Detection of pituitary antibodies by immunofluorescence: Approach and results in patients with pituitary diseases

Adriana Ricciuti, Alessandra De Remigis, Melissa A. Landek-Salgado, Ludovica De Vincentiis, Federica Guaraldi, Isabella Lupi, Shintaro Iwama, Gary S Wand, Roberto Salvatori, Patrizio P Caturegli

Research output: Contribution to journalArticle

Abstract

Context: 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, they have yielded variable results and are thus considered of limited clinical utility. Objectives: Our objectives were to analyze all publications reporting pituitary antibodies by immunofluorescence for detecting the major sources of variability, to experimentally test these sources and devise an optimized immunofluorescence protocol, and to assess prevalence and significance of pituitary antibodies in patients with pituitary diseases. Study Design and Outcome Measures: We first evaluated the effect of pituitary gland species, section fixation, autofluorescence quenching, blockade of unwanted antibody binding, and use of purified IgG on the performance of this antibody assay. We then measured crosssectionally 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. Results: 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 .001) but did not discriminate among pituitary diseases when reported dichotomously. However, when expressed according to their cytosolic staining, a granular pattern was highly predictive of pituitary autoimmunity (P <.0001). Conclusion: We report a comprehensive study of pituitary antibodies by immunofluorescence and provide a method and an interpretation scheme that should be useful for identifying and monitoring patients with pituitary autoimmunity.

Original languageEnglish (US)
Pages (from-to)1758-1766
Number of pages9
JournalJournal of Clinical Endocrinology and Metabolism
Volume99
Issue number5
DOIs
StatePublished - 2014

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Pituitary Diseases
Fluorescent Antibody Technique
Antibodies
Autoimmunity
Publications
Staining and Labeling
Patient monitoring
Physiologic Monitoring
Signal-To-Noise Ratio
Pituitary Gland
Quenching
Assays
Signal to noise ratio
Immunoglobulin G
Outcome Assessment (Health Care)

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Endocrinology
  • Biochemistry, medical
  • Endocrinology, Diabetes and Metabolism

Cite this

Detection of pituitary antibodies by immunofluorescence : Approach and results in patients with pituitary diseases. / Ricciuti, Adriana; De Remigis, Alessandra; Landek-Salgado, Melissa A.; De Vincentiis, Ludovica; Guaraldi, Federica; Lupi, Isabella; Iwama, Shintaro; Wand, Gary S; Salvatori, Roberto; Caturegli, Patrizio P.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 99, No. 5, 2014, p. 1758-1766.

Research output: Contribution to journalArticle

Ricciuti, Adriana ; De Remigis, Alessandra ; Landek-Salgado, Melissa A. ; De Vincentiis, Ludovica ; Guaraldi, Federica ; Lupi, Isabella ; Iwama, Shintaro ; Wand, Gary S ; Salvatori, Roberto ; Caturegli, Patrizio P. / Detection of pituitary antibodies by immunofluorescence : Approach and results in patients with pituitary diseases. In: Journal of Clinical Endocrinology and Metabolism. 2014 ; Vol. 99, No. 5. pp. 1758-1766.
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abstract = "Context: 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, they have yielded variable results and are thus considered of limited clinical utility. Objectives: Our objectives were to analyze all publications reporting pituitary antibodies by immunofluorescence for detecting the major sources of variability, to experimentally test these sources and devise an optimized immunofluorescence protocol, and to assess prevalence and significance of pituitary antibodies in patients with pituitary diseases. Study Design and Outcome Measures: We first evaluated the effect of pituitary gland species, section fixation, autofluorescence quenching, blockade of unwanted antibody binding, and use of purified IgG on the performance of this antibody assay. We then measured crosssectionally 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. Results: 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 .001) but did not discriminate among pituitary diseases when reported dichotomously. However, when expressed according to their cytosolic staining, a granular pattern was highly predictive of pituitary autoimmunity (P <.0001). Conclusion: We report a comprehensive study of pituitary antibodies by immunofluorescence and provide a method and an interpretation scheme that should be useful for identifying and monitoring patients with pituitary autoimmunity.",
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AU - De Remigis, Alessandra

AU - Landek-Salgado, Melissa A.

AU - De Vincentiis, Ludovica

AU - Guaraldi, Federica

AU - Lupi, Isabella

AU - Iwama, Shintaro

AU - Wand, Gary S

AU - Salvatori, Roberto

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