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Flow Cytometric Analyses of the Subclasses of Red Cell IgG Antibodies
Author(s) -
Lynen R.,
Neuhaus R.,
Schwarz D.W.M.,
Simson G.,
Riggert J.,
Mayr W.R.,
Köhler M.
Publication year - 1995
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/j.1423-0410.1995.tb01682.x
Subject(s) - subclass , antibody , red cell , flow cytometry , autoantibody , polyclonal antibodies , immunology , coombs test , autoimmune hemolytic anemia , medicine , hemolytic disease of the newborn (abo) , lymphoma , microbiology and biotechnology , biology , fetus , pregnancy , genetics
We report on flow cytometric IgG subclass determinations of red cell antibodies using polyclonal FITC‐labeled antibodies. The limit of detection of this method was 1 ng anti‐D per 1×10 7 red cells. The inter‐ and intra‐assay coefficients of variance were 8.2 and 2.3%, respectively. In 8 newborns with a positive direct antiglobulin test (DAT) in the gel centrifugation test (GCT), due to ABO antibodies, IgG1 was detected in all and IgG2 additionally in 4 of these cases. In 5 severe cases of hemolytic disease of the newborn (HDN) due to anti‐D, large amounts of IgG1 were found, and in 3 of these 5, IgG3 in combination with IgG1. In 8 mild or moderate HDN cases (4 anti‐D, 2 anti‐E, 1 anti‐Fy a , 1 anti‐Jk a ), phototherapy sufficed, and IgG1 was the only antibody. In 7 adult patients with malignant lymphoma and a positive DAT (GCT), only small amounts of IgG1 red cell autoantibodies could be demonstrated by flow cytometry. In 5 further patients with malignant lymphoma, a positive DAT, and severe hemolytic anemia, large amounts of IgG1 autoantibodies were found and IgG3 was also present in 3 of these cases. Flow‐cytometric determination of IgG subclasses may be a useful tool in immunohematology, since subclass determinations were possible in all of these cases. This method is suited for clinical routine and offers the possibility of sufficient standardization.