Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Burkholderia mallei and Burkholderia pseudomallei stimulate differential inflammatory responses from human alveolar type II cells (ATII) and macrophages

doi: 10.3389/fcimb.2012.00165

Figure Lengend Snippet: B. mallei ( BM ) and B. pseudomallei ( BP ) interaction with primary human ATII cells (ATII cells). ATII cells were grown on 3.0-um transwell filters to confluence prior to apical exposure of BM and BP at an MOI of approximately 100:1 for 4 h. Minimal contact of BM ( A and B ) with ATII cells was visualized via SEM. However, BP (C–E) adhesion to the apical surface of ATII cells was observed and the pathogen was entangled by the ATII microvilli [ D (white boxed area) and E ]. Further, quantitative analysis of attachment showed that BP significantly adhered better to ATII cells compared to BM ( p < 0.05; F ). Data is representative of triplicate samples of three experiments and is represented as mean ± SEM. * p < 0.05.

Article Snippet: Primary human type II alveolar epithelial (ATII) cells (ScienCell, San Diego, CA) were seeded on BioCoat Growth Factor Reduced Matrigel®-coated transwell filters (BD; Franklin Lakes, NJ) and maintained using Bronchial Epithelial Medium enhanced with the Bullet kit supplements provided by the manufacturer (BEGM; Lonza, Basel, Switzerland) supplemented with 10% charcoal-stripped FBS (Hyclone; Logan, UT) and 10 ng/mL of keratinocyte growth factor (KGF; PeproTech, Rockhill, UT).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Burkholderia mallei and Burkholderia pseudomallei stimulate differential inflammatory responses from human alveolar type II cells (ATII) and macrophages

doi: 10.3389/fcimb.2012.00165

Figure Lengend Snippet: BM and BP -induced time-dependent secretion of cytokines from ATII cells and MDM. ATII cells and MDM were infected with BM or BP (MOI 100:1) for 6 or 20 h. Supernatant was collected and analyzed for cytokine secretion. Un-stimulated cells served as baseline controls. BP stimulated higher levels of IL-6 (A) and TNF-α (B) from MDM as well as IL-6 (D) and IL-8 (E) from ATII cells at 6 and 20 h post-infection compared to BM . However, BM -induced significantly higher IL-10 secretion from MDM compared to BP at 6 h post-infection (C) . Additionally, IL-10 was significantly secreted from BM-ATII cells compared to BP at both 6 and 20 h post-infection (F) . Data is representative of two experiments containing duplicate conditions/experiment and is represented as mean ± SEM. * denotes significance of p < 0.05.

Article Snippet: Primary human type II alveolar epithelial (ATII) cells (ScienCell, San Diego, CA) were seeded on BioCoat Growth Factor Reduced Matrigel®-coated transwell filters (BD; Franklin Lakes, NJ) and maintained using Bronchial Epithelial Medium enhanced with the Bullet kit supplements provided by the manufacturer (BEGM; Lonza, Basel, Switzerland) supplemented with 10% charcoal-stripped FBS (Hyclone; Logan, UT) and 10 ng/mL of keratinocyte growth factor (KGF; PeproTech, Rockhill, UT).

Techniques: Infection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Burkholderia mallei and Burkholderia pseudomallei stimulate differential inflammatory responses from human alveolar type II cells (ATII) and macrophages

doi: 10.3389/fcimb.2012.00165

Figure Lengend Snippet: Phagocytosis and intracellular replication of BM and BP . MDM and ATII cells were infected with BM or BP (MOI of 100:1). As determined by CFUs, phagocytosis (accessed at 1 h) and intracellular replication (accessed at 3 h post-infection) of BP by MDM was significantly higher compared to BM (A) . Alternatively, neither BM nor BP invaded or replicated to high numbers in ATII cells (B) . Data is representative of two experiments containing duplicate conditions/experiment and is represented as mean ± SEM. * denotes significance of p < 0.05 and ** p < 0.01.

Article Snippet: Primary human type II alveolar epithelial (ATII) cells (ScienCell, San Diego, CA) were seeded on BioCoat Growth Factor Reduced Matrigel®-coated transwell filters (BD; Franklin Lakes, NJ) and maintained using Bronchial Epithelial Medium enhanced with the Bullet kit supplements provided by the manufacturer (BEGM; Lonza, Basel, Switzerland) supplemented with 10% charcoal-stripped FBS (Hyclone; Logan, UT) and 10 ng/mL of keratinocyte growth factor (KGF; PeproTech, Rockhill, UT).

Techniques: Infection

Journal: Respiratory Research

Article Title: YAP as a potential therapeutic target for myofibroblast formation in asthma

doi: 10.1186/s12931-025-03115-x

Figure Lengend Snippet: YAP was increased in protein abundance and nuclear translocated in ATII cells in human asthmatic and HDM-treated mouse lungs. Representative immunofluorescence images for a human and b mouse samples (white stars indicate YAP nuclear translocation in ATII cells). c Statistical analysis to show the higher YAP staining intensity in human asthmatic and HDM-treated mouse samples as compared with the controls. d Western blots to show the upregulation of YAP and non-pS127 YAP (active form of YAP) in HDM-treated mouse lung lysates. n = 5 mice/group, * P < 0.05, as compared with the saline group

Article Snippet: The human primary ATII cells were purchased from iCell Bioscience Inc. (Shanghai, China), and maintained in the iCell primary epithelial cell culture medium (PriM-iCell-001, iCell).

Techniques: Quantitative Proteomics, Immunofluorescence, Translocation Assay, Staining, Western Blot, Saline

Journal: Respiratory Research

Article Title: YAP as a potential therapeutic target for myofibroblast formation in asthma

doi: 10.1186/s12931-025-03115-x

Figure Lengend Snippet: CTGF protein abundance was increased in human asthmatic and HDM-treated mouse lungs, and localized around ATII cells. Representative immunofluorescence images for CTGF and pro-SPC staining in a human and b mouse lung samples. c Statistical analysis showed that the CTGF staining intensity was significantly higher in the human asthmatic and HDM group, as compared with non-asthmatic and saline groups. d Representative blots and statistical analysis to show that CTGF protein abundance was significantly upregulated in HDM-treated mouse lung lysates (n = 7 for asthmatic and n = 8 for non-asthmatic samples; n = 5 mice/group, * P < 0.05, as compared with non-asthmatic or mouse saline group)

Article Snippet: The human primary ATII cells were purchased from iCell Bioscience Inc. (Shanghai, China), and maintained in the iCell primary epithelial cell culture medium (PriM-iCell-001, iCell).

Techniques: Quantitative Proteomics, Immunofluorescence, Staining, Saline

Journal: Respiratory Research

Article Title: YAP as a potential therapeutic target for myofibroblast formation in asthma

doi: 10.1186/s12931-025-03115-x

Figure Lengend Snippet: YAP was differentially regulated in the bronchial epithelium and alveolar epithelial cells. a Representative immunofluorescence images and b statistical analysis to show that the alveolar cells number and non-pS127 YAP positive alveolar cells ratio were increased in HDM-induced mouse lungs (n = 5 mice/group, * P < 0.05, as compared with the saline group). Representative western blots and statistical analysis for c human primary ATII cells, and d BEAS-2B cells (n = 3–7, * P < 0.05, as compared with 0 min)

Article Snippet: The human primary ATII cells were purchased from iCell Bioscience Inc. (Shanghai, China), and maintained in the iCell primary epithelial cell culture medium (PriM-iCell-001, iCell).

Techniques: Immunofluorescence, Saline, Western Blot

Journal: Aging Cell

Article Title: Telomerase modRNA Offers a Novel RNA ‐Based Approach to Treat Human Pulmonary Fibrosis

doi: 10.1111/acel.70240

Figure Lengend Snippet: Dose‐dependent increase of hTERT mRNA and telomerase activity in vitro after modRNA hTERT transfection. (A) Schematic presentation of the modRNA hTERT template. (B) The modRNA production is divided into three main steps: the sequence design, then in vitro transcription (IVT) where the modRNA is generated and lastly the purification step. (C) Schematic describing experimental setup used to validate transfection efficiency of modRNA GFP and functionality of modRNA hTERT. (D) Transmission electron microscopy of ATII cells. Arrowheads indicate lamellar body like structures. Nucleus is depicted as n . (E) Fold change of hTERT mRNA expression in ATII cells transfected with modRNA hTERT in a concentration range of 0.25–2 μg/mL compared to modRNA GFP 2 μg/mL control (modRNA control) after 24 and 48 h ( n = 3). (F) TRAP assay revealed reactivation of telomerase activity in ATII cells after 24 and 48 h in dose‐dependent effect. Positive (pos.) control = human induced pluripotent stem cells, negative (neg.) control = HUVEC cells, LB = lysis buffer, 1 = cell lysate, Δ = heat‐inactivated lysate. (G) Quantification of TRAP assays indicated a dose‐dependent increase of telomerase activity compared to modRNA control ( n = 3). (H) No prolonged immune reaction was detected after 48 h modRNA treatment in ATII cells compared to untreated control ( n = 3). * p < 0.05; *** p < 0.001; Two‐way ANOVA, Dunnett's multiple comparisons test.

Article Snippet: Human primary ATII cells (EP71AL Epithelix) were obtained from non‐smoking donors with no reported pathology (Epithelix Sarl, Geneva, Switzerland).

Techniques: Activity Assay, In Vitro, Transfection, Sequencing, Generated, Purification, Transmission Assay, Electron Microscopy, Expressing, Concentration Assay, Control, TRAP Assay, Lysis

Journal: Aging Cell

Article Title: Telomerase modRNA Offers a Novel RNA ‐Based Approach to Treat Human Pulmonary Fibrosis

doi: 10.1111/acel.70240

Figure Lengend Snippet: Repetitive modRNA hTERT treatment increased proliferation capacity and enhanced cell viability in ATII cells. (A) Population doubling level (PDL) demonstrated an increase after single modRNA hTERT treatment (1×; at day 0), which was further increased upon double modRNA hTERT treatment (2×; at day 0 and 11) Arrowheads indicate the time point of modRNA hTERT treatment performed in ATII cells ( n = 3). Geisser–Greenhouse's epsilon; Two‐way ANOVA; Dunnett's multiple comparison test. (B) BrdU assay indicated toward an increase in proliferation in ATII cells after modRNA hTERT treatment ( n = 5). (C) Immunofluorescence staining of Ki67, pH3 and γH2AX, (green) revealed an increase in proliferation and a decrease in DNA damage after modRNA hTERT transfection in ATII cells ( n = 3). (D) Representative images of qFISH staining in ATII cells after modRNA hTERT treatment. Brighter telomere spots (yellow, Tel Cy3), representing longer telomeres, were visible after 2× treatment with modRNA hTERT compared to 1× treatment and modRNA control group. Nucleus is depicted in blue (Hoechst). (E) Telomeric spots analysis revealed an increase of telomere length compared to modRNA control group ( n ≥ 95 nuclei per group out of 3 biological replicates were imaged). Welch ANOVA; Games‐Howell's multiple comparisons test. * p < 0.05; ** p < 0.01; *** p < 0.001; One‐way ANOVA or Two‐way ANOVA, Dunnett's multiple comparisons test.

Article Snippet: Human primary ATII cells (EP71AL Epithelix) were obtained from non‐smoking donors with no reported pathology (Epithelix Sarl, Geneva, Switzerland).

Techniques: Comparison, BrdU Staining, Immunofluorescence, Staining, Transfection, Control