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Updated: Aug 8, 2026
Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 24, 2014
[Cytoplasmic staining pattern characteristic for anti Jo-1 antibody in HEp-2 cells]
Y Nagano1, S Yoshinoya, A Ohkubo
1Department of Laboratory Medicine, Faculty of Medicine, University of Tokyo.
Insights
Anti-Jo-1 antibodies, crucial for diagnosing inflammatory myopathies, typically exhibit cytoplasmic staining on HEp-2 cells. This study clarifies their pattern, aiding accurate diagnosis in polymyositis and dermatomyositis patients.
Area of Science:
- Immunology
- Autoimmunity
- Cellular Biology
Context:
- Anti-Jo-1 antibodies are associated with polymyositis and dermatomyositis (PM/DM).
- Previous studies reported variable HEp-2 cell staining patterns (cytoplasmic and nuclear) for anti-Jo-1 antibodies.
- The precise immunofluorescence staining pattern of anti-Jo-1 antibodies on HEp-2 cells requires confirmation.
Purpose:
- To confirm the characteristic immunofluorescence staining pattern of anti-Jo-1 antibodies on HEp-2 cells.
- To resolve discrepancies in reported staining patterns observed in indirect immunofluorescence assays.
- To establish a reliable diagnostic marker for anti-Jo-1 associated autoimmune conditions.
Summary:
- Eight patient sera positive for anti-Jo-1 antibodies (via double immunodiffusion) were tested on HEp-2 cells.
- Immunoblotting identified a 50 kDa band, corresponding to the Jo-1 antigen, in all tested sera.
- Eluted antibodies targeting the 50 kDa band produced fine granular cytoplasmic staining on HEp-2 cells, confirming the characteristic pattern.
Impact:
- Clarifies the definitive cytoplasmic staining pattern of anti-Jo-1 antibodies on HEp-2 cells.
- Aids in accurate interpretation of immunofluorescence assays for diagnosing PM/DM.
- Highlights the potential for confounding nuclear staining from other autoantibodies.
Abstract:
We wished to confirm the staining pattern of HEp-2 cells by the anti Jo-1 antibody, because we found antibodies in serum with positive anti Jo-1 antibody which showed either a fibrilar cytoplasmic staining or a nuclear speckled staining pattern in indirect immuno fluorescence+ examinations using HEp-2 cells. Sera available from eight patients with PM DM (polymyositis and/or dermatomyositis) showed positive anti Jo-1 antibody in the double immuno-diffusion technique but had various staining patterns of HEp-2 cells in the immunofluorescent examination. We examined these eight sera with the immuno-blotting method utilizing whole cell extract of HeLa cells, and found the 50 kDa band from all sera tested, to which Jo-1 antigen had been reported to move. We eluted the antibody which formed the 50 kDa band from the nitrocellulose membrane and applied it on HEp-2 cells. This maneuver gave us the fine granular cytoplasmic staining of anti Jo-1 antibody on HEp-2 cells. We therefore concluded that the anti Jo-1 antibody should have cytoplasmic staining on HEp-2 cells although observers might miss it due to other types of associated antinuclear antibodies.
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