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Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 24, 2014
Three human chromosomal autoantigens are recognized by sera from patients with anti-centromere antibodies
Insights
Anti-centromere antibodies (ACA) are highly specific to centromere proteins (CENP)-A, CENP-B, and CENP-C in patients with Raynaud's disease. Antibody to CENP-B strongly correlates with ACA detection.
Area of Science:
- Immunology
- Cell Biology
- Rheumatology
Background:
- Anti-centromere antibodies (ACA) are associated with Raynaud's phenomenon.
- The specific antigens targeted by ACA in patients with Raynaud's disease require further characterization.
Purpose of the Study:
- To identify and characterize the specific chromosomal antigens recognized by ACA in patients with Raynaud's syndrome or disease.
- To determine the uniformity of the ACA response within this patient population.
Main Methods:
- Sera from 39 ACA-positive patients with Raynaud's and 123 controls were used.
- Highly purified human mitotic chromosomes were screened using SDS-PAGE and immunoblotting.
- Specific centromere proteins (CENP)-A, (CENP)-B, and (CENP)-C were identified as targets.
Main Results:
- ACA-positive sera recognized three distinct chromosomal antigens: (CENP)-B (100%), (CENP)-A (97.4%), and (CENP)-C (94.9%).
- None of the control sera recognized these specific antigens.
- Antibody to (CENP)-B demonstrated a 100% correlation with anti-centromere staining via indirect immunofluorescence.
Conclusions:
- The ACA response in patients with Raynaud's disease is highly uniform, primarily targeting (CENP)-A, (CENP)-B, and (CENP)-C.
- (CENP)-B is a highly specific marker for ACA detection in this patient group.
- These findings enhance the understanding of autoimmune responses in Raynaud's disease.
Abstract:
We have identified 39 individuals with anti-centromere antibodies (ACA) in our patient population, all of whom have Raynaud's syndrome or disease. We have used sera from the ACA-positive patients and from 123 controls (22 normal individuals and 101 additional patients with either Raynaud's disease or Raynaud's syndrome plus an associated connective tissue disease) to screen the proteins of highly purified human (HeLa) mitotic chromosomes by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting. Three antigens were recognized by the sera from the ACA-positive patients. These were centromere protein (CENP)-B (80,000 mol wt--recognized by all ACA-positive sera), CENP-A (17,000 mol wt--recognized by 38 of 39 ACA-positive sera), and CENP-C (140,000 mol wt--recognized by 37 of 39 ACA-positive sera). None of these antigens were recognized by any of the 123 control sera, although binding was occasionally seen to other chromosomal antigens. Therefore the ACA response is highly uniform in our patient population. Antibody to CENP-B shows a 100% correlation with anti-centromere staining by indirect immunofluorescence.
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