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Blood, 1 June 2001, Vol. 97, No. 11, pp. 3513-3520
IMMUNOBIOLOGY
Molecular characterization and expression of a novel human
leukocyte cell-surface marker homologous to mouse
Ly-9
Miguel Angel de la Fuente,
Victoria Tovar,
Neus Villamor,
Nuria Zapater,
Pilar Pizcueta,
Elias Campo,
Jaime Bosch, and
Pablo Engel
From the Immunology Unit, Department of Cellular
Biology and Pathology, University of Barcelona Medical School; Liver
Unit, Hematopathology Section, Laboratory of Anatomic Pathology, and
Department of Hematology, Hospital Clínic; and Institut
d'Investigacions Biomèdiques August Pi i Sunyer, Barcelona,
Spain.
Ly-9 is a mouse cell-surface glycoprotein that is selectively
expressed on thymocytes and on mature T and B lymphocytes. Ly-9 belongs
to the CD2 subset of the immunoglobulin superfamily, an emerging family of cell signaling receptors. Recently, a
partial human Ly-9 complementary DNA (cDNA) sequence has been
described. Full-length cDNA clones were isolated that included the
initiation codon, the sequence encoding the full signal peptide, and 14 amino acids more in the cytoplasmic domain than in the previously
reported clone. The predicted extracellular domain of human Ly-9
contains 4 immunoglobulinlike domains, similar to those in mouse Ly-9. Northern blot analysis revealed that the human Ly-9 messenger RNA (2.6 kb) is expressed predominantly in lymph node, spleen, thymus, and
peripheral blood leukocytes. Four monoclonal antibodies (mAbs) were
raised against human Ly-9 by immunizing mice with the pre-B-cell line
300.19 stably transfected with human Ly-9 full-length cDNA. These mAbs
strongly stained the surfaces of cells transfected with human Ly-9 cDNA
but not of untransfected cells. Human Ly-9 expression was restricted to
T and B lymphocytes and thymocytes, with the highest levels of
expression on CD4+CD8 and
CD4CD8+ thymocytes. Monocytes, granulocytes,
platelets, and red blood cells were uniformly negative for Ly-9. These
mAbs immunoprecipitated major polypeptides of 120 kd from the
transfected cells and 120 kd and 100 kd from B-cell line Daudi,
probably because of the cell-surface-expressed isoforms. These data
demonstrate that human Ly-9 is a new marker for the study of normal and
malignant leukocytes.
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