12/15-Lipoxygenase deficiency protects mice from allergic airways inflammation and increases secretory IgA levels Amanda R. Hajek, BS, Alexa R. Lindley,

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12/15-Lipoxygenase deficiency protects mice from allergic airways inflammation and increases secretory IgA levels Amanda R. Hajek, BS, Alexa R. Lindley, BS, Silvio Favoreto, PhD, Roderick Carter, BS, Robert P. Schleimer, PhD, Douglas A. Kuperman, PhD Journal of Allergy and Clinical Immunology Volume 122, Issue 3, Pages 633-639.e3 (September 2008) DOI: 10.1016/j.jaci.2008.06.021 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 12/15-LO deficiency protects mice from allergic sensitization induced by airways exposure to allergen. Levels of transcripts encoding 12/15-LO in the lungs of wild-type (WT) and STAT6−/−(S6−/−) mice are shown (A). Levels of ovalbumin (Ova)–specific IgE (B), ovalbumin-specific IgG1 (C), and ovalbumin-specific IgG2a (D) in serum and transcripts encoding IL-4, IL-13, IFN-γ, Muc5ac, and Gob-5 (E) in lungs of wild-type (WT) and 12/15-LO−/−(KO) mice are also shown. ∗P < .05 versus PBS. †P < .05 versus ovalbumin. Journal of Allergy and Clinical Immunology 2008 122, 633-639.e3DOI: (10.1016/j.jaci.2008.06.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 12/15-LO deficiency does not protect mice from allergic sensitization induced by intraperitoneal exposure to allergen. Levels of ovalbumin (Ova)–specific IgE (A), ovalbumin-specific IgG1 (B), and ovalbumin-specific IgG2a (C) in serum and transcripts encoding IL-4, IL-13, and IFN-γ (D) in the lungs of wild-type (WT) and 12/15-LO−/−(KO) mice are shown. ∗P < .05 versus PBS. †P < .05 versus ovalbumin. Journal of Allergy and Clinical Immunology 2008 122, 633-639.e3DOI: (10.1016/j.jaci.2008.06.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 12/15-LO deficiency increases lung IgA levels but does not alter lung permeability. Levels of BSA (A), albumin (B), IgG (C), and IgA (D) in BALF are shown. WT, Wild-type mice; KO, 12/15-LO−/− mice. ∗P < .001 versus wild-type mice. Journal of Allergy and Clinical Immunology 2008 122, 633-639.e3DOI: (10.1016/j.jaci.2008.06.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 12/15-LO deficiency and 12/15-LO inhibition increases SIgA levels. Levels of SC (A) and IgA (B) in BALF and IgA in serum (C) from wild-type (WT), 12/15-LO−/−(KO), and AKR/J (a), C57Bl/6 (c), BALB/C (b), and 129/J (1) mice are shown. Levels of IgA in BALF from wild-type mice treated with a pharmacologic inhibitor of 12/15-LO are also shown (D). ∗P < .05 versus all other groups. Journal of Allergy and Clinical Immunology 2008 122, 633-639.e3DOI: (10.1016/j.jaci.2008.06.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 12/15-LO deficiency increases mucosal-specific production of IgA. Levels of IgA in 5 consecutive bronchoalveolar lavage washings (A) and IgA in supernatants from repeatedly washed blood-free homogenized lungs (B) are shown. ∗P < 0.001 versus wild-type mice. Journal of Allergy and Clinical Immunology 2008 122, 633-639.e3DOI: (10.1016/j.jaci.2008.06.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 12/15-LO deficiency increases expression of transcripts encoding the mucosal IgA system. A, Representative (n = 6) levels of IgA transcripts detected in lung tissue from wild-type (WT; upper row) and 12/15-LO−/− (KO; lower row) mice. Levels of transcripts encoding J chain (B), TACI (C), BCMA (D), pIgR (E), and Clara cell secretory protein (CCSP; F) in lung tissue are also shown. ∗P < .001 versus wild-type mice. Journal of Allergy and Clinical Immunology 2008 122, 633-639.e3DOI: (10.1016/j.jaci.2008.06.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12/15-LO deficiency decreases HETE and increases CysLT levels 12/15-LO deficiency decreases HETE and increases CysLT levels. Levels of 12(S)-HETE (A) and 15(S)-HETE (B) and the combined levels of the CysLTs, including LTC4, LTD4, and LTE4(C), detected in BALF are shown. ∗P < .001 versus wild-type mice. Journal of Allergy and Clinical Immunology 2008 122, 633-639.e3DOI: (10.1016/j.jaci.2008.06.021) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions