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Abstract: Endocan, a 50 kDa soluble proteoglycan, also called endothelial cell-specific molecule-1 (ESM-1), is involved in many major cellular activities and has been reported to be overexpressed in inflammatory conditions. This study aims to determine ESM-1 levels in gingival crevicular fluid (GCF) samples from individuals with periodontitis to determine the correlation between the levels of lymphocyte-function-associated antigen-1 (LFA-1), intercellular adhesion molecule-1 (ICAM-1), and clinical findings of periodontitis. This study enrolled 27 individuals diagnosed with Stage III-Grade C generalized periodontitis and 16 individuals as healthy controls. Bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment loss (CAL) were calculated. Enzyme-linked immunosorbent assay (ELISA) test was used for detecting the levels of ESM-1, ICAM-1, and LFA-1 in GCF samples. PPD, BOP, CAL, and GCF volumes were significantly increased in patients with periodontitis in comparison to the control group (p < 0.001). The total amount of ESM-1, ICAM-1, and LFA-1 levels in GCF were increased in the periodontitis group (p < 0.001). ESM-1 level correlated with PPD, BOP, and CAL (p < 0.05). ICAM-1 level correlated with BOP and CAL (p < 0.05). LFA-1 level correlated with PPD and CAL (p < 0.05). Our data indicate that ESM-1, ICAM-1, and LFA-1 levels are increased in GCF of patients with periodontitis. These molecules could be associated with the pathogenesis and progression of periodontal disease.
Subject(s)
Humans , Periodontitis , Chronic Periodontitis , Proteoglycans , Enzyme-Linked Immunosorbent Assay , Lymphocyte Function-Associated Antigen-1 , Gingival Crevicular Fluid , Intercellular Adhesion Molecule-1 , Neoplasm ProteinsABSTRACT
Objective To observe aerobic exercise on blood lipid and intercellular adhesion molecule-1 (ICAM-1),lymphocyte function-associated antigen-1 (LFA-1) in hyperlipidemia rats.Methods 120 male Sprague Dawley (SD) rats were randomized into the 4 groups (n =30):normal diet group (control group),high fat group (HF group),HF + SBC-115076 group [SBC-115076,8 mg/kg · w)] and HF + aerobic exercise group.Rats in HF group and HF + SBC-115076 group did not exercise during feeding and lived in normal cage.Rats in HF + SBC-115076 group recieved PCSK9 inhibitor SBC-115076 (8 mg/kg) injection once a week for 8 weeks.HF + aerobic exercise group underwent load-free swimming training 6 days a week for 8 weeks.The rats were sacrificed 8 weeks later.The serum levels of triglyceride (TG),total cholesterol (TC),low density lipoprotein (LDL),high density lipoprotein (HDL),ICAM-1 and LFA-1 were measured by apical blood sampling.Hematoxylin-eosin (HE) staining of thoracic aorta to observe pathological changes of aorta.Results After modeling,there was no significant difference in the food intake levels of the hyperlipidemia rats (P > 0.05).The body weight of HF rats and HF + SBC-115076 rats increased significantly than HF + aerobic exercise rats (P < 0.01).The levels of serum lipid profile,ICAM-1,and LFA-1 were significantly different between HF rats and control rats.Compared with HF rats,serum levels of TG,TC,LDL,ICAM-1 and LFA-1 in HF + SBC-115076 group and HF + aerobic exercise group were significantly lower and HDL levels were significantly higher.HE staining showed that the intimal thickening was significantly improved in HF + aerobic exercise group with less endothelial damage.Conclusions Aerobic exercise can reduce the levels of serum TG,TC,LDL and increase HDL level in rats with hyperlipidemia.It can improve the intimal thickening and endothelial damage caused by high-fat diet by reducing the expression levels of serum ICAM-1 and LFA-1,and has anti-atherosclerosis effect.
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It has been widely accepted that Faecalibacterium prausnitzii (Fp) induces the differentiation of Treg cells.Lymphocyte function-associated antigen-1 (LFA-1) is also involved in the differentiation of Treg cells.Aims: To investigate the effect of Fp on Treg cells and cytokines in colitis mice with LFA-1 knockout (LFA-1-/-).Methods: Twenty wild type mice and twenty LFA-1-/-mice with same genetic background were randomly divided into wild type control group, wild type treatment group, LFA-1-/-control group and LFA-1-/-treatment group.Colitis model was induced by drinking DSS solution.Mice in the two treatment groups were intragastrically administrated with Fp.General status and histopathological score were assessed.Percentages of Treg cells in spleen and mesenteric lymph nodes were measured by flow cytometry.Serum levels of IL-10 and TGF-β1 were measured by ELISA.mRNA expressions of IL-10 and TGF-β1 in colonic tissue were detected by real time PCR.Results: Compared with corresponding control groups, histopathological score was significantly decreased in wild type treatment group (P<0.05);percentages of Treg cells in spleen and mesenteric lymph nodes were significantly increased (P<0.05), serum levels of IL-10 and TGF-β1 were significantly increased (P<0.01), expression of TGF-β1 mRNA was significantly increased in wild type treatment group and LFA-1-/-treatment group (P<0.05);expression of IL-10 mRNA was significantly decreased in LFA-1-/-treatment group (P<0.01).Compared with wild type treatment group, serum level of TGF-β1 was significantly decreased (P<0.05), and mRNA expressions of IL-10 and TGF-β1 were significantly decreased in LFA-1-/-treatment group (P<0.05).Conclusions: Fp can up-regulate the percentages of Treg cells and enhance the secretion of anti-inflammatory cytokines IL-10 and TGF-β1 in LFA-1-/-mice.The therapeutic efficacy for colitis in wild type mice is superior to that in LFA-1-/-mice, which may be related to the inhibition of function of Treg cells and secretion of cytokines due to LFA-1 knockout.
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Objective To explore the activate process of lymphocyte function-associated antigen 1 (LFA-1) triggered by chemokine under shear stresses. Methods Jurkat cells were perfused over ICAM-1 in the parallel-plate flow under 10-30 mPa shear stresses. The effects of soluble and immobilized Chemokines on transient adhesion behavior of Jurkat cells were observed and analyzed to obtain their tether characteristics. Results The immobilized CXCL12 could mediate brief tether (0.13-0.2 s) of Jurkat cells under flow. Only immobilized CXCL12 could effectively activate LFA-1 on Jurkat cells to bind ICAM-1, and then enhance cell adhesion fraction and greatly prolong the tether time (0.8-1.2 s). Two distinct activation states of LFA-1/ICAM-1 were reflected by their dissociation rate k1 (1.09-1.24 s-1) and k2 (0.28-0.7 s-1), respectively. The shear stress would affect the transient adhesion behavior of cells through regulation of k2 and β (the contribution ratio of high affinity to total tether time). Conclusions Shear stress can rapidly trigger LFA-1 activation in 0.2 s through G protein coupled receptors induced by chemokine CXCL12, and further regulate the whole adhesion process of leukocyte. These research findings will contribute to further understanding the integrin activation mechanism of chemokine-force cooperative regulation.
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Objective To explore the activate process of lymphocyte function-associated antigen 1 (LFA-1) triggered by chemokine under shear stresses.Methods Jurkat cells were perfused over ICAM-1 in the parallel-plate flow under 10-30 mPa shear stresses.The effects of soluble and immobilized Chemokines on transient adhesion behavior of Jurkat cells were observed and analyzed to obtain their tether characteristics.Results The immobilized CXCL12 could mediate brief tether (0.13-0.2 s) of Jurkat cells under flow.Only immobilized CXCL12 could effectively activate LFA-1 on Jurkat cells to bind ICAM-1,and then enhance cell adhesion fraction and greatly prolong the tether time (0.8-1.2 s).Two distinct activation states of LFA-1/ICAM-1 were reflected by their dissociation rate k1 (1.09-1.24 s-1) and k2 (0.28-0.7 s-1),respectively.The shear stress would affect the transient adhesion behavior of cells through regulation of k2 andβ (the contribution ratio of high affinity to total tether time).Conclusions Shear stress can rapidly trigger LFA-1 activation in 0.2 s through G protein coupled receptors induced by chemokine CXCL12,and further regulate the whole adhesion process of leukocyte.These research findings will contribute to further understanding the integrin activation mechanism of chemokine-force cooperative regulation.
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Objective To reveal the mechanical regulation mechanism for activation of lymphocyte function-associated antigen 1 (LFA-1). Methods The LFA-1 expressed on Jurkat cell surface was pre-activated by Mg2+ from quiescent-to intermediate-affinity state, and the tether events of Jurkat cells under different wall shear stresses (4.5-10 mPa) were observed and analyzed by flow chamber experiment. Meanwhile, a probabilistic model of integrin affinity jumping was established. Results The affinity jumping model was well fitted with the data obtained from flow chamber experiment. Under flowing loads, LFA-1 from intermediate to high-affinity state was observed, with prolonging of the adhesion bonds. The probability of tether event was 15%-26%. LFA-1 at high-affinity state contributed a significant fraction (about 26%-40%) of the bond lifetime. The off-rate of LFA-1 at high-affinity state was slower by 19%-65% as compared to that at intermediate-affinity state. Dissociating of ICAM-1 from LFA-1 was force-dependent and governed either by slip-bond at intermediate-affinity state or by catch-slip bond at high-affinity state. Conclusions The force-induced activation of LFA-1 mediates the slower rolling and firm adhesion of the cells. This research finding will further the understanding of inflammatory response events of circulating leukocytes, and contribute to the discovery of new antibody drug targets for the associated diseases.
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Objective To check the neutrophil count and the expression of cell surface adhesion molecule,lymphocyte function associated antigen?1(LFA?1)in peripheral neutrophils in severe preeclamptic patients and normal pregnant women in order to investigate the correlation of neutrophil activation with preeclampsia. Methods Totally 28 pregnant women in the department of gynecology and obstetrics in Shengjing Hospital from No?vember 2013 to January 2014 were included in the study,and divided into the severe preeclampsia group(n=14),and the control group(normal pregnant women,n=14). There was no statistically significant difference in age and gestational weeks between the two groups. The expression of LFA?1 in peripheral neutrophils was detected by flow cytometry in the two groups. Mean blood pressure(MBP)was measured in the severe preeclamptic group,and the correlation of MBP with expression of LFA?1 was analyzed. Results The neutrophil count was 8.40±2.23 ×109/L in the severe pre?eclamptic group,significantly higher than(6.71±1.58)×109/L in the control group,(P<0.05). The positive rate of LFA?1 in peripheral neutrophils was 63.25±38.025%in the severe preeclamptic patients,significantly higher than 8.32±38.65%in the control group(P<0.05). The expression lev?el of LFA?1 in peripheral neutrophils was significantly correlated with mean blood pressure in severe preeclamptic patients(r=0.64,P=0.013). Conclusion The neutrophil count and the expression of LFA?1 in peripheral neutrophils were significantly increased in severe preeclamptic pa?tients compared to normal pregnant women,and significantly correlated with severity of preeclampsia,suggesting that neutrophil activation participat?ed in the pathogenesis of preeclampsia.
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BacKground:Lymphocyte function-associated antigen-1( LFA-1 ) pIays a cruciaI roIe in the pathogenesis of infIammatory boweI disease by reguIating the activation and function of T ceIIs. Aims:To investigate the effect of LFA-1 deficiency( LFA-1-/-)on differentiation of mice na?ve T ceIIs into Th17 ceIIs in vitro. Methods:LFA-1-/- mice were breeded and the progeny genome DNA was extracted from the taiIs for genotyping by PCR. CD4+CD62L+ na?ve T ceIIs were separated from spIenic mononucIear ceIIs of LFA-1-/- progeny mice and wiId type ( WT ) C57BL/6J mice, respectiveIy,by magnetic-activated ceII sorting( MACS),and then the purity of separated ceIIs was determined. Na?ve T ceIIs obtained were cuItured in different inducing systems[ transforming growth factor-β( TGF-β),TGF-β + interIeukin-6 (IL-6),and TGF-β + IL-6 + IL-23]in vitro for Th17 ceII differentiation;ceIIs in each inducing system were coIIected for anaIyzing the ratio of Th17 ceIIs by fIow cytometry,and the expressions of Th17-specific transcription factor ROR-γt and Th17-specific marker IL-17A were measured by qRT-PCR and ELISA methods. Results:AII fifteen progeny mice were identified as LFA-1-/- genotype. Purity of CD4+CD62L+ na?ve T ceIIs separated by MACS was above 95%. Th17 ceIIs couId be induced by Iow-dose TGF-βcombined with IL-6,and the differentiation ratio was increased obviousIy when IL-23 was added. In inducing system containing TGF-β,IL-6 and IL-23,na?ve T ceIIs from LFA-1-/- mice produced more Th17 ceIIs than those from WT mice(17. 2% ± 1. 4% vs. 5. 7% ± 0. 2%,P<0. 001),expressions of ROR-γt mRNA and IL-17A mRNA were up-reguIated(P<0. 001),and IL-17A concentration in ceII cuIture supernatant was increased(P<0. 01). Conclusions:Deficiency of LFA-1 promotes the differentiation of mice na?ve T ceIIs into Th17 ceIIs in vitro.
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Objective To investigate the molecular mechanism underlying lymphocyte functionassociated antigen-1 (LFA-1) / intercellular adhesion molecule-1 (ICAM-1) mediated anti-neoplastic effects of cytokine induced killer (CIK) cells. Methods Lymphocytes isolated from peripheral blood of children leukemia were induced with interferon-gamma (IFN-y), anti-CD3 monoclonal antibody (CD3McAb) and interleukin-2 (IL-2) and co-cultured with dendrite cells (DC) to generate DC-CIK cells. When treated with LFA-1 monoclonal antibody, cytotoxicity of DC-CIK cells against leukemia cell lines was measured by the MTT assay, while RT-PCR and Western blotting were used to determine mRNA and protein expressions of GATA-3 and T-bet in DC-CIK cells, respectively. IL-12, IFN-γ and tumor necrosis factor-α (TNF-α) levels released by DC-CIK cells were quantified by ELISA. Results Induced DC-CIK cells were regular, round and transparent with variable cell volume and cellular aggregation. When treated with mouse anti-human LFA-1 monoclonal antibody, the cytotoxicity decreased mostly towards B95 cells under administration of 20 μg/ml LFA-1 monoclonal antibody in comparison with the control group(t =10.138, P <0.05). It led to a highest elevation of GATA-3 mRNA and protein levels (t =16.386, P < 0.05; t =22.652, P < 0.05) and a most decrease of T-bet mRNA and protein levels (t =17.728, P <0.05; t =17.452, P <0.05) under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group. The expression levels of IL-12,IFN-γ, and TNF-o in supernatant were the lowest under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group (t =21.621, P <0.05; t =13.739, P <0.05; t =15.278, P <0.05).Conclusion GATA-3 and T-bet were implicated in the LFA-1/ICAM-1 mediated anti-neoplastic effects of DC-CIK cells via activation of the Th1 pathway, with high secretion of Th1 cytokines, such as IL-12, IFN-γ and TNF-α.
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The basic route and mechanism for diapedesis has not yet to be fully defined. Here we present evidence that "cell-cell separation" between endothelial cells (ECs) may provide a route for leukocyte diapedesis. We unexpectedly found that extensive interaction between peripheral blood leukocytes and ECs that were activated by TNF-alpha induced the opening of EC contacts and, surprisingly, resulted in cell-cell separation. This event was specific to the intercellular adhesion molecules-1 (ICAM-1)/leukocyte function-associated antigen-1 interaction, as demonstrated by the following: (1) ICAM-1 expression correlated with increased EC contraction; and (2) the blocking of ICAM-1 selectively inhibited EC separation. Thus, we suggest that "cell-cell separation" could be a mechanism for diapedesis in situations that may require massive leukocyte infiltration.
Subject(s)
Humans , Cell Movement , Cells, Cultured , Endothelial Cells/cytology , Flow Cytometry , Fluorescent Antibody Technique , Intercellular Adhesion Molecule-1/metabolism , Leukocytes/cytology , Lymphocyte Function-Associated Antigen-1/metabolismABSTRACT
The basic route and mechanism for diapedesis has not yet to be fully defined. Here we present evidence that "cell-cell separation" between endothelial cells (ECs) may provide a route for leukocyte diapedesis. We unexpectedly found that extensive interaction between peripheral blood leukocytes and ECs that were activated by TNF-alpha induced the opening of EC contacts and, surprisingly, resulted in cell-cell separation. This event was specific to the intercellular adhesion molecules-1 (ICAM-1)/leukocyte function-associated antigen-1 interaction, as demonstrated by the following: (1) ICAM-1 expression correlated with increased EC contraction; and (2) the blocking of ICAM-1 selectively inhibited EC separation. Thus, we suggest that "cell-cell separation" could be a mechanism for diapedesis in situations that may require massive leukocyte infiltration.
Subject(s)
Humans , Cell Movement , Cells, Cultured , Endothelial Cells/cytology , Flow Cytometry , Fluorescent Antibody Technique , Intercellular Adhesion Molecule-1/metabolism , Leukocytes/cytology , Lymphocyte Function-Associated Antigen-1/metabolismABSTRACT
Objective To investigate the expression of lymphocyte function-associated antigen-1 (LFA-1)on peripheral blood mononuclear cells(PBMCs)from patients with SLE and its relation to the occurrence and development of SLE.Methods PBMCS were obtained from 30 patients with SLE and 24 normal human controls.Atier labeled with anti-CD3-FITC and anti-CD11 a-PE,PBMCS were detected for the expression of CD11a by flow cytometric analysis.Results The positivity rate of CD11a was significantly higher in patients with SLE and those with active SLE than in the normal controls (73.74%±7.89%and 77.11%±7.46%vs 68.21%±4.58%.both P<0.05).Furthermore,increased positivity rate of CD11a was observed in patients with active SLE compared with those with stable SLE (77.11%±7.46%vs 69.33%±6.27%,P<0.05).but there was no significant difieFence between the latter group and normal controls(P>0.05).In addition.the expression level of CD11a was positively correlated with SLE disease activity index (r=0.64,P<0.0 1)in patients.Conclusion The overexpression of LFA-1 may take part in the occurrence of SLE and correlate with the activity of SLE.
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Objective To explore the neuroimmunomedulation mechanism of ICAM-1 and LFA-1 in children with febrile seizures (FS).Methods 40 children with FS were dividedinto simple FS(SFs)groupin20 cases and complex FS(CFS)groupin20 cases,and 30 health children matched with regard to age and sex were enrolled into control group.The real-time fluorescence quantitative PCR wag used to detect the expression of PBMC ICAM-1 mRNA.At the same time,the PBMC LFA-1 mRNA expression wfs studied with Send-QuantitativeRT-PCR analysis.Results The levels of PBMC ICAM-1 mRNA in SFS group were significantly higher than those in control group and CF$group(P<0.05).The levels ofPBMC ICAM-1 mRNA showed downtrend between CFS group and control group.but there was no statistical difference between the two groups(P>0.05).The levels of PBMC LFA-1 mRNA grey-scales in SFS group were significantly higher than those in control group and CFS group(P<0.05).In addition,the levels of PBMC LFA-1 mRNA in CFS group showed downtrend than those in control group,but there wti8 no statistical difference between the two groups(P>0.05).Conclusions The gene expression levels of PBMC ICAM-I/LFA-I in SFS group were different from those in CFS group.Inflammable immunopathology damage induced by ICAM-1/LFA-1 may play an important role in the pathogenesis of SFS.On the contrary,ICAM-1/LFA-1 may have seme neuroprotective effects on the pathogenesis of CFS.
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Objective To study the methylation of CpG motifs and expression of lymphocyte function-associated antigen-1 (LFA-1) mRNA in patients with systemic lupus erythematosus (SLE), and evaluate their associations. Methods The peripheral blood lymphocytes were collected from 26 patients with SLE and 17 normal individuals. The methylation of CpG motifs was detected by the 5-methylcytosine antibody and flow eytometry, and the expression of LFA-1 mRNA was analyzed by RT-PCR. Results Methylation of CpG motifs in patients with SLE was lower than the control subjects (P<0.05), and a negative correlation existed between methylation of CpG motifs and SLEDAI (P<0.05). While expression of LFA-1 mRNA in patients with SLE was higher than that in the control group (P<0.05), and a positive correlation could be detected between the expression of LFA-1 mRNA and SLEDAI (P<0.05). There was a negative correlation between methylation of CpG motifs and expression of LFA-1 mRNA (P<0.05) in patients with SLE. Conclusion Hypomethylation of CpG motifs does exist in patients with SI,E, and is correlated with high expression of LFA-1, therefore, epige-netics plays an important role in the pathogenesis of SLE.
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Objective: To study the effects and significances of lipopolysaccharide preconditioning on the activities of NF-?B and the expression of ICAM-1/LFA-1 in rats graft liver ischemia/reperfusion injury. Methods: Male Sprague-Dawley rats were divided into three groups: sham operation group(Sham group),orthotopic liver transplantation group (OLT group) and LPS preconditioning group (LPS group). Only dissecting hepatoduodenal ligament was perfomed in Sham group. Experiments of OLT were performed by two-cuff method in OLT group and LPS group. The activities of NF-?B and the expression of ICAM-1/LFA-1 in hepatic tissue ,the levels of ALT,AST in inferior caval vein blood were detected at 0,60,180 min after dissecting of hepatoduodenal ligament in Sham group and after portal vein reperfusion in OLT group and LPS group. Results: Compared with those in Sham group at the different time points respectively,the activities of NF-?B and the expression of ICAM-1/LFA-1 were higher in OLT group and LPS group(P0.05) at 0 min after reperfusion,they were evidently higher in OLT group than in LPS group (P
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Objective To observe the expression of leukocyte-function-associated antigen-1(LFA-1)immunoreaction in children with febrile seizures(FS),and explore the neuroimmunomodulation mechanisms in the pathogenesis of FS.Methods Adopting flow cytometry(FCM),the levels of LFA-1 contained in blood serum and peripheral blood mononuclear cells(PBMC)of 60 cases [simple FS(SFS)30 cases;complex FS(CFS)30 cases] with febrile convulsion were analyzed,and compared with those in a normal group(30 cases).Out of 60 children with FS group,the LFA-1 mRNA in 20 cases with SFS and 20 cases with CFS was analyzed,and LFA-1 mRNA in19 health children taken out from control group(30 cases)was analyzed.The real-time PCR was used to detect the expression of PBMC LFA-1 mRNA.Results The expression of LFA-1 in the surface of PBMC of the 3 groups,the highest LFA-1 level was in the SFS group(50.89?21.36),the lowest LFA-1 level was in the CFS group(34.35?11.45),and control group was(41.39?16.30).Significant differences were found in 3 groups(Pa
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To investigate the change in renal CD11a and myeloperoxidase (MPO) contents and renal function during ischemia-reperfusion injury (IRI) of rat kidney, as well as the role of lymphocyte function associated antigen-1 (LFA-1) played in renal IRI, we utilized a rat model of renal IRI to detect the renal tissue contents of CDlla and MPO and renal functioa The results showed that the levels of CD11a and MPO were very low in normal renal tissue, but increased significantly in ischemia reperfusion. Renal failure was presented in rats with IRI. The levels in the ischemia-60min-reperfusion group were higher than those in the ischemia-30min-reperfusion group in the same time points. Our conclusion is the levels of CD11a and MPO in rat renal tissue increase significantly during renal IRI. LFA-1-mediated leukocyte adherence plays an important role in renal IRI. Up-regulation of CDlla further elucidates the mechanism of relation between IRI and acute rejection: more heavier IRI is, more possible acute rejection would be.
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AIM: To investigate the effects of oxidized high-density lipoprotein (oxHDL) on membrane fluidity and the expression of lymphocyte function associated antigen(LFA-1) of monocyte. METHODS: The membrane fluidity of THP-1 cells was assayed by fluorescence anisotropy with DPH (1,6-dipheny-1,3,5-hexatriene), a fluorescent probe; The LFA-1 expression on THP-1 cells were assayed by flow cytometry with indirect immunofluorescence.RESULTS: The membrane fluidity of THP-1 cells was reduced by 45% and 52% respectively ( P