Does the renal expression of Toll-like receptors play a role in patients with IgA nephropathy?

  • Hana Ciferska
  • Eva Honsova
  • Alena Lodererova
  • Zdenka Hruskova
  • Michaela Neprasova
  • Jan Vachek
  • Miloslav Suchanek
  • Tomas Zima
  • Rosanna Coppo
  • Vladimir Tesar
  • Jan Novak
  • Dita MaixnerovaEmail author
Original Article


The onset of IgA nephropathy (IgAN), characterized by glomerular deposition of IgA-containing immune complexes, is often associated with synpharyngitic hematuria. Innate immune responses mediated by Toll-like receptors (TLR) may play a role in IgAN onset and/or progression. Here, we assessed the expression of TLR 4, 7, 8, and 9 in renal-biopsy specimens from patients with IgAN, with different degree of proteinuria and eGFR, compared with normal-kidney and disease-control tissues (ANCA-associated vasculitis). Renal-biopsy specimens from 34 patients with IgAN and 7 patients with ANCA-associated vasculitis were used. In addition, we used 15 healthy portions of renal-tissue specimens from kidneys after nephrectomy for cancer as control specimens. Expression of TLR 4, 7, 8, and 9 was assessed using immunohistochemical staining of paraffin-embedded renal-biopsy tissue specimens with specific antibodies and evaluated semiquantitatively by light microscopy. Linear discriminant analysis (LDA) was used to test whether intrarenal staining of TLR 4, 7, 8, and 9 distinguished patients with IgAN from controls or correlated with eGFR and/or proteinuria. eGFR was calculated using the creatinine-based formula. Moreover, the biopsies from patients with IgAN were scored according to the Oxford Classification. LDA showed that staining for TLR 4, 7, 8, and 9 was more intense in specimens from IgAN patients compared to normal kidney tissues. The intensity of intrarenal staining of TLRs discriminated four groups of IgAN patients with different eGFR and proteinuria and MEST scoring.


Toll-like receptors Renal biopsy Renal insufficiency IgA nephropathy ANCA-associated vasculitis 



Study was supported by a project of the Ministry of Health of the Czech Republic for conceptual research development by organization 023728 and by Grants LH15168, PROGRES Q25/LF1 and DRO VFN 64165 from the Ministry of Health of the Czech Republic. MS acknowledges the assistance provided by the Research Infrastructure NanoEnviCz supported by the Ministry of Education, Youth and Sports of the Czech Republic under Project No. LM2015073. JN was supported in part by Grants DK078244 and DK082753 from the National Institutes of Health.

Author contributions

HC, DM, ZH, JV and VT conceived and planned the experiments and worked on the manuscript. HC and AL carried out the experiments. EH and MS processed the experimental data, performed the analysis and the evaluation of the results. TZ, MN, RC and JN aided in interpreting the results and drafter and edited the manuscript. All authors discussed the results and commented on the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the ethical committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants involved in the study.

Supplementary material

40620_2019_640_MOESM1_ESM.docx (17 kb)
Supplementary material 1 (DOCX 16 kb)
40620_2019_640_MOESM2_ESM.tiff (36 kb)
Supplementary material 2 (TIFF 36 kb)
40620_2019_640_MOESM3_ESM.tiff (38 kb)
Supplementary material 3 (TIFF 38 kb)


  1. 1.
    Wyatt RJ, Julian BA (2013) IgA nephropathy. N Engl J Med 20(368):2402–2414. CrossRefGoogle Scholar
  2. 2.
    Kiryluk K, Li Y, Sanna-Cherchi S, Rohanizadegan M, Suzuki H, Eitner F et al (2012) Geographic differences in genetic susceptibility to IgA nephropathy: GWAS replication study and geospatial risk analysis. PLoS Genet 8:e1002765. CrossRefGoogle Scholar
  3. 3.
    Berthoux F, Suzuki H, Thibaudin L, Yanagawa H, Maillard N, Mariat C et al (2012) Autoantibodies targeting galactose-deficient IgA1 associate with progression of IgA nephropathy. J Am Soc Nephrol 23(9):1579–1587. CrossRefGoogle Scholar
  4. 4.
    Hastings MC, Moldoveanu Z, Suzuki H, Berthoux F, Julian BA, Sanders JT et al (2013) Biomarkers in IgA nephropathy: relationship to pathogenetic hits. Expert Opin Med Diagn 7(6):615–627. CrossRefGoogle Scholar
  5. 5.
    Caliskan Y, Kiryluk K (2014) Novel biomarkers in glomerular disease. Adv Chronic Kidney Dis 21(2):205–216. CrossRefGoogle Scholar
  6. 6.
    Himanshu K, Kiyoshi T, Shizuo A (2013) Toll-Like Receptors. In: The Encyclopedia of biological chemistry. (Waltham, MA, USA:Elsevier), 396-401Google Scholar
  7. 7.
    González-Guerrero C, Cannata-Ortiz P, Guerri C, Egido J, Ortiz A, Ramos AM (2017) TLR4-mediated inflammation is a key pathogenic event leading to kidney damage and fibrosis in cyclosporine nephrotoxicity. Arch Toxicol 91(4):1925–1939. CrossRefGoogle Scholar
  8. 8.
    Muto M, Manfroi B, Suzuki H, Joh K, Nagai M, Wakai S et al (2017) Toll-like receptor 9 stimulation induces aberrant expression of a proliferation-inducing ligand by tonsillar germinal center B cells in IgA nephropathy. J Am Soc Nephrol 28(4):1227–1238. CrossRefGoogle Scholar
  9. 9.
    Liu Y, Liu H, Peng Y, Liu F et al (2014) New insights into the pathogenesis of IgA nephropathy: do Toll like receptor 9-B cell activation factor-IgA class switching recombination signaling axis induce IgA hyper-production? Ren Fail 36(6):970–973. CrossRefGoogle Scholar
  10. 10.
    Li W, Peng X, Liu Y, Liu H, Liu F, He L et al (2014) TLR9 and BAFF: their expression in patients with IgA nephropathy. Mol Med Rep 10(3):1469–1474. CrossRefGoogle Scholar
  11. 11.
    Coppo R, Camilla R, Amore A, Peruzzi L, Daprà V, Loiacono E et al (2010) Toll-like receptor 4 expression is increased in circulating mononuclear cells of patients with immunoglobulin A nephropathy. Clin Exp Immunol 159(1):73–81. CrossRefGoogle Scholar
  12. 12.
    Roberts IS, Cook HT, Troyanov S, Alpers CE, Amore A, Barratt J et al (2009) The Oxford classification of IgA nephropathy: pathology definitions, correlations, and reproducibility. Kidney Int 76(5):546–556. CrossRefGoogle Scholar
  13. 13.
    Vandeginste BGM (1998) Handbook of chemometrics and qualimetrics, part B. Elsevier, WalthamGoogle Scholar
  14. 14.
    Program XLSTAT (
  15. 15.
    Saito A, Komatsuda A, Kaga H, Togashi M, Okuyama S, Wakui H et al (2016) Different expression patterns of Toll-like receptor mRNAs in blood mononuclear cells of IgA nephropathy and IgA vasculitis with nephritis. Tohoku J Exp Med 240(3):199–208. CrossRefGoogle Scholar
  16. 16.
    Nakata J, Suzuki Y, Suzuki H, Sato D, Kano T, Yanagawa H et al (2014) Changes in nephritogenic serum galactose-deficient IgA1 in IgA nephropathy following tonsillectomy and steroid therapy. PLoS One 9(2):e89707. CrossRefGoogle Scholar
  17. 17.
    Donadio ME, Loiacono E, Peruzzi L, Amore A, Camilla R, Chiale F et al (2014) Toll-like receptors, immunoproteasome and regulatory T cells in children with Henoch-Schönlein purpura and primary IgA nephropathy. Pediatr Nephrol 29(9):1545–1551. CrossRefGoogle Scholar
  18. 18.
    Zou JN, Xiao J, Hu SS, Fu CS, Zhang XL, Zhang ZX et al (2017) Toll-like receptor 4 signaling pathway in the protective effect of pioglitazone on experimental immunoglobulin A nephropathy. Chin Med J 130(8):906–913. CrossRefGoogle Scholar
  19. 19.
    Vergano L, Loiacono E, Albera R, Coppo R, Camilla R, Peruzzi L et al (2015) Can tonsillectomy modify the innate and adaptive immunity pathways involved in IgA nephropathy? J Nephrol 28(1):51–58. CrossRefGoogle Scholar
  20. 20.
    Anders HJ, Andersen K, Stecher B (2013) The intestinal microbiota, a leaky gut, and abnormal immunity in kidney disease. Kidney Int 83(6):1010–1016. CrossRefGoogle Scholar
  21. 21.
    Suzuki H, Suzuki Y, Narita I, Aizawa M, Kihara M, Yamanaka T et al (2008) Toll-like receptor 9 affects severity of IgA nephropathy. J Am Soc Nephrol 19(12):2384–2395. CrossRefGoogle Scholar
  22. 22.
    Sallustio F, Cox SN, Serino G, Curci C, Pesce F, De Palma G et al (2015) Genome-wide scan identifies a copy number variable region at 3p21.1 that influences the TLR9 expression levels in IgA nephropathy patients. Eur J Hum Genet 23(7):940–948. CrossRefGoogle Scholar
  23. 23.
    Imaizumi T, Aizawa T, Segawa C, Shimada M, Tsuruga K, Kawaguchi S et al (2015) Toll-like receptor 3 signaling contributes to the expression of a neutrophil chemoattractant, CXCL1 in human mesangial cells. Clin Exp Nephrol 19(5):761–770. CrossRefGoogle Scholar
  24. 24.
    Zhu FG, Jiang W, Bhagat L, Wang D, Yu D, Tang JX et al (2013) A novel antagonist of Toll-like receptors 7, 8 and 9 suppresses lupus disease-associated parameters in NZBW/F1 mice. Autoimmunity 46(7):419–428. CrossRefGoogle Scholar
  25. 25.
    Kimura J, Ichii O, Miyazono K, Nakamura T, Horino T, Otsuka-Kanazawa S et al (2014) Overexpression of Toll-like receptor 8 correlates with the progression of podocyte injury in murine autoimmune glomerulonephritis. Sci Rep 4:7290. CrossRefGoogle Scholar

Copyright information

© Italian Society of Nephrology 2019

Authors and Affiliations

  • Hana Ciferska
    • 1
  • Eva Honsova
    • 2
  • Alena Lodererova
    • 2
  • Zdenka Hruskova
    • 3
  • Michaela Neprasova
    • 3
  • Jan Vachek
    • 3
  • Miloslav Suchanek
    • 4
  • Tomas Zima
    • 5
  • Rosanna Coppo
    • 6
  • Vladimir Tesar
    • 3
  • Jan Novak
    • 7
  • Dita Maixnerova
    • 3
    Email author
  1. 1.Institute of Rheumatology, 1st Faculty of MedicineCharles UniversityPragueCzech Republic
  2. 2.Department of PathologyInstitute of Clinical and Experimental MedicinePragueCzech Republic
  3. 3.Department of Nephrology, 1st Faculty of Medicine and General University HospitalCharles UniversityPrague 2Czech Republic
  4. 4.Faculty of EnvironmentJan Evangelista Purkyne University in Usti nad LabemUsti nad LabemCzech Republic
  5. 5.Institute of Clinical Biochemistry and Laboratory Diagnostics, 1st Faculty of Medicine and General University HospitalCharles UniversityPragueCzech Republic
  6. 6.Fondazione Ricerca MolinetteRegina Margherita HospitalTurinItaly
  7. 7.Department of MicrobiologyUniversity of Alabama at BirminghamBirminghamUSA

Personalised recommendations