Natural Monoclonal Antibody to Oxidized Low-Density Lipoprotein and Aggregatibacter actinomycetemcomitans

  • Chunguang Wang
  • Sohvi Hörkkö
Part of the Methods in Molecular Biology book series (MIMB, volume 1643)


Natural antibodies are produced by B lymphocytes without exogenous antigenic exposure and are present at the time of birth. They usually bind to conserved epitopes on antigens of different chemical compositions. We cloned and characterized a natural mouse monoclonal IgM antibody (Aa_Mab) by selecting the binding to malondialdehyde acetaldehyde (MAA) adducts on low-density lipoprotein (LDL). The data showed that the Aa_Mab cross-reacted with Aggregatibacter actinomycetemcomitans (Aa) bacteria, an important oral pathogen in periodontitis associated with atherosclerosis. Surprisingly, the binding molecule of Aa bacteria to the Aa_Mab was Aa chaperonin 60 or HSP60, a protein that is not only responsible for maintaining cellular proteins conformation, but also functions as a potent virulence factor prompting bone resorption in periodontitis and as a putative pathogenic factor in atherosclerosis.

Key words

Natural antibody Oxidized low-density lipoprotein Malondialdehyde acetaldehyde Aggregatibacter actinomycetemcomitans Chaperonin 60 



This study was funded by Sigrid Juselius Foundation and Finnish Foundation for Cardiovascular Research.


  1. 1.
    Grönwall C, Vas J, Silverman GJ (2012) Protective Roles of Natural IgM Antibodies. Front Immunol 3:66. doi: 10.3389/fimmu.2012.00066 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Panda S, Zhang J, Tan NS, Ho B, Ding JL (2013) Natural IgG antibodies provide innate protection against ficolin-opsonized bacteria. EMBO J 32(22):2905–2919. doi: 10.1038/emboj.2013.199 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Pabst O (2012) New concepts in the generation and functions of IgA. Nat Rev Immunol 12(12):821–832. doi: 10.1038/nri3322 CrossRefPubMedGoogle Scholar
  4. 4.
    Tsiantoulas D, Diehl CJ, Witztum JL, Binder C (2014) B cells and humoral immunity in atherosclerosis. Circ Res 114(11):1743–1756. doi: 10.1161/circresaha.113.301145 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Grönwall C, Silverman GJ (2014) Natural IgM: beneficial autoantibodies for the control of inflammatory and autoimmune disease. J Clin Immunol 34(Suppl 1):12–21. doi: 10.1007/s10875-014-0025-4 CrossRefGoogle Scholar
  6. 6.
    Lewis MJ, Malik TH, Ehrenstein MR, Boyle JJ, Botto M, Haskard DO (2009) Immunoglobulin M is required for protection against atherosclerosis in low-density lipoprotein receptor-deficient mice. Circulation 120(5):417–426. doi: 10.1161/circulationaha.109.868158 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Kyaw T, Tay C, Krishnamurthi S, Kanellakis P, Agrotis A, Tipping P et al (2011) B1a B lymphocytes are atheroprotective by secreting natural IgM that increases IgM deposits and reduces necrotic cores in atherosclerotic lesions. Circ Res 109(8):830–840. doi: 10.1161/circresaha.111.248542 CrossRefPubMedGoogle Scholar
  8. 8.
    Karvonen J, Päivänsalo M, Kesäniemi YA, Hörkkö S (2003) Immunoglobulin M type of autoantibodies to oxidized low-density lipoprotein has an inverse relation to carotid artery atherosclerosis. Circulation 108(17):2107–2112. doi: 10.1161/01.CIR.0000092891.55157.A7 CrossRefPubMedGoogle Scholar
  9. 9.
    Tsimikas S, Miyanohara A, Hartvigsen K, Merki E, Shaw PX, Chou MY et al (2011) Human oxidation-specific antibodies reduce foam cell formation and atherosclerosis progression. J Am Coll Cardiol 58(16):1715–1727. doi: 10.1016/j.jacc.2011.07.017 CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Wang C, Turunen SP, Kummu O, Veneskoski M, Lehtimäki J, Nissinen AE et al (2013) Natural antibodies of newborns recognize oxidative stress-related malondialdehyde acetaldehyde adducts on apoptotic cells and atherosclerotic plaques. Int Immunol 25(10):575–587. doi: 10.1093/intimm/dxt022 CrossRefPubMedGoogle Scholar
  11. 11.
    Turunen SP, Kummu O, Harila K, Veneskoski M, Soliymani R, Baumann M et al (2012) Recognition of Porphyromonas gingivalis gingipain epitopes by natural IgM binding to malondialdehyde modified low-density lipoprotein. PLoS One 7(4):e34910. doi: 10.1371/journal.pone.0034910 CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Wang C, Kankaanpää J, Kummu O, Turunen SP, Akhi R, Bergmann U et al (2016) Characterization of a natural mouse monoclonal antibody recognizing epitopes shared by oxidized low-density lipoprotein and chaperonin 60 of Aggregatibacter actinomycetemcomitans. Immunol Res. doi: 10.1007/s12026-015-8781-7 Google Scholar
  13. 13.
    Demmer RT, Desvarieux M (2006) Periodontal infections and cardiovascular disease: the heart of the matter. J Am Dent Assoc 137(Suppl):14S–20S. quiz 38S. 137/suppl_2/14S [pii]CrossRefPubMedGoogle Scholar
  14. 14.
    Lockhart PB, Bolger AF, Papapanou PN, Osinbowale O, Trevisan M, Levison ME et al (2012) Periodontal disease and atherosclerotic vascular disease: does the evidence support an independent association? A scientific statement from the American Heart Association. Circulation 125(20):2520–2544. doi: 10.1161/CIR.0b013e31825719f3 CrossRefPubMedGoogle Scholar
  15. 15.
    Pussinen PJ, Jousilahti P, Alfthan G, Palosuo T, Asikainen S, Salomaa V (2003) Antibodies to periodontal pathogens are associated with coronary heart disease. Arterioscler Thromb Vasc Biol 23(7):1250–1254. doi: 10.1161/01.ATV.0000072969.71452.87 CrossRefPubMedGoogle Scholar
  16. 16.
    Hyvärinen K, Mäntylä P, Buhlin K, Paju S, Nieminen MS, Sinisalo J et al (2012) A common periodontal pathogen has an adverse association with both acute and stable coronary artery disease. Atherosclerosis 223(2):478–484. doi: 10.1016/j.atherosclerosis.2012.05.021 CrossRefPubMedGoogle Scholar
  17. 17.
    Calandrini CA, Ribeiro AC, Gonnelli AC, Ota-Tsuzuki C, Rangel LP, Saba-Chujfi E et al (2014) Microbial composition of atherosclerotic plaques. Oral Dis 20(3):e128–e134. doi: 10.1111/odi.12205 CrossRefPubMedGoogle Scholar
  18. 18.
    Buhlin K, Holmer J, Gustafsson A, Hörkkö S, Pockley AG, Johansson A et al (2015) Association of periodontitis with persistent, pro-atherogenic antibody responses. J Clin Periodontol. doi: 10.1111/jcpe.12456 PubMedGoogle Scholar
  19. 19.
    Turunen SP, Kummu O, Wang C, Harila K, Mattila R, Sahlman M et al (2015) Immunization with malondialdehyde-modified low-density lipoprotein (LDL) reduces atherosclerosis in LDL receptor-deficient mice challenged with Porphyromonas gingivalis. Innate Immun 21(4):370–385. doi: 10.1177/1753425914542444 CrossRefPubMedGoogle Scholar
  20. 20.
    Pussinen PJ, Vilkuna-Rautiainen T, Alfthan G, Mattila K, Asikainen S (2002) Multiserotype enzyme-linked immunosorbent assay as a diagnostic aid for periodontitis in large-scale studies. J Clin Microbiol 40(2):512–518CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Greenfield EA (2014) Generating monoclonal antibodies. In: Greenfield EA (ed) Antibodies: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 201–221Google Scholar
  22. 22.
    Wang Z, Raifu M, Howard M, Smith L, Hansen D, Goldsby R et al (2000) Universal PCR amplification of mouse immunoglobulin gene variable regions: the design of degenerate primers and an assessment of the effect of DNA polymerase 3′ to 5′ exonuclease activity. J Immunol Methods 233(1–2):167–177CrossRefPubMedGoogle Scholar

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© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Medical Microbiology and Immunology, Research Unit of Biomedicine, Faculty of MedicineUniversity of OuluOuluFinland
  2. 2.Medical Research Center and Nordlab OuluUniversity Hospital and University of OuluOuluFinland

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