Dietary Vitamin D Mitigates Coronavirus-Induced Lung Inflammation and Damage in Mice
Gabriel Campolina-Silva, Ana Cláudia Dos Santos Pereira Andrade, Manoela Couto, Paloma G Bittencourt-Silva, Celso M Queiroz-Junior, Larisse De Souza B Lacerda, Ian De Meira Chaves, Leonardo C De Oliveira, Fernanda Martins Marim, Cleida A Oliveira, Glauber S F Da Silva, Mauro Martins Teixeira, Vivian Vasconcelos Costa
Viruses, doi:10.3390/v15122434
The COVID-19 pandemic caused by the SARS-CoV-2 (β-CoV) betacoronavirus has posed a significant threat to global health. Despite the availability of vaccines, the virus continues to spread, and there is a need for alternative strategies to alleviate its impact. Vitamin D, a secosteroid hormone best known for its role in bone health, exhibits immunomodulatory effects in certain viral infections. Here, we have shown that bioactive vitamin D (calcitriol) limits in vitro replication of SARS-CoV-2 and murine coronaviruses MHV-3 and MHV-A59. Comparative studies involving wild-type mice intranasally infected with MHV-3, a model for studying β-CoV respiratory infections, confirmed the protective effect of vitamin D in vivo. Accordingly, mice fed a standard diet rapidly succumbed to MHV-3 infection, whereas those on a vitamin D-rich diet (10,000 IU of Vitamin D 3 /kg) displayed increased resistance to acute respiratory damage and systemic complications. Consistent with these findings, the vitamin D-supplemented group exhibited lower viral titers in their lungs and reduced levels of TNF, IL-6, IL-1β, and IFN-γ, alongside an enhanced type I interferon response. Altogether, our findings suggest vitamin D supplementation ameliorates β-CoV-triggered respiratory illness and systemic complications in mice, likely via modulation of the host's immune response to the virus.
by MHV-3. While these findings open promising avenues for further exploration and emphasize the importance of continued research in this area, it is imperative for future studies to broaden the scope of this study by encompassing in vivo infection with SARS-CoV-2. This imperative step will provide a more comprehensive understanding of vitamin D's prospects in addressing COVID-19. As result, we argue that our findings should not be construed in the context of the current clinical landscape of studies on vitamin D and COVID-19. Instead, they may serve as a springboard for future preclinical research aimed at exploring the intricate relationship between Vitamin D and β-CoV infections.
Supplementary Materials: The following supporting information can be downloaded at: https:// www.mdpi.com/article/10.3390/v15122434/s1, Figure S1 and Raw data_Figures 1-6. Informed Consent Statement: Not applicable.
Conflicts of Interest: The authors declare no conflict of interest.
References
Agrawal, Gupta, Agrawal, Vitamin D Supplementation Reduces Airway Hyperresponsiveness and Allergic Airway Inflammation in a Murine Model, Clin. Exp. Allergy,
doi:10.1111/cea.12102Alon, Sportiello, Kozlovski, Kumar, Reilly et al., Leukocyte Trafficking to the Lungs and beyond: Lessons from Influenza for COVID-19, Nat. Rev. Immunol,
doi:10.1038/s41577-020-00470-2Amrein, Scherkl, Hoffmann, Neuwersch-Sommeregger, Köstenberger et al., Vitamin D deficiency 2.0: An update on the current status worldwide, Eur. J. Clin. Nutr,
doi:10.1038/s41430-020-0558-yAndrade, Campolina-Silva, Queiroz-Junior, De Oliveira, Lacerda et al., A Biosafety Level 2 Mouse Model for Studying Betacoronavirus-Induced Acute Lung Damage and Systemic Manifestations, J. Virol,
doi:10.1128/jvi.01276-21Araujo, Machado, Amgarten, Malta, De Araujo et al., SARS-CoV-2 Isolation from the First Reported Patients in Brazil and Establishment of a Coordinated Task Network, Mem. Inst. Oswaldo Cruz,
doi:10.1590/0074-02760200342Arora, Patel, Nicol, Field, Restori et al., Vitamin D and the Ability to Produce 1,25(OH)2D Are Critical for Protection from Viral Infection of the Lungs, Nutrients,
doi:10.3390/nu14153061Bastard, Rosen, Zhang, Michailidis, Hoffmann et al., Autoantibodies against Type I IFNs in Patients with Life-Threatening COVID-19, Science,
doi:10.1126/science.abd4585Bikle, Christakos, New Aspects of Vitamin D Metabolism and Action-Addressing the Skin as Source and Target, Nat. Rev. Endocrinol,
doi:10.1038/s41574-019-0312-5Blanco-Melo, Nilsson-Payant, Liu, Uhl, Hoagland et al., Imbalanced Host Response to SARS-CoV-2 Drives Development of COVID-19, Cell,
doi:10.1016/j.cell.2020.04.026Butler-Laporte, Nakanishi, Mooser, Morrison, Abdullah et al., Vitamin D and COVID-19 Susceptibility and Severity in the COVID-19 Host Genetics Initiative: A Mendelian Randomization Study, PLoS Med,
doi:10.1371/journal.pmed.1003605Campolina-Silva, Maria, Mahecha, Oliveira, Reduced Vitamin D (VDR) Expression and Plasma Vitamin D Levels Are Associated with Aging-Related Prostate Lesions, Prostate,
doi:10.1002/pros.23498Castillo, Costa, Barrios, Díaz, Miranda et al., Effect of Calcifediol Treatment and Best Available Therapy versus Best Available Therapy on Intensive Care Unit Admission and Mortality among Patients Hospitalized for COVID-19: A Pilot Randomized Clinical Study, J. Steroid Biochem. Mol. Biol,
doi:10.1016/j.jsbmb.2020.105751Cervantes-Barragán, Kalinke, Züst, König, Reizis et al., Type I IFN-Mediated Protection of Macrophages and Dendritic Cells Secures Control of Murine Coronavirus Infection, J. Immunol,
doi:10.4049/jimmunol.182.2.1099Charoenngam, Holick, Immunologic Effects of Vitamin on Human Health and Disease, Nutrients,
doi:10.3390/nu12072097Chen, Sung, Chuang, Lai, Lee et al., Vitamin D3 Decreases TNF-α-Induced Inflammation in Lung Epithelial Cells through a Reduction in Mitochondrial Fission and Mitophagy, Cell Biol. Toxicol,
doi:10.1007/s10565-021-09629-6Chu, Chan, -W.; Yuen, Shuai, Yuan et al., Comparative Tropism, Replication Kinetics, and Cell Damage Profiling of SARS-CoV-2 and SARS-CoV with Implications for Clinical Manifestations, Transmissibility, and Laboratory Studies of COVID-19: An Observational Study, Lancet Microbe,
doi:10.1016/S2666-5247(20)30004-5Coussens, Wilkinson, Hanifa, Nikolayevskyy, Elkington et al., Vitamin D Accelerates Resolution of Inflammatory Responses during Tuberculosis Treatment, Proc. Natl. Acad. Sci,
doi:10.1073/pnas.1200072109De Albuquerque, Baig, Ma, Zhang, He et al., MurineHepatitis Virus Strain 1 Produces a Clinically Relevant Model of Severe Acute Respiratory Syndrome in A/J Mice, J. Virol,
doi:10.1128/JVI.00747-06Dimitrov, Barbier, Ismailova, Wang, Dmowski et al., Vitamin D-Regulated Gene Expression Profiles: Species-Specificity and Cell-Specific Effects on Metabolism and Immunity, Endocrinology,
doi:10.1210/endocr/bqaa218Ding, Ning, Liu, Lai, Leibowitz et al., Fulminant Hepatic Failure in Murine Hepatitis Virus Strain 3 Infection: Tissue-Specific Expression of a Novel Fgl2 Prothrombinase, J. Virol,
doi:10.1128/JVI.72.4.3504a-3504a.1998Dissanayake, De Silva, Sumanatilleke, De Silva, Gamage et al., Prognostic and Therapeutic Role of Vitamin D in COVID-19: Systematic Review and Meta-Analysis, J. Clin. Endocrinol. Metab,
doi:10.1210/clinem/dgab892Fischer, Hall, Agrawal, Vitamin D Supplementation Reduces Induction of Epithelial-Mesenchymal Transition in Allergen Sensitized and Challenged Mice, PLoS ONE,
doi:10.1371/journal.pone.0149180Garcia, De Moraes, Rodrigues, Gilioli, De Oliveira-Filho et al., Coding-Complete Genome Sequence of Murine Hepatitis Virus, Microbiol. Resour. Announc,
doi:10.1128/MRA.00248-21Gombart, Borregaard, Koeffler, Human Cathelicidin Antimicrobial Peptide (CAMP) Gene Is a Direct Target of the Vitamin D Receptor and Is Strongly Up-regulated in Myeloid Cells by 1,25-dihydroxyvitamin D 3, FASEB J,
doi:10.1096/fj.04-3284comGong, Worley, Carver, Goldstein, Deng, Neutrophils Drive Pulmonary Vascular Leakage in MHV-1 Infection of Susceptible A/J Mice, Front. Immunol,
doi:10.3389/fimmu.2022.1089064Hadjadj, Yatim, Barnabei, Corneau, Boussier, Impaired Type I Interferon Activity and Inflammatory Responses in Severe COVID-19 Patients, Science,
doi:10.1126/science.abc6027Hafezi, Saheb Sharif-Askari, Saheb Sharif-Askari, Ali Hussain Alsayed, Alsafar et al., Vitamin D Enhances Type I IFN Signaling in COVID-19 Patients, Sci. Rep,
doi:10.1038/s41598-022-22307-9Hirai, Ohtsuka, Ikeda, Taniguchi, Blau et al., Role of Mouse Hepatitis Virus (MHV) Receptor Murine CEACAM1 in the Resistance of Mice to MHV Infection: Studies of Mice with Chimeric MCEACAM1a and MCEACAM1b, J. Virol,
doi:10.1128/JVI.02680-09Jolliffe, Camargo, Sluyter, Aglipay, Aloia et al., Vitamin D supplementation to prevent acute respiratory infections: A systematic review and meta-analysis of aggregate data from randomised controlled trials, Lancet Diabetes Endocrinol,
doi:10.1016/S2213-8587(21)00051-6Jolliffe, Holt, Greenig, Talaei, Perdek et al., Effect of a Test-and-Treat Approach to Vitamin D Supplementation on Risk of All Cause Acute Respiratory Tract Infection and COVID-19: Phase 3 Randomised Controlled Trial (CORONAVIT), BMJ,
doi:10.1136/bmj-2022-071230Karki, Sharma, Tuladhar, Williams, Zalduondo et al., Synergism of TNF-α and IFN-γ Triggers Inflammatory Cell Death, Tissue Damage, and Mortality in SARS-CoV-2 Infection and Cytokine Shock Syndromes, Cell,
doi:10.1016/j.cell.2020.11.025Kim, Park, Cho, Yoon, Song et al., Low Serum 25-Hydroxyvitamin D Level: An Independent Risk Factor for Tuberculosis?, Clin. Nutr,
doi:10.1016/j.clnu.2013.11.014Körner, Majjouti, Alejandre Alcazar, Mahabir, Of Mice and Men: The Coronavirus Mhv and Mouse Models as a Translational Approach to Understand Sars-Cov-2, Viruses,
doi:10.3390/v12080880Li, Pei, Chen, Song, Zhang et al., Substantial Undocumented Infection Facilitates the Rapid Dissemination of Novel Coronavirus (SARS-CoV-2), Science,
doi:10.1126/science.abb3221Liu, Liu, Xiang, Pu, Xiong et al., Neutrophil-to-Lymphocyte Ratio Predicts Critical Illness Patients with 2019 Coronavirus Disease in the Early Stage, J. Transl. Med,
doi:10.1186/s12967-020-02374-0Lucas, Wong, Klein, Castro, Silva et al., Longitudinal Analyses Reveal Immunological Misfiring in Severe COVID-19, Nature,
doi:10.1038/s41586-020-2588-yMaj, Trynda, Maj, Gębura, Bogunia-Kubik et al., Differential Response of Lung Cancer Cell Lines to Vitamin D Derivatives Depending on EGFR, KRAS, P53 Mutation Status and VDR Polymorphism, J. Steroid Biochem. Mol. Biol,
doi:10.1016/j.jsbmb.2019.105431Malireddi, Bynigeri, Mall, Connelly, Pruett-Miller et al., Inflammatory Cell Death, PANoptosis, Screen Identifies Host Factors in Coronavirus Innate Immune Response as Therapeutic Targets, Commun. Biol,
doi:10.1038/s42003-023-05414-9Marim, Silveira, Lima, Zamboni, A Method for Generation of Bone Marrow-Derived Macrophages from Cryopreserved Mouse Bone Marrow Cells, PLoS ONE,
doi:10.1371/journal.pone.0015263Meng, Li, Liu, Xiao, Tang et al., The role of vitamin D in the prevention and treatment of SARS-CoV-2 infection: A meta-analysis of randomized controlled trials, Clin. Nutr,
doi:10.1016/j.clnu.2023.09.008Mok, Ng, Ahidjo, Aw, Chen et al., Evaluation of In Vitro and In Vivo Antiviral Activities of Vitamin D for SARS-CoV-2 and Variants, Pharmaceutics,
doi:10.3390/pharmaceutics15030925Murai, Fernandes, Sales, Pinto, Goessler et al., Effect of a Single High Dose of Vitamin D3 on Hospital Length of Stay in Patients with Moderate to Severe COVID-19: A Randomized Clinical Trial, JAMA-J. Am. Med. Assoc,
doi:10.1001/jama.2020.26848Oh, Riek, Darwech, Funai, Shao et al., Deletion of Macrophage Vitamin D Receptor Promotes Insulin Resistance and Monocyte Cholesterol Transport to Accelerate Atherosclerosis in Mice, Cell Rep,
doi:10.1016/j.celrep.2015.02.043Otto, Rathkolb, Oestereicher, Lengger, Moerth et al., Clinical Chemistry Reference Intervals for C57BL/6J, C57BL/6N, and C3HeB/FeJ Mice (Mus Musculus), J. Am. Assoc. Lab. Anim. Sci
Pan, Shen, Yu, Ge, Chen et al., SARS-CoV-2 N Protein Promotes NLRP3 Inflammasome Activation to Induce Hyperinflammation, Nat. Commun,
doi:10.1038/s41467-021-25015-6Pelaia, Tinello, Vatrella, De Sarro, Pelaia, Lung under Attack by COVID-19-Induced Cytokine Storm: Pathogenic Mechanisms and Therapeutic Impications, Ther. Adv. Respir. Dis,
doi:10.1177/1753466620933508Petrelli, Luciani, Perego, Dognini, Colombelli et al., Therapeutic and prognostic role of vitamin D for COVID-19 infection: A systematic review and meta-analysis of 43 observational studies, J. Steroid Biochem. Mol. Biol,
doi:10.1016/j.jsbmb.2021.105883Qiao, Lau, Xie, Poon, Chan et al., SARS-CoV-2 infection induces inflammatory bone loss in golden Syrian hamsters, Nat. Commun,
doi:10.1038/s41467-022-30195-wQueiroz-Junior, Santos, Gonçalves, Brito, Barrioni et al., Acute Coronavirus Infection Triggers a TNF-Dependent Osteoporotic Phenotype in Mice, Life Sci,
doi:10.1016/j.lfs.2023.121750Ray, Personius, Dungan, Dhillon, Hershberger, Vitamin D3 Intake Modulates Diaphragm but Not Peripheral Muscle Force in Young Mice, J. Appl. Physiol,
doi:10.1152/japplphysiol.00643.2015Sarhan, Warda, Ibrahim, Schaalan, Fathy, Evaluation of Infliximab/Tocilizumab versus Tocilizumab among COVID-19 Patients with Cytokine Storm Syndrome, Sci. Rep,
doi:10.1038/s41598-023-33484-6Sharma, Peng, Qing, Hilliard, Kim et al., Distinct Roles of Type I and Type III Interferons during a Native Murine β Coronavirus Lung Infection, J. Virol,
doi:10.1128/JVI.01241-21Taylor, Nadeau, Abbasi, Lachance, Nguyen et al., The Ultimate QPCR Experiment: Producing Publication Quality, Reproducible Data the First Time, Trends Biotechnol,
doi:10.1016/j.tibtech.2018.12.002Tebben, Singh, Kumar, Vitamin D-Mediated Hypercalcemia: Mechanisms, Diagnosis, and Treatment, Endocr. Rev,
doi:10.1210/er.2016-1070Toro, Arevalo, Pereira-Gómez, Sabater, Zizzi et al., Coronavirus Pathogenesis in Mice Explains the SARS-CoV-2 Multi-Organ Spread by Red Blood Cells Hitch-Hiking, medRxiv,
doi:10.1101/2023.03.29.23287591Verone-Boyle, Shoemaker, Attwood, Morrison, Makowski et al., Diet-Derived 25-Hydroxyvitamin D3 Activates Vitamin D Receptor Target Gene Expression and Suppresses EGFR Mutant Non-Small Cell Lung Cancer Growth in Vitro and in Vivo, Oncotarget,
doi:10.18632/oncotarget.6493Vieira-Alves, Alves, Souza, Melo, Coimbra Campos et al., TNF/INOS/NO Pathway Mediates Host Susceptibility to Endothelial-Dependent Circulatory Failure and Death Induced by Betacoronavirus Infection, Clin. Sci,
doi:10.1042/CS20220663Villasis-Keever, López-Alarcón, Miranda-Novales, Zurita-Cruz, Barrada-Vázquez et al., Efficacy and Safety of Vitamin D Supplementation to Prevent COVID-19 in Frontline Healthcare Workers. A Randomized Clinical Trial, Arch. Med. Res,
doi:10.1016/j.arcmed.2022.04.003Wang, Nestel, Bourdeau, Nagai, Wang et al., Cutting Edge: 1,25-Dihydroxyvitamin D3 Is a Direct Inducer of Antimicrobial Peptide Gene Expression, J. Immunol,
doi:10.4049/jimmunol.173.5.2909Wang, Wang, Li, Chen, Han et al., Human Cathelicidin Inhibits SARS-CoV-2 Infection: Killing Two Birds with One Stone, ACS Infect. Dis,
doi:10.1021/acsinfecdis.1c00096White, Emerging Roles of Vitamin D-Induced Antimicrobial Peptides in Antiviral Innate Immunity, Nutrients,
doi:10.3390/nu14020284Xu, Li, Cao, Wu, Chen, Tumor Necrosis Factor α (TNF-α) Receptor-I Is Required for TNF-α-Mediated Fulminant Virus Hepatitis Caused by Murine Hepatitis Virus Strain-3 Infection, Immunol. Lett,
doi:10.1016/j.imlet.2013.11.008Yang, Du, Chen, Zhao, Yang et al., Coronavirus MHV-A59 Infects the Lung and Causes Severe Pneumonia in C57BL/6 Mice, Virol. Sin,
doi:10.1007/s12250-014-3530-yZeng, Xie, Feng, Xu, Han et al., Specific Inhibition of the NLRP3 Inflammasome Suppresses Immune Overactivation and Alleviates COVID-19 like Pathology in Mice, eBioMedicine,
doi:10.1016/j.ebiom.2021.103803Zhang, Li, Yang, Wang, Effect of vitamin D supplementation on COVID-19 patients: A systematic review and meta-analysis, Front. Nutr,
doi:10.3389/fnut.2023.1131103Zhang, Liu, Moncada-Velez, Chen, Ogishi et al., Inborn Errors of Type I IFN Immunity in Patients with Life-Threatening COVID-19, Science,
doi:10.1126/science.abd4570Zheng, Williams, Subbarao Malireddi, Karki, Banoth et al., Impaired NLRP3 Inflammasome Activation/Pyroptosis Leads to Robust Inflammatory Cell Death via Caspase-8/RIPK3 during Coronavirus Infection, J. Biol. Chem,
doi:10.1074/jbc.RA120.015036Zhu, Zhu, Gu, Zhan, Chen et al., Association Between Vitamin D and Influenza: Meta-Analysis and Systematic Review of Randomized Controlled Trials, Front. Nutr,
doi:10.3389/fnut.2021.799709
