PI3Kβ Plays a Critical Role in Neutrophil Activation by Immune Complexes

Suhasini Kulkarni, Cassian Sitaru, Zoltan Jakus, Karen E. Anderson, George Damoulakis, Keith Davidson, Misa Hirose, Jatinder Juss, David Oxley, Tamara A. M. Chessa, Faruk Ramadani, Herve Guillou, Anne Segonds-Pichon, Anja Fritsch, Gavin E. Jarvis, Klaus Okkenhaug, Ralf Ludwig, Detlef Zillikens, Attila Mocsai, Bart Vanhaesebroeck, Len R. Stephens,* Phillip T. Hawkins*

*To whom correspondence should be addressed. E-mail: phillip.hawkins{at}bbsrc.ac.uk (P.T.H.); len.stephens{at}bbsrc.ac.uk (L.R.S.)

This PDF file includes:

• Materials and Methods
• Fig. S1. Analysis of p110 and p85 protein abundance in p110 mutant mice.
• Fig. S2. βKD BMNs are defective in their ability to produce ROS in response to ICs.
• Fig. S3. The PI3Kδ inhibitor, IC87114, and the PI3Kγ inhibitor, AS605240, exhibit off-target activities in IC-mediated mouse BMN responses.
• Fig. S4. Inhibition of p110β does not affect the surface expression of FcγR or β2 integrin on resting or stimulated neutrophils.
• Fig. S5. Lack of p110β activity does not markedly affect BMN adhesion or spreading on ICs.
• Fig. S6. PI3Kβ, but not PI3Kδ or PI3Kγ, activity is required for ROS production by BMNs in response to adhesion to integrin ligands.
• Fig. S7. Normal rabbit anti-mcVII-IgG and C3 deposition in mouse skin, and serum IgG amounts in wild-type and βKO mice.
• Fig. S8. Skin blistering in βKO mouse skin upon local reconstitution with wild-type mouse neutrophils.
• Fig. S9. PI3Kβ is important in promoting ROS formation in response to autoimmune sera from EBA patients.
• Fig. S10. Immunoreactivity of normal human or EBA patient sera toward recombinant human collagen VII (hcVII) as assessed by enzyme-linked immunosorbent assay (ELISA).
• Fig. S11. Combined deficiency of p110β and p110δ activities confers substantial protection from arthritic symptoms in the K/BxN model of autoimmune arthritis.
• Fig. S12. Combined deficiency of p110β and p110δ activities confers substantial protection from joint swelling in the K/BxN model of autoimmune arthritis.
• Fig. S13. Normal Akt phosphorylation and antibody production in mice with lymphocytic deletion of p110β.
• Fig. S14. LTB₄-dependent ROS production and Akt phosphorylation in BMNs.
• Fig. S15. PI3Kβ is cooperatively engaged by FcγRs and the GPCR, BLT1, to drive sustained neutrophil responses.
• Table S1. Hematological analysis of βKO mouse peripheral blood.
• Table S2. Isoforms of p110 identified in p85 immunoprecipitates by mass spectrometry.
  

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© 2011 American Association for the Advancement of Science