Journal:

Article Title: Accelerated chemokine receptor 7-mediated dendritic cell migration in Runx3 knockout mice and the spontaneous development of asthma-like disease

doi: 10.1073/pnas.0504787102

Figure Lengend Snippet: Dysregulated expression of CCR7 in Runx3 KO DC. (A) Impaired TGFβ-dependent inhibition of CCR7 transcription in KO BMDC. WT and Runx3 KO BMDC were grown in the presence or absence of TGFβ (10 ng/ml). At day 6, cells were induced to undergo maturation by LPS, and, at day 7, RNA was prepared and analyzed by RT-PCR. (B) Impaired TGFβ-dependent inhibition of surface expression of CCR7 in KO BMDC. WT and Runx3 KO BMDC were grown and treated as in A. At day 7, cells were analyzed by FACS by using anti-CCR7 antibodies (goat anti-mouse CI0131, Capralogics). FSChighCD11c+ DC were gated, and their CCR7 expression was determined. Reduction in the level of surface CCR7 and in the number of cells expressing it was noted only in WT and not in Runx3 KO BMDC. (C and D) Increased CCR7 expression on alveolar and LN DC of Runx3 KO mice. BAL and peripheral LN cells of KO and WT mice (n = 3) were obtained and analyzed. (C) Alveolar DC (FSChigh/CD11chigh) were gated and analyzed for CCR7 expression. Of note, expression of CCR7 on the DC subpopulation CD11c+/CD11b+ present only in KO lungs is shown along with that of KO CD11c+/CD11b- DC. (D) FSChigh/CD11chigh DC of axillary and thoracic LN were gated and analyzed for CCR7 expression.

Article Snippet: At day 7, cells were analyzed by FACS by using anti-CCR7 antibodies (goat anti-mouse CI0131, Capralogics).

Techniques: Expressing, Inhibition, Reverse Transcription Polymerase Chain Reaction

Journal:

Article Title: Accelerated chemokine receptor 7-mediated dendritic cell migration in Runx3 knockout mice and the spontaneous development of asthma-like disease

doi: 10.1073/pnas.0504787102

Figure Lengend Snippet: Elevated CCR7 mediated in vivo trafficking of alveolar DC to the draining LN in the KO mice. (A and B) Runx3 KO (n = 7) and WT (n = 5) mice were treated by intranasal administration of CFSE to label in vivo the respiratory DC. When indicated, WT mice were treated with LPS (n = 4), and Runx3 KO mice were treated with anti-CCR7 antibody (n = 4) or with buffer only (n = 4). KO mice (n = 4) and LPS-treated WT mice (n = 3) were also treated by inhalation of Ciglitazone. Eighteen hours later, mice were killed, and single-cell suspensions of BAL, thoracic LN, and axillary LN were prepared and analyzed by FACS. (A) FSChigh/CD11c+ DC were gated (R1 and R2). Shown is representative side scatter (SSC) versus CFSE staining of DC populations in BAL and LN after the various treatments. (B) Migration index of alveolar DC to thoracic LN represents the ratio between the percentage of CFSE+ cells within the CD11c+ population in the thoracic LN and the respective value in BAL cells. Results are presented as mean ± SEM. Analysis of variance showed that the migration index of untreated KO DC was significantly higher than that of WT (*, P = 0.016). Notably, the anti-CCR7-treated KO DC migration index was similar to basal migration of WT, and Ciglitazone significantly (*, P = 0.03) reduced the migration of KO DC.

Article Snippet: At day 7, cells were analyzed by FACS by using anti-CCR7 antibodies (goat anti-mouse CI0131, Capralogics).

Techniques: In Vivo, Staining, Migration

Journal: BMC Infectious Diseases

Article Title: Evaluation of envelope domain III-based single chimeric tetravalent antigen and monovalent antigen mixtures for the detection of anti-dengue antibodies in human sera

doi: 10.1186/1471-2334-11-64

Figure Lengend Snippet: Comparative analysis of DENV-specific antibodies in patient sera in indirect ELISAs using DENV EDIII-based coating antigens . Anti-DENV IgG (panels A-D) and IgM (panels E-H) antibodies were detected using a mixture of four monovalent EDIIIs (panels A & E), EDIII-T (panels B & F), or b-EDIII-T (panels C, D, G & H) as the coating antigen. The antigens were coated on either polystyrene (panels A, B, C, E, F & G) or streptavidin (panels D & H) plates. The S/Co values of individual DENV-positive and -negative sera, clustered together, are indicated by filled and unfilled circles, respectively. The horizontal line in each panel indicates the cut-off corresponding to S/Co ratio of 1.

Article Snippet: Sera were diluted 1:80 in IgM assay diluent (IgG assay diluents lacking goat serum and KCl) and pre-incubated with Serion Rf-Absorbent as recommended by the manufacturer (Virion/Serion GmbH, Würzburg, Germany) to remove IgG antibodies before using in the assay.

Techniques: