Elsevier

Immunology Letters

Volume 78, Issue 3, 1 October 2001, Pages 161-168
Immunology Letters

Humoral immune response directed against LEDGF in patients with VKH

https://doi.org/10.1016/S0165-2478(01)00243-7Get rights and content

Abstract

Vogt–Koyanagi–Harada disease is an autoimmune systemic disorder. In Vogt–Koyanagi–Harada disease, inflammatory disorders occur in multiple organs containing melanocytes, including uvea (resulting in acute bilateral panuveitis), skin (resulting in vitiligo and alopecia), central nervous system (resulting in meningitis) and inner ears (resulting in hearing loss and tinnitus). These inflammatory aspects are attributed to the destruction of melanocytes through immunological mechanisms. Studies have been carried out to elucidate the exact etiology and target autoantigen in Vogt–Koyanagi–Harada disease, but much remains to be investigated. Identification of target autoantigen is important to understand the etiology of autoimmune diseases, and for development of antigen-specific immuno-modulation therapy. To identify the target autoantigens in Vogt–Koyanagi–Harada disease, we made use of an immunoscreening of a bovine uveal cDNA expression library with serum samples obtained from patients with Vogt–Koyanagi–Harada disease. We identified an immunoreactive cDNA clone that encodes bovine lens epithelium derived growth factor. mRNA of human lens epithelium derived growth factor was determined by reverse transcription-polymerase chain reaction and it was expressed in human uvea, retina and melanocytes. Immunoglobulin G (IgG) autoantibodies were quantitated in an enzyme-linked immunosorbent assay, using recombinant human lens epithelium derived growth factor. The prevalence of IgG anti-lens epithelium derived growth factor autoantibodies in patients with Vogt–Koyanagi–Harada disease was significantly higher than that in healthy controls (66.7% versus 21.6%, P<0.001). On the other hand, the prevalence of the autoantibody in patients with panuveitis of other etiology, Behçet's disease and sarcoidosis, was almost same as that in healthy controls. These results suggest that the humoral immune response agonist lens epithelium derived growth factor is not a mere secondary phenomena caused by uveal tissue damage.

Introduction

Vogt–Koyanagi–Harada disease (VKH) is an autoimmune systemic disorder. In VKH, inflammatory disorders occur in multiple organs containing melanocytes, including uvea (resulting in acute bilateral panuveitis), skin (resulting in vitiligo and alopecia), central nervous system (resulting in meningitis), and inner ears (resulting in hearing loss and tinnitus). These inflammatory aspects are attributed to destruction of melanocytes through immunological mechanisms. In the eyes of VKH patients, T cells infiltrate diffusely throughout the choroid, whereas B lymphocytes are scattered [1]. In the skin lesion, lymphocytic infiltration was observed and infiltrating mononuclear cells primarily consisted of T lymphocytes with a smaller number of B lymphocytes [2]. Melanin-laden macrophages were specifically found in cerebrospinal fluid of patients with early stage VKH [3]. Melanoma-specific and HLA-DR-restricted T helper 1 (Th1) lymphocyte cell lines and CD4+ T cell line recognizing the tyrosinase peptide carrying the HLA-DR4 (DRB1*0405)-binding peptide motif were established from the patients [4], [5]. On the other hand, CD8+ T cell clones recognizing a MART-1 peptide in the context of HLA-A2 were also established from the patients [6]. These findings suggest that Th1-type autoreactive T cells may be involved in a development of VKH. The strong association between HLA-DR4 (DRB1*0405)-DQ4 (DQA1*0302-DQB1*0401) haplotype and the susceptibility to VKH has been noted in the Japanese and Brazilian patients; hence, HLA-linked genetic background may be related to the development of VKH [7], [8], [9], [10].

Identification of target autoantigen is important to elucidate the etiology of autoimmune diseases, and for development of antigen-specific immuno-modulation therapy. In experimental autoimmune uveitis, S-antigen and interphotoreceptor retinoid-binding protein are pathogenic autoantigens [11], [12]. However the relationship between these autoantigens and human autoimmune uveitis is uncertain. Studies have been carried out to elucidate the target autoantigen in VKH. For example, the tyrosinase family proteins were reported as the target autoantigen in VKH because peptides derived from these proteins stimulated proliferation of the lymphocytes from patients with VKH, and immunization of rats with these peptides induced autoimmunity resembled to human VHK [13], [14]. But much remains to be investigated to elucidate the exact etiology of VKH. To identify target autoantigens in autoimmune diseases, serological analysis of recombinant cDNA expression libraries (SEREX) had been done. SEREX is an immunoscreening method that makes use of procaryotically expressed cDNA libraries prepared from target organ of the autoimmune disease and sera from patients. This strategy proved effective to identify disease-related autoantigens, including type 1 diabetes, SLE, etc. [15], [16]. We used this approach to identify the target autoantigen of VKH and identified autoantigen UACA (uveal autoantigen with coiled coil domains and ankyrin repeats) [17]. But this novel UACA protein appears to be a possible target autoantigen shared by VKH, Behçet's disease (BD) and sarcoidosis that cause different types of panuveitis. In the current study, we used same approach to identify the autoantigen specific to VKH.

Section snippets

Patients

Thirty-six patients (age 19–80 years; 15 men and 21 women) with VKH were the subjects investigated in this study. All these patients underwent complete ophthalmologic and related examinations to determine the clinical diagnosis. Thirty-two patients (age 15–64 years; 20 men and 12 women) with BD complicated with panuveitis, seven patients (age 30–62 years; six men and one woman) with sympathetic ophthalmia (SO), 16 patients (age 16–67 years; two men and 14 women) with sarcoidosis complicated

Identification of immunoreactive cDNA clones

A cDNA expression library (1.5×106 pfu in primary clones) was prepared from bovine uvea, and 1.0×105 to ∼2.0×105 phage plaques were immunoscreened with each of serum obtained from four different VKH patients. A total of 6.1×105 phage plaques were screened. Twelve positive immunoreactive clones were identified in serum from one patient, JS (age 45 years, female), whereas sera from three other VKH patients did not react at detectable levels. These clones were purified, and their partial DNA

Discussion

We used the SEREX method to search for autoantigens associated with VKH. As it was impossible to obtain sufficient amounts of RNA from human uvea in Japan and because we expected there to be immunological similarities in protein population between bovine and human uvea, we used mRNA from bovine uvea to generate a cDNA expression library.

It is well established that specific autoantibodies are present in sera of patients with autoimmune diseases in which autoreactive T cells play major roles in

Acknowledgements

The authors are grateful to Dr Yuichi Obata (The Laboratory of Immunology, Aichi Cancer Center Research Institute), Dr Yutaka Kawakami (Division of Cellular Signaling, Institute for Advanced Medical Research, Keio University Graduate School of Medical Sciences), Dr Atsushi Irie (Division of Immunogenetics, Department of Neuroscience and Immunology, Kumamoto University Graduate School of Medical Sciences) and Dr Akira Kimura (Department of Ophthalmology Kumamoto University School of Medicine)

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