A20 and the noncanonical NF-κB pathway are key regulators of neutrophil recruitment during fetal ontogeny

Newborns are at high risk of developing neonatal sepsis, particularly if born prematurely. This has been linked to divergent requirements the immune system has to fulfill during intrauterine compared with extrauterine life. By transcriptomic analysis of fetal and adult neutrophils, we shed new light on the molecular mechanisms of neutrophil maturation and functional adaption during fetal ontogeny. We identified an accumulation of differentially regulated genes within the noncanonical NF-κB signaling pathway accompanied by constitutive nuclear localization of RelB and increased surface expression of TNF receptor type II in fetal neutrophils, as well as elevated levels of lymphotoxin α in fetal serum. Furthermore, we found strong upregulation of the negative inflammatory regulator A20 (Tnfaip3) in fetal neutrophils, which was accompanied by pronounced downregulation of the canonical NF-κB pathway. Functionally, overexpressing A20 in Hoxb8 cells led to reduced adhesion of these neutrophil-like cells in a flow chamber system. Conversely, mice with a neutrophil-specific A20 deletion displayed increased inflammation in vivo. Taken together, we have uncovered constitutive activation of the noncanonical NF-κB pathway with concomitant upregulation of A20 in fetal neutrophils. This offers perfect adaption of neutrophil function during intrauterine fetal life but also restricts appropriate immune responses particularly in prematurely born infants.

[1]  M. Sperandio,et al.  Perinatal development of innate immune topology , 2021, eLife.

[2]  A. Mócsai,et al.  In Vivo Functions of Mouse Neutrophils Derived from HoxB8-Transduced Conditionally Immortalized Myeloid Progenitors , 2020, The Journal of Immunology.

[3]  M. Uhlén,et al.  Dramatic changes in blood protein levels during the first week of life in extremely preterm infants , 2020, Pediatric Research.

[4]  A. Mócsai,et al.  Neutrophils as emerging therapeutic targets , 2020, Nature Reviews Drug Discovery.

[5]  Erin E. Gill,et al.  Dynamic molecular changes during the first week of human life follow a robust developmental trajectory , 2019, Nature Communications.

[6]  Ryan R Brinkman,et al.  Dynamic molecular changes during the first week of human life follow a robust developmental trajectory , 2019, Nature Communications.

[7]  J. Mikes,et al.  Stereotypic Immune System Development in Newborn Children , 2018, Cell.

[8]  B. Walzog,et al.  A Fundamental Role of Myh9 for Neutrophil Migration in Innate Immunity , 2018, The Journal of Immunology.

[9]  M. Kool,et al.  A20/Tumor Necrosis Factor α-Induced Protein 3 in Immune Cells Controls Development of Autoinflammation and Autoimmunity: Lessons from Mouse Models , 2018, Front. Immunol..

[10]  F. Geissmann,et al.  Yolk sac macrophage progenitors traffic to the embryo during defined stages of development , 2018, Nature Communications.

[11]  D. Baltimore,et al.  30 Years of NF-κB: A Blossoming of Relevance to Human Pathobiology , 2017, Cell.

[12]  Jamie Perin,et al.  Global, regional, and national causes of under-5 mortality in 2000–15: an updated systematic analysis with implications for the Sustainable Development Goals , 2016, The Lancet.

[13]  H. R. Razzaghian,et al.  An Immunological Perspective on Neonatal Sepsis. , 2016, Trends in molecular medicine.

[14]  M. Sperandio,et al.  Extracellular MRP8/14 is a regulator of β2 integrin-dependent neutrophil slow rolling and adhesion , 2015, Nature Communications.

[15]  A. Waisman,et al.  Catchup: a mouse model for imaging-based tracking and modulation of neutrophil granulocytes , 2015, Nature Methods.

[16]  A. Burlingame,et al.  A20 restricts ubiquitination of pro-interleukin-1β protein complexes and suppresses NLRP3 inflammasome activity. , 2015, Immunity.

[17]  S. Turvey,et al.  Impaired NLRP3 inflammasome activity during fetal development regulates IL‐1β production in human monocytes , 2015, European journal of immunology.

[18]  Manolis Kellis,et al.  The NF-κB genomic landscape in lymphoblastoid B cells. , 2014, Cell reports.

[19]  G. van Loo,et al.  Negative regulation of the NLRP3 inflammasome by A20 protects against arthritis , 2014, Nature.

[20]  L. Catrysse,et al.  A20 in inflammation and autoimmunity. , 2014, Trends in immunology.

[21]  C. McCall,et al.  RelB: an outlier in leukocyte biology , 2013, Journal of leukocyte biology.

[22]  M. Oyama,et al.  Involvement of A20 in the molecular switch that activates the non-canonical NF-кB pathway , 2013, Scientific Reports.

[23]  D. Finkelstein,et al.  Generation of hematopoietic progenitor cell lines with myeloid and lymphoid potential , 2013, Nature Methods.

[24]  D. Frommhold,et al.  Ontogenetic regulation of leukocyte recruitment in mouse yolk sac vessels. , 2013, Blood.

[25]  Jing Wang,et al.  WEB-based GEne SeT AnaLysis Toolkit (WebGestalt): update 2013 , 2013, Nucleic Acids Res..

[26]  D. Frommhold,et al.  Neutrophil and endothelial adhesive function during human fetal ontogeny , 2013, Journal of leukocyte biology.

[27]  A. Ma,et al.  A20: linking a complex regulator of ubiquitylation to immunity and human disease , 2012, Nature Reviews Immunology.

[28]  A. Butler,et al.  The developing human preterm neonatal immune system: a case for more research in this area. , 2012, Clinical immunology.

[29]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[30]  Noula Shembade,et al.  Regulation of NF-κB signaling by the A20 deubiquitinase , 2012, Cellular and Molecular Immunology.

[31]  E. Huang,et al.  Dendritic cell expression of A20 preserves immune homeostasis and prevents colitis and spondyloarthritis , 2011, Nature Immunology.

[32]  M. Kool,et al.  A20 (TNFAIP3) deficiency in myeloid cells triggers erosive polyarthritis resembling rheumatoid arthritis , 2011, Nature Genetics.

[33]  P. Gregersen,et al.  Association of a functional variant downstream of TNFAIP3 with systemic lupus erythematosus , 2011, Nature Genetics.

[34]  Rafael A. Irizarry,et al.  A framework for oligonucleotide microarray preprocessing , 2010, Bioinform..

[35]  G. Berbers,et al.  Transplacental Transport of IgG Antibodies Specific for Pertussis, Diphtheria, Tetanus, Haemophilus influenzae Type b, and Neisseria meningitidis Serogroup C Is Lower in Preterm Compared With Term Infants , 2010, The Pediatric infectious disease journal.

[36]  D. Ray,et al.  Functional evaluation of TNFAIP3 (A20) in rheumatoid arthritis. , 2010, Clinical and experimental rheumatology.

[37]  C. Liu,et al.  The ubiquitin modifying enzyme A20 restricts B cell survival and prevents autoimmunity. , 2010, Immunity.

[38]  E. Adams,et al.  African-Derived Genetic Polymorphisms in TNFAIP3 Mediate Risk for Autoimmunity , 2010, The Journal of Immunology.

[39]  M. Sperandio,et al.  Delayed but functional neutrophil extracellular trap formation in neonates. , 2009, Blood.

[40]  G. van Loo,et al.  The ubiquitin-editing enzyme A20 (TNFAIP3) is a central regulator of immunopathology. , 2009, Trends in immunology.

[41]  A. Weyrich,et al.  Comment on Chapter 5 , 1999 .

[42]  C. Dinarello,et al.  Immunological and inflammatory functions of the interleukin-1 family. , 2009, Annual review of immunology.

[43]  Ofer Levy,et al.  Innate immunity of the newborn: basic mechanisms and clinical correlates , 2007, Nature Reviews Immunology.

[44]  S. Yan,et al.  Human newborn polymorphonuclear neutrophils exhibit decreased levels of MyD88 and attenuated p38 phosphorylation in response to lipopolysaccharide. , 2007, Clinical and investigative medicine. Medecine clinique et experimentale.

[45]  A. Prusa,et al.  Immaturity of infection control in preterm and term newborns is associated with impaired toll-like receptor signaling. , 2007, The Journal of infectious diseases.

[46]  Bryan Heit,et al.  Intraluminal crawling of neutrophils to emigration sites: a molecularly distinct process from adhesion in the recruitment cascade , 2006, The Journal of experimental medicine.

[47]  K. Blum,et al.  Keystones in lymph node development , 2006, Journal of anatomy.

[48]  G. Wang,et al.  Quantitative production of macrophages or neutrophils ex vivo using conditional Hoxb8 , 2006, Nature Methods.

[49]  D. Tamandl,et al.  Monocyte Toll-Like Receptor 4 Expression and LPS-Induced Cytokine Production Increase during Gestational Aging , 2005, Pediatric Research.

[50]  Bing Zhang,et al.  WebGestalt: an integrated system for exploring gene sets in various biological contexts , 2005, Nucleic Acids Res..

[51]  K. Ley,et al.  P-selectin Glycoprotein Ligand-1 Mediates L-Selectin–dependent Leukocyte Rolling in Venules , 2003, The Journal of experimental medicine.

[52]  F. Weih,et al.  RelB is required for Peyer's patch development: differential regulation of p52–RelB by lymphotoxin and TNF , 2003, The EMBO journal.

[53]  W. Poole,et al.  Late-onset sepsis in very low birth weight neonates: the experience of the NICHD Neonatal Research Network. , 2002, Pediatrics.

[54]  M. Pillinger,et al.  The neutrophil: function and regulation in innate and humoral immunity. , 2001, Clinical immunology.

[55]  A. Ma,et al.  Failure to regulate TNF-induced NF-kappaB and cell death responses in A20-deficient mice. , 2000, Science.

[56]  T. Graf,et al.  Insertion of enhanced green fluorescent protein into the lysozyme gene creates mice with green fluorescent granulocytes and macrophages. , 2000, Blood.

[57]  R. Beyaert,et al.  The cytokine‐inducible zinc finger protein A20 inhibits IL‐1‐induced NF‐κB activation at the level of TRAF6 , 1999, FEBS letters.

[58]  M. Tcharmtchi,et al.  P-Selectin support of neonatal neutrophil adherence under flow: contribution of L-selectin, LFA-1, and ligand(s) for P-selectin. , 1998, Blood.

[59]  D. Goeddel,et al.  The tumor necrosis factor-inducible zinc finger protein A20 interacts with TRAF1/TRAF2 and inhibits NF-kappaB activation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[60]  F. Weih,et al.  Multiorgan inflammation and hematopoietic abnormalities in mice with a targeted disruption of RelB, a member of the NF-κB/Rel family , 1995, Cell.

[61]  A. Krikos,et al.  Transcriptional activation of the tumor necrosis factor alpha-inducible zinc finger protein, A20, is mediated by kappa B elements. , 1992, The Journal of biological chemistry.