Surfactant protein A alters endosomal trafficking of influenza A virus in macrophages

Influenza A virus infection (IAV) often leads to acute lung injury that impairs breathing and can lead to death, with disproportionate mortality in children and the elderly. Surfactant Protein A (SP-A) is a calcium-dependent opsonin that binds a variety of pathogens to help control pulmonary infections by alveolar macrophages. Alveolar macrophages play critical roles in host resistance and susceptibility to IAV infection. The effect of SP-A on IAV infection and antiviral response of macrophages, however, is not understood. Here, we report that SP-A attenuates IAV infection in a dose-dependent manner at the level of endosomal trafficking, resulting in infection delay in a model macrophage cell line. The ability of SP-A to suppress infection was independent of its glycosylation status. Binding of SP-A to hemagglutinin did not rely on the glycosylation status or sugar binding properties of either protein. Incubation of either macrophages or IAV with SP-A slowed endocytic uptake rate of IAV. SP-A interfered with binding to cell membrane and endosomal exit of the viral genome as indicated by experiments using isolated cell membranes, an antibody recognizing a pH-sensitive conformational epitope on hemagglutinin, and microscopy. Lack of SP-A in mice enhanced IFNβ expression, viral clearance and reduced mortality from IAV infection. These findings support the idea that IAV is an opportunistic pathogen that co-opts SP-A to evade host defense by alveolar macrophages. Our study highlights novel aspects of host-pathogen interactions that may lead to better understanding of the local mechanisms that shape activation of antiviral and inflammatory responses to viral infection in the lung.

[1]  C. Casals,et al.  Signaling Pathways That Mediate Alveolar Macrophage Activation by Surfactant Protein A and IL-4 , 2022, Frontiers in Immunology.

[2]  A. Lavie,et al.  pH-Dependent Mechanisms of Influenza Infection Mediated by Hemagglutinin , 2021, Frontiers in Molecular Biosciences.

[3]  Nikolaos M. Nikolaidis,et al.  Viral Evasion of Innate Immune Defense: The Case of Resistance of Pandemic H1N1 Influenza A Virus to Human Mannose-Binding Proteins , 2021, Frontiers in Microbiology.

[4]  S. Bhattacharyya,et al.  Respiratory Pathogen Coinfections in SARS-CoV-2-Positive Patients in Southeastern Wisconsin: A Retrospective Analysis , 2021, Microbiology spectrum.

[5]  Chunhua Song,et al.  Genomic and epigenomic adaptation in SP-R210 (Myo18A) isoform-deficient macrophages. , 2021, Immunobiology.

[6]  Guohua Yang,et al.  Relationship between hemagglutinin stability and influenza virus persistence after exposure to low pH or supraphysiological heating , 2021, PLoS pathogens.

[7]  Y. Yamauchi,et al.  How Influenza Virus Uses Host Cell Pathways during Uncoating , 2021, Cells.

[8]  J. Huskens,et al.  Multivalent Affinity Profiling: Direct Visualization of the Superselective Binding of Influenza Viruses , 2021, ACS nano.

[9]  C. Russell Hemagglutinin Stability and Its Impact on Influenza A Virus Infectivity, Pathogenicity, and Transmissibility in Avians, Mice, Swine, Seals, Ferrets, and Humans , 2021, Viruses.

[10]  Yu Chen,et al.  Coinfection with influenza A virus enhances SARS-CoV-2 infectivity , 2021, Cell Research.

[11]  H. Achdout,et al.  Increased lethality in influenza and SARS-CoV-2 coinfection is prevented by influenza immunity but not SARS-CoV-2 immunity , 2021, Nature Communications.

[12]  J. Madsen,et al.  SP-A and SP-D: Dual Functioning Immune Molecules With Antiviral and Immunomodulatory Properties , 2021, Frontiers in Immunology.

[13]  N. Behrendt,et al.  The collagen receptor uPARAP/Endo180 regulates collectins through unique structural elements in its FNII domain , 2020, The Journal of Biological Chemistry.

[14]  Tina Meischel,et al.  Influenza A virus interactions with macrophages: Lessons from epithelial cells , 2020, Cellular microbiology.

[15]  M. Kataoka,et al.  Serial Section Array Scanning Electron Microscopy Analysis of Cells from Lung Autopsy Specimens following Fatal A/H1N1 2009 Pandemic Influenza Virus Infection , 2019, Journal of Virology.

[16]  S. Kellokumpu Golgi pH, Ion and Redox Homeostasis: How Much Do They Really Matter? , 2019, Front. Cell Dev. Biol..

[17]  E. Makarov,et al.  Full-length human surfactant protein A inhibits influenza A virus infection of A549 lung epithelial cells: A recombinant form containing neck and lectin domains promotes infectivity. , 2019, Immunobiology.

[18]  Linden J. Gearing,et al.  Unique Transcriptional Architecture in Airway Epithelial Cells and Macrophages Shapes Distinct Responses following Influenza Virus Infection Ex Vivo , 2019, Journal of Virology.

[19]  Shamus P. Keeler,et al.  Influenza A Virus Infection Causes Chronic Lung Disease Linked to Sites of Active Viral RNA Remnants , 2018, The Journal of Immunology.

[20]  James B. Munro,et al.  Direct Visualization of the Conformational Dynamics of Single Influenza Hemagglutinin Trimers , 2018, Cell.

[21]  K. Hartshorn,et al.  The Role and Molecular Mechanism of Action of Surfactant Protein D in Innate Host Defense Against Influenza A Virus , 2018, Front. Immunol..

[22]  Y. Kawaoka,et al.  A Defect in Influenza A Virus Particle Assembly Specific to Primary Human Macrophages , 2018, mBio.

[23]  Y. Kawasawa,et al.  GM-CSF overexpression after influenza a virus infection prevents mortality and moderates M1-like airway monocyte/macrophage polarization , 2018, Respiratory Research.

[24]  Caitlin E. Mullarkey,et al.  Alveolar macrophages are critical for broadly-reactive antibody-mediated protection against influenza A virus in mice , 2017, Nature Communications.

[25]  Troy D. Cline,et al.  Influenza virus replication in macrophages: balancing protection and pathogenesis. , 2017, The Journal of general virology.

[26]  T. Abraham,et al.  SH3GLB2/endophilin B2 regulates lung homeostasis and recovery from severe influenza A virus infection , 2017, Scientific Reports.

[27]  Alex K. Heer,et al.  Frontline Science: Coincidental null mutation of Csf2rα in a colony of PI3Kγ−/− mice causes alveolar macrophage deficiency and fatal respiratory viral infection , 2017, Journal of leukocyte biology.

[28]  Guangshun Wang,et al.  Collectins, H-ficolin and LL-37 reduce influence viral replication in human monocytes and modulate virus-induced cytokine production , 2017, Innate immunity.

[29]  S. Schultz-Cherry,et al.  Influenza Virus Overcomes Cellular Blocks To Productively Replicate, Impacting Macrophage Function , 2016, Journal of Virology.

[30]  N. Reiling,et al.  Surfactant Protein A Enhances Constitutive Immune Functions of Clathrin Heavy Chain and Clathrin Adaptor Protein 2. , 2016, American journal of respiratory cell and molecular biology.

[31]  Eleonore Fröhlich,et al.  Measurements of Deposition, Lung Surface Area and Lung Fluid for Simulation of Inhaled Compounds , 2016, Front. Pharmacol..

[32]  P. Reading,et al.  DC-SIGN and L-SIGN Are Attachment Factors That Promote Infection of Target Cells by Human Metapneumovirus in the Presence or Absence of Cellular Glycosaminoglycans , 2016, Journal of Virology.

[33]  P. Reading,et al.  Endocytic function is critical for influenza A virus infection via DC-SIGN and L-SIGN , 2016, Scientific Reports.

[34]  P. Reading,et al.  The C-type Lectin Langerin Functions as a Receptor for Attachment and Infectious Entry of Influenza A Virus , 2015, Journal of Virology.

[35]  E Scott Halstead,et al.  Lethal influenza infection: Is a macrophage to blame? , 2015, Expert review of anti-infective therapy.

[36]  D. Desmecht,et al.  Hyporeactivity of Alveolar Macrophages and Higher Respiratory Cell Permissivity Characterize DBA/2J Mice Infected by Influenza A Virus. , 2015, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[37]  N. Christensen,et al.  SP-R210 (Myo18A) Isoforms as Intrinsic Modulators of Macrophage Priming and Activation , 2015, PloS one.

[38]  Rongying Zhang,et al.  The effect of the size of fluorescent dextran on its endocytic pathway , 2015, Cell biology international.

[39]  Christine E. Becker,et al.  Influenza virus-induced lung injury: pathogenesis and implications for treatment , 2015, European Respiratory Journal.

[40]  M. Colonna,et al.  OSCAR Is a Receptor for Surfactant Protein D That Activates TNF-α Release from Human CCR2+ Inflammatory Monocytes , 2015, The Journal of Immunology.

[41]  A. García-Sastre,et al.  The NS1 protein: a multitasking virulence factor. , 2015, Current topics in microbiology and immunology.

[42]  T. Stehle,et al.  The sweet spot: defining virus–sialic acid interactions , 2014, Nature Reviews Microbiology.

[43]  K. Karjalainen,et al.  Transient ablation of alveolar macrophages leads to massive pathology of influenza infection without affecting cellular adaptive immunity , 2014, European journal of immunology.

[44]  H. Haagsman,et al.  Leukocyte‐associated Ig‐like receptor‐1 is a novel inhibitory receptor for surfactant protein D , 2014, Journal of leukocyte biology.

[45]  Steven F. Baker,et al.  Influenza A Virus Attenuation by Codon Deoptimization of the NS Gene for Vaccine Development , 2014, Journal of Virology.

[46]  E. López-Granados,et al.  Surfactant protein A genetic variants associate with severe respiratory insufficiency in pandemic influenza A virus infection , 2014, Critical Care.

[47]  Alex K. Heer,et al.  Alveolar Macrophages Are Essential for Protection from Respiratory Failure and Associated Morbidity following Influenza Virus Infection , 2014, PLoS pathogens.

[48]  M. Pohl,et al.  Entry of influenza A virus: host factors and antiviral targets. , 2014, The Journal of general virology.

[49]  T. Irimura,et al.  The Macrophage Galactose-Type Lectin Can Function as an Attachment and Entry Receptor for Influenza Virus , 2013, Journal of Virology.

[50]  S Yuan,et al.  Drugs to cure avian influenza infection – multiple ways to prevent cell death , 2013, Cell Death and Disease.

[51]  K. Ku,et al.  The severe pathogenicity of alveolar macrophage-depleted ferrets infected with 2009 pandemic H1N1 influenza virus. , 2013, Virology.

[52]  P. Thomas,et al.  Depletion of Alveolar Macrophages during Influenza Infection Facilitates Bacterial Superinfections , 2013, The Journal of Immunology.

[53]  Javier G. Magadán,et al.  Influenza A Virus Hemagglutinin Trimerization Completes Monomer Folding and Antigenicity , 2013, Journal of Virology.

[54]  J. Yewdell To dream the impossible dream: universal influenza vaccination. , 2013, Current opinion in virology.

[55]  Yoshihiro Kawaoka,et al.  Receptor binding by a ferret-transmissible H5 avian influenza virus , 2013, Nature.

[56]  J. Yewdell,et al.  Defining influenza A virus hemagglutinin antigenic drift by sequential monoclonal antibody selection. , 2013, Cell host & microbe.

[57]  J. Yewdell,et al.  Reassortment Complements Spontaneous Mutation in Influenza A Virus NP and M1 Genes To Accelerate Adaptation to a New Host , 2013, Journal of Virology.

[58]  F. Aeffner,et al.  Influenza A H1N1 induces declines in alveolar gas exchange in mice consistent with rapid post‐infection progression from acute lung injury to ARDS , 2012, Influenza and other respiratory viruses.

[59]  Huy A. Nguyen,et al.  Pulmonary surfactant protein A and surfactant lipids upregulate IRAK-M, a negative regulator of TLR-mediated inflammation in human macrophages. , 2012, American journal of physiology. Lung cellular and molecular physiology.

[60]  D. Burke,et al.  A review of influenza haemagglutinin receptor binding as it relates to pandemic properties. , 2012, Vaccine.

[61]  G. Kochs,et al.  Altered receptor specificity and fusion activity of the haemagglutinin contribute to high virulence of a mouse-adapted influenza A virus. , 2012, The Journal of general virology.

[62]  T. Kuiken,et al.  Distribution patterns of influenza virus receptors and viral attachment patterns in the respiratory and intestinal tracts of seven avian species , 2012, Veterinary Research.

[63]  A. García-Sastre,et al.  Attenuated Influenza Virus Construct with Enhanced Hemagglutinin Protein Expression , 2012, Journal of Virology.

[64]  Jonathan W. Yewdell,et al.  Fitness costs limit influenza A virus hemagglutinin glycosylation as an immune evasion strategy , 2011, Proceedings of the National Academy of Sciences.

[65]  J. Tichelaar,et al.  GM-CSF modulates pulmonary resistance to influenza A infection. , 2011, Antiviral research.

[66]  Andrew G. Brooks,et al.  Specific Sites of N-Linked Glycosylation on the Hemagglutinin of H1N1 Subtype Influenza A Virus Determine Sensitivity to Inhibitors of the Innate Immune System and Virulence in Mice , 2011, The Journal of Immunology.

[67]  P. Woo,et al.  Structural basis and sequence co-evolution analysis of the hemagglutinin protein of pandemic influenza A/H1N1 (2009) virus , 2011, Experimental biology and medicine.

[68]  J. Whitsett,et al.  GM-CSF in the lung protects against lethal influenza infection. , 2011, American journal of respiratory and critical care medicine.

[69]  J. Davis,et al.  Susceptibility to infection and inflammatory response following influenza virus (H1N1, A/PR/8/34) challenge: role of macrophages. , 2011, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[70]  V. Booth,et al.  Lung Surfactant Protein A (SP-A) Interactions with Model Lung Surfactant Lipids and an SP-B Fragment , 2011, Biochemistry.

[71]  A. Sáenz,et al.  Surfactant protein A (SP-A)-tacrolimus complexes have a greater anti-inflammatory effect than either SP-A or tacrolimus alone on human macrophage-like U937 cells. , 2011, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[72]  V. Sender,et al.  Pulmonary Surfactant Protein A Enhances Endolysosomal Trafficking in Alveolar Macrophages through Regulation of Rab7 , 2011, The Journal of Immunology.

[73]  M. Tate,et al.  N-Linked Glycosylation Facilitates Sialic Acid-Independent Attachment and Entry of Influenza A Viruses into Cells Expressing DC-SIGN or L-SIGN , 2010, Journal of Virology.

[74]  B. Geisbrecht,et al.  Surfactant Protein A (SP-A)-mediated Clearance of Staphylococcus aureus Involves Binding of SP-A to the Staphylococcal Adhesin Eap and the Macrophage Receptors SP-A Receptor 210 and Scavenger Receptor Class A* , 2010, The Journal of Biological Chemistry.

[75]  Bali Pulendran,et al.  Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus , 2010, Proceedings of the National Academy of Sciences.

[76]  Michael Shaw,et al.  Clinical aspects of pandemic 2009 influenza A (H1N1) virus infection. , 2010, The New England journal of medicine.

[77]  U. Holmskov,et al.  Viral aggregating and opsonizing activity in collectin trimers. , 2010, American journal of physiology. Lung cellular and molecular physiology.

[78]  Z. Sever-Chroneos,et al.  Pulmonary Surfactant: An Immunological Perspective , 2009, Cellular Physiology and Biochemistry.

[79]  Tasleem Samji Influenza A: Understanding the Viral Life Cycle , 2009, The Yale journal of biology and medicine.

[80]  C. Stamme,et al.  Role of clathrin-mediated endocytosis of surfactant protein A by alveolar macrophages in intracellular signaling. , 2009, American journal of physiology. Lung cellular and molecular physiology.

[81]  Lucy A. Perrone,et al.  H5N1 and 1918 Pandemic Influenza Virus Infection Results in Early and Excessive Infiltration of Macrophages and Neutrophils in the Lungs of Mice , 2008, PLoS pathogens.

[82]  Sung Whan Cho,et al.  Alveolar Macrophages Are Indispensable for Controlling Influenza Viruses in Lungs of Pigs , 2008, Journal of Virology.

[83]  K. Hartshorn,et al.  Inhibition of hemagglutination activity of influenza A viruses by SP-A1 and SP-A2 variants expressed in CHO cells , 2007, Medical Microbiology and Immunology.

[84]  David F. Smith,et al.  Receptor binding specificity of recent human H3N2 influenza viruses , 2007, Virology Journal.

[85]  B. Trapnell,et al.  PU.1 Redirects Adenovirus to Lysosomes in Alveolar Macrophages, Uncoupling Internalization from Infection1 , 2007, The Journal of Immunology.

[86]  L. Schlesinger,et al.  Endocytic pathway for surfactant protein A in human macrophages: binding, clathrin-mediated uptake, and trafficking through the endolysosomal pathway. , 2006, American journal of physiology. Lung cellular and molecular physiology.

[87]  David E. Swayne,et al.  Pathogenicity of Influenza Viruses with Genes from the 1918 Pandemic Virus: Functional Roles of Alveolar Macrophages and Neutrophils in Limiting Virus Replication and Mortality in Mice , 2005, Journal of Virology.

[88]  N. Bovin,et al.  Receptor-binding properties of swine influenza viruses isolated and propagated in MDCK cells. , 2005, Virus research.

[89]  J. Shabanowitz,et al.  Identification of the Surfactant Protein A Receptor 210 as the Unconventional Myosin 18A* , 2005, Journal of Biological Chemistry.

[90]  J. Goerke,et al.  Pulmonary Collectins Modulate Strain-Specific Influenza A Virus Infection and Host Responses , 2004, Journal of Virology.

[91]  T. Kodama,et al.  Pulmonary Surfactant Protein A Augments the Phagocytosis of Streptococcus pneumoniae by Alveolar Macrophages through a Casein Kinase 2-dependent Increase of Cell Surface Localization of Scavenger Receptor A* , 2004, Journal of Biological Chemistry.

[92]  D. Voelker,et al.  Pulmonary Surfactant Protein A Up-Regulates Activity of the Mannose Receptor, a Pattern Recognition Receptor Expressed on Human Macrophages1 , 2002, The Journal of Immunology.

[93]  J. Whitsett,et al.  Absence of SP-A modulates innate and adaptive defense responses to pulmonary influenza infection. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[94]  S. Hawgood,et al.  Surfactant protein-A--deficient mice display an exaggerated early inflammatory response to a beta-resistant strain of influenza A virus. , 2002, American journal of respiratory cell and molecular biology.

[95]  J. Whitsett,et al.  GM-CSF regulates alveolar macrophage differentiation and innate immunity in the lung through PU.1. , 2001, Immunity.

[96]  J. Whitsett,et al.  GM-CSF regulates protein and lipid catabolism by alveolar macrophages. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[97]  M. Peppelenbosch,et al.  Macrophages Present Pinocytosed Exogenous Antigen Via MHC Class I Whereas Antigen Ingested by Receptor-Mediated Endocytosis Is Presented Via MHC Class II , 2000, The Journal of Immunology.

[98]  D. Bruttomesso,et al.  Different pathways of degradation of SP-A and saturated phosphatidylcholine by alveolar macrophages. , 2000, American journal of physiology. Lung cellular and molecular physiology.

[99]  P. Reading,et al.  Involvement of the Mannose Receptor in Infection of Macrophages by Influenza Virus , 2000, Journal of Virology.

[100]  J. Whitsett,et al.  Surfactant protein A (SP-A) gene targeted mice. , 1998, Biochimica et biophysica acta.

[101]  K. Edwards,et al.  SP-A enhances uptake of bacillus Calmette-Guérin by macrophages through a specific SP-A receptor. , 1997, The American journal of physiology.

[102]  B. Benaissa-Trouw,et al.  Surfactant protein A, but not surfactant protein D, is an opsonin for influenza A virus phagocytosis by rat alveolar macrophages , 1997, European journal of immunology.

[103]  P. Reading,et al.  Changes in the hemagglutinin molecule of influenza type A (H3N2) virus associated with increased virulence for mice , 1997, Archives of Virology.

[104]  A. Fisher,et al.  Surfactant protein A is degraded by alveolar macrophages. , 1996, The American journal of physiology.

[105]  J. Whitsett,et al.  Purification of a Cell-surface Receptor for Surfactant Protein A* , 1996, The Journal of Biological Chemistry.

[106]  F. Ratjen,et al.  Age-dependency of surfactant phospholipids and surfactant protein A in bronchoalveolar lavage fluid of children without bronchopulmonary disease. , 1996, The European respiratory journal.

[107]  V. Shepherd,et al.  Differential regulation of the mannose and SP-A receptors on macrophages. , 1995, The American journal of physiology.

[108]  J. Wright,et al.  Degradation of surfactant lipids and surfactant protein A by alveolar macrophages in vitro. , 1995, The American journal of physiology.

[109]  M. Harmsen,et al.  Interactions of surfactant protein A with influenza A viruses: binding and neutralization. , 1995, The Journal of infectious diseases.

[110]  S. Thiel,et al.  Binding of human collectins (SP-A and MBP) to influenza virus. , 1994, The Biochemical journal.

[111]  G. Dranoff,et al.  Involvement of granulocyte-macrophage colony-stimulating factor in pulmonary homeostasis. , 1994, Science.

[112]  K. Sakai,et al.  Heterogeneity of immunohistochemical staining with pulmonary surfactant protein A among fractionated alveolar macrophages which involves metabolism of pulmonary surfactant. , 1992, Cellular and molecular biology.

[113]  J. Swanson,et al.  M-CSF-induced macropinocytosis increases solute endocytosis but not receptor-mediated endocytosis in mouse macrophages. , 1992, Journal of cell science.

[114]  J. Wright,et al.  Human pulmonary surfactant protein (SP-A), a protein structurally homologous to C1q, can enhance FcR- and CR1-mediated phagocytosis. , 1989, The Journal of biological chemistry.

[115]  J. Yewdell,et al.  Monoclonal antibodies localize events in the folding, assembly, and intracellular transport of the influenza virus hemagglutinin glycoprotein , 1988, Cell.

[116]  K. Drickamer,et al.  The major lung surfactant protein, SP 28-36, is a calcium-dependent, carbohydrate-binding protein. , 1987, The Journal of biological chemistry.

[117]  R. Crystal,et al.  Estimation of volume of epithelial lining fluid recovered by lavage using urea as marker of dilution. , 1986, Journal of applied physiology.

[118]  J. Yewdell,et al.  Monoclonal antibodies detect different forms of influenza virus hemagglutinin during viral penetration and biosynthesis , 1985, Journal of virology.