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Differentiation of nasal airway cell cultures and impact of HRV and influenza infection on mediators of airway inflammation (IL-1β and IL-6) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and <t>H1N1</t> for 24 and 48 h, as described in the methods. (A) hNEC differentiation was confirmed by qPCR using four independent differentiation genes that are not expressed in non-ciliated basal undifferentiated cells (FOX-J1, CCP-110, CDHR3) and for MUC5AC as described in Methods. (B–E) Proinflammatory cytokines interleukin 1 beta (IL-1β) (B,C) and IL-6 (D,E) were determined by multiplex Luminex immunoassay in both apical (B,D) and basal (C,E) chambers of air-liquid interface (ALI) inserts as described in methods. All panels show summary data as mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparisons between the uninfected control group and each experimental group were performed using a two-tailed Mann–Whitney test and are indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test, and the result is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway differentiation and inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.
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Differentiation of nasal airway cell cultures and impact of HRV and influenza infection on mediators of airway inflammation (IL-1β and IL-6) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and <t>H1N1</t> for 24 and 48 h, as described in the methods. (A) hNEC differentiation was confirmed by qPCR using four independent differentiation genes that are not expressed in non-ciliated basal undifferentiated cells (FOX-J1, CCP-110, CDHR3) and for MUC5AC as described in Methods. (B–E) Proinflammatory cytokines interleukin 1 beta (IL-1β) (B,C) and IL-6 (D,E) were determined by multiplex Luminex immunoassay in both apical (B,D) and basal (C,E) chambers of air-liquid interface (ALI) inserts as described in methods. All panels show summary data as mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparisons between the uninfected control group and each experimental group were performed using a two-tailed Mann–Whitney test and are indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test, and the result is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway differentiation and inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.
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Differentiation of nasal airway cell cultures and impact of HRV and influenza infection on mediators of airway inflammation (IL-1β and IL-6) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and <t>H1N1</t> for 24 and 48 h, as described in the methods. (A) hNEC differentiation was confirmed by qPCR using four independent differentiation genes that are not expressed in non-ciliated basal undifferentiated cells (FOX-J1, CCP-110, CDHR3) and for MUC5AC as described in Methods. (B–E) Proinflammatory cytokines interleukin 1 beta (IL-1β) (B,C) and IL-6 (D,E) were determined by multiplex Luminex immunoassay in both apical (B,D) and basal (C,E) chambers of air-liquid interface (ALI) inserts as described in methods. All panels show summary data as mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparisons between the uninfected control group and each experimental group were performed using a two-tailed Mann–Whitney test and are indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test, and the result is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway differentiation and inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.
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Differentiation of nasal airway cell cultures and impact of HRV and influenza infection on mediators of airway inflammation (IL-1β and IL-6) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 and 48 h, as described in the methods. (A) hNEC differentiation was confirmed by qPCR using four independent differentiation genes that are not expressed in non-ciliated basal undifferentiated cells (FOX-J1, CCP-110, CDHR3) and for MUC5AC as described in Methods. (B–E) Proinflammatory cytokines interleukin 1 beta (IL-1β) (B,C) and IL-6 (D,E) were determined by multiplex Luminex immunoassay in both apical (B,D) and basal (C,E) chambers of air-liquid interface (ALI) inserts as described in methods. All panels show summary data as mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparisons between the uninfected control group and each experimental group were performed using a two-tailed Mann–Whitney test and are indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test, and the result is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway differentiation and inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Journal: Frontiers in Microbiology

Article Title: Ex vivo modeling of nasal epithelial airway inflammation in respiratory viral infections

doi: 10.3389/fmicb.2026.1819581

Figure Lengend Snippet: Differentiation of nasal airway cell cultures and impact of HRV and influenza infection on mediators of airway inflammation (IL-1β and IL-6) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 and 48 h, as described in the methods. (A) hNEC differentiation was confirmed by qPCR using four independent differentiation genes that are not expressed in non-ciliated basal undifferentiated cells (FOX-J1, CCP-110, CDHR3) and for MUC5AC as described in Methods. (B–E) Proinflammatory cytokines interleukin 1 beta (IL-1β) (B,C) and IL-6 (D,E) were determined by multiplex Luminex immunoassay in both apical (B,D) and basal (C,E) chambers of air-liquid interface (ALI) inserts as described in methods. All panels show summary data as mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparisons between the uninfected control group and each experimental group were performed using a two-tailed Mann–Whitney test and are indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test, and the result is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway differentiation and inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Article Snippet: Human rhinovirus 16 strain 11757 (Cat# VR-283), and Influenza A virus (H1N1) strain A/PR/8/34 (Cat# VR-95) were procured from ATCC.

Techniques: Infection, Ex Vivo, Multiplex Assay, Luminex, Control, Two Tailed Test, MANN-WHITNEY, Comparison, Cell Culture

Impact of HRV and influenza infection on mediators of airway inflammation (IL-8, IL-10) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 and 48 h, as described in the methods. (A–D) Proinflammatory cytokines interleukin 8 (IL-8 (A,B) and IL-10 (C,D) were determined by multiplex Luminex immunoassay in both the apical (A,C) and basal (B,D) chambers of air-liquid interface (ALI) inserts, as described in the methods. All panels show summary data as the mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparison between the uninfected control group and each experimental group was performed using a two-tailed Mann–Whitney test and is indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test and is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Journal: Frontiers in Microbiology

Article Title: Ex vivo modeling of nasal epithelial airway inflammation in respiratory viral infections

doi: 10.3389/fmicb.2026.1819581

Figure Lengend Snippet: Impact of HRV and influenza infection on mediators of airway inflammation (IL-8, IL-10) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 and 48 h, as described in the methods. (A–D) Proinflammatory cytokines interleukin 8 (IL-8 (A,B) and IL-10 (C,D) were determined by multiplex Luminex immunoassay in both the apical (A,C) and basal (B,D) chambers of air-liquid interface (ALI) inserts, as described in the methods. All panels show summary data as the mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparison between the uninfected control group and each experimental group was performed using a two-tailed Mann–Whitney test and is indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test and is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Article Snippet: Human rhinovirus 16 strain 11757 (Cat# VR-283), and Influenza A virus (H1N1) strain A/PR/8/34 (Cat# VR-95) were procured from ATCC.

Techniques: Infection, Ex Vivo, Multiplex Assay, Luminex, Comparison, Control, Two Tailed Test, MANN-WHITNEY, Cell Culture

Impact of HRV and influenza infection on mediators of airway inflammation (TNF-α, VEGF) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 and 48 h, as described in the methods. (A–D) Proinflammatory cytokines, tumor necrosis factor alpha (TNF-α) (A,B) and Vascular endothelial growth factor (VEGF) (C,D) , were determined by multiplex Luminex immunoassay in both the apical (A,C) and basal (B,D) chambers of air-liquid interface (ALI) inserts, as described in the methods. All panels show summary data as the mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparison between the uninfected control group and each experimental group was performed using a two-tailed Mann–Whitney test and is indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test and is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Journal: Frontiers in Microbiology

Article Title: Ex vivo modeling of nasal epithelial airway inflammation in respiratory viral infections

doi: 10.3389/fmicb.2026.1819581

Figure Lengend Snippet: Impact of HRV and influenza infection on mediators of airway inflammation (TNF-α, VEGF) in an ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 and 48 h, as described in the methods. (A–D) Proinflammatory cytokines, tumor necrosis factor alpha (TNF-α) (A,B) and Vascular endothelial growth factor (VEGF) (C,D) , were determined by multiplex Luminex immunoassay in both the apical (A,C) and basal (B,D) chambers of air-liquid interface (ALI) inserts, as described in the methods. All panels show summary data as the mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparison between the uninfected control group and each experimental group was performed using a two-tailed Mann–Whitney test and is indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test and is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway inflammation in human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Article Snippet: Human rhinovirus 16 strain 11757 (Cat# VR-283), and Influenza A virus (H1N1) strain A/PR/8/34 (Cat# VR-95) were procured from ATCC.

Techniques: Infection, Ex Vivo, Multiplex Assay, Luminex, Comparison, Control, Two Tailed Test, MANN-WHITNEY, Cell Culture

Impact of HRV and influenza infection on secretion of proinflammatory cytokines in apical versus basal surface in ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 (A) and 48 (B) h, as described in the methods. The mean value of each protein measurement in each apical cell culture supernatant was normalized to the mean value of protein content in the same basal cell culture supernatant of the same ALI well. All panels show summary data as the mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparison between the uninfected control group and each experimental group was performed using a two-tailed Mann–Whitney test and is indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test, and the result is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway inflammation in apical versus basal surfaces of human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Journal: Frontiers in Microbiology

Article Title: Ex vivo modeling of nasal epithelial airway inflammation in respiratory viral infections

doi: 10.3389/fmicb.2026.1819581

Figure Lengend Snippet: Impact of HRV and influenza infection on secretion of proinflammatory cytokines in apical versus basal surface in ex vivo model of nasal airways. Human nasal epithelial cells (HNECs) from three human donors were prepared and infected with HRV and H1N1 for 24 (A) and 48 (B) h, as described in the methods. The mean value of each protein measurement in each apical cell culture supernatant was normalized to the mean value of protein content in the same basal cell culture supernatant of the same ALI well. All panels show summary data as the mean ± standard error of the mean (SEM) from at least 3 biological replicates per donor. Three HNEC ALI cultures were prepared per human donor, and data from 3 donors are shown. Each data point represents the average of at least two technical replicates per one biological sample. Statistical comparison between the uninfected control group and each experimental group was performed using a two-tailed Mann–Whitney test and is indicated by a blue asterisk. A statistical comparison between two non-control experimental groups was performed using a two-tailed Mann–Whitney test, and the result is indicated by a black asterisk. The p -value for each comparison is shown above each column (* p < 0.05, ** p < 0.01, *** p < 0.001). Alt Text: Graph and data showing measures of nasal airway inflammation in apical versus basal surfaces of human nasal airway cell culture models infected with influenza or human rhinovirus over 48 h compared to uninfected cell cultures.

Article Snippet: Human rhinovirus 16 strain 11757 (Cat# VR-283), and Influenza A virus (H1N1) strain A/PR/8/34 (Cat# VR-95) were procured from ATCC.

Techniques: Infection, Ex Vivo, Cell Culture, Comparison, Control, Two Tailed Test, MANN-WHITNEY