Journal: AIDS Research and Human Retroviruses

Article Title: Defective Plasmacytoid Dendritic Cell-NK Cell Cross-Talk in HIV Infection

doi: 10.1089/aid.2008.0311

Figure Lengend Snippet: HIV gp120-mediated interference of pDC-NK cell cross-talk involves direct binding to the integrin α4β7 receptor on NK cells. We performed experiments of pDC-NK cell interactions in the presence or absence of trimeric HIV gp120 as described below. (A) PBMCs from healthy normal volunteers were treated with trimeric HIV gp120 (1 μg/ml) for 1 h at 37°C. The cells were washed and treated with CpG (1 μg/ml) overnight. CD69 expression on NK cells was determined by flow cytometry as described in Materials and Methods. Exposure to HIV gp120 markedly reduces the ability of NK cells to be activated, while gp120 by itself does not activate NK cells. (B) Purified pDCs and NK cells from healthy normal volunteers were isolated as described in Materials and Methods. Both pDCs and NK cells were exposed to trimeric HIV gp120 (1 μg/ml) for 60 min. Afterward the cells were washed and cocultured as follows. HIV untreated pDCs and NK cells with or without CpG (1 μg/ml), HIV gp120 treated or untreated pDCs with HIV gp120, and untreated or treated NK cells with or without CpG (1 μg/ml). The suppression of CD69 expression is calculated as a percentage of CD69 expression seen on NK cells with experimental conditions appropriate to that seen on NK cells cocultured with pDCs treated with CpG. Results indicate both cell types are affected by exposure to HIV gp120. (C) We performed experiments similar to those described in (B); when NK cells were pretreated with α4 integrin antibody that blocks HIV gp120 binding to NK cells, there was a complete reversal of the suppressive effects of HIV gp120.

Article Snippet: The α 4 integrin monoclonal antibody (HP2/1) was purchased from Santa Cruz Biotechnology Inc., Santa Cruz, CA.

Techniques: Binding Assay, Expressing, Flow Cytometry, Purification, Isolation

Journal: Stem cells international

Article Title: Hypoxia Inducible Factor-1α Regulates the Migration of Bone Marrow Mesenchymal Stem Cells via Integrin α 4.

doi: 10.1155/2016/7932185

Figure Lengend Snippet: Figure 4: The effect of integrin 𝛼4 inhibition on BM-MSC migration and activities of MMPs under hypoxia. (a) The mRNA expression of integrin 𝛼4 in BM-MSCs was suppressed by transfection of integrin 𝛼4 siRNA. 18S rRNA was used as the loading control. (b) BM-MSC migration was significantly increased after siITGA4 transfection. Invasiveness of BM-MSCs was assessed by invasion assay (left). BM-MSCs invaded through the inserts were counted for quantification (right). (c) Enzymatic activities of MMP-9 and MMP-2 in BM-MSCs after siITGA4 transfection were determined by zymography (left). Quantification of enzymatic activities of MMP-9 (middle) and MMP-2 (right). ∗𝑃< 0.05 (compared with siITGA4 nontransfected group) and #𝑃< 0.05 (compared with normoxic group). MMP: matrix metalloproteinase; siITGA4: integrin 𝛼4 siRNA.

Article Snippet: BM-MSCs were treated with integrin α 4 siRNA (siITGA4) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mock to inhibit integrin α 4 expression according to the manufacturer’s protocol under normoxic or hypoxic conditions.

Techniques: Inhibition, Migration, Expressing, Transfection, Control, Invasion Assay, Zymography

Journal: Stem cells international

Article Title: Hypoxia Inducible Factor-1α Regulates the Migration of Bone Marrow Mesenchymal Stem Cells via Integrin α 4.

doi: 10.1155/2016/7932185

Figure Lengend Snippet: Figure 5: Interaction between integrin 𝛼4 and HIF-1𝛼and its effect on expression of Rho GTPases under hypoxia. (a) Protein expression levels of integrin 𝛼4, HIF-1𝛼, ROCK1, and Rac1/2/3 were assessed by Western blotting. GAPDH was used as the loading control. (b) HIF-1𝛼and integrin 𝛼4 were localized with immunofluorescence in BM-MSCs after siITGA4 transfection under normoxic or hypoxic conditions. Blue: DAPI; green: HIF-1𝛼; red: integrin 𝛼4. Scale bar = 80 𝜇m (400x original magnification). ∗𝑃< 0.05 (compared with siITGA4 nontransfected group) and #𝑃< 0.05 (compared with normoxic group). DAPI: 4,6-diamidino-2-phenylindole; HIF-1𝛼: hypoxia-inducible factor-1𝛼; ITGA4: integrin 𝛼4; ROCK1: Rho-associated kinase 1; siITGA4: integrin 𝛼4 siRNA.

Article Snippet: BM-MSCs were treated with integrin α 4 siRNA (siITGA4) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mock to inhibit integrin α 4 expression according to the manufacturer’s protocol under normoxic or hypoxic conditions.

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Transfection

Journal:

Article Title: Islet ?-Cell-Specific T Cells Can Use Different Homing Mechanisms to Infiltrate and Destroy Pancreatic Islets

doi: 10.2353/ajpath.2007.060142

Figure Lengend Snippet: Expression of homing receptors on T cells is determined by the site of T-cell activation. A: Proliferation of OT-I cells in antigen-draining lymph nodes after injection of 4 × 106 carboxyfluorescein succinimidyl ester-labeled OT-I cells into RIP-mOVA or RIP-OVAlo mice. The latter mice were given OVA orally (3 mg) or subcutaneously (0.2 mg). OT-I cells were collected from lymph nodes (PaLN, pancreatic; MLN, mesenteric; IngLN, inguinal lymph node) 48 hours (RIP-mOVA) or 60 hours (RIP-OVAlo) later. B: Expression of α4-, β1-, and α4β7 integrin and of L-selectin and P-selectin binding activity (P-selectin ligand) on OT-II cells proliferating in PaLN and KLN (kidney LN) of RIP-mOVA mice and on OT-II cells proliferating in MLN and PLN of RIP-OVAlo mice. C: OT-I cells simultaneously transferred into the same recipient mice were analyzed for the expression of the same adhesion molecules.

Article Snippet: Cells were stained with Alexa 647-conjugated anti-CD4 and PerCP-Cy5.5-conjugated anti-CD8 and with phycoerythrin (PE)-conjugated anti-integrin α4 monoclonal antibody (mAb) (R1-2), anti-α4β7 heterodimer (DATK-32), or with biotinylated anti-integrin β1 chain mAb (Ha2/5), or anti-CD62L mAb (MEL-14), or with isotype-matched control antibodies, all from Becton, Dickinson and Company, San Jose, CA.

Techniques: Expressing, Activation Assay, Injection, Labeling, Binding Assay, Activity Assay

Journal:

Article Title: Islet ?-Cell-Specific T Cells Can Use Different Homing Mechanisms to Infiltrate and Destroy Pancreatic Islets

doi: 10.2353/ajpath.2007.060142

Figure Lengend Snippet: Expression of homing receptors α4 and α4β7 integrin and L-selectin in islets. OT-II (0.3 × 106) and OT-I (0.2 × 106) cells were co-transferred into RIP-mOVA mice and into RIP-OVAlo mice given oral or subcutaneous OVA. After 10 days, mice were sacrificed and pancreata analyzed by fluorescence microscopy. Homing receptors (red fluorescence) were detected with PE-conjugated or biotinylated antibodies and SA-PE. T cells were detected with anti-CD4 and anti-CD8 antibodies directly conjugated to fluorescein isothiocyanate (green fluorescence). Original magnifications, ×200.

Article Snippet: Cells were stained with Alexa 647-conjugated anti-CD4 and PerCP-Cy5.5-conjugated anti-CD8 and with phycoerythrin (PE)-conjugated anti-integrin α4 monoclonal antibody (mAb) (R1-2), anti-α4β7 heterodimer (DATK-32), or with biotinylated anti-integrin β1 chain mAb (Ha2/5), or anti-CD62L mAb (MEL-14), or with isotype-matched control antibodies, all from Becton, Dickinson and Company, San Jose, CA.

Techniques: Expressing, Fluorescence, Microscopy

Journal: Frontiers in Immunology

Article Title: The Microenvironment in Barrett’s Esophagus Tissue Is Characterized by High FOXP3 and RALDH2 Levels

doi: 10.3389/fimmu.2018.01375

Figure Lengend Snippet: Antibodies for FACS staining.

Article Snippet: CD49d (α4-integrin)-FITC , clone BU49, 1:20 , EuroBiosciences, Friesoythe, Germany.

Techniques: Staining