Method for detection of anti-HLA antibodies
AssigneeUniversity of Virginia Patent Foundation
InventorPeter I. Lobo
Life of this patent
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Abstract
An improved method for detecting even low titer HLA antibodies in proposed organ transplant recipients employs flow cytometry crossmatching (FCXM) on pronase-treated B-cells and T-cells of the donor. Two-color FCXM is preferably employed. The peripheral blood lymphocytes, after pronase digestion, are maintained under conditions to suppress Fcy R receptor regeneration, combined with sera of the proposed transplant recipient, and tested against control sera, using fluorescent reporting complexing agents. Use of pronase digested lymphocytes permits the assay to distinguish between normal or irrelevant IgG B cell binding and immunologically important HLA antibody binding.
Claims5
A method for detecting Human Leukocyte Antigen (HLA) antibodies in the blood of a mammal, which antibodies are reactive with donor tissues from a donor, comprising:
- isolating lymphocytes from said donor,
- enzymatically digesting Fc.gamma. R receptors of said lymphocytes and maintaining said treated lymphocytes under conditions which suppress regeneration of said receptors,
- combining said treated lymphocytes with sera from said mammal, and conjugating any antibody-bound material with a fluorescent reporting agent to obtain a test sample,
- subjecting said test sample to flow cytometry cross-matching to obtain an image of detected fluorescence, and analyzing said image to determine the presence of HLA antibodies.
Dependencies are inferred from the claim text.