TY - JOUR
T1 - Tissue-based SARS-CoV-2 detection in fatal COVID-19 infections
T2 - Sustained direct viral-induced damage is not necessary to drive disease progression
AU - El Jamal, Siraj M.
AU - Pujadas, Elisabet
AU - Ramos, Irene
AU - Bryce, Clare
AU - Grimes, Zachary M.
AU - Amanat, Fatima
AU - Tsankova, Nadejda M.
AU - Mussa, Zarmeen
AU - Olson, Sara
AU - Salem, Fadi
AU - Miorin, Lisa
AU - Aydillo, Teresa
AU - Schotsaert, Michael
AU - Albrecht, Randy A.
AU - Liu, Wen Chun
AU - Marjanovic, Nada
AU - Francoeur, Nancy
AU - Sebra, Robert
AU - Sealfon, Stuart C.
AU - García-Sastre, Adolfo
AU - Fowkes, Mary
AU - Cordon-Cardo, Carlos
AU - Westra, William H.
N1 - Funding Information:
Competing interests: The AG-S laboratory has received research support from Pfizer, Pharmamar, Blade Therapeutics, Avimex, Dynavax, Kenall Manufacturing, ImmunityBio, Nanocomposix, Senhwa Biosciences, and 7Hills Pharma. AG-S has consulting agreements for the following companies involving cash and/or stock: Vivaldi Biosciences, Contrafect, 7Hills Pharma, Avimex, Vaxalto, Accurius, and Esperovax.I“n loving memory of Dr. Mary Fowkes”. This research was partly funded by CRIP (Center for Research for Influenza Pathogenesis), a NIAID supported Center of Excellence for Influenza Research and Surveillance (CEIRS, contract # HHSN272201400008C); by NIAID grant U19AI142733 and NCI grant U54CA260560; by the generous support of the JPB Foundation and the Open Philanthropy Project (research grant 2020-215611 (5384); and by anonymous donors to A.G-S.; partial funding by NIH grant R01NS106229-02S2, United States We thank Dr. Florian Kramer for providing the infected cell lines and Jason Reidy for providing the EM picture.
Funding Information:
This research was partly funded by CRIP (Center for Research for Influenza Pathogenesis) , a NIAID supported Center of Excellence for Influenza Research and Surveillance (CEIRS, contract # HHSN272201400008C); by NIAID grant U19AI142733 and NCI grant U54CA260560 ; by the generous support of the JPB Foundation and the Open Philanthropy Project (research grant 2020-215611 (5384); and by anonymous donors to A.G-S.; partial funding by NIH grant R01NS106229-02S2, United States We thank Dr. Florian Kramer for providing the infected cell lines and Jason Reidy for providing the EM picture.
Publisher Copyright:
© 2021 The Authors
PY - 2021/8
Y1 - 2021/8
N2 - Coronavirus disease 2019 (COVID-19) is an ongoing pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although viral infection is known to trigger inflammatory processes contributing to tissue injury and organ failure, it is unclear whether direct viral damage is needed to sustain cellular injury. An understanding of pathogenic mechanisms has been handicapped by the absence of optimized methods to visualize the presence and distribution of SARS-CoV-2 in damaged tissues. We first developed a positive control cell line (Vero E6) to validate SARS-CoV-2 detection assays. We then evaluated multiple organs (lungs, kidneys, heart, liver, brain, intestines, lymph nodes, and spleen) from fourteen COVID-19 autopsy cases using immunohistochemistry (IHC) for the spike and the nucleoprotein proteins, and RNA in situ hybridization (RNA ISH) for the spike protein mRNA. Tissue detection assays were compared with quantitative polymerase chain reaction (qPCR)-based detection. SARS-CoV-2 was histologically detected in the Vero E6 positive cell line control, 1 of 14 (7%) lungs, and none (0%) of the other 59 organs. There was perfect concordance between the IHC and RNA ISH results. qPCR confirmed high viral load in the SARS-CoV-2 ISH-positive lung tissue, and absent or low viral load in all ISH-negative tissues. In patients who die of COVID-19-related organ failure, SARS-CoV-2 is largely not detectable using tissue-based assays. Even in lungs showing widespread injury, SARS-CoV-2 viral RNA or proteins were detected in only a small minority of cases. This observation supports the concept that viral infection is primarily a trigger for multiple-organ pathogenic proinflammatory responses. Direct viral tissue damage is a transient phenomenon that is generally not sustained throughout disease progression.
AB - Coronavirus disease 2019 (COVID-19) is an ongoing pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although viral infection is known to trigger inflammatory processes contributing to tissue injury and organ failure, it is unclear whether direct viral damage is needed to sustain cellular injury. An understanding of pathogenic mechanisms has been handicapped by the absence of optimized methods to visualize the presence and distribution of SARS-CoV-2 in damaged tissues. We first developed a positive control cell line (Vero E6) to validate SARS-CoV-2 detection assays. We then evaluated multiple organs (lungs, kidneys, heart, liver, brain, intestines, lymph nodes, and spleen) from fourteen COVID-19 autopsy cases using immunohistochemistry (IHC) for the spike and the nucleoprotein proteins, and RNA in situ hybridization (RNA ISH) for the spike protein mRNA. Tissue detection assays were compared with quantitative polymerase chain reaction (qPCR)-based detection. SARS-CoV-2 was histologically detected in the Vero E6 positive cell line control, 1 of 14 (7%) lungs, and none (0%) of the other 59 organs. There was perfect concordance between the IHC and RNA ISH results. qPCR confirmed high viral load in the SARS-CoV-2 ISH-positive lung tissue, and absent or low viral load in all ISH-negative tissues. In patients who die of COVID-19-related organ failure, SARS-CoV-2 is largely not detectable using tissue-based assays. Even in lungs showing widespread injury, SARS-CoV-2 viral RNA or proteins were detected in only a small minority of cases. This observation supports the concept that viral infection is primarily a trigger for multiple-organ pathogenic proinflammatory responses. Direct viral tissue damage is a transient phenomenon that is generally not sustained throughout disease progression.
KW - COVID-19
KW - Coronavirus
KW - Diffuse alveolar damage
KW - Nucleoprotein
KW - RNA in situ hybridization
KW - SARS-CoV-2
KW - Spike protein
UR - http://www.scopus.com/inward/record.url?scp=85109212132&partnerID=8YFLogxK
U2 - 10.1016/j.humpath.2021.04.012
DO - 10.1016/j.humpath.2021.04.012
M3 - Article
C2 - 33961839
AN - SCOPUS:85109212132
VL - 114
SP - 110
EP - 119
JO - Human Pathology
JF - Human Pathology
SN - 0046-8177
ER -