Anti-platelet factor 4/heparin antibodies in patients with Hantaan virus infection.

Background: Hemorrhagic fever with renal syndrome (HFRS) induced by Hantaan virus infection and heparin-induced thrombocytopenia (HIT) are associated with symptoms such as thrombocytopenia and thrombosis. However, related molecules, such as anti-platelet factor 4 (PF4)/heparin antibodies, in patients with HFRS have not been evaluated.

Objectives: To test plasma levels of anti-PF4/heparin antibodies and study the possible role of these antibodies in HFRS pathogenesis.

CD Nomenclature 2015: Human Leukocyte Differentiation Antigen Workshops as a Driving Force in Immunology.

CD (cluster of differentiation) Ags are cell surface molecules expressed on leukocytes and other cells relevant for the immune system. CD nomenclature has been universally adopted by the scientific community and is officially approved by the International Union of Immunological Societies and sanctioned by the World Health Organization. It provides a unified designation system for mAbs, as well as for the cell surface molecules that they recognize.

MicroRNA-141 regulates the expression level of ICAM-1 on endothelium to decrease myocardial ischemia-reperfusion injury.

A growing number of studies have suggested microRNAs (miRNAs) are involved in the modulation of myocardial ischemia-reperfusion (MI/R) injury; however, the role of endogenous miRNAs targeting endothelial cells (ECs) and its interaction with ICAM-1 in the setting of MI/R remain poorly understood. Our microarray results showed that miR-146a, miR-146b-5p, miR-155*, miR-155, miR-497, and miR-451 were significantly upregulated, whereas, miR-141 and miR-564 were significantly downregulated in the ECs challenged with TNF-α for 6 h. Real-time PCR analyses additionally validated that the expression levels of miR-146a, miR-155*, and miR-141 were consistent with the microarray results.

Construction and evaluation of DNA vaccine encoding Hantavirus glycoprotein N-terminal fused with lysosome-associated membrane protein.

Background: Hantaviral diseases can have a high case fatality rate within the absence of broadly effective antiviral treatments or vaccines. We developed a DNA vaccine targeting the Hantavirus glycoprotein N-terminal (Gn) to major histocompatibility complex class II compartment by fusing the antigen with lysosome-associated membrane protein 1 (LAMP1), which altered antigen presenting pathway and activated the CD4+ T cells.

Methods: The segments of Gn and LAMP1 were cloned into vector pVAX1, and recombinant plasmid was constructed by inserting Gn sequence into LAMP1, between luminal and the transmembrane/cytoplasmic domains.

Improved implant osseointegration of a nanostructured titanium surface via mediation of macrophage polarization.

The use of endosseous implanted materials is often limited by undesirable effects that may be due to macrophage-related inflammation. The purpose of this study was to fabricate a nanostructured surface on a titanium implant to regulate the macrophage inflammatory response and improve the performance of the implant. Anodization at 5 and 20 V as well as UV irradiation were used to generate hydrophilic, nanostructured TiO2 surfaces (denoted as NT5 and NT20, respectively).

MiR-568 inhibits the activation and function of CD4⁺ T cells and Treg cells by targeting NFAT5.

CD4(+) T cells play critical roles in orchestrating adaptive immune responses. Their activation and proliferation are critical steps that occur before they execute their biological functions. Despite the important role of this process, the underlying molecular events are not fully understood.

Development and evaluation of a sandwich ELISA method for the detection of human CD306.

CD306, also known as soluble leukocyte-associated immunoglobulin-like receptor-2 (LAIR-2), is a member of an immunoglobulin superfamily with the shared characteristic of an immunoglobulin-like C2-type domain. CD306 is speculated to be secretory and has 84% similarity with the extracellular domain of CD305, which binds to the same ligands as CD306. However, data on its distribution are absent due to the lack of an efficient method to detect it.

Down regulation of TRAIL and FasL on NK cells by Cyclosporin A in renal transplantation patients.

TNF-related apoptosis-inducing ligand (TRAIL) and FasL can participate in cell mediated cytotoxicity via their death domain-mediated apoptotic signaling in the host-versus-graft disease occurred after renal transplantation. However, the effect of Cyclosporin A (CsA) commonly used as a drug to prevent and to treat renal transplant rejection, on these molecules have not been fully determined. In the present study, we found that with CsA administration, the expression of TRAIL and FasL predominantly on NK cells from renal transplantation patients was increased at day 5 after operation and went down to normal level on day 13.

Tamoxifen represses miR-200 microRNAs and promotes epithelial-to-mesenchymal transition by up-regulating c-Myc in endometrial carcinoma cell lines.

Although tamoxifen (TAM), a selective estrogen receptor modulator, has been widely used in the treatment of hormone-responsive breast cancer, its estrogen-like effect increases the risk of endometrial cancer. However, the molecular mechanisms of TAM-induced endometrial carcinoma still remain unclear. In this report, we explored the role of microRNAs (miRNAs) in TAM-induced epithelial-mesenchymal transition (EMT) in ECC-1 and Ishikawa endometrial cancer cell lines and found miR-200 is involved in this process via the regulation of c-Myc.

TSA suppresses miR-106b-93-25 cluster expression through downregulation of MYC and inhibits proliferation and induces apoptosis in human EMC.

Histone deacetylase (HDAC) inhibitors are emerging as a novel class of anti-tumor agents and have manifested the ability to decrease proliferation and increase apoptosis in different cancer cells. A significant number of genes have been identified as potential effectors responsible for the anti-tumor function of HDAC inhibitor. However, the molecular mechanisms of these HDAC inhibitors in this process remain largely undefined.

[Cloning and identification of the variable region gene of mouse anti-human BAFF mAb].

Aim: To obtain the variable region gene sequence of heavy and light chain of mouse anti-human BAFF monoclonal antibody (mAb) on base of BAFF mAb which was cloned in our laboratory.

Methods: The total RNA was extracted from mouse anti-human BAFF mAb hybridoma cell line FMMUB(4);, and then the RNA was reverse transcribed into cDNA. Specific primers were designed to amplify the targeted gene.

HER2 interacts with CD44 to up-regulate CXCR4 via epigenetic silencing of microRNA-139 in gastric cancer cells.

Background & Aims: Human epidermal growth factor receptor 2 (HER2) (neu/ERBB2) is overexpressed on many types of cancer cells, including gastric cancer cells; HER2 overexpression has been associated with metastasis and poor prognosis. We investigated the mechanisms by which HER2 regulates cell migration and invasion.

Methods: HER2 expression or activity was reduced in gastric cancer cell lines using small interfering RNAs or the monoclonal antibody, trastuzumab.

The effect of direct translocation across endosomes on the cytotoxicity of the recombinant protein e23sFv-Fdt-casp6 to HER2 positive gastric cancer cells.

HER2-positive cancers represent a class of malignancies with high metastasis and poor prognosis. We previously generated the e23sFv-PEA II-casp6 recombinant, which contains an anti-HER2 single-chain antibody (e23sFv), a Pseudomonas exotoxin A translocation domain (PEA II), and a constitutively active caspase-6 (casp6), and demonstrated its potent selective anti-tumor activities. In this study, we generated a smaller-sized recombinant e23sFv-Fdt-casp6, in which the PEA II domain was replaced with the furin cleavage sequence from diphtheria toxin (Fdt), and explored its translocation pathway and specific killing mechanism.

Human activated CD4(+) T lymphocytes increase IL-2 expression by downregulating microRNA-181c.

MicroRNAs, a large family of small regulatory RNAs, are posttranscriptional gene regulators that bind mRNA in a sequence-specific manner, thereby controlling diverse aspects of cell function, including immune reaction. In this study, we screened and identified a group of differentially expressed miRNAs in naive and activated CD4(+) T cells. Among the miRNAs studied, miR-181c was proven to have the potential to regulate CD4(+) T cell activation.

[The application of quantitative detection of cystatin C in evaluation of renal function after renal transplantation].

Aim: To establish ELISA method for quantitate the concentration of cystatin C (cys C) and to monitor the renal function of patients before and after renal transplantation.

Methods: Hybridomas secreting monoclonal antibodies (mAbs) against human cys C were produced and sandwich ELISA kit for quantitatively detecting cys C was established. Then the concentrations of serum cystatin C (Scys C) and urine cystatin C (Ucys C) from normal controls and 23 patients undergoing renal transplantation were detected and their relationship with serum creatinine (SCR) was analyzed.

HER2-mediated upregulation of MMP-1 is involved in gastric cancer cell invasion.

HER2 overexpression is associated with metastasis-the main cause of death in individuals with gastric cancer. In this study, we demonstrated that vector-based shRNA significantly knocked down the expression of HER2 and considerably inhibited both the migration and invasion of gastric cancer cells. HER2 knockdown resulted in the downregulation of the expression of MMP-1, while HER2 overexpression improved the transcription of MMP-1 through the activation of an MMP-1 promoter.

[Correlation between major histocompatibility complex class I-related chain A gene alleles and graft rejection in small intestine, liver and kidney transplantation].

Objective: To investigate the correlation between major histocompatibility complex (MHC) class I chain-related gene A (MICA) gene alleles matching rates and graft rejection in small intestine, liver and kidney transplantation.

Methods: Genome DNA were extracted from blood samples or pathological sections collected from donors and recipients of living-related transplantation, included 4 cases of small bowel transplantation, 5 cases of liver transplantation and 6 cases of kidney transplantation. The correlation between MICA alleles matching rates and acute graft rejection was analyzed following 13 MICA alleles determination by polymerase chain reaction based on sequence-specific primers (PCR-SSP).

Murine monoclonal antibodies generated against mouse/rat hemokinin-1.

The mouse/rat hemokinin-1 (m/rHK-1) was discovered nearly 9 years ago. This molecule is a peptide comprising 11 amino acids. The m/rHK-1 was found to be mainly expressed in central immune organs like bone marrow, and was proven to have lymphopoietic roles in B and T lymphocyte development.

Cancer/testis antigen CT45: analysis of mRNA and protein expression in human cancer.

CT45 is a cancer/testis gene that we previously identified by massively parallel signature sequencing. Encoded by a multigene family on chromosome X, CT45 showed restricted mRNA expression to normal testis and various cancers. In this study, monoclonal antibodies were generated against recombinant CT45 protein, and CT45 protein expression in normal and tumor tissues was evaluated by immunohistochemical analysis.

[Application of flow microbeads assay in detection of specific antibodies and cytokines in serum from patients with hemorrhagic fever with renal syndrome].

Aim: To establish a flow microbeads assay (FMA) to examine the level of hemorrhagic fever with renal syndrome virus (HFRS V.) specific IgM, IgG and cytokines in HFRS patients.

Methods: Serum samples from 28 cases of HFRS and 20 healthy controls were studied.

[Application of Epstein-Barr virus-transformed B lymphoblastic cells in identification of CTL epitopes specific for Hantaan virus].

Aim: To establish Epstein-Barr virus (EBV)-transformed B lymphoblastic cell lines (B-LCL) to present peptides as antigen-presenting cells (APC) and stimulate short-cultured T cells secreting IFN-gamma, by which the T cell epitopes are identified.

Methods: PBMCs from patients with hemorrhagic fever with renal syndrome (HFRS) were transformed using EBV from supernatant of B95-8 cells. ELISPOT assay was then employed to evaluate the IFN-gamma production of short-cultured G9L-specific CD8(+) T cells stimulated with peptide-pulsed autologous B-LCL cells.

Knockdown of human bid gene expression enhances survival of CD8+ T cells.

Tumor cells have developed immune evasion mechanisms such as considerably heterogenous FasL expression on their surface via which they could induce apoptosis of tumor-specific cytotoxic T lymphocytes (CTLs) in the immune system. Meanwhile, the competition of normal immune cells with tumor cells results in relative growth factors shortage for growth and proliferation of nontumor cells, which improves a susceptibility to early apoptosis of CTL. In an attempt to develop strategies for prolonging the survival of adoptively transferred T cells in a hostile pro-apoptotic tumor microenvironment, we used synthetic siRNA and vector-based shRNA to suppress the expression of Bid in human uterocervical carcinoma HeLa cells, followed by the further achievement of Bid gene silencing in human primary cells-CD8(+) lymphocytes via retrovirus-delivered siRNAs.

[Change in T cell-mediated immunity and its relationship with high mobility group box-1 protein levels in extensively burned patients].

Objective: To investigate the change in T cell-mediated immunity and its relationship with plasma high mobility group box-1 protein (HMGB1) levels in severely burned patients.

Methods: Thirty-five extensively burned patients (> 30% total body surface area) were included in this study, and were divided into MODS group (n = 13) and non-MODS group (n = 22). The blood samples were collected on post burn days 1, 3, 5, 7, 14, 21 and 28.

Application of sandwich ELISA for detecting tag fusion proteins in high throughput.

Based on a series of mAbs against four frequently used tags--the human Ig Fc fragment, GST, maltose-binding protein, and thioredoxin--we developed corresponding sandwich enzyme-linked immunosorbent assay (ELISA) to detect these tag fusion proteins. As a supplement for Western blot, the successfully established ELISA was specific, sensitive, quantitative, easy to perform, time-saving, and last but not least, suitable for high-throughput screening of tag fusion proteins. Determination of soluble tag fusion proteins expressed by various systems with the sandwich ELISA developed in the present study could be a valuable and promising tool for the wide application of tag-protein fusion systems in the rapidly growing field of proteomics research.

[The relationship between dysfunction of cell-mediated immunity and prognosis in severely burned patients].

Objective: To investigate the relationship between dysfunction of cell-mediated immunity and prognosis in severely burned patients.

Methods: Twenty-six patients with total burn surface area larger than 70% total body surface area (TBSA) were included in the present study, and they were divided into non-survival group (14 cases) and survival group (12 cases). Eleven healthy volunteers served as controls.