PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of molmedLink to Publisher's site
 
Mol Med. 1997 February; 3(2): 145–156.
PMCID: PMC2230052

Normal human epidermal keratinocytes express in vitro specific molecular forms of (pro)filaggrin recognized by rheumatoid arthritis-associated antifilaggrin autoantibodies.

Abstract

BACKGROUND: The so-called antikeratin antibodies and the antiperinuclear factor are the most specific serological markers of rheumatoid arthritis (RA). They were recently shown to be largely the same autoantibodies and to recognize human epidermal filaggrins and profilaggrin-related proteins of buccal epithelial cells (collectively referred to as (pro)filaggrin). MATERIALS AND METHODS: To further characterize the target antigens, we investigated their expression by normal human epidermal keratinocytes cultured in differentiating conditions, using immunofluorescence and immunoblotting with RA sera and three different monoclonal antibodies to (pro)filaggrin. RESULTS: On the cornified, stratified epithelial sheets obtained in vitro, RA sera with anti(pro)filaggrin autoantibodies (AFA) produced granular staining of the stratum granulosum and diffuse staining of the stratum corneum. The antigens recognized by RA sera strictly colocalized with (pro)filaggrin in keratohyalin granules. Following sequential extraction of the proteins from the epithelial sheets, the RA sera and the three monoclonal antibodies to (pro)filaggrin, recognized a series of low-salt-soluble molecules, including a neutral/acidic isoform of filaggrin and several proteins with sizes and pI intermediates between this isoform and profilaggrin. They also recognized urea-soluble high-molecular-weight profilaggrin-related molecules. CONCLUSIONS: These results show that in vitro epidermal keratinocytes express various molecular forms of (pro) filaggrin that bear epitopes targeted by AFA of RA sera, and that some of these are absent from epidermis. Moreover, these epitopes, which are present on the keratohyalin granules of buccal epithelial cells but not on those of epidermal cells, are present on the granules of the cultured keratinocytes. This work completes the molecular characterization of the proteins targeted by AFA.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Wolfe F, Cathey MA, Roberts FK. The latex test revisited. Rheumatoid factor testing in 8,287 rheumatic disease patients. Arthritis Rheum. 1991 Aug;34(8):951–960. [PubMed]
  • Steiner G, Hartmuth K, Skriner K, Maurer-Fogy I, Sinski A, Thalmann E, Hassfeld W, Barta A, Smolen JS. Purification and partial sequencing of the nuclear autoantigen RA33 shows that it is indistinguishable from the A2 protein of the heterogeneous nuclear ribonucleoprotein complex. J Clin Invest. 1992 Sep;90(3):1061–1066. [PMC free article] [PubMed]
  • Ferris J, Cooper S, Roessner K, Hochberg M. Antibodies to denatured type II collagen in rheumatoid arthritis: negative association with IgM rheumatoid factor. J Rheumatol. 1990 Jul;17(7):880–884. [PubMed]
  • NIENHUIS RL, MANDEMA E. A NEW SERUM FACTOR IN PATIENTS WITH RHEUMATOID ARTHRITIS; THE ANTIPERINUCLEAR FACTOR. Ann Rheum Dis. 1964 Jul;23:302–305. [PMC free article] [PubMed]
  • Young BJ, Mallya RK, Leslie RD, Clark CJ, Hamblin TJ. Anti-keratin antibodies in rheumatoid arthritis. Br Med J. 1979 Jul 14;2(6182):97–99. [PMC free article] [PubMed]
  • Quismorio FP, Jr, Kaufman RL, Beardmore T, Mongan ES. Reactivity of serum antibodies to the keratin layer of rat esophagus in patients with rheumatoid arthritis. Arthritis Rheum. 1983 Apr;26(4):494–499. [PubMed]
  • Miossec P, Youinou P, Le Goff P, Moineau MP. Clinical relevance of antikeratin antibodies in rheumatoid arthritis. Clin Rheumatol. 1982 Sep;1(3):185–189. [PubMed]
  • Kirstein H, Mathiesen FK. Antikeratin antibodies in rheumatoid arthritis. Methods and clinical significance. Scand J Rheumatol. 1987;16(5):331–338. [PubMed]
  • Vincent C, Serre G, Lapeyre F, Fournié B, Ayrolles C, Fournié A, Soleilhavoup JP. High diagnostic value in rheumatoid arthritis of antibodies to the stratum corneum of rat oesophagus epithelium, so-called 'antikeratin antibodies'. Ann Rheum Dis. 1989 Sep;48(9):712–722. [PMC free article] [PubMed]
  • Sebbag M, Simon M, Vincent C, Masson-Bessière C, Girbal E, Durieux JJ, Serre G. The antiperinuclear factor and the so-called antikeratin antibodies are the same rheumatoid arthritis-specific autoantibodies. J Clin Invest. 1995 Jun;95(6):2672–2679. [PMC free article] [PubMed]
  • Simon M, Vincent C, Haftek M, Girbal E, Sebbag M, Gomès-Daudrix V, Serre G. The rheumatoid arthritis-associated autoantibodies to filaggrin label the fibrous matrix of the cornified cells but not the profilaggrin-containing keratohyalin granules in human epidermis. Clin Exp Immunol. 1995 Apr;100(1):90–98. [PubMed]
  • Serre G, Vincent C, Fournié B, Lapeyre F, Soleilhavoup JP, Fournié A. Anticorps anti-stratum corneum d'oesophage de rat, auto-anticorps anti-kératines épidermiques et anti-épiderme dans la polyarthrite rhumatoïde et différentes affections rhumatologiques. Intérêt diagnostique, aspects fondamentaux. Rev Rhum Mal Osteoartic. 1986 Nov;53(11):607–614. [PubMed]
  • Johnson GD, Carvalho A, Holborow EJ, Goddard DH, Russell G. Antiperinuclear factor and keratin antibodies in rheumatoid arthritis. Ann Rheum Dis. 1981 Jun;40(3):263–266. [PMC free article] [PubMed]
  • Youinou P, Le Goff P, Colaco CB, Thivolet J, Tater D, Viac J, Shipley M. Antikeratin antibodies in serum and synovial fluid show specificity for rheumatoid arthritis in a study of connective tissue diseases. Ann Rheum Dis. 1985 Jul;44(7):450–454. [PMC free article] [PubMed]
  • Paimela L, Gripenberg M, Kurki P, Leirisalo-Repo M. Antikeratin antibodies: diagnostic and prognostic markers for early rheumatoid arthritis. Ann Rheum Dis. 1992 Jun;51(6):743–746. [PMC free article] [PubMed]
  • Kurki P, Aho K, Palosuo T, Heliövaara M. Immunopathology of rheumatoid arthritis. Antikeratin antibodies precede the clinical disease. Arthritis Rheum. 1992 Aug;35(8):914–917. [PubMed]
  • Girbal E, Sebbag M, Gomès-Daudrix V, Simon M, Vincent C, Serre G. Characterisation of the rat oesophagus epithelium antigens defined by the so-called 'antikeratin antibodies', specific for rheumatoid arthritis. Ann Rheum Dis. 1993 Oct;52(10):749–757. [PMC free article] [PubMed]
  • Simon M, Girbal E, Sebbag M, Gomès-Daudrix V, Vincent C, Salama G, Serre G. The cytokeratin filament-aggregating protein filaggrin is the target of the so-called "antikeratin antibodies," autoantibodies specific for rheumatoid arthritis. J Clin Invest. 1993 Sep;92(3):1387–1393. [PMC free article] [PubMed]
  • Lynley AM, Dale BA. The characterization of human epidermal filaggrin. A histidine-rich, keratin filament-aggregating protein. Biochim Biophys Acta. 1983 Apr 14;744(1):28–35. [PubMed]
  • Lonsdale-Eccles JD, Haugen JA, Dale BA. A phosphorylated keratohyalin-derived precursor of epidermal stratum corneum basic protein. J Biol Chem. 1980 Mar 25;255(6):2235–2238. [PubMed]
  • Scott IR, Harding CR. Studies on the synthesis and degradation of a high molecular weight, histidine-rich phosphoprotein from mammalian epidermis. Biochim Biophys Acta. 1981 Jun 29;669(1):65–78. [PubMed]
  • Simon M, Sebbag M, Haftek M, Vincent C, Girbal-Neuhauser E, Rakotoarivony J, Sommé G, Schmitt D, Serre G. Monoclonal antibodies to human epidermal filaggrin, some not recognizing profilaggrin. J Invest Dermatol. 1995 Sep;105(3):432–437. [PubMed]
  • Serre G, Mils V, Haftek M, Vincent C, Croute F, Réano A, Ouhayoun JP, Bettinger S, Soleilhavoup JP. Identification of late differentiation antigens of human cornified epithelia, expressed in re-organized desmosomes and bound to cross-linked envelope. J Invest Dermatol. 1991 Dec;97(6):1061–1072. [PubMed]
  • Rothblat GH, Arbogast LY, Ouellette L, Howard BV. Preparation of delipidized serum protein for use in cell culture systems. In Vitro. 1976 Aug;12(8):554–557. [PubMed]
  • Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, Healey LA, Kaplan SR, Liang MH, Luthra HS, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988 Mar;31(3):315–324. [PubMed]
  • Dale BA, Gown AM, Fleckman P, Kimball JR, Resing KA. Characterization of two monoclonal antibodies to human epidermal keratohyalin: reactivity with filaggrin and related proteins. J Invest Dermatol. 1987 Mar;88(3):306–313. [PubMed]
  • Asselineau D, Dale BA, Bernard BA. Filaggrin production by cultured human epidermal keratinocytes and its regulation by retinoic acid. Differentiation. 1990 Dec;45(3):221–229. [PubMed]
  • Asselineau D, Darmon M. Retinoic acid provokes metaplasia of epithelium formed in vitro by adult human epidermal keratinocytes. Differentiation. 1995 Apr;58(4):297–306. [PubMed]
  • Kautsky MB, Fleckman P, Dale BA. Retinoic acid regulates oral epithelial differentiation by two mechanisms. J Invest Dermatol. 1995 Apr;104(4):546–553. [PubMed]
  • Hoet RM, Voorsmit RA, Van Venrooij WJ. The perinuclear factor, a rheumatoid arthritis-specific autoantigen, is not present in keratohyalin granules of cultured buccal mucosa cells. Clin Exp Immunol. 1991 Apr;84(1):59–65. [PubMed]
  • Resing KA, Johnson RS, Walsh KA. Characterization of protease processing sites during conversion of rat profilaggrin to filaggrin. Biochemistry. 1993 Sep 28;32(38):10036–10045. [PubMed]
  • Resing KA, Walsh KA, Haugen-Scofield J, Dale BA. Identification of proteolytic cleavage sites in the conversion of profilaggrin to filaggrin in mammalian epidermis. J Biol Chem. 1989 Jan 25;264(3):1837–1845. [PubMed]
  • Resing KA, al-Alawi N, Blomquist C, Fleckman P, Dale BA. Independent regulation of two cytoplasmic processing stages of the intermediate filament-associated protein filaggrin and role of Ca2+ in the second stage. J Biol Chem. 1993 Nov 25;268(33):25139–25145. [PubMed]

Articles from Molecular Medicine are provided here courtesy of The Feinstein Institute for Medical Research at North Shore LIJ