Journal of Leukocyte Biology
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(Journal of Leukocyte Biology. 2006;79:46-58.)
© 2006 by Society for Leukocyte Biology

A study of CD33 (SIGLEC-3) antigen expression and function on activated human T and NK cells: two isoforms of CD33 are generated by alternative splicing

Trinidad Hernández-Caselles*,1, María Martínez-Esparza*, Ana B. Pérez-Oliva*, Ana M. Quintanilla-Cecconi*, Ana García-Alonso{dagger}, D. María Rocío Alvarez-López{dagger} and Pilar García-Peñarrubia*

* Departamento de Bioquímica y Biología Molecular (B) e Inmunología, Facultad de Medicina, Campus de Espinardo, Murcia, Spain; and
{dagger} Servicio de Inmunología, Hospital Universitario Virgen de la Arrixaca, Murcia, Spain

1 Correspondence: Departamento de Bioquímica y Biología Molecular (B) e Inmunología, Facultad de Medicina, Campus de Espinardo, Murcia 30100, Spain. E-mail: trini{at}um.es

The expression of CD33, a restricted leukocyte antigen considered specific for myeloid lineage, has been studied extensively on lymphoid cells. We demonstrated that wide subsets of mitogen- or alloantigen-activated human T and natural killer (NK) cells express CD33 at protein and nucleic acid levels. CD33+ and CD33– T and NK cell populations showed identical surface expression of activation markers such as CD25, CD28, CD38, CD45RO, or CD95. Myeloid and lymphoid CD33 cDNA were identical. However, lymphoid CD33 protein had lower molecular weight, suggesting cell type-specific, post-translational modifications. Additionally, reverse transcriptase-polymerase chain reaction and Northern blot analysis showed an unknown CD33 isoform (CD33m) expressed on all CD33+ cell lines or T cell clones tested. CD33m was identical to CD33 (CD33M) in the signal peptide, the immunoglobulin (Ig) domain C2, the transmembrane, and the cytoplasmic regions but lacked the extracellular ligand-binding variable Ig-like domain encoded by the second exon. CD33m mRNA was mostly detected on NKL and myeloid cell lines but poorly expressed on B cell lines and T lymphocytes. The CD33m extracellular portion was successfully expressed as a soluble fusion protein on transfected human cells, suggesting a functional role on cell membranes. Cross-linking of CD33 diminished the cytotoxic activity of NKL cells against K562 and P815 target cells, working as an inhibitory receptor on NK cells. These data demonstrate that CD33 expression is not restricted to the myeloid lineage and could exist as two different splicing variants, which could play an important role in the regulation of human lymphoid and myeloid cells.

Key Words: T lymphocytes • cytotoxicity






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