PMID- 35877763 OWN - NLM STAT- MEDLINE DCOM- 20220808 LR - 20231020 IS - 1553-7374 (Electronic) IS - 1553-7366 (Print) IS - 1553-7366 (Linking) VI - 18 IP - 7 DP - 2022 Jul TI - IFN-gamma-independent control of M. tuberculosis requires CD4 T cell-derived GM-CSF and activation of HIF-1alpha. PG - e1010721 LID - 10.1371/journal.ppat.1010721 [doi] LID - e1010721 AB - The prevailing model of protective immunity to tuberculosis is that CD4 T cells produce the cytokine IFN-gamma to activate bactericidal mechanisms in infected macrophages. Although IFN-gamma-independent CD4 T cell based control of M. tuberculosis infection has been demonstrated in vivo it is unclear whether CD4 T cells are capable of directly activating macrophages to control infection in the absence of IFN-gamma. We developed a co-culture model using CD4 T cells isolated from the lungs of infected mice and M. tuberculosis-infected murine bone marrow-derived macrophages (BMDMs) to investigate mechanisms of CD4 dependent control of infection. We found that even in the absence of IFN-gamma signaling, CD4 T cells drive macrophage activation, M1 polarization, and control of infection. This IFN-gamma-independent control of infection requires activation of the transcription factor HIF-1alpha and a shift to aerobic glycolysis in infected macrophages. While HIF-1alpha activation following IFN-gamma stimulation requires nitric oxide, HIF-1alpha-mediated control in the absence of IFN-gamma is nitric oxide-independent, indicating that distinct pathways can activate HIF-1alpha during infection. We show that CD4 T cell-derived GM-CSF is required for IFN-gamma-independent control in BMDMs, but that recombinant GM-CSF is insufficient to control infection in BMDMs or alveolar macrophages and does not rescue the absence of control by GM-CSF-deficient T cells. In contrast, recombinant GM-CSF controls infection in peritoneal macrophages, induces lipid droplet biogenesis, and also requires HIF-1alpha for control. These results advance our understanding of CD4 T cell-mediated immunity to M. tuberculosis, reveal important differences in immune activation of distinct macrophage types, and outline a novel mechanism for the activation of HIF-1alpha. We establish a previously unknown functional link between GM-CSF and HIF-1alpha and provide evidence that CD4 T cell-derived GM-CSF is a potent bactericidal effector. FAU - Van Dis, Erik AU - Van Dis E AUID- ORCID: 0000-0003-4773-379X AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. FAU - Fox, Douglas M AU - Fox DM AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. FAU - Morrison, Huntly M AU - Morrison HM AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. FAU - Fines, Daniel M AU - Fines DM AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. FAU - Babirye, Janet Peace AU - Babirye JP AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. FAU - McCann, Lily H AU - McCann LH AD - School of Public Health, Division of Infectious Diseases and Vaccinology, University of California, Berkeley, Berkeley, California, United States of America. FAU - Rawal, Sagar AU - Rawal S AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. FAU - Cox, Jeffery S AU - Cox JS AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. FAU - Stanley, Sarah A AU - Stanley SA AUID- ORCID: 0000-0002-4182-9048 AD - Department of Molecular and Cell Biology, Division of Immunology and Pathogenesis, University of California, Berkeley, Berkeley, California, United States of America. AD - School of Public Health, Division of Infectious Diseases and Vaccinology, University of California, Berkeley, Berkeley, California, United States of America. LA - eng SI - Dryad/10.5061/dryad.s7h44j18x GR - R01 AI143722/AI/NIAID NIH HHS/United States GR - R01 AI113270/AI/NIAID NIH HHS/United States GR - U19 AI135990/AI/NIAID NIH HHS/United States GR - U19 AI106754/AI/NIAID NIH HHS/United States GR - P01 AI063302/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20220725 PL - United States TA - PLoS Pathog JT - PLoS pathogens JID - 101238921 RN - 0 (Hif1a protein, mouse) RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (IFNG protein, mouse) RN - 31C4KY9ESH (Nitric Oxide) RN - 82115-62-6 (Interferon-gamma) RN - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor) SB - IM MH - Animals MH - CD4-Positive T-Lymphocytes MH - Granulocyte-Macrophage Colony-Stimulating Factor MH - Hypoxia-Inducible Factor 1, alpha Subunit MH - Interferon-gamma MH - Mice MH - *Mycobacterium tuberculosis MH - Nitric Oxide MH - *Tuberculosis PMC - PMC9352196 COIS- The authors have declared that no competing interests exist. EDAT- 2022/07/26 06:00 MHDA- 2022/08/09 06:00 PMCR- 2022/07/25 CRDT- 2022/07/25 13:44 PHST- 2021/12/21 00:00 [received] PHST- 2022/07/01 00:00 [accepted] PHST- 2022/08/04 00:00 [revised] PHST- 2022/07/26 06:00 [pubmed] PHST- 2022/08/09 06:00 [medline] PHST- 2022/07/25 13:44 [entrez] PHST- 2022/07/25 00:00 [pmc-release] AID - PPATHOGENS-D-21-02577 [pii] AID - 10.1371/journal.ppat.1010721 [doi] PST - epublish SO - PLoS Pathog. 2022 Jul 25;18(7):e1010721. doi: 10.1371/journal.ppat.1010721. eCollection 2022 Jul.