normal human becs  (Cambrex)

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: RSV induces TSLP expression in primary AECs. A, NHBECs were exposed to UV-irradiated RSV or RSV with indicated virus titers for 12 hours, and TSLP mRNA was measured by real-time quantitative PCR for virus titer. B and C, NHBECs were exposed to UV-irradiated RSV or RSV (MOI, 1) for indicated time course. Cell or culture supernatant fluids were harvested, and TSLP mRNA was measured by real-time quantitative PCR (Fig 1, B). TSLP protein levels were measured by ELISA (Fig 1, C). D and E, NHBECs were exposed to RSV (MOI, 1) without or with anti-TNFα antibodies (10, 50, or 100 ng/mL) (Fig 1, E), anti–IL-1R (1, 10, or 50 µg/mL) (Fig 1, E) for 24 hours, and culture supernatant fluids were harvested. TSLP protein levels were analyzed by ELISA. Data represent means ± SDs of 3 independent measurements. Similar results were obtained for 5 independent experiments (Fig 1, A–E). *P ≤ .05.

Article Snippet: Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Expressing, Irradiation, Virus, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: RIG-I and IPS-1 activation mediates Paramyxovirus-induced TSLP expression. A and B, A549 cells were transfected with control plasmid or luciferase constructs containing the human TSLP promoter alone or in combination with expression vectors for DN-RIG-I (Fig 2, A) or IPS-1 (Fig 2, B) followed by infection with RSV (MOI, 1). C, Normal littermate control (NLC) or RIG-I KO MEFs were infected with or without RSV, and TSLP protein expression was measured by ELISA. n.d. indicates not detected. D, NHBECs were transfected with control or RIG-I siRNA, followed by RSV infection, and TSLP mRNA was assessed at 8 hours after infection. Data represent means ± SDs of 3 independent measurements. Similar results were obtained for 5 independent experiments (Fig 2, A–D). *P ≤ .05.

Article Snippet: Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Activation Assay, Expressing, Transfection, Control, Plasmid Preparation, Luciferase, Construct, Infection, Enzyme-linked Immunosorbent Assay

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: Human asthmatic epithelium produces greater levels of TSLP in response to RSV infection. ALI cultures were generated from primary BECs isolated from healthy or asthmatic children via bronchial brushing. Data show TSLP protein levels, measured by ELISA, in basolateral culture supernatant fluids after infection with either RSV Line 19 or RSV A2 or control at an MOI of 0.5 (n = 12 patients for all groups). P values were calculated with an ANOVA with Tukey posttest.

Article Snippet: Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Infection, Generated, Isolation, Enzyme-linked Immunosorbent Assay, Control

Journal: The Journal of allergy and clinical immunology

Article Title: Thymic stromal lymphopoietin is induced by respiratory syncytial virus–infected airway epithelial cells and promotes a type 2 response to infection

doi: 10.1016/j.jaci.2012.07.031

Figure Lengend Snippet: RSV-mediated expression of RIG-I and TLR3 in ALI cultures from healthy and asthmatic children. RIG-I and TLR3 expression (log2 scale) by BECs from healthy (open plots; n = 9) and asthmatic children (solid plots; n = 12) in response to RSV infection by A2 and Line 19 strains or exposure to control vero cell supernatant fluid (VC). RIG-I expression: ANOVA P < .001 between groups; asthma RSV A2 group 3-fold greater RIG-I expression than asthma VC group (*P < .001); asthma RSV 19 group 4-fold greater RIG-I expression than asthma VC group (*P < .001); no significant change in RIG-I expression by healthy cells with RSV A2 or RSV 19; no significant difference in RIG-I expression between asthmatic and healthy VC groups. TLR3 expression: ANOVA P = .003 between groups; healthy RSV A2 group 1.4-fold greater TLR3 expression than by healthy VC group (**P = .05); healthy RSV 19 group 1.8-fold greater TLR3 expression than by healthy VC group (***P = .03); asthma RSV A2 group 1.5-fold greater TLR3 expression than by asthma VC group (†P = .002); asthma RSV 19 group 1.8-fold greater TLR3 expression than by asthma VC group (††P = .002); no significant difference in TLR3 expression between asthmatic and healthy VC groups. RIG-I and TLR3 expression normalized to GAPDH. Bottom and top of box plots represent 25th and 75th percentiles, respectively; dotted band represents the median and whiskers represent minimums and maximums.

Article Snippet: Normal human BECs (NHBECs) were maintained in appropriate medium according to the manufacturer’s instruction (Lonza, Walkersville, Md).

Techniques: Expressing, Infection, Control