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M9480538.TXT
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1994-08-20
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Document 0538
DOCN M9480538
TI Detection of antiphospholipid antibodies by immunostaining on thin layer
chromatography plates.
DT 9410
AU Sorice M; Griggi T; Circella A; Garofalo T; d'Agostino F; Pittoni V;
Pontieri GM; Lenti L; Valesini G; Dipartimento di Medicina Sperimentale,
Universita La Sapienza,; Roma, Italy.
SO J Immunol Methods. 1994 Jul 12;173(1):49-54. Unique Identifier :
AIDSLINE MED/94308563
AB There is increasing interest in the role of antiphospholipid antibodies
in the so-called 'antiphospholipid antibody syndrome' (APS). The two
major methods currently employed for detecting the autoantibodies are
the solid phase ELISA and the LAI test (inhibition of phospholipid
dependent coagulation assay). In our study we have tested the
possibility of detecting antiphospholipid antibodies by immunostaining
on thin layer chromatography (TLC) plates, since this technique permits
the use of pure phospholipid molecules as antigen. Sera were collected
from 20 patients with SLE without APS, 20 patients with APS, 20 anti-HIV
positive subjects, ten patients with signs of APS but antiphospholipid
negative (ELISA), 20 patients with syphilis and 40 matched blood donors.
Results showed that only 72.3% of sera containing detectable levels of
aCL antibodies in solid phase ELISA were also positive for aCL in TLC
immunostaining; these discrepancies may be due to the presence of
antibodies reacting with a protein complexed with phospholipid (beta
2-glycoprotein-I) or, alternatively, to the different antigenic
presentation of phospholipids on chromatograms compared to the surface
of microtitre wells. Furthermore, aCL monoclonal antibody CAL-3, as well
as nine sera positive for aCL, also reacted with PS and PE. Previous
absorption of these sera with CL micelles completely abolished the
reactivity with PS and PE, demonstrating cross-reactivity among these
three phospholipids. In conclusion, our findings reveal that TLC
immunostaining is more specific, but less sensitive, than ELISA for the
detection of antiphospholipid antibodies in human sera.
DE Antibodies, Anticardiolipin/BLOOD Antibodies, Antiphospholipid/*BLOOD
Antibodies, Monoclonal Antigens Antiphospholipid Syndrome/IMMUNOLOGY
Chromatography, Thin Layer/*METHODS/STATISTICS & NUMER DATA Comparative
Study Enzyme-Linked Immunosorbent Assay/STATISTICS & NUMER DATA
Evaluation Studies Human HIV Infections/IMMUNOLOGY
Immunoassay/*METHODS/STATISTICS & NUMER DATA Immunosorbent Techniques
Lupus Erythematosus, Systemic/IMMUNOLOGY Phospholipids/IMMUNOLOGY
Sensitivity and Specificity Support, Non-U.S. Gov't
Syphilis/IMMUNOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).