Reducing progression of experimental lupus nephritis via inhibition of the B7/CD28 signaling pathway.

The aim of the present study was to evaluate the effects of the B7/cluster of differentiation (CD)28 signaling pathway on experimental lupus nephritis and examine the molecular mechanism involved by inhibiting the B7/CD28 signaling pathway. A lupus nephritis model in C57BL/6 J mice was induced via intraperitoneal injection of pristane. A recombinant B7‑1 short hairpin RNA (shRNA) lentivirus vector was constructed by synthesis and splicing.

[Construction of 3D model of CD28 chimeric antibody with its antigen docked].

Aim: To construct a 3D model of the chimeric antibodies (AntiCD28: ch-2F5) with corresponding antigen molecule docked to theoretically verify the rationality of the binding of antibody with its antigen and to provide a method of 3D identification between antigen and antibody and spatial structure analysis.

Methods: We analyzed the sequence by submitting it to http://www.ncbi.

[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.

[Study on expression of Treg cells in attenuated Schistosoma japonicum cercariae immunized BALB/c mice].

Objective: To study the expression differences of CD4+CD25+Foxp3+Treg cells between the attenuated cercariae immunized mice and the normal infected mice and discuss the immune protection mechanisms of the mice immunized with attenuated cercariae.

Methods: Forty female BALB/c mice were divided into 2 groups, group A, the attenuated cercariae immunized group (16 mices) and the group B, the normal cercariae infected group (16 mices), and the last 8 ones served as the blank control. The spleen cells and the ratios of PBMC's CD4+CD25+Foxp3+/CD4+CD25+T cells were compared between the attenuated cercariae immunized mice and normal mice injected by FCM and the Foxp3 expression levels in spleens and livers were assayed by IHC.

[Dynamic changes of early immune responses to attenuated Schistosoma japonicum cercariae induced in BALB/c mice].

Objective: To study the early immune activation and its dynamic changes between the attenuated cercariae immunized mice and the normal infected mice.

Methods: The dendritic cell surface molecules CD11c and T cell surface molecule CD25 expression differences and CD3+CD25+/CD3+ T ratio of the early spleen and/or lung of the attenuated cercariae immunized mice and normal mice were assayed and compared by FCM and IHC, and the immune activation and dynamics of T cells were analyzed.

Results: CD3+CD25+CD3+ T ratio in the spleen cells 7 days post-infection in the immunized group and the normal infected group were (19.

[Mouse anti-human CXCR3 mAb preparation and its biological function].

Aim: Obtain hybridoma cell line with continuing expression of mouse anti-human CXCR3 mAb, investigate expression characteristics of human CXCR3 and how CXCR3 signal transduction function on L929-huCXCR3 and colon carcinoma cell lines transfer and growth.

Methods: Taking L929-huCXCR3 cell with high expression of human CXCR3 membrane molecule as immunogen to immunize BALB/c mouse, we fused immunized mouse spleen cell with myeloma cell Sp2/0 of its same germ line, then used L929-huCXCR3 as screening cell and empty vector transfected cell L929-mock as negative control. Obtained hybridoma cell line with continuing secretion of anti-human CXCR3 mAb through flow cytometry.

[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.

A novel monoclonal antibody against human CD80 and its immune protection in a mouse lupus-like disease.

Blockade of the interactions between CD28/CTLA-4 and their ligands, CD80 (B7, B7.1)/CD86 (B70, B7.2), is an attractive means to induce antigen-specific peripheral tolerance in autoimmune disease and organ transplantation.

[Construction of murine CXCR3 gene transfected cell line, identifiction and study of its biological function].

Aim: To construct recombinant murine CXCR3 gene retroviral vector and obtain L929-mCXCR3 gene transfected cell line for stably expressing murine CXCR3. We further study on L929-mCXCR3 migration effect resulted from interaction by CXCR3 and its ligand IP-10.

Methods: One female BALB/c mouse (7 weeks) was injected with 0.

[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.

[Preparation of two mouse anti-human PD-1 monoclonal antibodies and identification of their biological characteristics].

Aim: To prepare mouse anti-human PD-1 monoclonal antibodies (mAbs) and identify their biological characteristics.

Methods: The BALB/c mice were immunized with the transfected cell line PD-1/L929. The cells were fused with Sp2/0 using monoclonal antibody techniques and the positive clones were screened by FCS with PD-1/L929.

Preparation and characterization of a novel chimeric antibody against human CD40 with the potential to inhibit Daudi cell proliferation.

5C11, a murine monoclonal antibody with a high specificity for human CD40 molecule, is a promising candidate for cancer targeting therapy. We have therefore attempted to construct a humanized antibody of 5C11 to minimize its immunogenicity for potential clinical use. A chimeric version of 5C11 (ch-5C11) was generated by transferring these mouse variable regions onto a human framework.

Effects of preemptive intravenous lornoxicam on the analgesic efficacy of epidural morphine and expression of chemokines in women undergoing hysterectomy.

Background: It is believed that preemptive IV lornoxicam treatment can reduce the consumption of other analgesics, improve analgesic efficacy, and ameliorate immune function during patient-controlled IV analgesia. However, the effects of preemptive IV lornoxicam treatment on the analgesic efficacy of patient-controlled epidural analgesia (PCEA) with morphine and on chemokine expression remain unknown.

Objective: The aim of this prospective, randomized, controlled study was to observe the effects of preemptive IV lornoxicam treatment on the analgesic efficacy of PCEA with morphine and on the expression of monocyte chemotactic protein-1 (MCP-1) and stromal cell-derived factor-1α (SDF-1α) in women undergoing hysterectomy.

Fine-tuned expression of programmed death 1 ligands in mature dendritic cells stimulated by CD40 ligand is critical for the induction of an efficient tumor specific immune response.

During maturation, murine myeloid dendritic cells (DCs) upregulated the expressions of CD11c, CD25, CD40, CD80, CD86, MHC II and programmed death 1 ligands 1 and 2 (PD-L1 and PD-L2). Differential expression patterns of PD-L1 and PD-L2 were found when DCs were triggered by CD40 ligand and TNF-alpha. PD-L1 expression was repressed and PD-L2 expression remained unchanged in mature CD40-ligated DCs, whereas TNF-alpha stimulated DCs kept high expression of PD-L1 and significantly enhanced PD-L2 expression on DCs.

[Construction of anti-CD28 single chain antibody genes and expression of the ScFv in BmN cells and the larvae of Bombyx mori].

The V(H) and V(L) gene fragments of anti-CD28 mAb were combined to form anti-CD28 ScFv gene by using TP-PCR method. Sequence analysis showed that 6 x His tag was added to it for the ease of purification and the V(H), and V(L) gene fragments were connected by a linker containing 15 amino acids which are biased by the baculovirus promoter, ph. Then ScFv gene fragment was inserted into baculovirus transfer vector pBacPAK8.

A novel anti-human syndecan-1 (CD138) monoclonal antibody 4B3: characterization and application.

Syndecan-1 (CD138), a member of integral membrane heparin sulfate proteoglycans, is an essential matrix receptor for maintaining the normal morphological phenotypes. In this study, we generated a specific mouse anti-human syndecan-1 monoclonal antibody (mAb) 4B3 and identified it by competition assay with the available syndecan-1 mAb (BB4). Stained by 4B3, the expression of syndecan-1 was detected on tumor cell lines, such as 8226, U266, XG-1, XG-2, Daudi and Jurkat.

[Expression of human-mouse chimeric antibody against CD40 in CHO cell line and characterization of its function].

Aim: To investigate the stable expression of a chimeric antibody against CD40 moleculeèch-5C11éin CHO and its biological activity.

Methods: Human-mouse chimeric antibody against CD40 recombinant plasmid and mock plasmid were transfected into CHO cell line through lipofectamine mediation. Human kappa chain and Fc fragment of ch-5C11 were characterized by FACS and Western blot.

[Construction of a recombinant baculovirus transfer vector with two promoters expressing the anti-human CD28 chimeric antibody by using TP-PCR method].

CD28, a cell surface glycoprotein, predominantly expressed on T cells, belongs to the Ig superfamily and provides critical co-stimulatory signals. The data which have published indicate that the monoclonal antibody against CD28 can decrease curative effects when it was applied in vivo for a long time. In order to avoid the human-anti-mouse action, anti-CD28 mAb must be humanized before it can be used in clinical study.

Apoptosis of multiple myeloma cells induced by agonist monoclonal antibody against human CD28.

CD28 is expressed abnormally on human multiple myeloma (MM) cells but the significance had not been identified until now. In this paper, we are suggesting that abnormal expression of CD28 might be a marker of tumour progression. We therefore took the approach of generating a hybridoma cell line capable of secreting agonist monoclonal antibody directed against human CD28 (agonist anti-CD28 mAb) and then determined the expression of CD28 molecules on the MM cell lines U266 and XG1.

Time courses of B7 family molecules expressed on activated T-cells and their biological significance.

B7 family molecules are mainly expressed on the outer membrane of antigen-presenting cells. Here, our results demonstrate that CD80, CD86, and PD-L1 molecules are also expressed on T-cells that have been activated by simultaneous exposure to anti-CD3 and anti-CD28 mAbs, but PD-L2 and GL50 molecules were not detectable during the first six days of culture that follow such stimulation. We have analysed the time course of B7 family molecule expression on activated T-cells.

A functional anti-human 4-1BB ligand monoclonal antibody that enhances proliferation of monocytes by reverse signaling of 4-1BBL.

4-1BB Ligand (4-1BBL), a transmembrane molecule, member of the tumor necrosis factor ligand superfamily, is an important costimulatory molecule in the immune response. In this study a functional anti-human 4-1BBL MAb 1F1 was obtained and the specificity of this MAb was verified by flow cytometry and Western blotting. This MAb effectively recognized the 4-1BBL molecule expressed on a series of malignant cell lines as well as on DC and monocytes and it inhibited the proliferation of T lymphocytes, costimulated by soluble 4-1BBL and agonist anti-human CD3 MAb.