Published in Cancer Detection and Prevention 2000; 24(1):33-45.

The CD44 Receptor of the Mouse LB T Cell Lymphoma: Analysis of the Isoform Repertoire and Ligand Binding Properties by Reverse-Transcriptase Polymerase Chain Reaction and AntisenseOligonucleotides

Shularnith Batya Wallach, MSc 1, Adam Friedmann, PhD, 2and David Naor, PhD

1The Lautenberg Center for General and Tumor Immunology, Hebrew University-Hadassah Medical School, POB 12272,Jerusalem 91120, IsraelDepartment of Genetics, Silberman 2Institute for Life Sciences, Hebrew University of Jerusalem, Jerusalem 91904, Israel

Address all correspondence to: David Naor, Ph.D., The Lautenberg Center for General and tumor tmmnnology, Hebrew University-Hadassah Medical Schoo!. POB 12272, Jernsalem 91120. Israel. Tel: 972-2-675 8722, Fax: 972-2-642 4653

ABSTRACT CD44 is a cell surface glycoprotein involved in cell migration and cell docking in target organs viainteractions with various ligands, including hyaluronic acid (HA), which is the principal ligand of this receptor.Alternative splicing generates many isoforms of CD44, including standard CD44 (CD44s) and CD44 variants(CD44v). LB T cell lymphoma, which predominantly expresses CD44s, acquires additional CD44v and HA binding capacity after activation with phorbol ester. The HA9 cell line, isolated from parental LB cells, expressesCD44v and constitutively binds HA. Downregulation of CD44v isoforms of HA9 cells, by CD44v specific anti-sense inhibited their ability to bind HA, indicating that CD44v, rather than CD44s, is associated with the activationstatus of this molecule. Using the reverse transcriptase polymerase chain reaction, we found that LB cells after infiltrating spleen and lymph nodes of BALB/c mice, contain an enriched repertoire of CD44v, implying that themetastatic cells acquired the activated form of this receptor.

KEY WORDS: antisense, oligonucleotides, CD44 variants, lymphoma dissemination, receptor activation, standard CD44, I cell lymphoma.

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