Mitochondria complex III-generated superoxide is essential for IL-10 secretion in macrophages

Joshua S. Stoolman; Rogan A. Grant; Leah K. Billingham; Taylor A. Poor; Samuel E. Weinberg; Madeline C. Harding; Ziyan Lu; Jason Miska; Marten Szibor; G. R.Scott Budinger; Navdeep S. Chandel

doi:10.1126/sciadv.adu4369

Mitochondria complex III-generated superoxide is essential for IL-10 secretion in macrophages

Joshua S. Stoolman, Rogan A. Grant, Leah K. Billingham, Taylor A. Poor, Samuel E. Weinberg, Madeline C. Harding, Ziyan Lu, Jason Miska, Marten Szibor, G. R.Scott Budinger^*, Navdeep S. Chandel

^*Corresponding author for this work

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

3 Scopus citations

Abstract

Mitochondrial electron transport chain (ETC) function modulates macrophage biology; however, mechanisms underlying mitochondria ETC control of macrophage immune responses are not fully understood. Here, we report that mutant mice with mitochondria ETC complex III (CIII)-deficient macrophages exhibit increased susceptibility to influenza A virus (IAV) and LPS-induced endotoxic shock. Cultured bone marrow-derived macrophages (BMDMs) isolated from these mitochondria CIII-deficient mice released less IL-10 than controls following TLR3 or TLR4 stimulation. Unexpectedly, restoring mitochondrial respiration without generating superoxide using alternative oxidase (AOX) was not sufficient to reverse LPS-induced endotoxic shock susceptibility or restore IL-10 release. However, activation of protein kinase A (PKA) rescued IL-10 release in mitochondria CIII-deficient BMDMs following LPS stimulation. In addition, mitochondria CIII deficiency did not affect BMDM responses to interleukin-4 (IL-4) stimulation. Thus, our results highlight the essential role of mitochondria CIII-generated superoxide in the release of anti-inflammatory IL-10 in response to TLR stimulation.

Original language	English (US)
Article number	eadu4369
Journal	Science Advances
Volume	11
Issue number	4
DOIs	https://doi.org/10.1126/sciadv.adu4369
State	Published - Jan 24 2025

Funding

We thank M. Szibor and H. T. Jacobs for the generous donation of the ROSA26SNAPf-AOXmice to our laboratory. This work was supported by the NIH Health Grant 2P01AG049665-06 (to N.S.C.), NIH Health Grant 5P01HL154998 (to N.S.C.), NIH Health Grant NI2T32AI083216-11 (to J.S.S.), NIH Health Grant 5T32HL076139-18 to (to T.A.P.), NIH Health Grant NCI CC SG P30 CA060553 (Northwestern University Center for Advanced Microscopy supported by awarded to the Robert H Lurie Comprehensive Cancer Center), NIH Health Grant NCI CC SG P30 CA060553 (H. Abdala-V alencia and the Metabolomics Core Facility), NIH Grant 1S10OD011996-01 (Northwestern University Flow Cytometry Core Facility), NIH Grant 5P01HL154998 (Influenza A virus was provided by NHLBI program project core), and Schmidt Science Fellows, in partnership with the Rhodes Trust (to R.A.G.).

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title = "Mitochondria complex III-generated superoxide is essential for IL-10 secretion in macrophages",

abstract = "Mitochondrial electron transport chain (ETC) function modulates macrophage biology; however, mechanisms underlying mitochondria ETC control of macrophage immune responses are not fully understood. Here, we report that mutant mice with mitochondria ETC complex III (CIII)-deficient macrophages exhibit increased susceptibility to influenza A virus (IAV) and LPS-induced endotoxic shock. Cultured bone marrow-derived macrophages (BMDMs) isolated from these mitochondria CIII-deficient mice released less IL-10 than controls following TLR3 or TLR4 stimulation. Unexpectedly, restoring mitochondrial respiration without generating superoxide using alternative oxidase (AOX) was not sufficient to reverse LPS-induced endotoxic shock susceptibility or restore IL-10 release. However, activation of protein kinase A (PKA) rescued IL-10 release in mitochondria CIII-deficient BMDMs following LPS stimulation. In addition, mitochondria CIII deficiency did not affect BMDM responses to interleukin-4 (IL-4) stimulation. Thus, our results highlight the essential role of mitochondria CIII-generated superoxide in the release of anti-inflammatory IL-10 in response to TLR stimulation.",

author = "Stoolman, {Joshua S.} and Grant, {Rogan A.} and Billingham, {Leah K.} and Poor, {Taylor A.} and Weinberg, {Samuel E.} and Harding, {Madeline C.} and Ziyan Lu and Jason Miska and Marten Szibor and Budinger, {G. R.Scott} and Chandel, {Navdeep S.}",

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doi = "10.1126/sciadv.adu4369",

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AU - Stoolman, Joshua S.

AU - Grant, Rogan A.

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AU - Poor, Taylor A.

AU - Weinberg, Samuel E.

AU - Harding, Madeline C.

AU - Lu, Ziyan

AU - Miska, Jason

AU - Szibor, Marten

AU - Budinger, G. R.Scott

AU - Chandel, Navdeep S.

PY - 2025/1/24

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N2 - Mitochondrial electron transport chain (ETC) function modulates macrophage biology; however, mechanisms underlying mitochondria ETC control of macrophage immune responses are not fully understood. Here, we report that mutant mice with mitochondria ETC complex III (CIII)-deficient macrophages exhibit increased susceptibility to influenza A virus (IAV) and LPS-induced endotoxic shock. Cultured bone marrow-derived macrophages (BMDMs) isolated from these mitochondria CIII-deficient mice released less IL-10 than controls following TLR3 or TLR4 stimulation. Unexpectedly, restoring mitochondrial respiration without generating superoxide using alternative oxidase (AOX) was not sufficient to reverse LPS-induced endotoxic shock susceptibility or restore IL-10 release. However, activation of protein kinase A (PKA) rescued IL-10 release in mitochondria CIII-deficient BMDMs following LPS stimulation. In addition, mitochondria CIII deficiency did not affect BMDM responses to interleukin-4 (IL-4) stimulation. Thus, our results highlight the essential role of mitochondria CIII-generated superoxide in the release of anti-inflammatory IL-10 in response to TLR stimulation.

AB - Mitochondrial electron transport chain (ETC) function modulates macrophage biology; however, mechanisms underlying mitochondria ETC control of macrophage immune responses are not fully understood. Here, we report that mutant mice with mitochondria ETC complex III (CIII)-deficient macrophages exhibit increased susceptibility to influenza A virus (IAV) and LPS-induced endotoxic shock. Cultured bone marrow-derived macrophages (BMDMs) isolated from these mitochondria CIII-deficient mice released less IL-10 than controls following TLR3 or TLR4 stimulation. Unexpectedly, restoring mitochondrial respiration without generating superoxide using alternative oxidase (AOX) was not sufficient to reverse LPS-induced endotoxic shock susceptibility or restore IL-10 release. However, activation of protein kinase A (PKA) rescued IL-10 release in mitochondria CIII-deficient BMDMs following LPS stimulation. In addition, mitochondria CIII deficiency did not affect BMDM responses to interleukin-4 (IL-4) stimulation. Thus, our results highlight the essential role of mitochondria CIII-generated superoxide in the release of anti-inflammatory IL-10 in response to TLR stimulation.

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Mitochondria complex III-generated superoxide is essential for IL-10 secretion in macrophages

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