ECE2014 Oral Communications Pituitary Basic (5 abstracts)
Detection of pituitary antibodies by immunofluorescence: Methodological approach and results in patients with pituitary diseases
Adriana Ricciuti 1, , Alessandra De Remigis 1 , Melissa A. Landek-Salgado 1 , Ludovica De Vincentiis 1 , Federica Guaraldi 4 , Isabella Lupi 5 , Shintaro Iwama 6 , Gary S. Wand 3 , Roberto Salvatori 3 & Patrizio Caturegli 1
1Dept of Pathology, Johns Hopkins University, Baltimore, Maryland, USA; 2Dept of Pharmacology, Gabriele D’Annunzio University, Chieti, Italy; 3Dept of Endocrinology, Johns Hopkins University, Baltimore, MD, USA; 4Dept of Endocrinolgy, University of Turin, Turin, Italy; 5Dept of Endocrinology, University of Pisa, Pisa, Italy; 6Dept of Endocrinology, Nagoya University, Nagoya, Japan.
Pituitary antibodies have been measured mainly to identify patients whose disease is caused or sustained by pituitary-specific autoimmunity. Although reported in over 100 publications, this antibody test has variable results and limited clinical utility. Our goals were to: identify major sources of variability; test them experimentally for establishing an optimized immunofluorescence protocol; assess prevalence and significance of pituitary antibodies in patients with pituitary diseases. We first evaluated the effect of pituitary gland species, section fixation, autofluorescence quenching, blockade of unwanted antibody binding, and use of purified immunoglobulins G on the performance of this antibody assay. We then measured cross-sectionally the prevalence of pituitary antibodies in 390 pituitary cases and 60 healthy controls, expressing results as present or absent and according to the (granular, diffuse, perinuclear, or mixed) staining pattern. Human pituitary was the best substrate to detect pituitary antibodies, and yielded an optimal signal-to-noise ratio when treated with Sudan black B to reduce autofluorescence. Pituitary antibodies were more common in cases (95 of 390, 24%) than controls (3 of 60, 5%, P=0.001) but did not discriminate among pituitary diseases when reported dichotomously. On the contrary, when expressed according to their cytosolic staining, a granular pattern was highly predictive of pituitary autoimmunity (P<0.0001). This study reports a comprehensive study of pituitary antibodies by immunofluorescence and provides a method and an interpretation scheme that should be useful for identifying and monitoring patients with pituitary autoimmunity.
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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