PURPOSE: Receptors for the Fc of IgG(FcvR) and a beta2 integrin molecule, CD11c/CD18 are important in clearance of microbes by granulocytes. We performed an in vitro study on the effect of recombinant human granulocyte colony-stimulating factor(rhG-CSF), or granulocyte-macrophage colony-stimulating factor(rhGM-CSF) on the expression of Fc R II, Fc R III, CD11c and CD18 and respiratory burst function of granulocytes. METHODS: Peripheral blood was collected from six healthy adults. The isolated granulocytes were stimulated with rhG-CSF, 250mg/mL, rhGM-CSF, 100ng/mL or both of them for 1, 3, 9, 24, and 48 hours. Using flow cytometry, we compared the expression of the Fc R II, Fc R III, CD11c and CD18 of granulocytes before and after stimulation. We also the studied respiratory burst of granulocytes with chemiluminescence assay. RESULTS: Fc R II and CD18 expression were not changed significantly after stimulation. Fc R III expression was decreased significantly after stimulation with rhG-CSF, rhGM-CSF, or both of them. CD11c expression was increased initially but was found to decrease significantly after 9 hours of stimulation. Granulocytes treated with rhG-CSF, rhGM-CSF, or both displayed enhanced respiratory burst. Our results suggest that exposure of granulocyte to growth factor results in granulocyte activation. CONCLUSION: We have shown that rhG-CSF and rhGM-CSF resulted in an activation of peripheral blood granulocytes immunophenotypically and functionally.
Adult ; Antigens, CD18* ; Flow Cytometry ; Granulocytes* ; Humans* ; Luminescence ; Receptors, IgG* ; Respiratory Burst*

The modulation of leukocyte cell surface adhesion molecules may influence the development of cellular events that determine the course of the inflammatory process. Direct interaction between activated T cells and monocytes resulted in a large production of IL-1beta by monocytes. In this reactions, adhesion molecules play an important part, yet the role of them in T-monocytes interaction remain unclear. This study was undertaken in an effort to elucidate, 1) the influence of 1.25(OH)2D3-induced differentiation on the monocyte responsiveness to direct contact with T lymphocytes, and 2) the role of adhesion molecules on the T-monocyte direct interaction. Initially, I observed that direct contact of monocyte cell line THP-1 with stimulated fixed T cell line HuT78 markedly induces IL-1beta production by THP-1. IL-1beta production was higher when THP-1 had been previously exposed to 1.25(OH)2D3 as compared to control, with alpha-1.25(OH)2D3 dose-dependent and exposure time-dependent manner. It was shown that 1.25(OH)2D3 also increased the expression of beta2 integrin adhesion receptor Mac-1(CD11b/CD18) dose- and time- dependently, but did not increase the expression of human leukocyte antigen-D(HLA-D) and intercellular adhesion molecule-1(ICAM-1). The IL-1beta producing activity of THP-1 cells correlated well with the ability to induce the Mac-1 expression on THP-1 surface. Monoclonal antibody raised against relevant cell surface glycoproteins on THP-1 were tested for their ability to block the response of THP-1 to T cells. Antibody to Mac-1 only partially blocked IL-1beta production by THP-1, whereas antibodies to ICAM-1 and HLA-D did not. These data indicate that regulation of Mac-1 expression on THP-1 cells can alter the responsiveness of these cells to contact by activated T cells, however other unknown structures on the THP-1 cells may be involved in this process also.
Antibodies ; Antigens, CD18* ; Cell Line ; HLA-D Antigens ; Humans ; Intercellular Adhesion Molecule-1 ; Leukocytes ; Membrane Glycoproteins ; Monocytes* ; T-Lymphocytes*

The beta2 integrins are expressed exclusively on leukocytes and participate in many immune and inflammatory processes. This subfamily comprises four heterodimeric glycoproteins with a common beta-subunit, designated beta2 (CD18). Spontaneous mutations of the CD18 gene result in leukocyte adhesion deficiency type I (LAD-I). Low level of CD18 expression has also been implicated in the pathogenesis of psoriasis. We here describe a child with recurrent skin infections without pus formation, persistent gingivitis and periodontitis. His blood counts showed persistent leukocytosis (neutrophilia). CD11b expression was defective on neutrophils, while that of CD18 was normal. So, our patient represents a mild variant of LAD-I with possible dysfunctional CD18. Moreover, he developed psoriasis with reduced CD18 expression on CD4+ T-cells. Psoriasiform dermatitis has been described before in association with LAD-I, however, clinically and histologically confirmed psoriasis in association with LAD-I has been described only in CD18 hypomorphic mice. Therefore, our patient represents the first clinically and histopathologically documented association between LAD-I and psoriasis in humans. It lends support to the role of beta2 integrins in the etiopathogenesis of psoriasis.
Animals ; Antigens, CD18 ; Child ; Dermatitis ; Gingivitis ; Glycoproteins ; Humans ; Leukocytes ; Leukocytosis ; Mice ; Neutrophils ; Periodontitis ; Psoriasis ; Skin ; Suppuration ; T-Lymphocytes

BACKGROUND: It has been known that integrins are not only simple glue which mediate cell-to-cell or cell to extra-cellular matrix but also function as signaling molecules at the surface of cell by conformational change. Because of the diversity of subunits and versatility of ligand recognition, integrins have been recognized as important molecules in kidney and the expression of integrin in normal human kidney has already been reported. METHODS: To know the significance of integrin expression in the pathogenesis of various renal diseases, we stained kidney specimens from the patients with membrano- proliferative glomerulonephritis(MPGN), minimal change nephrotic syndrome(MCNS) and membraneous nephropathy(MN) of lupus nephritis with 13 monoclonal antibodies of integrins. RESULTS: There was a significant change in the distribution of integrin subunits according to the types of glomerular diseases. Integrin subunit alpha2 was helpful in confirming the mesangial interposition as there was intense staining of subendotherial area in MPGN, while only the mesangium was stained with this unit in normal. In addition, an abundant staining of alpha4 and beta2 was observed and this finding indicated non-glomerular resident cell is participating in the MPGN, while in MN type of lupus nephritis, alpha2 staining was restricted to mesangium and alpha4 or beta2 integrin staining was negative. In the MN type of lupus nephritis, the remarkable finding was a ragged appearance in the subepithelial area, just below the glomerular basement membrane, which was visible in alpha3 and alphaV integrin staining. In MCNS, there was no difference in the staining pattern of integrin. CONCLUSION: Integrin subunits can be useful in evaluating the specific cell types which is actively involved in pathogenesis and these results suggest that alteration of integrin distribution can play an important role in the pathogenesis of glomerular diseases.
Adhesives ; Antibodies, Monoclonal ; Antigens, CD18 ; Glomerular Basement Membrane ; Glomerulonephritis ; Glomerulonephritis, Membranoproliferative ; Humans ; Integrin alphaV ; Integrins ; Kidney ; Lupus Nephritis

OBJECTIVETo explore the expression of polymorphonuclear leucocyte adhesive molecules CD11b/CD18 and to study the possible mechanism of Chinese herbal medicine (TCM) for activating blood circulation to remove stasis in preventing vascular diseases.

METHODSForty-nine patients with diabetes mellitus (DM) but with no complications of hypertension and nephropathy were randomly divided into the treated group (26 patients treated by TCM) and the control group (23 patients treated by conventional treatment). They were treated for 3 months. The changes of urinary albumin excretion rate (UAER), CD11b/CD18 expression and tumor necrosis factor-alpha (TNF-alpha) concentration before and after treatment were observed.

RESULTSThe CD11b/CD18 expression and TNF-alpha concentration in DM patients were higher than those of normal range (P < 0.01). After treatment, the UAER, CD11b/CD18 expression and TNF-alpha concentration lowered significantly in the treated group (P < 0.01), but unchanged in the control group. Correlation analysis showed that the lowering of UAER was positively correlated with decreasing of CD11b/CD18 (r = 0.64, P < 0.01) and TNF-alpha (r = 0.56, P < 0.01).

CONCLUSIONExpression of CD11b/CD18 increases in patients with DM type 2. The mechanism of Chinese herbal medicine for activating blood circulation to remove stasis in preventing vascular disease in possibly related with its effect in inhibiting CD11b/CD18 expression.

Aged ; CD11b Antigen ; biosynthesis ; blood ; CD18 Antigens ; biosynthesis ; blood ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phytotherapy

OBJECTIVETo investigate the changes in perioperative expression level of CD11/CD18 of neutrophils in children undergoing cardiac surgery with cardiopulmonary bypass (CPB).

METHODSThirty children patients with congenital heart disease underwent cardiac surgery with CPB (CPB group) and the control group consisted of 20 children who received thoracic or general surgery without CPB. Blood samples were drawn at the following time points: pre-surgery, 15 min after onset of CPB, immediately after CPB, 2 h after surgery and on the 1st, 2nd, 3rd postoperative day. D11/CD18 expression on neutrophils and serum concentration of IL-6 and IL-8 were analyzed by flow cytometry and enzyme-linked immunosorbent assay, respectively.

RESULTIn CPB group plasma levels of IL-6 and IL-8 increased significantly and peaked at 2 h after initiation of CPB (P<0.05), and descended to the after-anesthesia level at 3rd day after operation. In non-CPB group there was a similar trend of changes in IL-6 and IL-8, but to a much lesser extent. The level of CD11b/CD18 in CPB group began to increase significantly and peaked at 15 min after initiation of CPB (P <0.05), and descended to the after-anesthesia level at 2 h after operation. There was no significant changes of CD11b/CD18 in control group (P >0.05). No significant differences were detected at any time points with respect to expression of CD11a/CD18 and CD11c/CD18 in both groups (P >0.05).

CONCLUSIONCPB surgery of children can cause increasing of the CD11b/CD18 expression level of neutrophil but has no significant effect on CD11a/CD18 and CD11c/CD18. CD11b/CD18 may play an important role in the systemic inflammation induced by CPB.

CD11b Antigen ; blood ; CD18 Antigens ; blood ; Cardiopulmonary Bypass ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Heart Defects, Congenital ; blood ; surgery ; Humans ; Infant ; Male ; Neutrophils ; cytology ; metabolism

OBJECTIVETo observe the expression of adhesion molecular CD11b/CD18 in peripheral neutrophils and its relation with arteriosclerotic obliterans (ASO), and to study the effect of Xuefu Zhuyu Oral Liquid (XZOL) on it.

METHODSFlow cytometery analysis was used to detect the expression of CD11b/CD18 in peripheral neutrophils of 30 patients with ASO and 30 healthy subjects by direct immunofluroscent technique. Neutrophils were separated from whole blood of ASO patients and cultured, CD11b/CD18 were detected after the cultured cells were interfered with XZOL dilution at different time points (1h,6h,12h).

RESULTSThe expression of CD11b/CD18 in neutrophils in ASO patients was significantly higher than that in the healthy subjects (P < 0.05) and stepped in keeping with the severity of the disease. It was significantly lowered in the treated group 6 and 12 h after XZOL intervention, showing significant difference as compared with that in the control group and the level 1 h after medication (P < 0.05).

CONCLUSIONCD11b/CD18 may involve in the pathogenesis of ASO and be related to the severity of arteriosclerosis. The possible mechanism of XZOL in treating and preventing ASO might be through reducing the expression of CD11b/CD18 in peripheral neutrophils to interfere the adhesive function of them.

Aged ; Arteriosclerosis Obliterans ; blood ; CD11b Antigen ; blood ; CD18 Antigens ; blood ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Neutrophils ; cytology ; metabolism

BACKGROUNDLeukocyte adhesion deficiency type 1 (LAD-1) is a rare, autosomal recessive inherited immunodeficiency disease characterized by recurrent severe bacterial infection, impaired pus formation, poor wound healing, associated with the mutation in the CD18 gene responsible for the ability of the leucocytes to migrate from the blood stream towards the site of inflammation. Correct and early diagnosis of LAD-1 is vital to the success of treatment and prevention of aggressive infections. The purpose of this study was to collect the clinical findings of the disease and to identify the genetic entity.

METHODSCD18 expression in the peripheral blood leukocytes from the patient, his parents and normal control was measured with flow cytometry. The entire coding regions of the CD18 gene were screened with direct sequencing genomic DNA.

RESULTSCD18 expression level on this patient's leukocyte surface was significantly decreased, with normal level in control group, his father and mother. Gene analysis revealed that this patient had a homozygous c.899A > T missense mutation in exon 8 of CD18 gene, causing the substitution of Asp to Val at the 300 amino acid. His parents were both heterozygous carriers while no such mutation was found in 50 normal controls.

CONCLUSIONThis study disclosed a novel point mutation Asp 300 Val located in a highly conserved region (HCR) of CD18 and confirmed the heterogeneity of the mutations causing LAD-1, indicating it was quite beneficial to establish correct and early diagnosis in children with severe LAD-1.

Asian Continental Ancestry Group ; CD18 Antigens ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Flow Cytometry ; Humans ; Leukocyte-Adhesion Deficiency Syndrome ; etiology ; genetics ; Male ; Point Mutation ; genetics ; Polymerase Chain Reaction

OBJECTIVETo determine the effect of albumin administration on lung injury in traumatic/hemorrhagic shock (T/HS) rats.

METHODSForty-eight adult Sprague-Dawley rats were divided into three groups randomly (n=16 in each group): Group A, Group B, Group C. In Group A, rats underwent laparotomy without shock. In Group B, rats undergoing T/HS were resuscitated with their blood plus lactated Ringer's (twice the volume of shed blood). In Group C, rats undergoing T/HS were resuscitated with their shed blood plus additional 3 ml of 5% human albumin. The expression of polymorphonuclear neutrophils CD18/CD11b in jugular vein blood was evaluated. The main lung injury indexes (the activity of myeloperoxidase and lung injury score) were measured.

RESULTSSignificant differences of the expression of CD18/11b and the severity degree of lung injury were founded between the three groups. (P<0.05). The expression of CD18/CD11b and the main lung injury indexes in Group B and Group C increased significantly compared with those in Group A (P<0.05). At the same time, the expression of CD18/CD11b and the main lung injury indexes in Group C decreased dramatically, compared those in Group B (P<0.05).

CONCLUSIONSThe infusion of albumin during resuscitation period can protect lungs from injury and decrease the expression of CD18/CD11b in T/HS rats.

Albumins ; therapeutic use ; Animals ; CD11b Antigen ; metabolism ; CD18 Antigens ; metabolism ; Disease Models, Animal ; Neutrophils ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; drug therapy ; etiology ; metabolism ; Shock, Hemorrhagic ; complications ; metabolism ; Treatment Outcome ; Wounds and Injuries ; complications ; metabolism

OBJECTIVETo observe the effects of WWOX on cell attachment in ovarian cancer, and to explore its mechanisms of action.

METHODSAttachment assay was used to assess the adhesion of wwox-transfected PEO1 cells and vector-transfected PEO1 cells that were constructed, as well as PEO1 parent cells. Alpha/beta integrin-mediated cell adhesion assays were designed to identify cells surface integrins in PEO1 clone cells. Integrin function blocking experiments were designed to further determine integrins in PEO1 clone cells according to the integrin that was selected in integrin expression profiling. FACS analysis was used to further detect the level of integrin alpha3 on the cell membrane.

RESULTSAttachment assay showed that adhesion of WWOX-transfected PEO1 cells to fibronectin was significantly slower than that in vector-transfected controls or PEO1 parent cells, cultured on the pre-coated fibronectin for 2 hours (P<0.01). The level of membranous integrins alpha2 and alpha3 in the WWOX-transfected PEO1 cells was significantly decreased, as compared with that in vector-transfected controls (P<0.05), but there was no association with the level of functioning integrins betal or beta2 in clone cells (P>0.05). The attachment assays were repeated after pre-incubating the cells with integrin alpha2 or alpha3 function-blocking antibodies. These results showed that blocking integrin alpha3 significantly reduced the binding to fibronectin of all the PEO1 clonal lines, as compared with cells pre-incubated with a non-specific IgG antibody (P<0.05). In contrast, preincubation with alpha2 blocking antibody had very little effect on fibronectin binding in these cells (P>0.05). FACS analysis showed that membranous integrin alpha3 expression revealed a marked reduction in WWOX-transfected cells than that in vector-transfected cells.

CONCLUSIONWWOX acts as an ovarian tumor suppressor by modulating the interaction between tumor cells and the extracellular matrix, decreasing integrin activity and adhesion of tumor cells to fibronectin. This suggests an important role for loss of WWOX tumor suppressor in promoting attachment and adhesion of ovarian cancer cells on locoregional peritoneum, and further resulting in enhancing locoregional peritoneal tumor spread.

CD18 Antigens ; metabolism ; Cell Adhesion ; Female ; Fibronectins ; metabolism ; Humans ; Integrin alpha2 ; metabolism ; Integrin alpha3 ; metabolism ; Integrin beta1 ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; Oxidoreductases ; genetics ; metabolism ; Protein Binding ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism ; WW Domain-Containing Oxidoreductase