       Document 0982
 DOCN  M9540982
 TI    Carbohydrate receptor-mediated gene transfer to human T leukaemic cells.
 DT    9504
 AU    Thurnher M; Wagner E; Clausen H; Mechtler K; Rusconi S; Dinter A;
       Birnstiel ML; Berger EG; Cotten M; Institute of Physiology, University
       of Zurich, Switzerland.
 SO    Glycobiology. 1994 Aug;4(4):429-35. Unique Identifier : AIDSLINE
       MED/95128080
 AB    The mucin-type carbohydrate Tn cryptantigen (GalNAc alpha 1-O-Ser/Thr,
       where GalNAc is N-acetyl-D-galactosamine) is expressed in many
       carcinomas, in haemopoietic disorders including the Tn syndrome, and on
       human immunodeficiency virus (HIV) coat glycoproteins, but is not
       expressed on normal, differentiated cells because of the expression of a
       Tn-processing galactosyltransferase. Using Jurkat T leukaemic cells
       which express high levels of Tn antigen due to deficient Tn
       galactosylation, we have established the Tn antigen-mediated gene
       transfer and demonstrate the considerable efficiency of this approach.
       We used poly(L-lysine) conjugates of the monoclonal antibody 1E3
       directed against the Tn antigen to deliver the luciferase and
       beta-galactosidase reporter genes to Jurkat cells by receptor-mediated
       endocytosis. Addition of unconjugated 1E3 reduced transfection
       efficiency in a concentration-dependent manner and incubation with free
       GalNAc abolished DNA transfer completely, indicating that gene delivery
       is indeed mediated by the Tn antigen. Pre-treatment of Jurkat cells with
       Vibrio cholerae sialidase, which uncovers additional Tn antigens,
       resulted in an improvement of gene transfection. Both human and chicken
       adenovirus particles attached to the DNA/polylysine complex strongly
       augmented transgene expression. When the beta-galactosidase (lacZ) gene
       was delivered to Jurkat cells by Tn-mediated endocytosis, up to 60% of
       the cells were positive in the cytochemical stain using
       5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) as a
       chromogenic substrate. The efficiency of the transferrin
       receptor-mediated DNA uptake into Jurkat cells was comparatively low,
       although these cells were shown to express considerable amounts of
       transferrin receptor. We show here that a mucin-type carbohydrate
       antigen mediates highly efficient DNA uptake by endocytosis into Jurkat
       T cells. This method represents a 50-fold improvement of Jurkat cell
       transfection efficiency over other physical gene transfer techniques.
       Specific gene delivery to primary cancer cells exhibiting Tn epitopes
       may especially be desirable in immunotherapy protocols.
 DE    Adenoviridae/GENETICS  Animal  Antibodies, Monoclonal  Antigens,
       Tumor-Associated, Carbohydrate/*GENETICS/METABOLISM  DNA/GENETICS  Gene
       Expression  Gene Therapy  *Gene Transfer  Genes, Reporter  Genetic
       Vectors  Human  Lac Operon  Leukemia, T-Cell/*GENETICS  Support,
       Non-U.S. Gov't  Tumor Cells, Cultured/METABOLISM  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

