Immunofluorescence and flow cytometry were used to analyze the detection of membrane CD80 in Raji and Daudi by ch-4E5. After the co-culture of ch-4E5 with Raji and Daudi, respectively, at the final concentration of 10 mg/L, the expression of Fas and FasL was observated to be at the 0 h, 4 h, 10 h, 16 h, 24 h and 48 h by direct immunofluorescence and flow cytometry, and the blocking effect on cell growth of ch-4E5 was determined at 72 h by MTT assay.

[Dynamic changes of cytokines in G-CSF mobilized peripheral blood].

Objective: To investigate the level and significance of interleukin-8 (IL-8), soluble intercellular adhesion molecule (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) in patients peripheral blood (PB) during mobilization for peripheral blood stem cells harvesting.

Methods: The levels of IL-8, sICAM-1 and sVCAM-1 in patients were dynamically assayed by ELISA during the mobilization procedure and the number of CD(34)(+) cell, white blood cell (WBC) and platelet (BPC) by flow cytometric analysis and hematometry respectively. Colony formation was assayed by using semisolid methycellulose culture.