Detection of autoantibodies by indirect immunofluorescence and related techniques: the pathologist´s view

Main Article Content

Julius Rajčáni Lubica Korinkova Marian Bencat

Abstract

The increasing importance of autoimmunity based diseases attracts the interest of pathologists, for whom the wide acceptance of indirect fluorescent antibody (IFA) technique at autoantibody screening is relevant diagnostic approach. The IFA method has gained perfection by introducing the so called diagnostic mosaics, consisting of various tissue sections (primate and/or mammalian in origin), of Hep-2 cells, of microbeads (biochips) coated with specific purified antigen(s) and/or of transfected HEK cells expressing the target autoantigens. As result of this, the IFA technique dominates not only in the detection of anti-nuclear antibodies (ANA) and of anti-neutrophil cytoplasmic antibodies (ANCA), but is useful also in the search for serological markers in many other autoimmune disorders. Among diseases in question, the inflammatory bowel disease (GAB, PAB, ASCA), celiac disease (ARA, AEmA, AGA) and/or autoimmune hepatitis (AMA, ASMA, ABCA, anti-LKM1, SLA/LP) have emerged as important. Furthermore, antibodies to gastric parietal cells (APCA), to skin epidermal structures (anti-BP180, anti-Dsg1, anti-Dsg3), antibodies against neurons (anti-Hu, anti -Yo) and glial cells (anti-AGNA) and to several endocrine gland cells (AICA, SCA, OAB, anti-TPO, anti-TG) have recently gained attention. In addition to ANA, the detection of autoantibodies against skeletal or cardiac striated muscles (ASMA/ACMA) is of prognostic value in systemic sclerosis, Sjoegren´s syndrome, dilated cardiomyopathy, myositis and dermatomyositis. At last but not least, detection of anti-sperm antibodies (ASA) has become important due to their association with infertility occurring in the absence of mechanical obstruction of female reproductive system. We bring here an overview of reasonable applications of IFA, which, in combination with additional immunological techniques such as immunoblot (IB), allows a precise identification of several autoantibody target proteins. Though this paper stresses the advantages of IB technique, the enzyme linked immunosorbent assay (ELISA) cannot be neglected, since it is inevitable for certain diagnostic decisions (Table 1).

Article Details

How to Cite
RAJČÁNI, Julius; KORINKOVA, Lubica; BENCAT, Marian. Detection of autoantibodies by indirect immunofluorescence and related techniques: the pathologist´s view. Medical Research Archives, [S.l.], v. 2, n. 1, aug. 2015. ISSN 2375-1924. Available at: <https://esmed.org/MRA/mra/article/view/330>. Date accessed: 28 mar. 2024.
Keywords
autoantibody, autoantigen, autoimmune disease, serological diagnostic, indirect immunofluorescence, immunoblot, ELISA.
Section
Review Articles

References

References
Agmon-Levin, N. et 33 coauthors. 2013. International recommendations for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies. Ann Rheum Dis on line as doi: 10.1136/annhreumdis-2013-203863.

Alberts, B., Johnson, A, Lewis, J, Raff, M.,Roberts, K.M. and Walter, P. 2008. Sexual Reproduction: Meiosis, Germ cells and Fertilization, pp. 1269-1304 in: Molecular Biology of the Cell, 5th ed., GS Garland Science, Taylor and Francis Group, New York.

Amagai, M. 1999. Autoimmunity against desmosomal cadherins in pemphigus. J Dermatol Sci. June 20 (2): 92-102.

Andrade, EC., Chan, ERL., Raska, I. et al. 1991. Human autoantibody to the nuclear coiledbody: Immunological characterization and cDNA cloning of p80 coilin. J Exp Med 173: 1 407– 1 417.
Betterle, C. and Volpato, M. Adrenal and ovarian autoimmunity. 1998. Europ J Endocrinol 138, 16-25.

Bogdanos, DP., Baum, H. and Vergani, D. 2003. Antimitochondrial and other autoantibodies. Clin Liv Dis 7: 759–77.

Bogdanos, DP., Invernizzi, P., Ian R Mackay, IR. And Diego Vergani, D. 2008. Autoimmune liver serology: Current diagnostic and clinical challenges. World J Gastroenterol June 7; 14(21): 3374-3387.

Bradwell, A.R, Stokes, R.P, and Johnson, G.D. 1995. Atlas of Hep-2 patterns and laboratory techniques. AR Bradwell, Birmingham.

Callaghan, Jm., Khan, MA., Alderuccio, F., van Dreil, IF., Gleeson, PA., and Toth, BH. 1993. Alpha and beta subunits of the gastric H+/K+ATPase are concordantly targeted by parietal cell autoantibodies associated with autoimmune gastritis. Autoimmunity 16, 2896-295.

Calvani, M,. Jr., Parisi, G., Miotti, AM., Alessandri, C., Notarnicola, MA. 1992. Anti-endomysium antibodies: a new marker for the diagnosis and treatment of celiac disease. Pediatr Med Chir Jan-Feb;14(1): 33-6.

Coons, A.H., and Kaplan, M.H. 1950. J exp Med 91:1-10.

Csernok, E., Ahlquist, D., Ullrich, S., Gross, WL. 2002. A critical evaluation of commercial immunoassays for antineutrophil cytoplasmic antibodies directed against proteinase 3 and myeloperoxidase in Wegener’s granulomatosis and microscopic polyangiitis. Rheumatology 41: 1313-1317.

Dalakas, MC. 2011. Review: An update on inflammatory and autoimmune myopathies Neuropathol Appl Neurobiol Apr; 37(3): 226-42.

Dalmau, J., Lancaster, E., Martinez-Hernandez, E. et al. 2011. Clinical experience and laboratory nvestigations in patients with anti-NMDAR encephalitis. Lancet Neurol 10: 63-74.

Damoiseaux, J., van Rijsingen, M., Warnemunde, N. et al. 2012. Autoantibody detection in bullous pemphigoid: Clinical evaluation of the Europlus Dermatology Mosaic. J Immunol Meth 682, 76-80.

Daneshpazhooh, M., Chams-Davatchi, C., Khamesipour, A., Mansoori, P., Taheri, A., Firooz, A., Mortazavi, H., Esmaili, N., Dowlati,Y. 2007. Desmoglein 1 and 3 enzyme-linked immunosorbent assay in Iranian patients with pemphigus vulgaris: correlation with phenotype, severity, and disease activity. J Eur Acad Dermatol Venerol Nov; 21(10):1319-1324.

Darnell, RB. and Posner, JB. 2006. Paraneoplastic syndromes affecting the nervous system. Semin Oncol 33: 270-296.

Davis, MK., Andres, JM., Jolley, CD., Novak, DA., Haafiz, AB., González-Peralta, RP. 2007. Antibodies to Escherichia coli outer membrane porin C in the absence of anti-Saccharomyces cerevisiae antibodies and anti-neutrophil cytoplasmic antibodies are an unreliable marker of Crohn disease and ulcerative colitis. J Pediatr Gastroenterol Nutr Oct;45(4):409-13.

Davis, MK., Andres, JM., Jolley, CD., Novak, DA., Haafiz, AB., González-Peralta, RP. 2007. Antibodies to Escherichia coli outer membrane porin C in the absence of anti-Saccharomyces cerevisiae antibodies and anti-neutrophil cytoplasmic antibodies are an unreliable marker of Crohn disease and ulcerative colitis. J Pediatr Gastroenterol Nutr Oct; 45(4):409-13.

Dähnrich, C., Pares, A., Caballeria, L. et al. 2009. Detecting primary biliary cirrhosis-specific antimitochondrial antibodies. Clin Chem 55, 978-985.

Dellavance, A., Leser, PG., and Andrade, LE. 2007. Relevance of the immunofluorescence pattern for interpretation of the ANA test: the case of the dense fine speckled pattern. Rev Assoc Med Brasil 53: 439–445.

Dick, T., Mierau, R., Bartz-Bazzanella, P. et al. 2002. Coexistence of antitopoisomerase I and anticentromere antibodies in patients with systemic sclerosis. Ann Rheum Dis 61:121–127.

Dieterich, W., Ehnis, T., Bauer, M., Donner, P., Volta, U., Riecken, EO. and Schuppan, D. 1997. Identification of tissue transglutaminase as the autoantigen of celiac disease. Nat Med 3: 797-801.

Eber, T., Chapman, J., and Shoenfeld, Y. 2005. Anti-ribosomal P-protein and its role in psychiatric manifestations of systemic lupus erythematosus: myth or reality? Lupus 14:571-575.

Feltkamp, T.E. 1996. Antinuclear antibody determination in a routine laboratory. Ann Rheum Dis 55 : 723-7.

Fernando, M.M.A., Stevens, C.R., Walsh, C.R. et al. 2008. Defining the role of the MHC in autoimmunity: a review and pooled analysis. PLOS Genetics 4: 1–9.

Focarelli, R., Giuffrida, A., Capparelli, S., Scibona, M., Menchini-Fabris, F., Francavilla, F., Francavilla, S., Giovampaola, CD., and Rosati, F. 1998. Specific localization in the equatorial region of gp20, a 20 kDa sialylglycoprotein of the capacitated human spermatozoon acquired during epididymal transit which is necessary to penetrate zona-free hamster eggs. Molecular Human Reproduction vol.4 no.2 pp. 119–125.


Fujinami, RS., von Herrath, MG., Christen, U., and Whitton, JL. 2006. Molecular mimicry, bystander activation, or viral persistence: infections and autoimmune disease. Clin Microbiol Rev. 19(1), 80–94.

Geffriaud-Ricouard, C., Noël, LH., Chauveau, D., Houhou, S., Grünfeld, JP., and Lesavre, P. 1993. Clinical spectrum associated with ANCA of defined antigen specificities in 98 selected patients. Clin Nephrol 39: 125-36.

Gendron, M. 2001. Heterogeneity of human Ro ribonucleoprotein (RNPs). Nuclear retention of Ro RNPs containing the human hY5 RNA in human and mouse cells. Clin Exp Immunol 125: 162–158.

Gentile, F., Comte, M., and Formisano, S. 2004. Thyroglobulin as an autoantigen: What can we learn about immunopathogenicity from the correlation of antigenic properties with proteins structure? Immunology 112:13-25.

Geppert, A., Zeromski, J., Szczech, J. et al. 1999. Immunofluorescent analysis of antibodies against neurons in the case of paraneoplastic syndrome. Folia Neuropathol 37, 199-202.

Gerli, R., and Caponi, L. 2005. Antiribosomal P protein antibodies. Autoimmunity 38: 85–92.

Ghosh, P., Dwivedi, S., Naik, S. et al. 2007. Antinuclear antibodies by indirect immunofluorescence: optimumm screening dilution by diagnosis of systemic lupus erythematosus. Indian J Med Res 126, 34–38.

Goldman, M. Fluorescent antibody methods. Acad Press New York, 1968, 303 p.

Gomez, AM., Van Den Broeck, J., Vrolix, K., Janssen, SP., Lemmens, MA., Van Der Esch, E., Duimel, H., and Frederik, P. 2010. Antibody effector mechanisms in myasthenia gravis-pathogenesis at the neuromuscular junction. Autoimmunity 43 (5-6): 353-70.

Gunawardena, H., Betteridge, ZE., and McHug, NJ. 2009. Myositis-specific autoantibodies: their clinical and pathogenic significance in disease expression. Rheumatology 48:607–612 doi:10.1093.

Gutierez-Ramos, R., Gonzáles-Diaz, V., Pacheco-Tovar, MG. et al. 2008. A dermatomyositis and scleroderma overlap syndrome with remarkable high titer of anti-exosome antibody. Rheumatism 60: 296-300.

Hamaguchi, V. 2010. Autoantibody profiles in systemic sclerosis. J Dermatol 37, 42-53.

Hashimoto, T. 2003. Recent advances in the study of the pathophysiology of pemphigus. Arch Dermatol Res Apr; 295 Suppl 1:S2-11.

Hawkins, BR., McDonald, BR., and Dawkins, RL. 1977. Characterization of immunofluorescent heterophile antibodies which may be confused by autoantibodies. J clin Pathol 30: 299-307.

Henault, J., Tremblay, M., Clement, I., Raymond, Y., and Senecal, JC. 2004. Direct Binding of Anti–DNA Topoisomerase I Autoantibodies to the Cell Surface of Fibroblasts in Patients with Systemic Sclerosis. Arthr. Rheumat. 50, 3265-3274.


Hoffman, I.E.A., Peene, I., Veys, E.M., and De Keyser, F. 2002. Detection of Specific Antinuclear Reactivities in Patients with Negative Anti-nuclear Antibody Immunofluorescence Screening. Tests Clinical Chemistry 48 (12), 2171–2176.

Chang, S., Yang Liu, Moon, LM. et al. 2009. Anti-nuclear antibody reactivity in lupus may be partly hard-wired into the primary B-cell repertoir. Molecular Immunology 46: 3420–3426.

Chorzelski, TP., Sulej, J., Tchorzewska, H., Jablonska, S., Beutner, EH. and Kumar V. 1983. IgA class endomysium antibodies in dermatitis herpetiformis and coeliac disease. Ann NY Acad Sci 420: 325-334.

Ito, M., Haruma, K., Kaya, S. et al. 2002. Role of parietal cell autoantibody in Helicobacter pylori associated atrophic gastritis: evaluation in a country of high prevalence of atrophic gastritis. Scand J Gastroenterol 37, 287-293.
Jennings, J.S.R., and Howdle, PD. 2003. New Developments in Celiac Disease. Current Opinion in Gastroenterology 19, 112-124.

Jordon, RE., Kawana, S., and Fritz, KA. 1985. Immunopathologic mechanisms in pemphigus and bullous pemphigoid. J Invest Dermatol 85, (1 Suppl):72s-78s.

Kallenberg, CG., Mulder, AH., and Tervaert, JW. 1992. Antineutrophil cytoplasmic antibodies: a still-growing class of autoantibodies in inflammatory disorders. Am J Med. 93, 675-82.

Kamradt, T., and Mitchison, NA. 2001. Tolerance and autoimmunity. N Eng J Med 344, 655-664.

Kaplan, MM., and Gershwin, ME. 2005. Primary biliary cirrhosis. N Engl J Med 353: 1261-1273.

Kattah, NH., Kattah, MG., and Utz, PJ. 2010. The U1-snRNP complex: structural properties relating to autoimmune pathogenesis in rheumatic diseases. Immunol Rev January 233(1): 126–145. doi:10.1111/j.0105-2896.2009.00863.x.

Kavanaugh, A., Tomar, R., Reveille, J., Solomon, DH., and Homburger, HA. 2000. Guidelines for clinical use of the antinuclear antibody test and tests for specific autoantibodies to nuclear antigens. Arch Pathol Lab Med 124:71–81.

Kayaa, Z., Katusa, HA., and Roseb, NR. 2010. Cardiac Troponins and Autoimmunity: their role in the pathogenesis of myocarditis and of heart failure. Clin Immunol. 134: 80 -86.

Keech, CI., Howarth, S., Caotes, T. et al. 1996. Rapid sensitive detection of anti-Ro (SS-A) antibodies by indirect immmunofluorescence of 60kDa Ro Hep-2 transfectants. Pathology 28, 54–57.

Keren, DF. 2002. Antinuclear antibody testing. Clin Lab Med 22, 447–474.

Khanh, T. Ho. and John, D. and Reveille. 2003. The clinical relevance of autoantibodies in scleroderma. Arthritis Res Ther 5:80-93 (DOI 10.1186/ar628).

Kim, E.,Yamashita, M., Kimura, M., Honda, A., Kashiwabara, S-I., and Baba, T. 2008. Sperm penetration through cumulus mass and zona pellucida Int. J. Dev. Biol 52: 677-682.

Kivity, S., Giburd, B., Agmon-Levin, N. et al. 2012. A novel authomated indirect immunofluorescence autoantibody evaluation. Clin Rheumatol 31: 503-509.

Klein, R., Marx, A., Ströbel, P., Schalke, B., Nix, W., and Willcox, N. 2013. Autoimmune associations and autoantibody screening show focused recognition in patient subgroups with generalized myasthenia gravis. Hum Immunol Sep 74(9):1184-93.

Klotz, M., Blaes, F.J., Scheper, T., Meyer, W. et al. 2002. Differentiation of autoantibodies in patients with paraneoplastic neurological syndrome. Autoimmunity Rev 1:51-56.

Koenig, M., Frityler, M.J., Targiff, I.N. et al. 2007. Heterogeneity of autoantibodies in 100 patients with autoimmune myositis: insight into clinical features and outcomes. Arthririts Res Ther 9, R78.

Kuhn, G., Jarzabek-Chorzelska, M., Blaszczyk, M. et al. 1989. The Scl-70 antibody and its clinical significance. Dermatol. Monatschr 175: 133–147.

Kuna, A.T. 2013. Serological markers of inflammatory bowel disease. Biochemia Medica 3: 23-42.

La Marca, A., Brozzetti, A., Sighinolfi, G., Marzotti, S., Volpe, A., and Falorni, A. 2010. Primary ovarian insufficiency: autoimmune causes. Curr Opin Obstet Gynecol Aug;22(4):277-282.

Lancaster, E, Lai, M., Peng, X. et al. Antibodies to the GABA(B) receptor in limbic encephalitis with seazures: case series and characterization of the antigen. Lancet Neurol 9, 67-76, 2010.

Lassoued, K., Guilly, M.N., Danon, F. et al. 1988. Antinuclear antibodies specific for lamins. Characterization and clinical significance. Ann Intern Med 108: 929–833.

Lauer, B., Schannwell, M., Kühl, U., Strauer, B.E. and Schultheiss HP. 2000. Antimyosin autoantibodies are associated with deterioration of systolic and diastolic left ventricular function in patients with chronic myocarditis. J Am Coll Cardiol 35(1):11-8.

Leffler, D. and Kelly, CP. 2006. Update on the evaluation and diagnosis of celiac disease. Curr Opin Allergy Clin Immunol 6:191–196.

Lin, M.S., Mascaró, J.M. Jr, Liu, Z, España, A., and Diaz, L.A. 1997. The desmosome and hemidesmosome in cutaneous autoimmunity. Clin Exp Immunol Jan 107 Suppl 1:9-15.

Lindor, KD., Gershwin, ME., Poupon, R., Kaplan, M., Bergasa, N.V., and Heathcote, E.J. 2009. Primary biliary cirrhosis. Hepatology 50 : 291-308.

Lorusso, L, Hart, I.K., Giometto, B. et al. 2004. Immunologicl features of neurological paraneoplastic syndromes. Int J Immunopathol Pharmacol 17: 135-144.

Love, L.A., Leff, R.L., Fraser, D.D., Targoff, I.N., Dalakas, M., Plotz, P.H., and Miller, F.W. 1991. A new approach to the classification of idiopathic inflammatory myopathy: myositis-specific autoantibodies define useful homogeneous patient groups. Medicine (Baltimore) Nov;70(6):360-74.

Maga, G., and Hubscher, U. 2003. Proliferating cell nuclear antigen (PCNA): a dancer with many partners. J Cell Sci 116, 3 051–3 060.

Mahler, M., and Raijmakers, R. 2007. Novel aspects of autoantibodies to the PM/Scl complex: clinical, genetic and diagnostic insights. Autoimmun Rev 8: 432–437.

Mahler, M., Kassenbrock, K., Szmyrka, M. et al. 2006. International multicenter evaluation of autoantibodies to ribosomal P proteins. Clin Vaccine Immunol 13: 77–83.

Mahler, M., Ngo, J.T., Schulte-Pelkum, J. et al. Limited reliability of the indirect immunofluorescence technique for the detection of anti-Rib-P antibodies. Arthritis Research & Therapy 2008, 10:R131 (doi:10.1186/ar2548).

Maiuri L, Ciacci C, Ricciardelli I, Vacca L, Raia V, Rispo A, Griffin M, Issekutz T, Quaratino S, and Londei M. 2005. Unexpected role of surface transglutaminase type II in celiac disease. Gastroenterology .Nov;129(5):1400-13.

Mascart-Lemone, F., Lambrechts, A. Serology of celiac disease: early diagnosis and therapeutic impact. Acta Gastroenterol Belg 58(5-6), 388-96, 1995.

Masumoto, H., Masukata, H., Muro, Y. et al. 1989. A human centromere antigen (CENP-B) interacts with a short specific sequence in alphoid DNA, a human centromeric satellite. J Cell Biol 109, 1 963–1 973.

McFarlane, IG., McFarlane, BM., Major, GN. et al. 1984. Identification of the asialoglycoprotein receptor (hepatic lectin) as a component of liver specific membrane. Clin Exp Immunol 55: 347-355.

Meriggioli, M.N., and Sanders, D.B. 2012. Muscle autoantibodies in myasthenia gravis: beyond diagnosis? Expert Rev Clin Immunol 8, 427-438.

Miglionni, P., Baldini, C., Rocci, V., and Bombaridien, S. 2005. Anti-Sm and anti-RNP antibodies. Autoimmunity 38, 47–54.

Mizutani, T., Shinoda, M., Tanaka, Y., Kuno, T., Hattori, A., Usui, T., Kuno, N., and Osaka, T. 2005. Autoantibodies against CYP2D6 and other drug-metabolizing enzymes in autoimmune hepatitis type 2. Drug Metab Rev 37(1):235-52.

Motomura, M., and Narita M.T. 2013. Autoantibodies in myasthenia gravis. Brain Nerve Apr;65(4):433-9.

Muratori, P., Muratori, L., Cassani, F. et al. 2002. Anti-multiple nuclear dots (anti-MND) and anti-Sp100 antibodies in hepatic and rhgeumatological disorders. Clin Exp Immunol 127: 172–175.

Nairn, R.C. 1969. Fluorescent protein tracing. ES Livingstone Ltd, IIIrd ed., Edinburgh-London, 503 pp.

Nakamura, R.M.I., Matsutani, M., and Barry, M. 2003. Advances in clinical laboratory tests for inflammatory bowel disease. Clin Chim Acta Sep;335(1-2):9-20.

Nandiwada, S.L., and Tebo, A.E. 2013.Testing for antireticulin antibodies in patients with celiac disease is obsolete: a review of recommendations for serologic screening and the literature. Clin Vaccine Immunol 20(4), 447-450.

Naz, R.K. 1999. Involvement of protein serine and threonine phosphorylaton in human sperm capacitation. Biol of Reproduction 60: 142-149.

Naz, R.K., Zhu, X.,and Kadam, A.L. 2000. Identification of Human Sperm Peptide Sequence Involved in Egg Binding for Immunocontraception. Biol of Reproduction 62: 318–324.

Nishikai, M., and Reichlin, M. 1980. Heterogenity of precipitating antibodies in polymyositis and dermatomyositis: characterization of the Jo-1 antibody system. Arthritis Rheum 23: 881–888.

Nožička, Z. 1991. Patologico-anatomic contribution to antibody detection in immunopathologic diseases. Sb. Ved Pr Lek Fak Karlovy Univerzity v Hradci Králové 34S:1-169.

Oddis, C.V., Okano, Y., Rudert, W.A. et al. 1992. Serum autoantibody to the nucleolar antigen PM-Scl. Clinical and immunogenic associations. Arthririts Rheum 35: 1 211–1 217.

Orgiazzi. J. 2000. Anti-TSH receptor autoantibodies in clinical practice. Endocrinol Met Clin. N. Amer 29: 339-355.
Ozaki, S. 2007. ANCA-associated vasculitis: diagnostic and therapeutic strategy. Allergol Int. 56: 87-96.

Pan Fan, Yu Hong, Dang EV et al. 2009. Eos mediates Foxp3-dependent gene silencing in regulatory T cells. Science. August 28; 325(5944): 1142–1146. doi:10.1126/science. 1176077.

Park, S.G., Mathur, R., Long, M. et al. 2010. T regulatory cells maintain intestinal homeostasis by suppressing γδ T cells. Immunity November 24; 33(5): 791–803.

Peene, I., Meheus, L., and Veyas , E.M. 2001. Detection and identification of antinuclear antibodies (ANA) in a large and consecutive cohort of serum samples referred to ANA testing. Ann Rheum Dis 60: 1191-1136.

Peterson, P., Uibo, J., Peranen, J., and Krohn, K. 1997. Immunoprecipitaion of steroidic enzyme autoantigens with autoimmune polyglandular syndrome type 1 sera: furher evidence for independent immunity to P450c17 and P450c21. Clin exp Immunol 107, 335-340.

Pintér, M., Pintérová, K.M., Drahosová, M., Rejchrt, S., Douda, T., Tachecí, I., Kopácová, M., and Bures, J. 2007. The role of antibodies ANCA, ASCA, ABBA in the diagnostic of idiopathic bowel inflammation. Cas Lek Cesk 146(11):863-7 (in Czech).

Pollock, W., Ban-Hock Toh. 1999. Routine immunofluorescence detection of Ro/SS-A autoantibody using Hep-2 cells transfected with human 60 kDa Ro/SS-A. J clin Pathol 52: 684–687.

Pruijn, G.J., Simons, F.H., and Venroij, W.J. 1997. Intracellular localization and nucleocytoplasmic transport of Ro-RNP components. Eur J Cell Biol 74: 123–132.

Radice, A., and Sinico, R.A. 2005. Antineutrophil cytoplasmic antibodies (ANCA). Autoimmunity 38 :103-110.
Rayavarapu, S., Coley, W., Kinder, T.B., and Nagaraju, K. 2013. Idiopathic inflammatory myopathies: pathogenic mechanisms of muscle weakness. Skelet Muscle 7 (1), 133- 138.

Reuter R, and Luhrman R. 1986. Immunization of mice with purified U1 small nuclear ribonucleoprotein(RNP) induces a pattern of antibody specificities characteristic of the anti-Sm and anti-RNP autoimmune response of patients with lupus erythematosus as measured by monoclonal antibodies. Proc Natl Acad Sci USA 83: 8 689–8 693.

Rigon, A., Soda, P., Zennaro, D., Iannello, G., and Afeltra, A. 2007. Indirect Immunofluorescence in Autoimmune Diseases: Assessment of Digital Images for Diagnostic Purpose. Cytometry Part B (Clinical Cytometry) 72B:472–477.

Roberts, W.K., and Darnell, R.B. 2004. Neuroimmunology of the paraneoplastic neurological degenerations. Curr Opinion Immunol 16, 616-222.

Roberts-Thompson, P.J., Nikoloutsopoulos, T., COX, S. et al. 2003. Antinuclear antibody testing in a regional immunopathology laboratory. Immunology Cell Biology 81, 409–412.

Rojana-Udomsart, A., Mitrpant, C., Bundell, C., Price, L., Luo, Y.B., Fabian, V., Wilton, S.D, Hollingsworth, P., and Mastaglia, F.L. 2013. Complement-mediated muscle cell lysis: a possible mechanism of myonecrosis in anti-SRP associated necrotizing myopathy (ASANM). J Neuroimmunol Nov 15; 264(1-2):65-70.

Rønningen, K.S., Keiding, N., Green, A. 2001. EURODIAB ACE Study Group. Europe and Diabetes. Correlations between the incidences of childhood-onset type 1 diabetes in Europe and HLA genotypes. Diabetologia Oct; 44 Suppl 3:B51–9.

Roslyn, M.B., Blatt, N.B., and Gary, D. 1997. Anti-DNA autoantibodies: the other DNA-binding proteins. Bioorganic Med Chemistry 5, 467–472.

Rubio-Tapia, A., and Murray, J. 2010. Celiac Disease. Curr Opin Gastroenterol 26(2): 116–122.

Rujner, J., Socha, J., Barra, E., Gregorek, H., Madaliński, K., Woźniewicz, B., Giera, B. Serum and salivary antigliadin antibodies and serum IgA anti-endomysium antibodies as a screening test for coeliac disease. Acta Paediatr. 85(7), 814-817, 1996.

Rutjes, S.A., Vres Egberts, W.T.M., Jongen, P. et al. 1997. Anti-Ro-52 antibodies frequently occur with anti-Jo-1 antibodies in sera from patients with idiopathic inflammatory myopathy. Clin Exp Immun 109: 32–40.

Sakaguchi, S. 2005. Naturally occurring focxp3-expressing CD25+/CD4+ regulatory T cells in immunological tolerance to self and non self. Natl Immunol 9: 345-352.

Satoh, M., Chan, E.K.L., Sobel, J.S. et al. 2007. Clinical implication of autoantibodies in patients with systemic rheumatic diseases. Exp Rev Clin Immunol 3: 721–738.

Saxena, R., Johansson, C., Byren, P., and Wieslander, J. 1992. Autoimmunity and glomerulonephritis. Postgrad Med J 68, 242-250.

Servais, G., Karmali, R., Guillaume, M.P. et al. 2009. Anti-DNA antibodies are not restricted to a specific pattern of fluorescence on Hep-2 cells. Clin Chem Lab Med 47: 543–549.

Sharp, G.C., Irvin, W.S., Tan, E.M. et al. 1972. Mixed connective tissue disease and apparently distinct rheumatic disease associated with a specific antibody to an extractable antigen. Am J Med 52: 148-159.

Shetty, J., Klotz, KL., Wolkovicz, M.J. et al. 2007. Radial spoke protein 44 (human meichroacidin) is an axonemal alloantigen of sperm and cilia. Gene 396: 93-107.

Shokat, K.M., and Goodnow, C.C. 1995. Antigen-induced B-cell death and elimination during germinal-centre immune responses. Nature 375 (6529), 334–338.

Schlumberger, W., Hornig, N., Lange, S., Probst, Ch., Komorowski, L., Fechner, K., Dähnrich, C., and Stöcker, W. 2014. Differential diagnosis of membranous nephropathy with autoantibodies to phospholipase A2 receptor 1. Autoimmunity Reviews 13: 108–113.

Schulte-Pelkum, J., Fritzler, M., and Mahler, M. 2009. Latest update on the Ro/SS-A autoantibody system. Autoimmun Rev 7: 832–837.

Simpson, N., Gatenby, P.A,, Wilson, A. et al. 2010. Expansion of Circulating T Cells Resembling Follicular Helper T Cells Is a Fixed Phenotype That Identifies a Subset of Severe Systemic Lupus Erythematosus. Arthritis and Rheumatism 62, (1), January: 234–244. DOI 10.1002/art.25032.

Sordet, C., Goetz, J., and Sibilia, J. 2006. Contribution of autoantibodies to the diagnosis and nosology of inflammatory muscle disease. Joint Bone Spine Dec;73(6):646-54.

Stechemesser, E., Klein, R., Berg, P.A. 1993. Characterization and clinical relevance of liver-pancreas autoantibodies in autoimmune hepatitis. Hepatology 18: 1-19.

Stoecker, W., Otte, M., Ulrich et al. 1984. Antibodies against exocrine pancreas and goblet cells in the diagnostic of Crohn´s disease and ulcerative colitis. D. Med. Wschr 109: 1963-1969 (in German).

Stoecker, W., Teegen, B. Probst C et al. 2009. CUZD and GP2 are the exocrine pancreas autoantigens in Crohn´s disease, pp.463-473. In: Conrad K et al. (ed.) From pathogenesis to Therapy of Autoimmune Diseases: Autoantigens, Autoantibodies, Auroimmunity. Pabst Sci Publ.

Suri, A. Sperm specific proteins-potential candidate molecules for fertility control. Review. Reproductive Biology and Endocrinology . 2:10-16, 2004.

Touloupi, E., Routsias, J.G., Tzioufas, A.G. 2005. Cross-recognition between histones and La/SSB may account for anti-DNA reactivity in SLE patients. Clin Exp Immunol 142: 172–179.

Tsiakalou, V., Tsangaridou, E., Polioudaki, H. et al. 2006. Optimized detection of circulating anti-nuclear envelope autoantibodies by immunofluorescence. BMC Immunology 7:20 doi:10.1186/1471-2172-7-20.


Tsui, F.W., and Siminovits, K.A. 1991. Anti-Jo-1 antibodies and immunopathogenesis of autoimmune myositis. Int Rev Immunol 7: 226-235.

VanGelder, H., and Charles-Schoeman, C. The heart in inflammatory myopathies. Rheum Dis Clin North Am 40(1), 1-10, 2014.

Vergani, D., Alvarez, F., Bianchi, F.B. et al. 2004. Liver autoimmune serology: a consensus statement from the committee for autoimmune serology of International Autoimmune hepatitis Group. J Hepatol 41: 677-683.

Verstegen, G., Duyk, M.C., Meeus, P. et al. 2009. Detection and identification of antinuclear antibodies (ANA) in a large commnity hospital. Acta Clin Belg 64: 317–323.

Villalta, D., Bizzaro, N., Da Re, M., Tozzoli, R., Komorowski, L., Tonutti, E. 2008. Diagnostic accuracy of four different immunological methods for the detection of ani-F actin autoantibodies in type I autoimmune hepatitis and other liver disorders. Autoimmunity 41:105-110.

Wayaku, T., Hasegawa, M., Kaji, K. et al. 2007. Antigen specificity of antihistone antibodies in connective tissue disease patients with anti-U1RNP antibodies. Rheumatol Int 28: 119–119.

Weithauser, A., Bobbert, P., Antoniak, S., Böhm, A., Rauch, B.H., Klingel, K., Savvatis, K., Kroemer, H.K., Tschope, C., Stroux, A., Zeichhardt, H., Poller, W., Mackman, N., Schultheiss, H.P., and Rauch, U. 2013. Protease-activated receptor-2 regulates the innate immune response to viral infection in a coxsackievirus B3-induced myocarditis. J Am Coll Cardiol 62(19): 1737-45.

Welt, C.K.L., Falorni, A., Taylor, A.E., Martin, K.A., Hall, J.E. 2005. Selective theca cell dysfunction in autoimmune oophoritis results in multifollicular development, decreased estradiol, and elevated inhibin B levels. J Clin Endocrinol Metab May;90(5):3069-3076.

Wiik, A.S., Hoyer-Madsen, M., Forslid, J., Charles, P., and Meyerovits, J. 2010. Antinuclear antibodies: A contemporary nomenclature using Hep-2 cells. J Autoimmun 35: 276–290.

Winter, W.E., Schatz, D.A. 2011. Autoimmune markers in diabetes. Clin Chem Feb; 57(2):168–175.