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T cell clones with normal or defective O‐galactosylation from a patient with permanent mixed‐field polyagglutinability
Author(s) -
Thurnher Martin,
Berger Eric G.,
Clausen Henrik,
Fierz Walter,
Lanzavecchia Antonio
Publication year - 1992
Publication title -
european journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.272
H-Index - 201
eISSN - 1521-4141
pISSN - 0014-2980
DOI - 10.1002/eji.1830220724
Subject(s) - biology , microbiology and biotechnology , sialic acid , antigen , t cell , cd19 , cd8 , cd5 , peanut agglutinin , monoclonal antibody , epitope , flow cytometry , population , t lymphocyte , antibody , biochemistry , lectin , immunology , immune system , demography , sociology
Abstract To delineate the extent of O‐galactosyltransferase deficiency within the lymphoid lineage, monoclonal antibody specific for the Thomsen‐Friedenreich (TF) antigen (Galβ1 → 3GalNAcα1‐O‐Ser/Thr) and its precursor the Tn antigen (GalNAcα1‐O‐Ser/Thr) were applied to the flow cytometric analysis of peripheral blood lymphocytes from a patient with permanent mixed‐field polyagglutinability (PMFP). We show that only a minor population of 4% expressed the Tn antigen which is in contrast to 93% of the patient's erythrocytes carrying the defect. Tn + lymphocytes mainly belonged to the CD3 + subset, but were also CD19 + or CD16 + . Both Tn + and TF + T cell clones from patient R. R. were established and shown to belong to the CD4 + or CD8 + antigenic subset. Three glycosyltransferase activities were determined in lysates from these clones: all Tn + clones were deficient in UDP‐Gal: GalNAcα1‐O‐Ser/Thr β1 → 3 galactosyl‐transferase (β3Gal‐T) activity; by contrast this activity was present in all lysates from TF‐expressing clones. UDP‐GalNAc:polypeptide α‐N‐acetyl‐galactosaminyltransferase (GalNAc‐T) and UDP‐Gal:GlcNAc‐Rβ1 → 4 galactosyl‐transferase (β4Gal‐T) exhibited similar activities in both Tn + and TF + T cell clones. As a consequence of defective O‐galactosylation in Tn + T cells, cell surface sialic acid of Tn + clones was reduced by > 50% when compared to TF + clones as demonstrated by sialic acid‐specific labeling using fluoresceinated Limax flavus agglutinin(LA) and flow cytometry. The Tn phenotype of T cell clones was stable for more than 1 year of continuous expansion in vitro. These data demonstrate that in PMFP, T cells may also be affected by the O‐galactosyltransferase deficiency which is accompanied by a substantial loss of cell surface sialic acid. However, the frequency of Tn + lymphocytes in peripheral blood from patient R.R. was strikingly low. These T cell clones should be useful to study the defect at a genetic level and the importance of O‐linked carbohydrates for proper T cell function.