S Wolf1,2,3, G F Perez1,2,3, L Mukharesh1,2,3, N Isaza1,2,4, D Preciado1,2,5, R J Freishtat1,2,6, D Pillai1,2,3, M C Rose1,2,3, G Nino1,2,3. 1. Center for Genetic Research Medicine, Children's National Medical Center, Washington, DC, USA. 2. Department of Pediatrics, George Washington University School of Medicine and Health Sciences, Washington, DC, USA. 3. Division of Pulmonary and Sleep Medicine, Children's National Medical Center, Washington, DC, USA. 4. Division of Neonatology, Children's National Medical Center, Washington, DC, USA. 5. Division of Pediatric Otorhinolaryngology, Children's National Medical Center, Washington, DC, USA. 6. Division of Emergency Medicine, Children's National Medical Center, Washington, DC, USA.
Abstract
BACKGROUND: Airway epithelial cells (AEC) are quite difficult to access in newborns and infants. It is critically important to develop robust life-extended models to conduct translational studies in this age group. We propose the use of a recently described cell culture technology (conditionally reprogrammed cells-CRC) to generate continuous primary cell cultures from nasal and bronchial AEC of young children. METHODS: We collected nasal and/or bronchial AEC from a total of 23 subjects of different ages including newborns/infants/toddlers (0-2 years; N = 9), school-age children (4-11 years; N = 6), and a group of adolescent/adult donors (N = 8). For CRC generation, we used conditioned medium from mitotically inactivated 3T3 fibroblasts and Rho-associated kinase (ROCK) inhibitor (Y-27632). Antiviral immune responses were studied using 25 key antiviral genes and protein production of type III epithelial interferon (IFN λ1) after double-stranded (ds) RNA exposure. RESULTS: CRC derived from primary AEC of neonates/infants and young children exhibited: (i) augmented proliferative capacity and life extension, (ii) preserved airway epithelial phenotype after multiple passages, (iii) robust immune responses characterized by the expression of innate antiviral genes and parallel nasal/bronchial production of IFN λ1 after exposure to dsRNA, and (iv) induction of airway epithelial inflammatory and remodeling responses to dsRNA (eg, CXCL8 and MMP9). CONCLUSION: Conditional reprogramming of AEC from young children is a feasible and powerful translational approach to investigate early-life airway epithelial immune responses in humans.
BACKGROUND: Airway epithelial cells (AEC) are quite difficult to access in newborns and infants. It is critically important to develop robust life-extended models to conduct translational studies in this age group. We propose the use of a recently described cell culture technology (conditionally reprogrammed cells-CRC) to generate continuous primary cell cultures from nasal and bronchial AEC of young children. METHODS: We collected nasal and/or bronchial AEC from a total of 23 subjects of different ages including newborns/infants/toddlers (0-2 years; N = 9), school-age children (4-11 years; N = 6), and a group of adolescent/adult donors (N = 8). For CRC generation, we used conditioned medium from mitotically inactivated 3T3 fibroblasts and Rho-associated kinase (ROCK) inhibitor (Y-27632). Antiviral immune responses were studied using 25 key antiviral genes and protein production of type III epithelial interferon (IFN λ1) after double-stranded (ds) RNA exposure. RESULTS: CRC derived from primary AEC of neonates/infants and young children exhibited: (i) augmented proliferative capacity and life extension, (ii) preserved airway epithelial phenotype after multiple passages, (iii) robust immune responses characterized by the expression of innate antiviral genes and parallel nasal/bronchial production of IFN λ1 after exposure to dsRNA, and (iv) induction of airway epithelial inflammatory and remodeling responses to dsRNA (eg, CXCL8 and MMP9). CONCLUSION: Conditional reprogramming of AEC from young children is a feasible and powerful translational approach to investigate early-life airway epithelial immune responses in humans.
Authors: Prescott G Woodruff; Barmak Modrek; David F Choy; Guiquan Jia; Alexander R Abbas; Almut Ellwanger; Laura L Koth; Joseph R Arron; John V Fahy Journal: Am J Respir Crit Care Med Date: 2009-05-29 Impact factor: 21.405
Authors: Nancy Palechor-Ceron; Frank A Suprynowicz; Geeta Upadhyay; Aleksandra Dakic; Tsion Minas; Vera Simic; Michael Johnson; Christopher Albanese; Richard Schlegel; Xuefeng Liu Journal: Am J Pathol Date: 2013-10-03 Impact factor: 4.307
Authors: Susan D Reynolds; Cydney Rios; Agata Wesolowska-Andersen; Yongbin Zhuang; Mary Pinter; Carrie Happoldt; Cynthia L Hill; Scott W Lallier; Gregory P Cosgrove; George M Solomon; David P Nichols; Max A Seibold Journal: Am J Respir Cell Mol Biol Date: 2016-09 Impact factor: 6.914
Authors: Hong Guo-Parke; Paul Canning; Isobel Douglas; Rémi Villenave; Liam G Heaney; Peter V Coyle; Jeremy D Lyons; Michael D Shields; Ultan F Power Journal: Am J Respir Crit Care Med Date: 2013-10-01 Impact factor: 21.405
Authors: Neil R Hackett; Renat Shaykhiev; Matthew S Walters; Rui Wang; Rachel K Zwick; Barbara Ferris; Bradley Witover; Jacqueline Salit; Ronald G Crystal Journal: PLoS One Date: 2011-05-04 Impact factor: 3.240
Authors: Colin R Butler; Robert E Hynds; Kate H C Gowers; Dani Do Hyang Lee; James M Brown; Claire Crowley; Vitor H Teixeira; Claire M Smith; Luca Urbani; Nicholas J Hamilton; Ricky M Thakrar; Helen L Booth; Martin A Birchall; Paolo De Coppi; Adam Giangreco; Christopher O'Callaghan; Sam M Janes Journal: Am J Respir Crit Care Med Date: 2016-07-15 Impact factor: 21.405
Authors: Xilei XuChen; Jered Weinstock; Maria Arroyo; Kyle Salka; Elizabeth Chorvinsky; Karima Abutaleb; Hector Aguilar; Ryan Kahanowitch; Carlos E Rodríguez-Martínez; Geovanny F Perez; Maria J Gutierrez; Gustavo Nino Journal: Front Pediatr Date: 2021-02-26 Impact factor: 3.418
Authors: Kelly M Martinovich; Thomas Iosifidis; Alysia G Buckley; Kevin Looi; Kak-Ming Ling; Erika N Sutanto; Elizabeth Kicic-Starcevich; Luke W Garratt; Nicole C Shaw; Samuel Montgomery; Francis J Lannigan; Darryl A Knight; Anthony Kicic; Stephen M Stick Journal: Sci Rep Date: 2017-12-21 Impact factor: 4.379
Authors: Nancy Palechor-Ceron; Ewa Krawczyk; Aleksandra Dakic; Vera Simic; Hang Yuan; Jan Blancato; Weisheng Wang; Fleesie Hubbard; Yun-Ling Zheng; Hancai Dan; Scott Strome; Kevin Cullen; Bruce Davidson; John F Deeken; Sujata Choudhury; Peter H Ahn; Seema Agarwal; Xuexun Zhou; Richard Schlegel; Priscilla A Furth; Chong-Xian Pan; Xuefeng Liu Journal: Cells Date: 2019-10-27 Impact factor: 7.666