High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier array
Dysregulated neutrophil recruitment drives many pulmonary diseases, but most preclinical screening methods are unsuited to evaluate pulmonary neutrophilia, limiting progress toward therapeutics. Namely, high-throughput therapeutic assays typically exclude critical neutrophilic pathophysiology, inclu...
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| Format: | Article |
| Language: | English |
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AIP Publishing LLC
2025-06-01
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| Series: | APL Bioengineering |
| Online Access: | http://dx.doi.org/10.1063/5.0220367 |
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| author | Hannah Viola Liang-Hsin Chen Seongbin Jo Kendra Washington Cauviya Selva Andrea Li Daniel Feng Vincent Giacalone Susan T. Stephenson Kirsten Cottrill Ahmad Mohammad Evelyn Williams Xianggui Qu Wilbur Lam Nga L. Ng Anne Fitzpatrick Jocelyn Grunwell Rabindra Tirouvanziam Shuichi Takayama |
| author_facet | Hannah Viola Liang-Hsin Chen Seongbin Jo Kendra Washington Cauviya Selva Andrea Li Daniel Feng Vincent Giacalone Susan T. Stephenson Kirsten Cottrill Ahmad Mohammad Evelyn Williams Xianggui Qu Wilbur Lam Nga L. Ng Anne Fitzpatrick Jocelyn Grunwell Rabindra Tirouvanziam Shuichi Takayama |
| author_sort | Hannah Viola |
| collection | DOAJ |
| description | Dysregulated neutrophil recruitment drives many pulmonary diseases, but most preclinical screening methods are unsuited to evaluate pulmonary neutrophilia, limiting progress toward therapeutics. Namely, high-throughput therapeutic assays typically exclude critical neutrophilic pathophysiology, including blood-to-lung recruitment, dysfunctional activation, and resulting impacts on the air-blood barrier. To meet the conflicting demands of physiological complexity and high throughput, we developed an assay of 96-well leukocyte recruitment in an air-blood barrier array (L-ABBA-96) that enables in vivo-like neutrophil recruitment compatible with downstream phenotyping by automated flow cytometry. We modeled acute respiratory distress syndrome (ARDS) with neutrophil recruitment to 20 ng/mL epithelial-side interleukin 8 and found a dose-dependent reduction in recruitment with physiologic doses of baricitinib, a JAK1/2 inhibitor recently Food and Drug Administration-approved for severe Coronavirus Disease 2019 ARDS. Additionally, neutrophil recruitment to patient-derived cystic fibrosis sputum supernatant induced disease-mimetic recruitment and activation of healthy donor neutrophils and upregulated endothelial e-selectin. Compared to 24-well assays, the L-ABBA-96 reduces required patient sample volumes by 25 times per well and quadruples throughput per plate. Compared to microfluidic assays, the L-ABBA-96 recruits two orders of magnitude more neutrophils per well, enabling downstream flow cytometry and other standard biochemical assays. This novel pairing of high-throughput in vitro modeling of organ-level lung function with parallel high-throughput leukocyte phenotyping substantially advances opportunities for pathophysiological studies, personalized medicine, and drug testing applications. |
| format | Article |
| id | doaj-art-cc84427a5d2c40dfbe7ea83dee02db46 |
| institution | OA Journals |
| issn | 2473-2877 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | AIP Publishing LLC |
| record_format | Article |
| series | APL Bioengineering |
| spelling | doaj-art-cc84427a5d2c40dfbe7ea83dee02db462025-08-20T02:38:29ZengAIP Publishing LLCAPL Bioengineering2473-28772025-06-0192026110026110-1210.1063/5.0220367High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier arrayHannah Viola0Liang-Hsin Chen1Seongbin Jo2Kendra Washington3Cauviya Selva4Andrea Li5Daniel Feng6Vincent Giacalone7Susan T. Stephenson8Kirsten Cottrill9Ahmad Mohammad10Evelyn Williams11Xianggui Qu12Wilbur Lam13Nga L. Ng14Anne Fitzpatrick15Jocelyn Grunwell16Rabindra Tirouvanziam17Shuichi Takayama18Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USASchool of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USACenter for CF & Airways Disease Research, Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USACenter for CF & Airways Disease Research, Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USACenter for CF & Airways Disease Research, Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USACenter for CF & Airways Disease Research, Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USADepartment of Mathematics and Statistics, Oakland University, Rochester, Michigan 48309, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USASchool of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USACenter for CF & Airways Disease Research, Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USADepartment of Pediatrics, Division of Critical Care Medicine, Emory University School of Medicine and Children's Healthcare of Atlanta at Arthur M. Blank Hospital, Atlanta, Georgia 30322, USACenter for CF & Airways Disease Research, Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USAParker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USADysregulated neutrophil recruitment drives many pulmonary diseases, but most preclinical screening methods are unsuited to evaluate pulmonary neutrophilia, limiting progress toward therapeutics. Namely, high-throughput therapeutic assays typically exclude critical neutrophilic pathophysiology, including blood-to-lung recruitment, dysfunctional activation, and resulting impacts on the air-blood barrier. To meet the conflicting demands of physiological complexity and high throughput, we developed an assay of 96-well leukocyte recruitment in an air-blood barrier array (L-ABBA-96) that enables in vivo-like neutrophil recruitment compatible with downstream phenotyping by automated flow cytometry. We modeled acute respiratory distress syndrome (ARDS) with neutrophil recruitment to 20 ng/mL epithelial-side interleukin 8 and found a dose-dependent reduction in recruitment with physiologic doses of baricitinib, a JAK1/2 inhibitor recently Food and Drug Administration-approved for severe Coronavirus Disease 2019 ARDS. Additionally, neutrophil recruitment to patient-derived cystic fibrosis sputum supernatant induced disease-mimetic recruitment and activation of healthy donor neutrophils and upregulated endothelial e-selectin. Compared to 24-well assays, the L-ABBA-96 reduces required patient sample volumes by 25 times per well and quadruples throughput per plate. Compared to microfluidic assays, the L-ABBA-96 recruits two orders of magnitude more neutrophils per well, enabling downstream flow cytometry and other standard biochemical assays. This novel pairing of high-throughput in vitro modeling of organ-level lung function with parallel high-throughput leukocyte phenotyping substantially advances opportunities for pathophysiological studies, personalized medicine, and drug testing applications.http://dx.doi.org/10.1063/5.0220367 |
| spellingShingle | Hannah Viola Liang-Hsin Chen Seongbin Jo Kendra Washington Cauviya Selva Andrea Li Daniel Feng Vincent Giacalone Susan T. Stephenson Kirsten Cottrill Ahmad Mohammad Evelyn Williams Xianggui Qu Wilbur Lam Nga L. Ng Anne Fitzpatrick Jocelyn Grunwell Rabindra Tirouvanziam Shuichi Takayama High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier array APL Bioengineering |
| title | High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier array |
| title_full | High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier array |
| title_fullStr | High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier array |
| title_full_unstemmed | High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier array |
| title_short | High-throughput quantitation of human neutrophil recruitment and functional responses in an air-blood barrier array |
| title_sort | high throughput quantitation of human neutrophil recruitment and functional responses in an air blood barrier array |
| url | http://dx.doi.org/10.1063/5.0220367 |
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