Classical complement pathway activation in the nasal tissue of patients with chronic rhinosinusitis

Griet A. Van Roey, Christopher C. Vanison, Jeffanie Wu, Julia H. Huang, Lydia A. Suh, Roderick G. Carter, James E. Norton, Stephanie Shintani-Smith, David B. Conley, Kevin C. Welch, Anju T. Peters, Leslie C. Grammer, Kathleen E. Harris, Kathryn E. Hulse, Atsushi Kato, Whitney W. Stevens, Robert C. Kern, Robert P. Schleimer, Bruce K. Tan*

*Corresponding author for this work

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background Complement plays a major role in inflammatory diseases, but its involvement and mechanisms of activation in patients with chronic rhinosinusitis (CRS) are not known. Objectives After earlier studies discovering autoantibodies in patients with CRS, we sought to investigate the nature, extent, and location of complement activation in nasal tissue of patients with CRS. Specifically, we were interested in whether antibody-mediated activation through the classical pathway was a major mechanism for complement activation in patients with CRS. Methods Nasal tissue was obtained from patients with CRS and control subjects. Tissue homogenates were analyzed for complement activation products (ELISA–C5b-9, C4d, activated C1, and C5a) and major complement-fixing antibodies (Luminex). Tissue sections were stained for C5b-9, C4d, and laminin. Antibodies were purified with protein A/G columns from nasal polyps (NP), matching patient serum, and control serum and assayed for basement membrane binding by means of ELISA. Results C5b-9 levels were significantly increased in NP tissue compared with uncinate tissue (UT) of patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and those with chronic rhinosinusitis without nasal polyps (CRSsNP; P <.01). Similarly, C4d levels were increased in NPs compared with UT of patients with CRSwNP, patients with CRSsNP, and control subjects (P <.05). Activated C1 levels were also increased in NP tissue compared with UT of patients with CRSsNP and control subjects (P <.05) and correlated with levels of C5a (P <.01), local immunoglobulins (especially IgM, P <.0001), and anti–double-stranded DNA IgG (P <.05). Immunofluorescence showed that C5b-9 and C4d deposition occurred linearly along the epithelial basement membrane. NP tissue extracts had significantly more anti–basement membrane antibodies than sera from patients with CRSwNP and control subjects (P <.0001). Conclusion Levels of C5b-9, C4d, and activated C1 were significantly increased locally in NP tissue. C5b-9 and C4d were almost universally deposited linearly along the basement membrane of NP tissue. Furthermore, activated C1 levels were best correlated with local immunoglobulin and C5a levels. Together, these data suggest that the classical pathway plays a major role in complement activation in patients with CRS.

Original languageEnglish (US)
Pages (from-to)89-100.e2
JournalJournal of Allergy and Clinical Immunology
Volume140
Issue number1
DOIs
StatePublished - Jul 1 2017

Fingerprint

Classical Complement Pathway
Nasal Polyps
Nose
Complement Membrane Attack Complex
Complement Activation
Basement Membrane
Antibodies
Immunoglobulins
Serum
Complement C5a
Tissue Extracts
Staphylococcal Protein A
Laminin
Autoantibodies
Fluorescent Antibody Technique
Immunoglobulin M

Keywords

  • Chronic rhinosinusitis
  • antibodies
  • basement membrane
  • classical complement pathway
  • complement

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

@article{e9425cf084ff4f74aee976b88493a13f,
title = "Classical complement pathway activation in the nasal tissue of patients with chronic rhinosinusitis",
abstract = "Background Complement plays a major role in inflammatory diseases, but its involvement and mechanisms of activation in patients with chronic rhinosinusitis (CRS) are not known. Objectives After earlier studies discovering autoantibodies in patients with CRS, we sought to investigate the nature, extent, and location of complement activation in nasal tissue of patients with CRS. Specifically, we were interested in whether antibody-mediated activation through the classical pathway was a major mechanism for complement activation in patients with CRS. Methods Nasal tissue was obtained from patients with CRS and control subjects. Tissue homogenates were analyzed for complement activation products (ELISA–C5b-9, C4d, activated C1, and C5a) and major complement-fixing antibodies (Luminex). Tissue sections were stained for C5b-9, C4d, and laminin. Antibodies were purified with protein A/G columns from nasal polyps (NP), matching patient serum, and control serum and assayed for basement membrane binding by means of ELISA. Results C5b-9 levels were significantly increased in NP tissue compared with uncinate tissue (UT) of patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and those with chronic rhinosinusitis without nasal polyps (CRSsNP; P <.01). Similarly, C4d levels were increased in NPs compared with UT of patients with CRSwNP, patients with CRSsNP, and control subjects (P <.05). Activated C1 levels were also increased in NP tissue compared with UT of patients with CRSsNP and control subjects (P <.05) and correlated with levels of C5a (P <.01), local immunoglobulins (especially IgM, P <.0001), and anti–double-stranded DNA IgG (P <.05). Immunofluorescence showed that C5b-9 and C4d deposition occurred linearly along the epithelial basement membrane. NP tissue extracts had significantly more anti–basement membrane antibodies than sera from patients with CRSwNP and control subjects (P <.0001). Conclusion Levels of C5b-9, C4d, and activated C1 were significantly increased locally in NP tissue. C5b-9 and C4d were almost universally deposited linearly along the basement membrane of NP tissue. Furthermore, activated C1 levels were best correlated with local immunoglobulin and C5a levels. Together, these data suggest that the classical pathway plays a major role in complement activation in patients with CRS.",
keywords = "Chronic rhinosinusitis, antibodies, basement membrane, classical complement pathway, complement",
author = "{Van Roey}, {Griet A.} and Vanison, {Christopher C.} and Jeffanie Wu and Huang, {Julia H.} and Suh, {Lydia A.} and Carter, {Roderick G.} and Norton, {James E.} and Stephanie Shintani-Smith and Conley, {David B.} and Welch, {Kevin C.} and Peters, {Anju T.} and Grammer, {Leslie C.} and Harris, {Kathleen E.} and Hulse, {Kathryn E.} and Atsushi Kato and Stevens, {Whitney W.} and Kern, {Robert C.} and Schleimer, {Robert P.} and Tan, {Bruce K.}",
year = "2017",
month = "7",
day = "1",
doi = "10.1016/j.jaci.2016.11.015",
language = "English (US)",
volume = "140",
pages = "89--100.e2",
journal = "Journal of Allergy and Clinical Immunology",
issn = "0091-6749",
publisher = "Mosby Inc.",
number = "1",

}

Classical complement pathway activation in the nasal tissue of patients with chronic rhinosinusitis. / Van Roey, Griet A.; Vanison, Christopher C.; Wu, Jeffanie; Huang, Julia H.; Suh, Lydia A.; Carter, Roderick G.; Norton, James E.; Shintani-Smith, Stephanie; Conley, David B.; Welch, Kevin C.; Peters, Anju T.; Grammer, Leslie C.; Harris, Kathleen E.; Hulse, Kathryn E.; Kato, Atsushi; Stevens, Whitney W.; Kern, Robert C.; Schleimer, Robert P.; Tan, Bruce K.

In: Journal of Allergy and Clinical Immunology, Vol. 140, No. 1, 01.07.2017, p. 89-100.e2.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Classical complement pathway activation in the nasal tissue of patients with chronic rhinosinusitis

AU - Van Roey, Griet A.

AU - Vanison, Christopher C.

AU - Wu, Jeffanie

AU - Huang, Julia H.

AU - Suh, Lydia A.

AU - Carter, Roderick G.

AU - Norton, James E.

AU - Shintani-Smith, Stephanie

AU - Conley, David B.

AU - Welch, Kevin C.

AU - Peters, Anju T.

AU - Grammer, Leslie C.

AU - Harris, Kathleen E.

AU - Hulse, Kathryn E.

AU - Kato, Atsushi

AU - Stevens, Whitney W.

AU - Kern, Robert C.

AU - Schleimer, Robert P.

AU - Tan, Bruce K.

PY - 2017/7/1

Y1 - 2017/7/1

N2 - Background Complement plays a major role in inflammatory diseases, but its involvement and mechanisms of activation in patients with chronic rhinosinusitis (CRS) are not known. Objectives After earlier studies discovering autoantibodies in patients with CRS, we sought to investigate the nature, extent, and location of complement activation in nasal tissue of patients with CRS. Specifically, we were interested in whether antibody-mediated activation through the classical pathway was a major mechanism for complement activation in patients with CRS. Methods Nasal tissue was obtained from patients with CRS and control subjects. Tissue homogenates were analyzed for complement activation products (ELISA–C5b-9, C4d, activated C1, and C5a) and major complement-fixing antibodies (Luminex). Tissue sections were stained for C5b-9, C4d, and laminin. Antibodies were purified with protein A/G columns from nasal polyps (NP), matching patient serum, and control serum and assayed for basement membrane binding by means of ELISA. Results C5b-9 levels were significantly increased in NP tissue compared with uncinate tissue (UT) of patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and those with chronic rhinosinusitis without nasal polyps (CRSsNP; P <.01). Similarly, C4d levels were increased in NPs compared with UT of patients with CRSwNP, patients with CRSsNP, and control subjects (P <.05). Activated C1 levels were also increased in NP tissue compared with UT of patients with CRSsNP and control subjects (P <.05) and correlated with levels of C5a (P <.01), local immunoglobulins (especially IgM, P <.0001), and anti–double-stranded DNA IgG (P <.05). Immunofluorescence showed that C5b-9 and C4d deposition occurred linearly along the epithelial basement membrane. NP tissue extracts had significantly more anti–basement membrane antibodies than sera from patients with CRSwNP and control subjects (P <.0001). Conclusion Levels of C5b-9, C4d, and activated C1 were significantly increased locally in NP tissue. C5b-9 and C4d were almost universally deposited linearly along the basement membrane of NP tissue. Furthermore, activated C1 levels were best correlated with local immunoglobulin and C5a levels. Together, these data suggest that the classical pathway plays a major role in complement activation in patients with CRS.

AB - Background Complement plays a major role in inflammatory diseases, but its involvement and mechanisms of activation in patients with chronic rhinosinusitis (CRS) are not known. Objectives After earlier studies discovering autoantibodies in patients with CRS, we sought to investigate the nature, extent, and location of complement activation in nasal tissue of patients with CRS. Specifically, we were interested in whether antibody-mediated activation through the classical pathway was a major mechanism for complement activation in patients with CRS. Methods Nasal tissue was obtained from patients with CRS and control subjects. Tissue homogenates were analyzed for complement activation products (ELISA–C5b-9, C4d, activated C1, and C5a) and major complement-fixing antibodies (Luminex). Tissue sections were stained for C5b-9, C4d, and laminin. Antibodies were purified with protein A/G columns from nasal polyps (NP), matching patient serum, and control serum and assayed for basement membrane binding by means of ELISA. Results C5b-9 levels were significantly increased in NP tissue compared with uncinate tissue (UT) of patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and those with chronic rhinosinusitis without nasal polyps (CRSsNP; P <.01). Similarly, C4d levels were increased in NPs compared with UT of patients with CRSwNP, patients with CRSsNP, and control subjects (P <.05). Activated C1 levels were also increased in NP tissue compared with UT of patients with CRSsNP and control subjects (P <.05) and correlated with levels of C5a (P <.01), local immunoglobulins (especially IgM, P <.0001), and anti–double-stranded DNA IgG (P <.05). Immunofluorescence showed that C5b-9 and C4d deposition occurred linearly along the epithelial basement membrane. NP tissue extracts had significantly more anti–basement membrane antibodies than sera from patients with CRSwNP and control subjects (P <.0001). Conclusion Levels of C5b-9, C4d, and activated C1 were significantly increased locally in NP tissue. C5b-9 and C4d were almost universally deposited linearly along the basement membrane of NP tissue. Furthermore, activated C1 levels were best correlated with local immunoglobulin and C5a levels. Together, these data suggest that the classical pathway plays a major role in complement activation in patients with CRS.

KW - Chronic rhinosinusitis

KW - antibodies

KW - basement membrane

KW - classical complement pathway

KW - complement

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