Mycobacterium tuberculosis and SARS‐CoV‐2 Co‐Infection: An Efficient Cell Model to Study Lung Host–Pathogen Interactions
Thays Maria Costa de Lucena, Débora Elienai de Oliveira Miranda, Juliana Vieira de Barros Arcoverde, Mariana Souza Bezerra Cavalcanti, Willyene Marilia Dantas, Lindomar José Pena, Michelle Christiane da Silva Rabello, Jaqueline de Azevêdo SilvaAbstract
In less than a year, SARS‐CoV‐2 managed to displace Mycobacterium tuberculosis (Mtb) as the leading cause of death worldwide due to a single infectious agent. Both pathogens affect the respiratory tract, mainly the lungs. However, the impact that a possible Mtb + SARS‐CoV‐2 co‐infection can have on the host response is still unknown. Herein we depict a rigorous system to evaluate the complex interaction between two simultaneous infections in vitro in a lung epithelial cell line (A549). Overall, the process includes eight steps: (1) Mtb culture, (2) cell maintenance, (3) preparation of viral stocks, (4) determination of infectious titers, (5) Mtb and SARS‐CoV‐2 co‐infection, (6) determination of intracellular bacterial load, (7) SARS‐CoV‐2 viability test, and (8) optional UV‐C validation assay for infectivity reduction. This comprehensive protocol will allow experimentalists to study the pathogenesis of co‐infection in vitro and facilitate collaborative work in the literature. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC.
Basic Protocol : In vitro coinfection model using Mtb and SARS‐CoV‐2 in human lung epithelial cells