Expression Profile Of Adhesion Molecules Research Articles

Expression profile of adhesion molecules in blastocyst vis-a-vis uterine epithelial cells

Models using in vitro produced buffalo embryos and in vitro cultured uterine epithelial cells (UECs) may be useful in understanding the intricacies of embryo-uterine cross talk. In the present study, buffalo UECs were obtained from slaughterhouse derived non-gravid uterus. UECs monolayer was treated with steroids (10pg/ml estradiol for 24h and 3.14 ng/ml progesterone for another 5 days). In vitro produced buffalo blastocysts were co-cultured over steroid treated UECs monolayer and at 72 h of co-culture, embryo attachment rate was higher in UECs treated with steroids (71.86% vs. 26.55%) while no attachment was observed on plastic surface. Naturally hatched or assisted hatched blastocysts were co-cultured over UECs monolayer treated with 3.14ng/ml progesterone (P4), or without any treatment for 72 h and the effect of co-culture on the expression profile of adhesion related biomolecules was analyed in UECs and blastocysts. Cultured UECs and blastocysts cultured in embryo culture media were considered as control. It was observed that the expression of MUC1 in UECs was significantly (p < 0.05) higher in control group than treatment groups. The relative mRNA abundance of integrins and osteopontin was significantly (p < 0.05) higher in UECs and blastocysts of treatment groups than control group. Expression of IFN-τ was significantly higher (p < 0.05) in embryos co-cultured with UECs than other treatment groups. It can be concluded that P4 supplementation is required for the modulation of adhesion molecules and co-culture of blastocysts and UECs together affect the expression of adhesion molecules both in blastocyts and in UECs.

Airborne particulate matter upregulates expression of early and late adhesion molecules and their receptors in a lung adenocarcinoma cell line

BackgroundEpidemiological evidence associates chronic exposure to particulate matter (PM) with respiratory damage and lung cancer. Inhaled PM may induce systemic effects including inflammation and metastasis. This study evaluated whether PM induces expression of adhesion molecules in lung cancer cells promoting interaction with monocytes. MethodsThe expression of early and late adhesion molecules and their receptors was evaluated in A549 (human lung adenocarcinoma) cells using a wide range of concentrations of PM2.5 and PM10. Then we evaluated cellular adhesion between A549 cells and U937 (human monocytes) cells after PM exposure. ResultsWe found higher expression of both early and late adhesion molecules and their ligands in lung adenocarcinoma cells exposed to PM2.5 and PM10 particles present in the air pollution at Mexico City from 0.03 μg/cm2 with a statistically significant difference (p ≤ 0.05). PM10 had stronger effect than PM2.5. Both PM also stimulated cellular adhesion between tumor cells and monocytes. ConclusionsThis study reveals a comprehensive expression profile of adhesion molecules and their ligands upregulated by PM2.5 and PM10 in A549 cells. Additionally these particles induced cellular adhesion of lung cancer cells to monocytes. This highlights possible implications of PM in two cancer hallmarks i.e. inflammation and metastasis, underlying the high cancer mortality associated with air pollution.

Human Bone Marrow-Resident Natural Killer Cells Have a Unique Transcriptional Profile and Resemble Resident Memory CD8+ T Cells.

Human lymphoid tissues harbor, in addition to CD56bright and CD56dim natural killer (NK) cells, a third NK cell population: CD69+CXCR6+ lymphoid tissue (lt)NK cells. The function and development of ltNK cells remain poorly understood. In this study, we performed RNA sequencing on the three NK cell populations derived from bone marrow (BM) and blood. In ltNK cells, 1,353 genes were differentially expressed compared to circulating NK cells. Several molecules involved in migration were downregulated in ltNK cells: S1PR1, SELPLG and CD62L. By flow cytometry we confirmed that the expression profile of adhesion molecules (CD49e−, CD29low, CD81high, CD62L−, CD11c−) and transcription factors (Eomeshigh, Tbetlow) of ltNK cells differed from their circulating counterparts. LtNK cells were characterized by enhanced expression of inhibitory receptors TIGIT and CD96 and low expression of DNAM1 and cytolytic molecules (GZMB, GZMH, GNLY). Their proliferative capacity was reduced compared to the circulating NK cells. By performing gene set enrichment analysis, we identified DUSP6 and EGR2 as potential regulators of the ltNK cell transcriptome. Remarkably, comparison of the ltNK cell transcriptome to the published human spleen-resident memory CD8+ T (Trm) cell transcriptome revealed an overlapping gene signature. Moreover, the phenotypic profile of ltNK cells resembled that of CD8+ Trm cells in BM. Together, we provide transcriptional and phenotypic data that clearly distinguish ltNK cells from both the CD56bright and CD56dim NK cells and substantiate the view that ltNK cells are tissue-resident cells, which are functionally restrained in killing and have low proliferative activity.

Immune Imbalance in Nasal Polyps of Caucasian Chronic Rhinosinusitis Patients Is Associated with a Downregulation of E-Selectin

Chronic rhinosinusitis with nasal polyps (CRSwNP) in Caucasians is a chronic Th2 inflammatory disease of the nasal and paranasal mucosa and the recruitment of leukocytes to the site of inflammation is poorly understood. We studied mRNA and protein expression profiles of adhesion molecules in nasal polyp and associated inferior turbinate tissues using molecular, biochemical, and immunohistological methods. Analysis showed a strongly decreased E-selectin expression in nasal polyps with a significant difference between eosinophil and neutrophil counts in nasal polyps and balanced counts in inferior turbinates. E-selectin expression is known to be downregulated in a Th2 milieu and has an essential role in immunosurveillance by locally activating neutrophil arrest and migratory function. A downregulation of E-selectin may come along with an immune imbalance in Caucasian nasal polyps due to a significant inhibition of neutrophil recruitment. Therefore, we suggest that an upregulation of E-selectin and the associated influx of neutrophils may play a significant role in the resolution of inflammation as well as for the pathophysiology of nasal polyps of Caucasian chronic rhinosinusitis patients.

A Direct Comparison of Expression Profiles of Adhesion Molecules on Naïve T Cells Between Cord Blood and Steady-State Bone Marrow Grafts of Healthy Donors

IntroductionWe compared the profiles of adhesion molecule expression on naïve T cells between umbilical cord blood (UCB) and steady-state bone marrow (SS-BM) grafts. MethodsThe expressions of 4 adhesion molecules, including very late antigen 4 (VLA-4), intercellular adhesion molecule-1 (ICAM-1), L-selectin, and lymophocyte function-associated antigen-1 (LFA-1) on naïve T cells in UCB (n = 25) and SS-BM (n = 10) were analyzed using flow cytometry. ResultsThe expressions of ICAM-1 and L-selectin on CD4+ T cells and CD8+ T cells in UCB were significantly lower than those on SS-BM (P < .05 for all). The expressions of VLA-4 and LFA-1 on CD8+ T cells in UCB were significantly lower than those of SS-BM (P = .002 and .047, respectively). Compared with SS-BM, we observed lower expression of ICAM-1 on naïve CD4+ and CD8+ T cells in UCB (P < .001 for all). The percentages of interferon (IFN)-γ positive cells among naïve CD4+ and CD8+ T-cell subsets were significantly lower in UCB, leading to ready polarization of naïve UCB T cells from a Th1 to Th2 phenotype versus those on SS-BM. ConclusionsOur results among UCB suggested lower intensities of ICAM-1 expression on naïve T cells and their easier polarization from Th1 to Th2 elements.

Addition of Hydroxyurea to Transfusion Programme to Treat Progressive Cerebral Vasculopathy

AbstractAbstract 4813Cerebro-vascular accidents (CVA) amongst paediatric patients with sickle cell disease (SCD) cause significant morbidity and mortality. Regular blood transfusions are used to prevent progressive cerebral vasculopathy. In primary stroke prevention, transcranial Doppler (TCD) studies are used to identify children at high risk of developing CVA and maintaining regular transfusion to keep HaemoglobinS (HbS) below 30% has shown to reduce this risk by 90%. However, about 10% children maintained on regular blood transfusions continue to have progressive vasculopathy despite maintaining HbS% below 30. There are few established options to try and prevent progressive vasculopathy despite adequate transfusion; possible approaches include intensifying the transfusion regime to maintain an even lower HbS%, revascularization surgery and correction of other thrombogenic risk factors. Hydroxyurea (HU) has been used in SCD to reduce the frequency of painful crises and acute chest syndrome. Its primary mode of action is thought to be to increase HbF levels but other mechanisms of action which may be beneficial include the reduction in white cell count, endothelium stabilization, improved expression profile of adhesion molecules, and nitric oxide donation. Herein we present our experience of combining treatment with HU to a transfusion programme in paediatric sickle cell patients with evidence of progressive cerebral vasculopathy whilst optimally transfused.4 paediatric patients with SCD were transfused as secondary stroke prevention, but had progressive cerebrovascular disease on magnetic resonance imaging (MRI/MRA). Following detailed discussions with the parents, HU was started in addition to the transfusion to try and arrest the progression. The dose of HU was gradually increased whilst monitoring counts, assessing tolerability with a view to achieving doses of at least 25mg/kg. All patients had regular clinical review, full blood counts, HbF and HbS percentages monitored baseline pre and post transfusion. All patients had annual TCD to assess cerebral circulation flow velocities and MRI/MRA. Iron chelation was commenced or continued as appropriate to their clinical circumstances for secondary iron overload.In this cohort, there were 2 females and 2 males, all of African descent. The mean age at diagnosis of first CVA was 5years (Range3-5yrs) with a mean steady state Hb of 7g/dl, mean HbF of 2.8% and mean HbS of 82.1%. MRI/MRA showed middle cerebral artery (MCA) cortical infarction in all patients at diagnosis but in addition showed varying degrees of areas of watershed infarction, small vessel disease with deep white matter ischemic change and moya-moya formation with small haemorrhages. The cohort remained on transfusions for a mean period of 55 months (Range 25–108) and received transfusions at a mean of 4 weekly intervals (Range 2–6 weeks). Mean Hb values post transfusion were 10.8g/dl with a mean HbF of 2% and HbS of 24.2%. TCD velocities in non-stenosed vessels showed a mean decrease in MCA by 26cm/sec (-11.8%), and in Internal carotid artery (ICA) by 14cm/sec (-8%). All 4 patients had evidence of clinical symptoms with new neurological deficits, and MRI/MRA scans showed a worsening picture of cerebral vasculopathy with increasing stenosis and new vessel formation. The mean duration of treatment with HU and transfusion was 18 months (Range 12–32months) with a mean Hb value of 11.4g/dl with a mean HbF of 5.8% and HbS of 14.6%. TCD on treatment with HU and transfusion showed a further mean reduction in flow velocity in MCA by 8cm/sec (-4.1%) in the non-stenosed vessels, and in ICA by 4cm/sec (-2.5%). MRI/MRA showed no evidence of progression. One patient had clinical symptoms of TIAs without further infarction and underwent neuro-revascularization.Our study shows that a carefully selected cohort of paediatric patients with advancing cerebral vasculopathy, may benefit from HU in addition to transfusion with modest but clinically significant improvement in HbF% and TCD profile over and above the substantial improvement seen after transfusions. It is possible that this added benefit is by way of HU reducing vascular endothelial damage, mild increases in HbF% and as yet, undefined mechanism. Further, larger studies are needed to assess the value of combining HU and transfusion, although our experience suggests that it is safe, well tolerated and potentially beneficial. Disclosures:No relevant conflicts of interest to declare.

Chondrogenic Transdifferentiation of Human Dermal Fibroblasts Stimulated with Cartilage-Derived Morphogenetic Protein 1

This study aimed to investigate the chondrogenic transdifferentiation potential of human dermal fibroblasts (DFs) by stimulation with cartilage-derived morphogenetic protein 1 (CDMP1). Using CDMP1 (100 ng/mL) we induced human DFs at passage 5 in both monolayer and micromass culture. Chondrogenic-specific markers were detected via immunochemistry, reverse transcription-polymerase chain reaction, and Western blot analysis in the collected specimens. The expression profile of adhesion molecules including integrin alpha5, beta1, and N-cadherin of DFs accompanying with chondrogenesis was further investigated. After 7 days of induction in monolayer culture, DFs acquired the polygonal chondrocyte-like shape with positive expression of chondrogenic-specific markers. Such a phenotypic transition of DFs was lost at 14 days. However, in micromass culture the chondrogenic transdifferentiation of DFs can be maintained even at 14 days. No chondrogenesis was detected in DFs without CDMP1 treatment under both culture conditions. By neutralization assay with blocking antibodies, it was further revealed that integrin alpha5 expression was in direct proportion to the degree of chondrogenic differentiation. Based on our findings, it can be ascertained that DFs are capable of transdifferentiating into a chondrogenic lineage by stimulation with CDMP1 in vitro. The integrin alpha5 mediating cell-cell and cell-matrix interactions is required for maintaining the chondrogenic phenotype of DFs.

Expression profiles of adhesion molecules on naïve T cells in bone marrow grafts of healthy donors treated with granulocyte colony-stimulating factor

Background and objectives Adhesion molecules on T cells were implicated in the process of leukocyte migration and GVHD. The aim of this study was to investigate the profiles of adhesion molecule expression on naïve T cells in bone marrow grafts of healthy donors treated with G-CSF. Methods The expression of four adhesion molecules, including VLA-4, ICAM-1, L-selectin, and LFA-1, on naïve T cells in G-CSF-primed bone marrow grafts (G-BM) from 35 healthy donors was analyzed using flow cytometry. Results G-CSF administration significantly decreased the expression of VLA-4, ICAM-1, L-selectin, and LFA-1 on naïve CD4 + and CD8 + T cells in bone marrow grafts. Treating healthy donors with G-CSF significantly increased the percentages of interleukin-4 (IL-4) positive cells in naïve CD4 + and CD8 + T cell subsets, leading to polarization of bone marrow naïve CD4 + and CD8 + T cells from Th1 to Th2 phenotype. Conclusions Our results suggest that CD4 + and CD8 + naïve T cells in G-BM have lower intensities of expression of several adhesion molecules. The clinical significance of these alteration warrants further study.

N-cadherin as a therapeutic target in cancer

During tumor progression, cancer cells undergo dramatic changes in the expression profile of adhesion molecules resulting in detachment from original tissue and acquisition of a highly motile and invasive phenotype. A hallmark of this change, also referred to as the epithelial–mesenchymal transition, is the loss of E- (epithelial) cadherin and the de novo expression of N- (neural) cadherin adhesion molecules. N-cadherin promotes tumor cell survival, migration and invasion, and a high level of its expression is often associated with poor prognosis. N-cadherin is also expressed in endothelial cells and plays an essential role in the maturation and stabilization of normal vessels and tumor-associated angiogenic vessels. Increasing experimental evidence suggests that N-cadherin is a potential therapeutic target in cancer. A peptidic N-cadherin antagonist (ADH-1) has been developed and has entered clinical testing. In this review, the authors discuss the biochemical and functional features of N-cadherin, its potential role in cancer and angiogenesis, and summarize the preclinical and clinical results achieved with ADH-1.

Involvement of alpha 1 and alpha 4 integrins in gut mucosal injury of graft-versus-host disease.

We have investigated the involvement of adhesion molecules in the lymphocyte infiltration associated with acute intestinal graft-versus-host disease (GVHD) induced by injection of C3H lymph node cells into irradiated (C3H x DBA/2)F1 mice. First we analyzed the expression profile of adhesion molecules including alpha 1, alpha 2, alpha 4, alpha 5, alpha 6, alpha L and beta 7 integrins, CD44 and L-selectin of lymphocytes from lymph nodes and gut mucosa in normal mice. In normal mice, intraepithelial lymphocytes (IEL) and lamina propria lymphocytes (LPL) uniquely showed increased expression of alpha 1, alpha 2 and beta 7 integrins, and decreased expression of L-selectin compared with that of lymphocytes of the lymph nodes and Peyer's patches. In mice with GVHD, IEL and LPL of donor lymph node cells origin underwent phenotypic changes characterized by the increased expression of alpha 1, alpha L and beta 7 integrins, and the loss of L-selectin. The expression profile of adhesion molecules on IEL and LPL of GVHD mice resembled that of normal mice except for the lack of alpha 2 integrin. Treatment of GVHD mice with anti-alpha 1, -alpha 4 or -beta 7 integrin antibody alone partially prevented the mucosal pathology of intestinal GVHD, whereas only mice treated with anti-alpha 1 showed reduced donor lymphocytic infiltration into the intestinal mucosa. In contrast, treatment with anti-alpha L or anti-CD44 antibody did not affect the intestinal GVHD. Furthermore, dual blockade of both alpha 1 and alpha 4 integrins completely inhibited the mucosal pathology and donor lymphocyte infiltration of intestinal GVHD.(ABSTRACT TRUNCATED AT 250 WORDS)