The interactions between blood cells and blood cells or blood cells and endothelium of blood vessel are mainly mediated by adhesion molecules. The role of adhesion molecules is diverse in vivo, which involved in adhesion, migration, differentiation and signal transduction of blood cells. The function of adhesion molecules is necessary to maintain the normal structure and fulfill many physiological processes of these cells. Therefore, the abnormal or deficiency of their expression and function will disrupts normal physiological processes and results in clinical disease. In this paper, several generic classes of adhesion molecules, including the integrins, the selectins, the immunoglobulin superfamily and others are introduced, and a lot of related physiopathological status, such as inflammation, hemostasis, arteriosclerosis, thrombosis and stem cell homing are discussed. The studies on the adhesion molecules of blood cells will contribute not only to understand the pathogenesis of some disorders, but also to search new targets in diagnosis and treatment of these diseases.
Animals ; Antibodies, Monoclonal ; therapeutic use ; Arteriosclerosis ; etiology ; Blood Cells ; chemistry ; Cell Adhesion Molecules ; antagonists & inhibitors ; physiology ; Humans ; Inflammation ; etiology ; Integrins ; physiology ; Selectins ; physiology ; Thrombosis ; etiology

Hemostasis ; physiology ; Humans ; Research ; Thrombosis ; physiopathology

Apoptosis ; Blood Platelets ; cytology ; Humans

Mer is one member of Axl receptor tyrosine kinase family, and its ligand Gas6 can stimulate activity of Mer receptor tyrosine kinase after binding it, and then activate the downstream signal transduction pathway, Mer participates in cell inflammation, apoptosis, tumorigenesis, thrombosis and hemostasis. Rencet advances of study on Mer function were reviewed, and its potential prospects of antithrombosis and antitumor were discussed in this article.
Animals ; Fibrinolytic Agents ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Signal Transduction ; c-Mer Tyrosine Kinase

This study was aimed to construct a pEGFP-N1 vector of von Willebrand factor cleaving protease (ADAMTS13, a disintegrin and metalloprotease with a thrombospondin type 1 motifs 13) so as to pave the way for further studying its synthesis and secretion. Human full-length cDNA sequence of ADAMTS13 was acquired by polymerase chain reaction (PCR) with Phusion(®) High-Fidelity (NEB), then the PCR product was double digested with EcoRI and XhoI. After digestion, the ADAMTS13 cDNA sequence was purified and recombined with the pEGFP-N1 vector. The DNA sequence analysis showed that ADAMTS13 was ligated to the pEGFP-N1 vector correctly. After transient expression in HeLa cells, the expression of EGFP could be detected by fluorescent microscopy, and the expression of ADAMTS13 protein could be detected by SDS-PAGE and Western blot. It is concluded that the ADAMTS13-pEGFP-N1 vector is successfully constructed, and it can be widely used in further research on the mechanism of the synthesis and secretion of ADAMTS13.
ADAM Proteins ; genetics ; ADAMTS13 Protein ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HeLa Cells ; Humans ; Transfection

The aim of this research is to prepare a novel monoclonal antibody against granulocytes by the intraperitional routine procedure and evaluate the usefulness of (99)Tc(m) labelled anti-granulocyte monoclonal antibody (McAb) SZ-102 for the detection of experimental inflammatory areas in rabbits. It was characterized as IgG1 subclass by the double immunodiffusion analysis. The flow cytometry demonstrated that SZ-102 reacted selectively with human granulocytes, monocytes, and with their bone marrow precursors, while human lymphocytes, red blood cells and platelets remained negative. In addition, SZ-102 antigen was expressed on the macrophages of liver, lung, thymus, spleen and lymph node by immunohistochemical SP methods. It is suggested that McAb SZ-102 is mainly against granulocytes. SZ-102 was labelled with (99)Tc(m) using 2-iminothiolane modification McAb and (99)Tc(m) -glucoheptonate (GH) transchelation method. The experimental rabbit model of inflammatory areas was prepared,through injecting with (99)Tc(m)-SZ-102 by ear-edge vein, and then imaged by SPECT. (99)Tc(m) labelled murine IgG was used as a negative control. The inflammatory areas in rabbits were clearly imaged at 2 to 4 hour after injection of (99)Tc(m)-SZ-102, while the control group after injection of (99)Tc(m)-labeled murine IgG was negative. In conclusion, the McAb SZ-102 may be a potential agent for the diagnosis and localization of inflammatory areas of carcinomas and clinically concealed infectious diseases.
Animals ; Antibodies, Monoclonal ; Flow Cytometry ; Granulocytes ; immunology ; Humans ; Inflammation ; diagnostic imaging ; Mice ; Mice, Inbred BALB C ; Rabbits ; Radioimmunodetection ; Technetium

The aim of this study is to screen out the monoclonal antibody reactive to a kind of endothelial cell line (ECV304) from SZ-1, -2, -21, -22, -51, -65, -262 and explore its function of anti-angiogenesis. The inhibitory effects of monoclonal antibody reactive to ECV304 and human lung carcinom cells (A549) adhesion and migration to extracellular matrix proteins (i.e, fibronectin and collagen IV) were studied by ELISA, the inhibition of angiogenesis in vivo was analyzed by chick chorioallantoic membrane (CAM) assays, the percentage of apoptotic cells in A549 cells was assayed by flow cytometric Annexin-V-FITC/PI dual labeling technique. The results showed that SZ-21 exhibited inhibitory effects on human umbilical vein endothelial cell line (ECV304) and pulmonary cancer cell line (A549) adhesion and migration to extracellular matrix proteins (i.e, fibronectin and collagen IV). In addition, it disrupted ongoing angiogenesis on the chick chorioallantoic membrane (CAM) model. SZ-21 also induced apoptosis of the A549 cells. In conclusion, SZ-21 inhibits angiogenesis in vivo and in vitro by blocking integrin beta(3) and inducing apoptosis. SZ-21 recognized the sequence beta(3) 28 - 35 which is far away from the RGD ligation site on GPIIIa. Integrin may interact the extracellular matrix via recognition sites other than RGD sequence.
Allantois ; blood supply ; Angiogenesis Inhibitors ; pharmacology ; Animals ; Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Cell Adhesion ; drug effects ; Cell Line ; Cell Movement ; drug effects ; Chick Embryo ; Chorion ; blood supply ; Extracellular Matrix Proteins ; pharmacology ; Flow Cytometry ; Humans ; Integrin beta3 ; immunology ; Neovascularization, Physiologic ; drug effects ; Tumor Cells, Cultured

In order to study the value of sequential and quantitative analysis of chimerism in determination of optional time of donor lymphocyte infusion (DLI) and prediction of efficacy of DLI, six patients with leukemias who relapsed or failed of engraftment were treated with DLI. Serial and quantitative analyses of donor chimerism (DC) both prior to and following DLI were performed by multiplex PCR amplification of STR markers (STR-PCR) and capillary electrophoresis with fluorescence detection. The results showed that at the time of relapse or graft rejection, STR-PCR indicated the decreasing donor chimerism in all six patients, at levels ranging from 27.3% to 85.7%. The declining value of DC (<90%) was detected in four patients at 26 days before relapse or graft rejection diagnosed clinically. Therefore the decrease of value of DC can be identified the high risk of relapse or graft failure and can be used to guide DLI implementation at early stage. In this study the clinical response were seen in two patients, the value of DC in these patients increased with convertion to a predominant donor profile (>90%) or converted to stable FDC shortly after DLI, while in the patients without clinical response, the level of DC decreased persistently or declined after transient increase. Three patients without response received second DLI. It is concluded that the monitoring of chimerism is proved to be a valuable to determine the optional time point of DLI and to early evaluate the efficacy of DLI. Furthermore, it can present a rational basis for treatment of intensification in the patients who did not respond to first-line DLI treatment.
Adolescent ; Adult ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphocyte Transfusion ; Recurrence ; Tissue Donors ; Transplantation Chimera

OBJECTIVETo construct the expression vectors of vWF73 and vWF114 fragments of von Willebrand factor (vWF) A2 domain, and to express glutathione S-transferase (GST) fusion proteins in E. coli, and to explore their values in measuring ADAMTS13 activity as substrates.

METHODSThe DNA fragments encoding vWF73 and vWF114 were generated using PCR and separately cloned into pGEX-6P-1, a Schistosoma japonicum GST fusion expression vector. The expression of GST-vWF73-H and GST-vWF114-H was induced in liquid culture, followed by purification with Ni-NTA agarose column. The cleavage of two GST fusion proteins by recombinant ADAMTS13 (rADAMTS13) or plasma from normal individuals and thrombotic thrombocytopenic purpura (TTP) patients were identified by Western blot. Based on an enzyme-linked immunosorbent assay (ELISA) with anti-GST and anti-His monoclonal antibodies, GST-vWF73-H and GST-vWF114-H were used to measure plasma ADAMTS13 activity as substrates.

RESULTSTwo small molecular substrates of ADAMTS13, GST-vWF73-H and GST-vWF114-H, are expressed and purified, which could be specifically cleaved by rADAMTS13 or plasma from healthy individuals, but not by plasma from congenital or idiopathic TTP patients. An ELISA assay was established to detect plasma ADAMTS13 activity using GST-vWF73-H and GST-vWF114-H as substrates.

CONCLUSIONSTwo GST fusion proteins in vWF A2 domain, vWF73 and vWF114, were expressed effectively using a prokaryotic expression system and could be used to detect ADAMTS13 activity as substrates.

ADAM Proteins ; blood ; genetics ; ADAMTS13 Protein ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; metabolism ; Glutathione Transferase ; metabolism ; Humans ; Male ; Purpura, Thrombotic Thrombocytopenic ; blood ; genetics ; metabolism ; von Willebrand Factor ; genetics ; metabolism