Detection of HLA antibodies by using flow cytometry and latex beads coated with HLA antigens.
BACKGROUND
Detection of HLA class I antibodies in sera is needed in various clinical situations. The standard method for detecting HLA class I antibodies is the complement-dependent lymphocytotoxicity (CDC) assay, but solid-phase assays are now available.


METHODS
This study assessed the ability of a flow cytometric assay using latex beads coated with HLA class I antigens to detect HLA class I-specific antibodies. The CDC assay was compared with the pooled-bead assay for the detection of HLA class I antibodies. Thirty-one randomly selected serum samples previously tested by CDC assay were tested with pooled beads and analyzed by flow cytometry. Twenty-seven additional serum samples, chosen by clinical criteria and CDC assay results, were tested against the pooled beads. Next, samples from six patients from whom three or more serum samples were drawn on consecutive days were tested with both methods. Finally, serum samples that were proved positive by both methods were tested with selected beads coated with antigens from a single person.


RESULTS
Among the randomly selected serum samples, there was 90-percent agreement between the two assays. There was 96-percent agreement between the two assays of the 27 samples that were selected by clinical criteria and CDC assay results. Testing the sera with individual beads suggested that the HLA class I antibodies react with beads expressing the corresponding HLA antigen and beads expressing antigens in the same cross-reactive group.


CONCLUSIONS
The pooled-bead assay can be used as an alternative method for detecting HLA class I antibodies. However, if the specificity of the HLA class I antibody is required, another assay must be used.
Department of Transfusion Medicine, Warren G. Magnuson Clinical Center,
L A
LA
Moses
D F
DF
Stroncek
K M
KM
Cipolone
F M
FM
Marincola
D000906:Antibodies / Q000032:analysis* / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D006680:HLA Antigens / Q000276:immunology*; D006801:Humans; D007840:Latex; D008863:Microspheres; D012680:Sensitivity and Specificity
D000906:Antibodies; D006680:HLA Antigens; D007840:Latex