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SARS-CoV-2 productively infects primary human immune system cells in vitro and in COVID-19 patients
Marjorie C. Pontelli1,†,* , Ítalo A. Castro1,†,* , Ronaldo B. Martins1 , Leonardo La Serra1 , Flávio P. Veras2 , Daniele C. Nascimento2 , Camila M. Silva2 , Ricardo S. Cardoso1 , Roberta Rosales3 , Rogério Gomes4 , Thais M. Lima1 , Juliano P. Souza1 , Brenda C. Vitti1 , Diego B. Caetité2 , Mikhael H. F. de Lima2 , Spencer D. Stumpf5 , Cassandra E. Thompson5 , Louis-Marie Bloyet5 , Juliana E. Toller-Kawahisa2 , Marcela C. Giannini2,6 , Letícia P. Bonjorno2,6 , Maria I. F. Lopes2,6 , Sabrina S. Batah7 , Li Siyuan7 , Rodrigo Luppino-Assad2,6 , Sergio C. L. Almeida2,6 , Fabiola R. Oliveira2,6 , Maíra N. Benatti2,6 , Lorena L. F. Pontes4 , Rodrigo C. Santana2,6 , Fernando C. Vilar2,6 , Maria Auxiliadora-Martins2,6 , Pei-Yong Shi5 , Thiago M. Cunha2 , Rodrigo T. Calado4 , José C. Alves-Filho2 , Dario S. Zamboni2,3 , Alexandre T. Fabro7 , Paulo Louzada-Junior2,6 , Rene D. R. Oliveira2,6 , Sean P. J. Whelan8 , Fernando Q. Cunha2 , Eurico Arruda1,3,*
1Virology Research Center, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo 14049-900, Brazil
2Center of Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo 14049-900, Brazil
3Department of Cell and Molecular Biology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo 14049-900, Brazil
4Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo 14049-900, Brazil
5Department of Biochemistry & Molecular Biology, the University of Texas Medical Branch, Galveston, TX 77555, USA
6Divisions of Clinical Immunology, Infectious Diseases and Intensive Care Unit, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo 14049-900, Brazil
7Department of Pathology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo 14049-900, Brazil
8Department of Molecular Microbiology, Washington University in St. Louis, Saint Louis, MO 63110, USA
These authors contributed equally to this work.
*Correspondence to:Marjorie C. Pontelli , Email:cmarjorie@wustl.edu Ítalo A. Castro , Email:italo@wustl.edu Eurico Arruda , Email:eaneto@fmrp.usp.br
J Mol Cell Biol, Volume 14, Issue 4, April 2022, mjac021,  https://doi.org/10.1093/jmcb/mjac021
Keyword: SARS-CoV-2, COVID-19, lymphocytopenia, peripheral blood mononuclear cell (PBMC), lymphocytes, monocytes, apoptosis

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is associated with a hyperinflammatory state and lymphocytopenia, a hallmark that appears as both signature and prognosis of disease severity outcome. Although cytokine storm and a sustained inflammatory state are commonly associated with immune cell depletion, it is still unclear whether direct SARS-CoV-2 infection of immune cells could also play a role in this scenario by harboring viral replication. We found that monocytes, as well as both B and T lymphocytes, were susceptible to SARS-CoV-2 infection in vitro, accumulating double-stranded RNA consistent with viral RNA replication and ultimately leading to expressive T cell apoptosis. In addition, flow cytometry and immunofluorescence analysis revealed that SARS-CoV-2 was frequently detected in monocytes and B lymphocytes from coronavirus disease 2019 (COVID-19) patients. The rates of SARS-CoV-2-infected monocytes in peripheral blood mononuclear cells from COVID-19 patients increased over time from symptom onset, with SARS-CoV-2-positive monocytes, B cells, and CD4+ T lymphocytes also detected in postmortem lung tissue. These results indicated that SARS-CoV-2 infection of blood-circulating leukocytes in COVID-19 patients might have important implications for disease pathogenesis and progression, immune dysfunction, and virus spread within the host.