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Abstract
Background
Autoimmune thrombocytopenia (AITP) is characterized by autoantibody-induced platelet destruction. Although several studies have shown that pathogenic autoantibodies are mainly IgG directed platelet glycoproteins (GP), a platelet GP specific test is not available in clinical laboratories. The aim of this study was to evaluate the clinical usefulness of a Modified Antigen Capture Enzyme-linked immunosorbent assay (MACE) test in the diagnosis of AITP.
Methods
We investigated fifty-seven patients who showed a platelet count lower than 100×109/L and underwent a bone marrow examination. They were classified into primary AITP (P-AITP) (n=21), secondary AITP (S-AITP) (n=15), and non-immune thrombocytopenia (NITP) (n=21) by bone marrow findings and clinical diagnosis. Platelet GP (IIb/IIIa, Ia/IIa, Ib/IX, IV)-specific antibodies and anti-HLA class I antibody were detected by MACE test.
Results
Among 57 samples, platelet GP specific antibodies were detected in 8 (22.2%) of 36 patients with AITP and 1 (4.8%) of 21 patients with NITP. The specificities were as follows: GP IIb/IIIa (n=4), GP Ia/IIa (n=5), GP Ib/IX (n=3) and GPIV (n=2). Of the nine patients with platelet GP specific antibodies, four (44.4%) had more than two platelet GP specific antibodies. The sensitivity, specificity, positive predictive value and negative predictive values of the MACE test for AITP were 22.2%, 95.2%, 88.9%, 41.7%, respectively. A previous transfusion history was associated with a higher detection rate of anti-HLA class I antibodies (P<0.05).
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Fig. 1.
Detection of platelet specific IgG antibodies to platelet glycoproteins (GP) IIb/IIIa, Ia/IIa, Ib/IX, IV or HLA class I (GP IIb/IIIa in this example) by MACE assay. Washed platelets were first incubated with human plasma (containing anti-platelet GP IIb/IIIa in this sample). The antibody-sensitized platelets were then lysed and the supernatant was added to a microplate precoated with murine monoclonal antibodies (MoAb) specific to platelet GP or HLA class I. Any human IgG bound to the GP or HLA class I antigen were detected with a alkaline phosphatase (ALP) conjugated antihuman IgG and substrate solution.
Table 1.
Main features of 57 patients with AITP and NITP
|
AITP |
NITP | |||
|---|---|---|---|---|
| P-AITP | S-AITP | Total | ||
| N | 21 | 15 | 36 | 21 |
| Male:Female | 1.1 | 2.0 | 1.4 | 1.6 |
| Age (years)* | 52.3±1.9 | 38.6±9.0 | 49.2±12.7 | 63.0±16.0 |
| Platelets count*, ×109/L | 37.0±31.7 | 57.1±27.0 | 45.3±31.4 | 56.5±23.7 |
| Previous transfusion, N (%) | 4 (19.0) | 7 (46.7) | 11 (30.6) | 12 (57.1) |
| Platelet GP-specific antibody, no (%) | 5 (23.8) | 3 (20.0) | 8 (22.2) | 1 (4.8) |
| GP IIb/IIIa | 2 | 2 | 4 | 0 |
| GP Ia/IIa | 2 | 2 | 4 | 1 |
| GP Ib/IX | 2 | 1 | 3 | 0 |
| GP IV | 1 | 1 | 2 | 0 |
| Anti-HLA class I antibody, no (%) | 11 (52.4) | 9 (60.0) | 20 (55.6) | 21 (100) |
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