B-lymphocytes as Key Players in Chemical-Induced Asthma.docVIP

B-lymphocytes as Key Players in Chemical-Induced Asthma Vanessa De Vooght , Vincent Carlier , Fien C. Devos , Steven Haenen , Erik Verbeken , Benoit Nemery , 1 2 1 1 3 1 Peter H. M. Hoet , Jeroen A. J. Vanoirbeek 1 1* 1 Laboratory for Occupational and Environmental Toxicology, Leuven, Belgium, 2 Center for Molecular and Vascular Biology, Leuven, Belgium, 3 Translational Cell Tissue Research, KU Leuven, Leuven, Belgium Abstract T-lymphocytes and B-lymphocytes are key players in allergic asthma, with B-lymphocytes producing antigen-specific immunoglobulins E (IgE). We used a mouse model of chemical-induced asthma and transferred B-lymphocytes from sensitized animals into na?ve wild type mice, B-lymphocyte knock-out (B-KO) mice or severe combined immunodeficiency (SCID) mice. On days 1 and 8, BALB/c mice were dermally sensitized with 0.3% toluene diisocyanate (TDI) (20μl/ear). On day 15, mice were euthanized and the auricular lymph nodes isolated. B- lymphocytes (CD19 ) were separated from the whole cell suspension and 175,000 cells were injected in the tail vein + of na?ve wild type, B-KO or SCID mice. Three days later, the mice received a single oropharyngeal challenge with 0.01% TDI (20μl) or vehicle (acetone/olive oil (AOO)) (controls). Airway reactivity to methacholine and total and differential cell counts in the bronchoalveolar lavage (BAL) fluid were measured 24 hours after challenge. B- lymphocytes of AOO or TDI-sensitized mice were characterized for the expression of surface markers and production of cytokines. We found that transfer of B-cells obtained from mice dermally sensitized to toluene diisocyanate (TDI) into na?ve wild type mice, B-