Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis

Marissa A. Guttenberg; Aaron T. Vose; Anastasiya Birukova; Kaitlyn Lewars; R. Ian Cumming; Michaela C. Albright; Jasper I. Mark; Claudia J. Salazar; Suchitra Swaminathan; Zhan Yu; Yuliana V. Sokolenko; Elsie Bunyan; Michael J. Yaeger; Michael B. Fessler; Loretta G. Que; Kymberly M. Gowdy; Alexander V. Misharin; Robert M. Tighe

doi:10.1165/rcmb.2023-0390OC

Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis

Marissa A. Guttenberg, Aaron T. Vose, Anastasiya Birukova, Kaitlyn Lewars, R. Ian Cumming, Michaela C. Albright, Jasper I. Mark, Claudia J. Salazar, Suchitra Swaminathan, Zhan Yu, Yuliana V. Sokolenko, Elsie Bunyan, Michael J. Yaeger, Michael B. Fessler, Loretta G. Que, Kymberly M. Gowdy, Alexander V. Misharin, Robert M. Tighe^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Lung inflammation, caused by acute exposure to ozone (O₃), one of the six criteria air pollutants, is a significant source of morbidity in susceptible individuals. Alveolar macrophages (AMØs) are the most abundant immune cells in the normal lung, and their number increases after O₃ exposure. However, the role of AMØs in promoting or limiting O₃-induced lung inflammation has not been clearly defined. In this study, we used a mouse model of acute O₃ exposure, lineage tracing, genetic knockouts, and data from O₃-exposed human volunteers to define the role and ontogeny of AMØs during acute O₃ exposure. Lineage-tracing experiments showed that 12, 24, and 72 hours after exposure to O₃ (2 ppm) for 3 hours, all AMØs were of tissue-resident origin. Similarly, in humans exposed to filtered air and O₃ (200 ppb) for 135 minutes, we did not observe at ∼21 hours postexposure an increase in monocyte-derived AMØs by flow cytometry. Highlighting a role for tissue-resident AMØs, we demonstrate that depletion of tissue-resident AMØs with clodronate-loaded liposomes led to persistence of neutrophils in the alveolar space after O₃ exposure, suggesting that impaired neutrophil clearance (i.e., efferocytosis) leads to prolonged lung inflammation. Moreover, depletion of tissue-resident AMØs demonstrated reduced clearance of intratracheally instilled apoptotic Jurkat cells, consistent with reduced efferocytosis. Genetic ablation of MerTK (MER proto-oncogene, tyrosine kinase), a key receptor involved in efferocytosis, also resulted in impaired clearance of apoptotic neutrophils after O₃ exposure. Overall, these findings underscore the pivotal role of tissue-resident AMØs in resolving O₃-induced inflammation via MerTK-mediated efferocytosis.

Original language	English (US)
Pages (from-to)	493-506
Number of pages	14
Journal	American journal of respiratory cell and molecular biology
Volume	70
Issue number	6
DOIs	https://doi.org/10.1165/rcmb.2023-0390OC
State	Published - Jun 2024

Funding

Supported by the National Institute of Environmental Health Sciences of the National Institutes of Health under award number T32ES021432 (to M.A.G., Duke University Program in Environmental Health); National Institute of Environmental Health Sciences grant Z01 ES102005 (to M.B.F.); and National Institutes of Health grants R01ES027574 (to R.M.T.), R01ES028829 (to K.M.G.), R01ES034350 (to R.M.T. and A.V.M.), and U19AI135964, P01AG049665, P01HL154998, R01HL153312, R01HL158139, R01ES034350, and R21AG075423 (to A.V.M.). The authors acknowledge contributions of the Duke Cancer Institute Flow Cytometry Facility and the Rodent Inhalation Core at Duke University, Durham, NC. Flow cytometric analysis of the human BAL specimens was performed at Northwestern University Flow Cytometry Core Facility. Northwestern University Flow Cytometry Core Facility is supported by National Cancer Institute Cancer Center Support Grant P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. Cell sorting was performed on a BD FACSAria SORP cell sorter purchased through the support of National Institutes of Health grant 1S10OD011996-01. Supported by the National Institute of Environmental Health Sciences of the National Institutes of Health under award number T32ES021432 (to M.A.G., Duke University Program in Environmental Health); National Institute of Environmental Health Sciences grant Z01 ES102005 (to M.B.F.); and National Institutes of Health grants R01ES027574 (to R.M.T.), R01ES028829 (to K.M.G.), R01ES034350 (to R.M.T. and A.V.M.), and U19AI135964, P01AG049665, P01HL154998, R01HL153312, R01HL158139, R01ES034350, and R21AG075423 (to A.V.M.). Acknowledgment: The authors acknowledge contributions of the Duke Cancer Institute Flow Cytometry Facility and the Rodent Inhalation Core at Duke University, Durham, NC. Flow cytometric analysis of the human BAL specimens was performed at Northwestern University Flow Cytometry Core Facility. Northwestern University Flow Cytometry Core Facility is supported by National Cancer Institute Cancer Center Support Grant P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. Cell sorting was performed on a BD FACSAria SORP cell sorter purchased through the support of National Institutes of Health grant 1S10OD011996-01.

Keywords

MerTK
efferocytosis
macrophages
ozone

ASJC Scopus subject areas

Molecular Biology
Pulmonary and Respiratory Medicine
Clinical Biochemistry
Cell Biology

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Guttenberg, M. A., Vose, A. T., Birukova, A., Lewars, K., Cumming, R. I., Albright, M. C., Mark, J. I., Salazar, C. J., Swaminathan, S., Yu, Z., Sokolenko, Y. V., Bunyan, E., Yaeger, M. J., Fessler, M. B., Que, L. G., Gowdy, K. M., Misharin, A. V., & Tighe, R. M. (2024). Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis. American journal of respiratory cell and molecular biology, 70(6), 493-506. https://doi.org/10.1165/rcmb.2023-0390OC

@article{91dd6763a7d04426ba2d890782bfce8a,

title = "Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis",

abstract = "Lung inflammation, caused by acute exposure to ozone (O3), one of the six criteria air pollutants, is a significant source of morbidity in susceptible individuals. Alveolar macrophages (AM{\O}s) are the most abundant immune cells in the normal lung, and their number increases after O3 exposure. However, the role of AM{\O}s in promoting or limiting O3-induced lung inflammation has not been clearly defined. In this study, we used a mouse model of acute O3 exposure, lineage tracing, genetic knockouts, and data from O3-exposed human volunteers to define the role and ontogeny of AM{\O}s during acute O3 exposure. Lineage-tracing experiments showed that 12, 24, and 72 hours after exposure to O3 (2 ppm) for 3 hours, all AM{\O}s were of tissue-resident origin. Similarly, in humans exposed to filtered air and O3 (200 ppb) for 135 minutes, we did not observe at ∼21 hours postexposure an increase in monocyte-derived AM{\O}s by flow cytometry. Highlighting a role for tissue-resident AM{\O}s, we demonstrate that depletion of tissue-resident AM{\O}s with clodronate-loaded liposomes led to persistence of neutrophils in the alveolar space after O3 exposure, suggesting that impaired neutrophil clearance (i.e., efferocytosis) leads to prolonged lung inflammation. Moreover, depletion of tissue-resident AM{\O}s demonstrated reduced clearance of intratracheally instilled apoptotic Jurkat cells, consistent with reduced efferocytosis. Genetic ablation of MerTK (MER proto-oncogene, tyrosine kinase), a key receptor involved in efferocytosis, also resulted in impaired clearance of apoptotic neutrophils after O3 exposure. Overall, these findings underscore the pivotal role of tissue-resident AM{\O}s in resolving O3-induced inflammation via MerTK-mediated efferocytosis.",

keywords = "MerTK, efferocytosis, macrophages, ozone",

author = "Guttenberg, {Marissa A.} and Vose, {Aaron T.} and Anastasiya Birukova and Kaitlyn Lewars and Cumming, {R. Ian} and Albright, {Michaela C.} and Mark, {Jasper I.} and Salazar, {Claudia J.} and Suchitra Swaminathan and Zhan Yu and Sokolenko, {Yuliana V.} and Elsie Bunyan and Yaeger, {Michael J.} and Fessler, {Michael B.} and Que, {Loretta G.} and Gowdy, {Kymberly M.} and Misharin, {Alexander V.} and Tighe, {Robert M.}",

note = "Publisher Copyright: Copyright {\textcopyright} 2024 by the American Thoracic Society.",

year = "2024",

month = jun,

doi = "10.1165/rcmb.2023-0390OC",

language = "English (US)",

volume = "70",

pages = "493--506",

journal = "American journal of respiratory cell and molecular biology",

issn = "1044-1549",

publisher = "American Thoracic Society",

number = "6",

}

Guttenberg, MA, Vose, AT, Birukova, A, Lewars, K, Cumming, RI, Albright, MC, Mark, JI, Salazar, CJ, Swaminathan, S, Yu, Z, Sokolenko, YV, Bunyan, E, Yaeger, MJ, Fessler, MB, Que, LG, Gowdy, KM, Misharin, AV & Tighe, RM 2024, 'Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis', American journal of respiratory cell and molecular biology, vol. 70, no. 6, pp. 493-506. https://doi.org/10.1165/rcmb.2023-0390OC

TY - JOUR

T1 - Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis

AU - Guttenberg, Marissa A.

AU - Vose, Aaron T.

AU - Birukova, Anastasiya

AU - Lewars, Kaitlyn

AU - Cumming, R. Ian

AU - Albright, Michaela C.

AU - Mark, Jasper I.

AU - Salazar, Claudia J.

AU - Swaminathan, Suchitra

AU - Yu, Zhan

AU - Sokolenko, Yuliana V.

AU - Bunyan, Elsie

AU - Yaeger, Michael J.

AU - Fessler, Michael B.

AU - Que, Loretta G.

AU - Gowdy, Kymberly M.

AU - Misharin, Alexander V.

AU - Tighe, Robert M.

PY - 2024/6

Y1 - 2024/6

N2 - Lung inflammation, caused by acute exposure to ozone (O3), one of the six criteria air pollutants, is a significant source of morbidity in susceptible individuals. Alveolar macrophages (AMØs) are the most abundant immune cells in the normal lung, and their number increases after O3 exposure. However, the role of AMØs in promoting or limiting O3-induced lung inflammation has not been clearly defined. In this study, we used a mouse model of acute O3 exposure, lineage tracing, genetic knockouts, and data from O3-exposed human volunteers to define the role and ontogeny of AMØs during acute O3 exposure. Lineage-tracing experiments showed that 12, 24, and 72 hours after exposure to O3 (2 ppm) for 3 hours, all AMØs were of tissue-resident origin. Similarly, in humans exposed to filtered air and O3 (200 ppb) for 135 minutes, we did not observe at ∼21 hours postexposure an increase in monocyte-derived AMØs by flow cytometry. Highlighting a role for tissue-resident AMØs, we demonstrate that depletion of tissue-resident AMØs with clodronate-loaded liposomes led to persistence of neutrophils in the alveolar space after O3 exposure, suggesting that impaired neutrophil clearance (i.e., efferocytosis) leads to prolonged lung inflammation. Moreover, depletion of tissue-resident AMØs demonstrated reduced clearance of intratracheally instilled apoptotic Jurkat cells, consistent with reduced efferocytosis. Genetic ablation of MerTK (MER proto-oncogene, tyrosine kinase), a key receptor involved in efferocytosis, also resulted in impaired clearance of apoptotic neutrophils after O3 exposure. Overall, these findings underscore the pivotal role of tissue-resident AMØs in resolving O3-induced inflammation via MerTK-mediated efferocytosis.

AB - Lung inflammation, caused by acute exposure to ozone (O3), one of the six criteria air pollutants, is a significant source of morbidity in susceptible individuals. Alveolar macrophages (AMØs) are the most abundant immune cells in the normal lung, and their number increases after O3 exposure. However, the role of AMØs in promoting or limiting O3-induced lung inflammation has not been clearly defined. In this study, we used a mouse model of acute O3 exposure, lineage tracing, genetic knockouts, and data from O3-exposed human volunteers to define the role and ontogeny of AMØs during acute O3 exposure. Lineage-tracing experiments showed that 12, 24, and 72 hours after exposure to O3 (2 ppm) for 3 hours, all AMØs were of tissue-resident origin. Similarly, in humans exposed to filtered air and O3 (200 ppb) for 135 minutes, we did not observe at ∼21 hours postexposure an increase in monocyte-derived AMØs by flow cytometry. Highlighting a role for tissue-resident AMØs, we demonstrate that depletion of tissue-resident AMØs with clodronate-loaded liposomes led to persistence of neutrophils in the alveolar space after O3 exposure, suggesting that impaired neutrophil clearance (i.e., efferocytosis) leads to prolonged lung inflammation. Moreover, depletion of tissue-resident AMØs demonstrated reduced clearance of intratracheally instilled apoptotic Jurkat cells, consistent with reduced efferocytosis. Genetic ablation of MerTK (MER proto-oncogene, tyrosine kinase), a key receptor involved in efferocytosis, also resulted in impaired clearance of apoptotic neutrophils after O3 exposure. Overall, these findings underscore the pivotal role of tissue-resident AMØs in resolving O3-induced inflammation via MerTK-mediated efferocytosis.

KW - MerTK

KW - efferocytosis

KW - macrophages

KW - ozone

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UR - http://www.scopus.com/inward/citedby.url?scp=85195061614&partnerID=8YFLogxK

U2 - 10.1165/rcmb.2023-0390OC

DO - 10.1165/rcmb.2023-0390OC

M3 - Article

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AN - SCOPUS:85195061614

SN - 1044-1549

VL - 70

SP - 493

EP - 506

JO - American journal of respiratory cell and molecular biology

JF - American journal of respiratory cell and molecular biology

IS - 6

ER -

Tissue-Resident Alveolar Macrophages Reduce Ozone-induced Inflammation via MerTK-mediated Efferocytosis

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