title: IgG, IgA and C3 deposits in the extra-thyroidal manifestations of autoimmune Graves' disease: their in vitro solubilization by intravenous immunoglobulin.
abstract: OBJECTIVE
  To study the involvement of antibodies in the extrathyroidal manifestations of autoimmune Graves' disease, we determined the presence of IgG, IgA and IgM antibodies and C3c in connective tissue samples from patients with Graves' disease and pretibial myxedema (PTM) or thyroid associated ophthalmopathy (TAO).
  
  
  METHODS
  Connective orbital tissue samples were obtained from 12 patients undergoing orbital decompression for TAO, and skin samples from lesions on the pretibial area were obtained in 7 patients with PTM. Sections from each tissue sample were stained with fluorescin-isothiocianate conjugated anti-human IgG, IgA, IgM and C3c and were examined by a fluorescence optical instrument. Other serial sections from each sample were incubated with human IgG solutions (concentration 6 mg/ml or 20 mg/ml), human albumin (40 mg/ml), PBS, myoglobin (40 mg/ml), or IgA (20 mg/ml), and were then processed by a standard direct immunofluorescence staining procedure.
  
  
  RESULTS
  Among the samples from TAO patients 8/12 (67%) were positive for IgG deposition, 4/9 (44%) were positive for IgA, 1/9 (11%) was positive for IgM and 4/9 (44%) were positive for C3c deposition. Orbital connective samples from 3 non-TAO patients were all negative. Among samples from PTM patients 4/7 (57%) were positive for IgG deposition, 3/ 4 (75%) were positive for IgA, 0/4 was positive for IgM and 3/7 (43%) were positive for C3c deposition. Skin samples from 5 control patients undergoing skin biopsy for non-autoimmune diseases were all negative. Incubation with human IgG (20 mg/ml) resulted in the complete disappearance of IgG and C3c deposition in all positive patients. No significant variation in IgG fluorescent staining after incubation with either 6 mg/ml of IgG solution, human albumin, PBS, myoglobin or IgA was observed.
  
  
  CONCLUSIONS
  The results of our study suggest that different classes of antibodies, mainly IgG and IgA, may be implicated in the disease process in autoimmune TAO and PTM. Activation of the complement cascade, via the classic or the alternative pathway, could take place in about 40% of these patients. IVIG in vitro may solubilize, by a specific mechanism, IgG and complement immune complex deposition in the extrathyroidal manifestations of autoimmune Grave's disease.
authors.affiliation: Institute of Clinical Medicine II, University of Pisa, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Antonelli
authors.forename: R
authors.initials: R
authors.lastname: Palla
authors.forename: L
authors.initials: L
authors.lastname: Casarosa
authors.forename: P
authors.initials: P
authors.lastname: Fallahi
authors.forename: L
authors.initials: L
authors.lastname: Baschieri
mesh_terms: D003176:Complement C3 / Q000032:analysis / Q000737:chemistry; D003238:Connective Tissue / Q000737:chemistry / Q000276:immunology; D005128:Eye Diseases / Q000276:immunology / Q000378:metabolism; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods; D006111:Graves Disease / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis / Q000737:chemistry; D016756:Immunoglobulins, Intravenous / Q000737:chemistry*; D066298:In Vitro Techniques; D007868:Leg Dermatoses / Q000276:immunology* / Q000378:metabolism; D009230:Myxedema / Q000276:immunology* / Q000378:metabolism; D012867:Skin / Q000737:chemistry / Q000276:immunology; D012871:Skin Diseases / Q000276:immunology / Q000378:metabolism; D012995:Solubility; D013967:Thyroiditis, Autoimmune / Q000276:immunology*
chemical_list: D003176:Complement C3; D007070:Immunoglobulin A; D016756:Immunoglobulins, Intravenous