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Updated: Jul 8, 2026
Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 23, 2014
[Dense cytoplasmic fluorescence on Hep-2 cells: do not overlook anti-PL]
M Krebs1, B Evrard, T Espinouze
1Laboratoire d'immunologie, CHU de Clermont-Ferrand, Hôtel-Dieu, boulevard Léon-Malfreyt, 63058 Clermont-Ferrand cedex 1, France.
Insights
This study highlights the importance of thoroughly investigating dense cytoplasmic fluorescence in Hep-2000 cells. It successfully identified anti-PL antibodies, aiding in the diagnosis of antisynthetase syndrome.
Area of Science:
- Immunology
- Autoimmunity
- Cellular fluorescence assays
Context:
- Dense cytoplasmic fluorescence on Hep-2000 cells can indicate various autoimmune conditions.
- Distinguishing between different autoantibodies is crucial for accurate diagnosis and patient management.
Purpose:
- To investigate dense cytoplasmic fluorescence on Hep-2000 cells for the presence of anti-PL antibodies.
- To differentiate anti-PL antibodies from other autoantibodies like anti-ribosomes, anti-JO1, and anti-SSA.
Summary:
- A study of twelve patients with dense cytoplasmic fluorescence on Hep-2000 cells was conducted.
- Testing excluded anti-ribosome and anti-JO1 antibodies, with sera sent for confirmation and anti-PL antibody detection.
- Results showed anti-PL7 (4 patients), anti-PL12 (3 patients), and anti-SSA antibodies (4 patients); one patient had no antibody detected.
Impact:
- Systematic exploration of dense cytoplasmic fluorescence efficiently diagnosed five new cases of antisynthetase syndrome, including three not clinically suspected.
- Improved recognition of anti-PL antibodies, preventing confusion with anti-SSA antibodies.
- Enhanced diagnostic capabilities for autoimmune myopathies.
Objective:
Any dense cytoplasmic fluorescence on Hep-2000 cells seen in the immunology laboratory of the Clermont-Ferrand teaching hospital was closely studied to determine the presence or not of anti-PL antibodies.
Patients And Methods:
From January 2006 to January 2007, twelve patients presented a dense cytoplasmic fluorescence on Hep-2000 cells. So we activated a Hep-2 cells'slide, a triple substrate's slide and a dot in order to exclude antiribosomes and anti-JO1 antibodies. After having excluded these antibodies, we sent the sera to the immunology laboratory of the South Lyon hospital to confirm the dense cytoplasmic fluorescence and to detect anti-PL antibodies.
Results:
Four patients presented anti-PL7 and three anti-PL12. For four other patients, the dense cytoplasmic fluorescence was only due to anti-SSA antibodies. Last for one patient, no antibody was found despite an evocative clinic of myositis.
Conclusion:
The systematically extensive exploration during the discovery of a dense cytoplasmic fluorescence proved very efficient, permitting to diagnose five new cases of antisynthetases syndrome including three not evoked by the clinician and to confirm two cases clinically known. This study permitted us to better recognize anti-PL from others dense cytoplasmic fluorescences and not to mix them up with particular anti-SSA.
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