Th17-relevant cytokines vary with sera of different ANA staining patterns

https://doi.org/10.1016/j.intimp.2013.03.001Get rights and content

Highlights

  • Th17-relevant cytokines (IL-17, IL-6, IL-21, IL-22, and IL-23) varied with different ANA-positive sera.

  • TGF-β showed no difference between ANA-positive sera and healthy donors.

  • IL-17 correlated with IL-6, IL-21, IL-22, and IL-23 (p < 0.01), but not with TGF-β in ANA-positive sera.

Abstract

Antinuclear antibodies (ANA) react with components located in the cell nucleus and cytoplasm. Differing ANA staining patterns may reflect the specificity of autoantibodies in sera and indicate some autoimmune diseases specifically, to some extent. Th17-relevant cytokines have been shown to be involved in a variety of autoimmune diseases, but not consistently. In this study, we investigated whether differences in Th17-relevant cytokines exist between different ANA pattern sera. Sera of 64 ANA-positive patients (12 homogeneous, 13 speckled particle, 11 nucleolar, 15 centromere, 6 peripheral nuclear) and 16 healthy donors were analyzed for IL-17, IL-6, IL-21, IL-22, IL-23 (p19), and TGF-β, and subsequently correlations between IL-17 and IL-6, IL-21, IL-22, IL-23, and TGF-β were analyzed. Results showed that these Th17-relevant cytokines varied with different ANA-positive sera compared with healthy donors, except TGF-β. Among them, IL-21 and IL-22 were higher with all ANA-positive sera and IL-17, IL-6, and IL-23 were higher with three or more ANA staining sera. No significant difference in these cytokines was seen between the different ANA staining sera except IL-17 levels in sera of peripheral nuclear staining positive subjects were higher than nucleolar. Additionally, in ANA-positive sera, IL-17 correlated with IL-6, IL-21, IL-22, and IL-23, but not with TGF-β. Thus, we demonstrated that Th17-relevant cytokines varied with different ANA staining pattern sera, suggesting that Th17-relevant cytokines play differing roles in autoimmune diseases.

Introduction

Antinuclear antibodies (ANA), reacting with components located in the cell nucleus and cytoplasm, were first described more than 50 years ago and detecting them has been suggested as an initial diagnostic step since then [1]. ANA determinations normally use indirect immunofluorescence (IIF) and HEp-2 cells (American Type Culture Collection CCL23, Rockville, MD, USA) as a substrate. ANA staining pattern is correlated with antibody specificity to some degree; that is, each cell domain is enriched with a distinct set of antigens [2], [3], [4]. In brief, the nuclear coarse speckled pattern is strongly associated with anti-Smith (anti-Sm), Ro/SSA, La/SSB, and anti-snRNP antibodies, the nuclear homogeneous pattern with antibodies to native DNA, nucleosomes, and histones, the centromeric type with CENP-A, CENP-B, and CENP-C, the peripheral nuclear type with lamins A, B, and C, and glycoprotein 210, and the nucleolar type with PM/Scl, nucleolin, fibrillarin, RNA polymerase I, and human upstream binding factor [2], [4], [5].

Furthermore, different antigens are associated with certain specific autoimmune diseases. Indeed, the presence of anti-double-stranded native DNA (anti-dsDNA) or anti-Sm antibodies is associated with systemic lupus erythematosus (SLE) [6], [7], anti-Ro/SSA or anti-La/SSB antibodies with Sjögren's syndrome [8], anti-Jo-1 antibodies with dermatopolymyositis [9], anti-centromere or anti-topoisomerase I (anti-Scl70) antibodies with cutaneous systemic sclerosis (SS) [10], and anti-uridine-rich 1 ribonucleoprotein (U1RNP) antibodies with mixed connective tissue disease (MCTD) (Table 2) [11], [12]. Thus, different ANA-HEp-2 IIF patterns may be associated with different autoimmune diseases, to some degree.

To date, accumulated evidence show that Th17 cell, and relevant cytokines, IL-17, IL-6, IL-21, IL-22, IL-23 (p19), and TGF-β, are closely related to autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, Sjögren's syndrome, psoriasis, inflammatory bowel disease, and allergies [13], [14]. However, there is also evidence that Th17 cell-relevant cytokines do not change completely consistent in all related autoimmune diseases. Thus, whether there is any connection between Th17-relevant cytokine expression and different ANA pattern sera remains to be determined. In this study, we investigated whether there was any difference in Th17 cell-relevant cytokines in different ANA pattern sera that might provide hints for exploring their effect in these diseases in future.

Section snippets

Serum samples

Serum samples from 826 subjects were collected from patients referred to the outpatient department at Gongli Clinical Hospital, Second Military Medicine University and Shuguang Hospital, Traditional Chinese Medicine University, Shanghai, China. All samples were collected according to the sample manipulation guidelines of the hospitals. ANA IIF determination was carried out and positive sera were selected. Among those, 64 cases were confirmed as being ANA-positive with different staining

ANA patterns

Sixty-four ANA positive sera were confirmed. Among them, there were 12 with a homogeneous pattern, 13 with a speckled particle pattern, 11 with a nucleolar pattern, 15 with a centromere pattern, and six of the peripheral nuclear type. Subject files are summarized in Table 1. All sera were confirmed to show a single type of staining pattern. Sera of healthy donors were confirmed to be ANA-negative. Representative fluorescence pictures are shown in Fig. 1.

Th17-relevant cytokines varied with different ANA pattern sera

The cytokines IL-17, IL-6, IL-21, IL-22,

Discussion

Various studies have shown that Th17-relevant cytokines, typically IL-17, IL-21, IL-22, IL-23, IL-6, and TGF-β, are associated with autoimmune diseases. In this study, we demonstrated that IL-17 was higher in sera with homogeneous, speckled particle, and peripheral nuclear pattern staining, primarily associated with SLE, Sjögren's syndrome, and systemic sclerosis (SS). Additionally, IL-17 in sera with peripheral nuclear staining was higher than in sera with nucleolar staining. Many data show

Conclusions

In this study, we analyzed concentrations of IL-17, IL-6, IL-21, IL-22, IL-23, and TGF-β in sera with different ANA staining patterns. Subsequently, we analyzed the correlation between IL-17 and IL-6, IL-21, IL-22, IL-23, and TGF-β. We confirmed that the concentrations of these Th17-relevant cytokines varied with different ANA-positive pattern sera. We also confirmed that IL-17 correlated with IL-6, IL-21, IL-22, and IL-23, but not TGF-β; thus, these results suggest that Th17-relevant cytokines

Contract grant sponsor

This paper was supported by

  • 1.

    Open Research Fund Program of Shanghai Key Laboratory of Female Reproductive Endocrine-Related Diseases (No. 10dz2220300).

  • 2.

    Science and Technology Development & Innovation Fund, Pudong New Area, Shanghai (No. PKJ2012-Y28).

  • 3.

    The Chinese National postdoctoral grant" (No. 2011 M500067).

Acknowledgment

We are grateful for the technical support provided by Weiwei Liu, Huashan Hospital, Fudan University.

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  • Cited by (3)

    • Relationship between certain T helper cytokines and ANA staining: Who is the helper?

      2015, International Immunopharmacology
      Citation Excerpt :

      It isn't very clear about cytokine variation tendency in different ANA-positive sera. In our recent study, Th17-relevant cytokines were shown to be varied differently in sera with different ANA staining sera, suggesting that these cytokines may play different roles in related autoimmune diseases [2]. In this study we go further to investigate whether other T helper relevant cytokines, including IL-12(p70), IFN-γ, IL-4, and IL-10, vary in different ANA sera.

    1

    Jinhui Hu and Wei Meng contributed equally to this work.

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