The human CXC chemokine receptor CXCR4 is activated by stromal cell-derived factor 1. The receptor is present in many cell types and regulates a variety of cellular functions, including chemotaxis, adhesion, hematopoiesis, and organogenesis. Human CXCR4 also serves as a cofactor for cell entry by certain strains of HIV-1 and HIV-2. In the mouse, alternative RNA splicing produces two transcripts encoding two CXCR4 isoforms, mCXCR4-A and mCXCR4-B, differing by the presence of two amino acids in the amino terminal portion of the longer protein, mCXCR4-B. Only one CXCR4 transcript, encoding the human counterpart of mCXCR4-A, is known in man. The involvement of the aminoterminal-most portion of CXCR4 in both ligand and HIV envelope protein recognition led us to determine whether a CXCR4 variant corresponding to mCXCR4-B is present in human tissues. To this end, the genomic organization and expression of the human CXCR4 gene was examined. Both the human and the mouse CXCR4 gene consist of two exons separated by an approximately 2.1 kbp intron between codons five and six and carry splice donor sites at the 5' end of their introns. These similarities notwithstanding, single nucleotide primer extension, reverse transcriptase PCR amplification, and sequencing of CXCR4 cDNA clones show that a splice variant of CXCR4 corresponding to mCXCR4-B is absent in man.