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Inusuale pattern immunofluoroscopico su cellule HEp-2

Unusual immunofluorescence pattern on HEp-2 cells

  • Caso Clinico
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La Rivista Italiana della Medicina di Laboratorio - Italian Journal of Laboratory Medicine

Riassunto

Premesse

La ricerca degli anticorpi anti-nucleo (ANA) in immunofluorescenza indiretta (IFI) su cellule HEp-2 rappresenta una tappa fondamentale nella diagnostica delle patologie autoimmuni. Riportiamo il caso di 5 pazienti, che all’IFI per la ricerca di ANA eseguita con kit Nova Lite™ HEp-2 (Inova Diagnostics, San Diego, CA), presentavano un pattern citoplasmatico particolare.

Metodi

I sieri analizzati appartengono a cinque pazienti (età compresa tra 45 e i 69 anni). I sieri sono stati testati per la ricerca degli anticorpi anti-antigeni nucleari estraibili con metodo ELISA (ENA: anti-Sm, RNP, SSA/Ro, SSB/La, Scl-70 e anti-Jo-1) (ENA Screen, Inova Diagnostics) per la ricerca di ANA su HEp-2 di altri produttori (MBL International, Nagoya, Giappone e Alphadia, Waver, Belgio), per la ricerca di AMA, ASMA, APCA, LKM e Lc-1 in IFI su triplo tessuto di roditore (Inova Diagnostics) e per la ricerca di anticorpi anti-Sm, RNP, SSA/Ro, SSB/La, Scl-70, Jo-1, PM/Scl, Ku, Cenp-A/B, PCNA, LKM-1, Lc-1, SLA, F-actina e AMA-M2 con metodica Dot Blot (Alphadia).

Risultati

Quattro pazienti presentavano un pattern nucleare speckled a titolo 1:80, mentre il quinto risultava negativo (<1:80), ma tutti presentavano un pattern citoplasmatico, con la presenza di una struttura rotondeggiante relativamente grande, presente in circa un terzo delle cellule. La ricerca di tutte le altre specificità anticorpali ricercate in ELISA, in IFI e in DotBlot ha dato esito negativo.

Conclusioni

Le informazioni cliniche sui 5 pazienti sono state ottenute dai rispettivi medici curanti; due pazienti non avevano una diagnosi definitiva di malattia, uno risultava affetto da ulcera duodenale e uno da gastrite cronica, entrambi in trattamento con inibitori della pompa protonica. Il quinto paziente era affetto da ipotiroidismo ed era trattato con terapia sostitutiva. Interessante appare l’osservazione di un tale pattern citoplasmatico non ascrivibile a una struttura citoplasmatica facilmente identificabile. Sono ovviamente necessari ulteriori studi per verificare se tale pattern abbia significatività clinica.

Summary

Background

The detection of antinuclear antibodies (ANA) by indirect immunofluorescence assay (IFA) on HEp-2 cells is a fundamental step in the diagnosis of autoimmune diseases. We report the case of five patients that, on IFA for detection ANA (Nova Lite™ HEp-2, Inova Diagnostics), presented an unusual cytoplasmatic pattern.

Methods

Serum samples belonging to 5 patients (3 women and 2 men, age 45–69 years). All the samples were analysed with ELISA for detection of antiextractable nuclear antigens (ENA) (anti-Sm, RNP, SSA/Ro, SSB/La, Scl-701 and anti-Jo-1) (ENA Screen, Inova Diagnostics). ANA detection was performed also with IFA using HEp-2 cells from different manufacturers (MBL and Alphadia). Furthermore we performed the detection of anti-mitochondrial antibodies, antismooth muscle antibodies, anti-gastric parietal cell antibodies, anti-liver/kidney microsomal and anti-liver cytosol antibodies on IFA on rat tissue (Inova Diagnostics), and the detection of anti-Sm, RNP, SSA/Ro, SSB/La, Scl-70, Jo-1, PM/Scl, Ku, Cenp-A/B, PCNA, LKM-1, Lc-1, SLA, F-actin and AMA-M2 with Dot Blot (Alphadia).

Results

Four patients showed a nuclear speckled pattern at 1:80 dilution, while the fifth patient was negative (<1:80), but all of them showed a cytoplasmic pattern with a round-shaped body of relatively large dimension in about one third of the cells. Detection of all other autoantibodies assayed was negative.

Conclusions

We contacted the clinicians of these five patients to obtaine clinical information. Two patients had no definite clinical diagnosis; one was affected by chronic gastritis and one was affected by duodenal ulcer; both the latter patients were in therapy with protonic pump inhibitors. The fifth patient suffered of hypothyroidism and was treated with substitutive therapy.

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Authors and Affiliations

  1. Laboratorio di Biochimica Clinica, Comprensorio Sanitario di Bolzano, Azienda Sanitaria dell’Alto Adige, Bolzano, Italy

    Massimo Daves, Silvestro Scala, Giovanni Cosio, Mirella Floreani & Evi Peer

  2. U.O. di Diagnostica Ematochimica, Azienda Ospedaliero-Universitaria di Parma, Parma, Italy

    Chiara Bonaguri & Giuseppe Lippi

  3. Laboratorio di Biochimica Clinica, Comprensorio Sanitario di Bolzano, Azienda Sanitaria dell’Alto Adige, Via L. Boehler 5, 39100, Bolzano, Italy

    Massimo Daves

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  1. Massimo Daves
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  2. Chiara Bonaguri
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  3. Silvestro Scala
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  4. Giovanni Cosio
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  5. Mirella Floreani
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  6. Evi Peer
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  7. Giuseppe Lippi
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Correspondence to Massimo Daves.

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Daves, M., Bonaguri, C., Scala, S. et al. Inusuale pattern immunofluoroscopico su cellule HEp-2. Riv Ital Med Lab 8, 54–57 (2012). https://doi.org/10.1007/s13631-012-0041-4

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  • DOI: https://doi.org/10.1007/s13631-012-0041-4

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