Abstract
We analyzed by bidimensional electrophoresis the acid-insoluble fraction of saliva from three classes of angioedema patients and a healthy control group, highlighting significant variations of several normalized spot volumes. Characterization of the corresponding proteins was performed by in-gel tryptic digestion of the spots, followed by high-resolution HPLC-ESI-MS/MS analysis of tryptic mixtures. By this strategy, 16 differentially-expressed proteins among two or more groups were identified. We found higher concentration of proteins involved in immune response (interleukin-1 receptor antagonist and annexin A1), and of moonlighting proteins acting as plasminogen receptors (glyceraldehyde-3-phosphate dehydrogenase, α-enolase, and annexin A2) in patients affected by the idiopathic non-histaminergic or hereditary angioedema with unknown origin with respect to healthy controls. These data provide new information on the molecular basis of these less characterized types of angioedema.
Similar content being viewed by others
Abbreviations
- AAE:
-
acquired angioedema
- ACN:
-
acetonitrile
- Abs:
-
antibodies
- C1-INH:
-
complement component 1 esterase inhibitor
- DTT:
-
Dithiotreitol
- EAACI:
-
European Academy of Allergy and Clinical Immunology
- E-FABP:
-
epidermal fatty acid-binding protein
- ENOA:
-
α-enolase, FA, formic acid
- FXII:
-
coagulation factor XII
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- HAWK:
-
Hereditary Angioedema International Working Group
- HAE:
-
hereditary angioedema
- HC:
-
healthy controls
- HMWK:
-
prekallikrein-high-molecular-weight-kininogen
- IAM:
-
iodoacetamide
- IH:
-
idiopathic histaminergic
- InH:
-
idiopathic non histaminergic
- IL-1:
-
interleukin 1
- IL-1Ra:
-
interleukin-1 receptor antagonist
- IPG:
-
immobilized pH gradient
- LC-MS:
-
liquid mass spectrometry
- LEI:
-
leukocyte elastase inhibitor
- MS/MS:
-
tandem mass spectrometry
- MW:
-
molecular weight; pI, isoelectric point
- PAGE:
-
polyacrylamide Gel Electrophoresis
- pIgR:
-
polymeric immunoglobulin receptor
- PIP:
-
prolactin inducible protein
- PLG:
-
plasminogen
- PTMs:
-
post-translational modifications
- SC:
-
secretory component
- SDS:
-
sodium dodecyl sulfate; #, spot
- TA:
-
tranexamic acid
- TFA:
-
trifluoroacetic acid
- TCA:
-
trichloroacetic acid
- U-HAE:
-
hereditary angioedema without an identified cause
- 2-DE:
-
2-dimensional electrophoresis
References
Cicardi M, Suffritti C, Perego F, Caccia S. Novelties in the Diagnosis and Treatment of Angioedema. J Investig Allergol Clin Immunol. 2016;26(4):212–21; quiz two pages after page 21. doi:https://doi.org/10.18176/jiaci.0087.
Orsenigo F, Giampietro C, Ferrari A, Corada M, Galaup A, Sigismund S, et al. Phosphorylation of VE-cadherin is modulated by haemodynamic forces and contributes to the regulation of vascular permeability in vivo. Nat Commun. 2012;3:1208. https://doi.org/10.1038/ncomms2199.
Cicardi M, Aberer W, Banerji A, Bas M, Bernstein JA, Bork K, et al. Classification, diagnosis, and approach to treatment for angioedema: consensus report from the hereditary angioedema international working group. Allergy. 2014;69(5):602–16. https://doi.org/10.1111/all.12380.
Bova M, Suffritti C, Bafunno V, Loffredo S, Cordisco G, Del Giacco S, et al. Impaired control of the contact system in hereditary angioedema with normal C1-inhibitor. Allergy. 2019;75:1394–403. https://doi.org/10.1111/all.14160.
Firinu D, Bafunno V, Vecchione G, Barca MP, Manconi PE, Santacroce R, et al. Characterization of patients with angioedema without wheals: the importance of F12 gene screening. Clin Immunol. 2015;157(2):239–48. https://doi.org/10.1016/j.clim.2015.02.013.
Dewald G, Bork K. Missense mutations in the coagulation factor XII (Hageman factor) gene in hereditary angioedema with normal C1 inhibitor. Biochem Biophys Res Commun. 2006;343(4):1286–9. https://doi.org/10.1016/j.bbrc.2006.03.092.
Bjorkqvist J, de Maat S, Lewandrowski U, Di Gennaro A, Oschatz C, Schonig K, et al. Defective glycosylation of coagulation factor XII underlies hereditary angioedema type III. J Clin Invest. 2015;125(8):3132–46. https://doi.org/10.1172/JCI77139.
de Maat S, Bjorkqvist J, Suffritti C, Wiesenekker CP, Nagtegaal W, Koekman A, et al. Plasmin is a natural trigger for bradykinin production in patients with hereditary angioedema with factor XII mutations. J Allergy Clin Immunol. 2016;138(5):1414–23 e9. https://doi.org/10.1016/j.jaci.2016.02.021.
Bafunno V, Firinu D, D'Apolito M, Cordisco G, Loffredo S, Leccese A, et al. Mutation of the angiopoietin-1 gene (ANGPT1) associates with a new type of hereditary angioedema. J Allergy Clin Immunol. 2018;141(3):1009–17. https://doi.org/10.1016/j.jaci.2017.05.020.
Bork K, Wulff K, Rossmann H, Steinmuller-Magin L, Braenne I, Witzke G, et al. Hereditary angioedema cosegregating with a novel kininogen 1 gene mutation changing the N-terminal cleavage site of bradykinin. Allergy. 2019;74(12):2479–81. https://doi.org/10.1111/all.13869.
Bork K, Wulff K, Steinmuller-Magin L, Braenne I, Staubach-Renz P, Witzke G, et al. Hereditary angioedema with a mutation in the plasminogen gene. Allergy. 2018;73(2):442–50. https://doi.org/10.1111/all.13270.
Firinu D, Loffredo S, Bova M, Cicardi M, Margaglione M, Del Giacco S. The role of genetics in the current diagnostic workup of idiopathic non-histaminergic angioedema. Allergy. 2019;74(4):810–2. https://doi.org/10.1111/all.13667.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680–5. https://doi.org/10.1038/227680a0.
Cabras T, Iavarone F, Manconi B, Olianas A, Sanna MT, Castagnola M, et al. Top-down analytical platforms for the characterization of the human salivary proteome. Bioanalysis. 2014;6(4):563–81. https://doi.org/10.4155/bio.13.349.
Haskill S, Martin G, Van Le L, Morris J, Peace A, Bigler CF, et al. cDNA cloning of an intracellular form of the human interleukin 1 receptor antagonist associated with epithelium. Proc Natl Acad Sci U S A. 1991;88(9):3681–5. https://doi.org/10.1073/pnas.88.9.3681.
Corradi A, Franzi AT, Rubartelli A. Synthesis and secretion of interleukin-1 alpha and interleukin-1 receptor antagonist during differentiation of cultured keratinocytes. Exp Cell Res. 1995;217(2):355–62. https://doi.org/10.1006/excr.1995.1097.
Arend WP. Cytokine imbalance in the pathogenesis of rheumatoid arthritis: the role of interleukin-1 receptor antagonist. Semin Arthritis Rheum. 2001;30(5 Suppl 2):1–6. https://doi.org/10.1053/sarh.2001.23693.
Vamvakopoulos J, Green C, Metcalfe S. Genetic control of IL-1beta bioactivity through differential regulation of the IL-1 receptor antagonist. Eur J Immunol. 2002;32(10):2988–96. https://doi.org/10.1002/1521-4141(2002010)32:10<2988::AID-IMMU2988>3.0.CO;2-9.
Perrier S, Darakhshan F, Hajduch E. IL-1 receptor antagonist in metabolic diseases: Dr Jekyll or Mr Hyde? FEBS Lett. 2006;580(27):6289–94. https://doi.org/10.1016/j.febslet.2006.10.061.
Dubost JJ, Perrier S, Afane M, Viallard JL, Roux-Lombard P, Baudet-Pommel M, et al. IL-1 receptor antagonist in saliva; characterization in normal saliva and reduced concentration in Sjogren's syndrome (SS). Clin Exp Immunol. 1996;106(2):237–42. https://doi.org/10.1046/j.1365-2249.1996.d01-824.x.
Joseph K, Tholanikunnel BG, Kaplan AP. Cytokine and estrogen stimulation of endothelial cells augments activation of the prekallikrein-high molecular weight kininogen complex: implications for hereditary angioedema. J Allergy Clin Immunol. 2017;140(1):170–6. https://doi.org/10.1016/j.jaci.2016.09.032.
Eiffert H, Quentin E, Decker J, Hillemeir S, Hufschmidt M, Klingmuller D, et al. The primary structure of human free secretory component and the arrangement of disulfide bonds. Hoppe Seylers Z Physiol Chem. 1984;365(12):1489–95.
Eiffert H, Quentin E, Wiederhold M, Hillemeir S, Decker J, Weber M, et al. Determination of the molecular structure of the human free secretory component. Biol Chem Hoppe Seyler. 1991;372(2):119–28. https://doi.org/10.1515/bchm3.1991.372.1.119.
Kaetzel CS. The polymeric immunoglobulin receptor: bridging innate and adaptive immune responses at mucosal surfaces. Immunol Rev. 2005;206:83–99. https://doi.org/10.1111/j.0105-2896.2005.00278.x.
Johansen FE, Kaetzel CS. Regulation of the polymeric immunoglobulin receptor and IgA transport: new advances in environmental factors that stimulate pIgR expression and its role in mucosal immunity. Mucosal Immunol. 2011;4(6):598–602. https://doi.org/10.1038/mi.2011.37.
Salemi M, Mandala V, Muggeo V, Misiano G, Milano S, Colonna-Romano G, et al. Growth factors and IL-17 in hereditary angioedema. Clin Exp Med. 2016;16(2):213–8. https://doi.org/10.1007/s10238-015-0340-y.
D'Acquisto F, Piras G, Rattazzi L. Pro-inflammatory and pathogenic properties of Annexin-A1: the whole is greater than the sum of its parts. Biochem Pharmacol. 2013;85(9):1213–8. https://doi.org/10.1016/j.bcp.2013.02.011.
Spurr L, Nadkarni S, Pederzoli-Ribeil M, Goulding NJ, Perretti M, D'Acquisto F. Comparative analysis of Annexin A1-formyl peptide receptor 2/ALX expression in human leukocyte subsets. Int Immunopharmacol. 2011;11(1):55–66. https://doi.org/10.1016/j.intimp.2010.10.006.
Lorenz K, Bader M, Klaus A, Weiss W, Gorg A, Hofmann T. Orosensory stimulation effects on human saliva proteome. J Agric Food Chem. 2011;59(18):10219–31. https://doi.org/10.1021/jf2024352.
Smith SF, Goulding NJ, Godolphin JL, Tetley TD, Roberts CM, Guz A, et al. An assay for the assessment of lipocortin 1 levels in human lung lavage fluid. J Immunol Methods. 1990;131(1):119–25. https://doi.org/10.1016/0022-1759(90)90241-m.
Dreier R, Schmid KW, Gerke V, Riehemann K. Differential expression of annexins I, II and IV in human tissues: an immunohistochemical study. Histochem Cell Biol. 1998;110(2):137–48. https://doi.org/10.1007/s004180050275.
D'Acunto CW, Gbelcova H, Festa M, Ruml T. The complex understanding of Annexin A1 phosphorylation. Cell Signal. 2014;26(1):173–8. https://doi.org/10.1016/j.cellsig.2013.09.020.
Dorovkov MV, Kostyukova AS, Ryazanov AG. Phosphorylation of annexin A1 by TRPM7 kinase: a switch regulating the induction of an alpha-helix. Biochemistry. 2011;50(12):2187–93. https://doi.org/10.1021/bi101963h.
Callera GE, He Y, Yogi A, Montezano AC, Paravicini T, Yao G, et al. Regulation of the novel Mg2+ transporter transient receptor potential melastatin 7 (TRPM7) cation channel by bradykinin in vascular smooth muscle cells. J Hypertens. 2009;27(1):155–66. https://doi.org/10.1097/hjh.0b013e3283190582.
Wang W, Creutz CE. Regulation of the chromaffin granule aggregating activity of annexin I by phosphorylation. Biochemistry. 1992;31(41):9934–9. https://doi.org/10.1021/bi00156a011.
Torriglia A, Martin E, Jaadane I. The hidden side of SERPINB1/leukocyte Elastase inhibitor. Semin Cell Dev Biol. 2017;62:178–86. https://doi.org/10.1016/j.semcdb.2016.07.010.
Veszeli N, Csuka D, Zotter Z, Imreh E, Jozsi M, Benedek S, et al. Neutrophil activation during attacks in patients with hereditary angioedema due to C1-inhibitor deficiency. Orphanet J Rare Dis. 2015;10:156. https://doi.org/10.1186/s13023-015-0374-y.
Jeffery CJ. Moonlighting proteins. Trends Biochem Sci. 1999;24(1):8–11. https://doi.org/10.1016/s0968-0004(98)01335-8.
Miles LA, Dahlberg CM, Plescia J, Felez J, Kato K, Plow EF. Role of cell-surface lysines in plasminogen binding to cells: identification of alpha-enolase as a candidate plasminogen receptor. Biochemistry. 1991;30(6):1682–91. https://doi.org/10.1021/bi00220a034.
Chauhan AS, Kumar M, Chaudhary S, Patidar A, Dhiman A, Sheokand N, et al. Moonlighting glycolytic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH): an evolutionarily conserved plasminogen receptor on mammalian cells. FASEB J. 2017;31(6):2638–48. https://doi.org/10.1096/fj.201600982R.
MacLeod TJ, Kwon M, Filipenko NR, Waisman DM. Phospholipid-associated annexin A2-S100A10 heterotetramer and its subunits: characterization of the interaction with tissue plasminogen activator, plasminogen, and plasmin. J Biol Chem. 2003;278(28):25577–84. https://doi.org/10.1074/jbc.M301017200.
Maas C. Plasminflammation-an emerging pathway to Bradykinin production. Front Immunol. 2019;10:2046. https://doi.org/10.3389/fimmu.2019.02046.
Joseph K, Tholanikunnel BG, Wolf B, Bork K, Kaplan AP. Deficiency of plasminogen activator inhibitor 2 in plasma of patients with hereditary angioedema with normal C1 inhibitor levels. J Allergy Clin Immunol. 2016;137(6):1822–9 e1. https://doi.org/10.1016/j.jaci.2015.07.041.
Ewald GA, Eisenberg PR. Plasmin-mediated activation of contact system in response to pharmacological thrombolysis. Circulation. 1995;91(1):28–36. https://doi.org/10.1161/01.cir.91.1.28.
Bork K, Wulff K, Witzke G, Hardt J. Treatment for hereditary angioedema with normal C1-INH and specific mutations in the F12 gene (HAE-FXII). Allergy. 2017;72(2):320–4. https://doi.org/10.1111/all.13076.
Du-Thanh A, Raison-Peyron N, Drouet C, Guillot B. Efficacy of tranexamic acid in sporadic idiopathic bradykinin angioedema. Allergy. 2010;65(6):793–5. https://doi.org/10.1111/j.1398-9995.2009.02234.x.
Defendi F, Charignon D, Ghannam A, Baroso R, Csopaki F, Allegret-Cadet M, et al. Enzymatic assays for the diagnosis of bradykinin-dependent angioedema. PLoS One. 2013;8(8):e70140. https://doi.org/10.1371/journal.pone.0070140.
van Geffen M, Cugno M, Lap P, Loof A, Cicardi M, van Heerde W. Alterations of coagulation and fibrinolysis in patients with angioedema due to C1-inhibitor deficiency. Clin Exp Immunol. 2012;167(3):472–8. https://doi.org/10.1111/j.1365-2249.2011.04541.x.
Cugno M, Hack CE, de Boer JP, Eerenberg AJ, Agostoni A, Cicardi M. Generation of plasmin during acute attacks of hereditary angioedema. J Lab Clin Med. 1993;121(1):38–43.
Pancholi V. Multifunctional alpha-enolase: its role in diseases. Cell Mol Life Sci. 2001;58(7):902–20. https://doi.org/10.1007/pl00000910.
Wygrecka M, Marsh LM, Morty RE, Henneke I, Guenther A, Lohmeyer J, et al. Enolase-1 promotes plasminogen-mediated recruitment of monocytes to the acutely inflamed lung. Blood. 2009;113(22):5588–98. https://doi.org/10.1182/blood-2008-08-170837.
Bae S, Kim H, Lee N, Won C, Kim HR, Hwang YI, et al. Alpha-Enolase expressed on the surfaces of monocytes and macrophages induces robust synovial inflammation in rheumatoid arthritis. J Immunol. 2012;189(1):365–72. https://doi.org/10.4049/jimmunol.1102073.
Takaoka Y, Goto S, Nakano T, Tseng HP, Yang SM, Kawamoto S, et al. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) prevents lipopolysaccharide (LPS)-induced, sepsis-related severe acute lung injury in mice. Sci Rep. 2014;4:5204. https://doi.org/10.1038/srep05204.
Miller VA, Madureira PA, Kamaludin AA, Komar J, Sharma V, Sahni G, et al. Mechanism of plasmin generation by S100A10. Thromb Haemost. 2017;117(6):1058–71. https://doi.org/10.1160/TH16-12-0936.
Marceau F, Bachelard H, Rivard GE, Hebert J. Increased fibrinolysis-induced bradykinin formation in hereditary angioedema confirmed using stored plasma and biotechnological inhibitors. BMC Res Notes. 2019;12(1):291. https://doi.org/10.1186/s13104-019-4335-8.
Germenis AE, Loules G, Zamanakou M, Psarros F, Gonzalez-Quevedo T, Speletas M, et al. On the pathogenicity of the plasminogen K330E mutation for hereditary angioedema. Allergy. 2018;73(8):1751–3. https://doi.org/10.1111/all.13324.
Donato R. S100: a multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles. Int J Biochem Cell Biol. 2001;33(7):637–68. https://doi.org/10.1016/s1357-2725(01)00046-2.
Wolk K, Witte E, Wallace E, Docke WD, Kunz S, Asadullah K, et al. IL-22 regulates the expression of genes responsible for antimicrobial defense, cellular differentiation, and mobility in keratinocytes: a potential role in psoriasis. Eur J Immunol. 2006;36(5):1309–23. https://doi.org/10.1002/eji.200535503.
Arba M, Iavarone F, Vincenzoni F, Manconi B, Vento G, Tirone C, et al. Proteomic characterization of the acid-insoluble fraction of whole saliva from preterm human newborns. J Proteome. 2016;146:48–57. https://doi.org/10.1016/j.jprot.2016.06.021.
Ihedioha OC, Shiu RP, Uzonna JE, Myal Y. Prolactin-inducible protein: from breast Cancer biomarker to immune modulator-novel insights from knockout mice. DNA Cell Biol. 2016;35(10):537–41. https://doi.org/10.1089/dna.2016.3472.
Acknowledgements
This research was supported by an unrestricted grant to Dr. Firinu by C.S.L. Behring, Italy (64330-2018/MLOI, 2018) for research in primary immunodeficiency and HAE. The authors acknowledge also the financial support of Cagliari University (FIR 2016) and Catholic University. The authors wish to thank Prof. Enzo Tramontano for the use of the ChemiDoc imaging system.
Authorship Contributions
Study conception and design: MTS, TC, DF, SDG; Cared for patients, extracted the clinical and lab data DF, GC, MA, SDG; Acquisition of data: MA, MTS, FI, FV; Analysis and interpretation of data: TC, IM, MC, MTS, DF; Drafting of manuscript: MTS, MA, TC, DF, GC, SDG; Critical revision: MC, IM, TC, FV, FI. All authors read and worked on the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Declaration of Competing Interest
The Authors declare they have no conflicts of interest.
Additional information
The Authors are available to share data upon request to the Authors and via public repository.
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(DOCX 23 kb)
Rights and permissions
About this article
Cite this article
Firinu, D., Arba, M., Vincenzoni, F. et al. Proteomic Analysis of the Acid-Insoluble Fraction of Whole Saliva from Patients Affected by Different Forms of Non-histaminergic Angioedema. J Clin Immunol 40, 840–850 (2020). https://doi.org/10.1007/s10875-020-00802-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10875-020-00802-w