title: Suppression of splenic lymphocyte function by 7,12-dimethylbenz[a]anthracene (DMBA) in vitro.
abstract: The effects of the immunosuppressive polycyclic aromatic hydrocarbon (PAH) 7,12-dimethylbenz[a]anthracene (DMBA) were studied directly by in vitro exposure of splenic lymphocytes. On the basis of evidence from prior studies that DMBA immunotoxicity in vivo may not be dependent upon induction of the Ah locus in mice, splenocytes from Ah-responsive B6C3F1, Ah-nonresponsive DBA/2N, and in C57BL/6J Ah-congenic mice (responsive B6-Ah(b)Ah(d) and nonresponsive B6-Ah(d)Ah(d) were exposed to xenobiotic in culture. For some experiments, B6C3F1 mice were pretreated with 200 nmol 2,3,7,8-tetrachlorodibenzop-dioxin (TCDD) to induce Ah-associated enzymatic activity prior to in vitro splenocyte exposure to DMBA. Humoral immunity assessed as splenic antibody plaque-forming cells measured after a 5-day in vitro immunization to sheep erythrocytes (SRBC) was suppressed up to 99% by continuous exposure to 20 microM DMBA, and was comparable between control mice having basal levels of hepatic monooxygenase activity and Ah-induced mice (TCDD-treated) having elevated enzyme activity. Similarly, cytotoxic T-lymphocyte generation against P815 target cells was suppressed up to 88 and 86% in 40 microM DMBA-exposed splenocytes from Ah-induced and noninduced mice, respectively. The mixed lymphocyte responsiveness (MLR) of B6C3F1, DBA/2N, B6-Ah(b)Ah(d), and B6-Ah(d)Ah(d) splenocytes exposed in vitro to 40 microM DMBA was suppressed 54, 72, 51, and 29%, respectively. However, the degree of suppression was not significantly different between the strains. The secretion of interleukin 2 (IL2) was also suppressed in splenocytes from both strains exposed to 40 microM DMBA in vitro. Studies which included benzo[a]pyrene (BaP) as a control xenobiotic known to demonstrate Ah dependence showed that the MLR of splenic lymphocytes from Ah-congenic mice was comparably suppressed following 40 microM DMBA exposure, whereas exposure to 40 microM BaP resulted in suppression of the MLR only in B6-Ah(b)Ah(d) splenocytes. In addition, mitogen-stimulated proliferation was inhibited in both B6C3F1 and DBA/2N splenocytes exposed to 40 microM DMBA, whereas 40 microM BaP inhibited only B6C3F1 splenocyte proliferation to LPS. These data suggest that DMBA may act on immunocytes by mechanisms largely independent of the Ah locus and associated metabolic processes.
authors.affiliation: Department of Cellular and Molecular Toxicology, Chemical Industry Institute of Toxicology, Research Triangle Park, North Carolina 27709.
authors.forename: L M
authors.initials: LM
authors.lastname: Thurmond
authors.forename: R V
authors.initials: RV
authors.lastname: House
authors.forename: L D
authors.initials: LD
authors.lastname: Lauer
authors.forename: J H
authors.initials: JH
authors.lastname: Dean
mesh_terms: D015127:9,10-Dimethyl-1,2-benzanthracene / Q000378:metabolism / Q000633:toxicity*; D000818:Animals; D000917:Antibody Formation / Q000187:drug effects; D005260:Female; D007111:Immunity, Cellular / Q000187:drug effects; D066298:In Vitro Techniques; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D008815:Mice, Inbred Strains; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug / Q000378:metabolism; D013154:Spleen / Q000187:drug effects* / Q000276:immunology
chemical_list: D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug; D015127:9,10-Dimethyl-1,2-benzanthracene