Measurement of alloantibody by flow cytometry.

Abstract:

:A new method based on flow cytometry has been developed to determine IgG alloantibodies in the serum of immunized rodents. The method utilizes target lymphoid cells, diluted serum and labeled anti-mouse or anti-rat IgG antibodies. In serum from highly immunized animals alloantibodies could be demonstrated up to a dilution of (3 x 10(4))-1 which makes the test approximately 300 times more sensitive than a simultaneously performed complement-dependent cytotoxicity assay (CDCA). Moreover a linear relationship between the amount of alloantibody and the log value of the mean fluorescence of the target cells was found. This linearity permits the comparison of alloantibody production between individual samples by comparing the mean fluorescence values. The reproducibility of the assay was excellent since the coefficients of variation for all dilutions were less than 15%. By using two-color fluorescence the method can discriminate alloantibodies directed against class I and class II MHC antigens. Finally, in addition to its high sensitivity and good reproducibility, the method was found to be at least twice as fast as CDCA.

journal_name

J Immunol Methods

authors

Leenaerts PL,De Ruysscher D,Vandeputte M,Waer M

doi

10.1016/0022-1759(90)90301-b

subject

Has Abstract

pub_date

1990-06-12 00:00:00

pages

73-9

issue

1

eissn

0022-1759

issn

1872-7905

pii

0022-1759(90)90301-B

journal_volume

130

pub_type

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