2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) suppresses B lymphocyte proliferation and immunoglobulin production. We previously reported that the aryl hydrocarbon receptor (AhR) complex, composed of the AhR ligand binding subunit and the Ah receptor nuclear translocator (ARNT), was constitutively present in nuclear extracts from two human B lymphocyte cell lines (Biochem. Biophys. Res. Commun. 212, 27-34, 1995). The present study compared the AhR complex in the IM-9 and PJS-91 human B lymphocyte and HepG2 human hepatoma cell lines. AhR mRNA levels in the two lymphocyte cell lines were substantially lower than those in HepG2 cells, as was immunoreactive AhR protein. In contrast, ARNT mRNA and protein were expressed at a high level in all three cell lines. TCDD induction of cytochrome P450 1A1 mRNA and protein was detected in only the PJS-91 lymphocyte cell line, and at a markedly lower level than that in HepG2 cells. In gel shift assays, the cytosolic DNA-binding AhR complex in IM-9 and PJS-91 cells was indistinguishable from that in HepG2 cells. In contrast, the nuclear DNA-binding AhR complex in IM-9 and PJS-91 cells consisted of several closely migrating species, one being recognized by an AhR antibody, while an ARNT antibody reacted with all species. Protein:DNA cross-linking analysis revealed the presence of a novel Mr 100,000 DNA-binding protein in nuclear extracts from IM-9 and PJS-91, but not HepG2, cells that was not recognized by either AhR or ARNT antibodies. These results show that IM-9 and PJS-91 human B cells constitutively express a distinct nuclear DNA-binding form of the AhR complex that may result from the presence of an additional protein or a structural variant of the AhR.