poster ro antibodies
DESCRIPTION
Congreso Internacional de Autoinmunidad Slovenia 2009TRANSCRIPT
EVALUATION OF ANTINUCLEAR RO/SS-A AND LA/SS-B ANTIBODIES BY MULTIPLEX TECHNOLOGY AT THE UNIVERSITY
HOSPITAL “12 DE OCTUBRE” (MADRID-SPAIN)
M. Talise1, V. Peréz1, M. Sevilla1, S. Lermo1, G. Badra2, L. Borque3, A. Serrano1 1Immunology Service, University Hospital 12 de Octubre, 2University Rey Juan Carlos, Madrid, 3San Pedro Hospital, Logroño, Spain
INTRODUCTION
Autoantibodies to Ro/SS-A and La/SS-B ribonucleoproteins are found
in autoimmune diseases such as primary Sjögren´s syndrome,
systemic lupus erythematousus (SLE) and rheumatoid arthritis1 (RA).
Two types of anti-Ro/SSA antibodies have been described, anti-
SSA-52 kDa and anti-SSA-60kDa, historically, these autoantibodies
were considered as a uniform autoantibody-system. However, recent
studies provided evidence that Ro60 and Ro52 are not part of a
stable macromolecular complex and that anti-Ro52 and anti-Ro60
(SS-A) antibodies have different clinical associations and each
specific to different antigens2, for this reason it is necessary to be
able to individually identify these antibodies in the autoimmunity
laboratory and the Bioplex TM 2200 automated system offers this
advantage; we compare this technique with the immunofluorescence
(IIF) and AtheNA Multi-lyte® ANA test system a multiplex fluorescent
microsphere immunoassay for the semi-quantitative of Ig G class
antibody to 9 separate analytes (DNA, SSA, SSB, Sm, RNP, Scl-70,
Jo-1, Centromere B, and Histone).
METHODOLOGY
We studied sera from 600 patients randomly admitted to the
autoimmunity section during May and June 2009, these sera were
obtained from patients referred from primary care physicians, general
internists, rheumatologists and other subspecialties such as
nephrology, dermatology, etc . Antinuclear antibodies (ANA´s ) were
determined by immunofluorescence (IIF) on HEp-2 cell, AtheNA Multi-
lyte® multiplex technique and BioPlex™2200 (by Bio-Rad Laboratories,
Hercules, CA) a fully automated Luminex based system developed for
analysis of 13 antibodies including anti-SSA-52 kDa and anti-SSA-60
kDa in a primary tube and SS-B, Sm, Sm/RNP, RNPA, RNP-68, Scl70,
C e n t r o m e r e - B , d s D N A , c h r o m a t i n , J o 1 , r i b o s o m a l P.
Immunofluorescence was considered positive from the dilution 1/ 80.
RESULTS Six hundred sera were evaluated, 33 sera were Anti-SSA/Ro52, 42 Anti-
SSA/Ro60, and 17 Anti-La/SS-B positive by Bioplex 2200; 37 sera were
positive to Anti SS-A/Ro and 11 AntiSS-B/La by AtheNA Multi-lyte® with a
global concordance of 96.5% and 98.33% respectively; by IIF we obtained
48 positive sera, 25 (52.08%) with coarse and fine speckled pattern, 18
(37.50%) homogeneous, 2 (4.20%) Centromere-B, 2 (4,20%) Nuclear dots.
Finally, 80.70% of the patients had at least one diagnosis of autoimmune
disease.
0
5
10
15
20
25
30
35
40
45
Anti-SSA Athena Anti-SSA Bioplex 2200
Anti-SSA/Ro52 Bioplex
Anti-SSA/Ro60 Bioplex
Anti-SSA Ro52 and Ro60 Bioplex
Anti-SSB/La Athena
Anti-SSB/La Bioplex 2200
We observed 15 discrepancies: Anti-SSA negative by Athena but
positive Anti-SSA/Ro52 and/or Anti-SSA/Ro60 by Bioplex 2200; of
which 12 were IIF positive and had some autoimmunity disease: 6
SLE; 2 Systemic Sclerosis, 2 autoimmune Hepatitis, 1 Sjögren´s
syndrome + Raynaud , 1 primary biliary cirrhosis, 1
dermatomyositis. Only three discrepancies were Anti-SSA Athena
positive, Anti-SSA Bioplex Negative and IIF negative, one had
rheumatoid arthritis.
On the other hand other 8 discrepancies were observed: Anti-
SSB Athena’s Negative but Bioplex 2200 Positive, of which 8 were
IIF positive and four had diagnosis of SLE, 2 RA, 2 Sjögren´s Sx.
Only two sera were Anti-SSB Athena positive but Bioplex 2200
negative, of which 1 was IIF positive and 1 had RA.
CONCLUSIONS
We conclude that BioPlex2200 offers a good percentage of
concordance with other methods, and it can be a good solution as
a multiplex approach to diagnose autoimmune diseases like
Sjögren´s syndrome , systemic lupus erythematousus and
rheumatoid arthritis because the separation of Ro / SSA 52 and 60
antigens conferred increased sensitivity, provides greater
specificity with a lower rate of false negatives.
REFERENCES 1. van Woerkom JM, Geertzema JG, Nikkels PG, et al. Expression of Ro/SS-A and La/
SS-B determined by immunohistochemistry in healthy, inflamed and autoimmune
diseased human tissues: a generalized phenomenon. Clin Exp Rheumatol. 2004 May-
Jun;22(3):285-92.
2. J. Schulte-Pelkum a,M. Fritzler , M. Mahler . Latest update on the Ro/SS-A
autoantibody system. Autoimmunity Reviews 8 (2009) 632–637