The document summarizes various mechanisms that Staphylococcus uses to evade the innate immune system, including proteins and molecules that inhibit complement activation, phagocytosis, chemotaxis, and cell activation. It lists the names and functions of several staphylococcal immune evasion proteins such as CHIPS, SSL proteins, FLIPr, SCIN, and aureolysin. The document also acknowledges former lab members and collaborators that have contributed to the research.

1. Staphylococcal Immune Evasion Jos van Strijp UMC Utrecht Netherlands Linkedin

2. Innate Immunity Antimicrobial peptides Complement Phagocytes EVOLUTIONARY CONSERVERED STRUCTURES Effectors Recognition

3. Innate Immunity Antimicrobial peptides Complement Phagocytes EVOLUTIONARY CONSERVERED STRUCTURES Effectors Recognition Flagellum LPS Lipoproteins TLR2-TLR6 TLR1-TLR2 TLR5 CD14 TLR4 MD2 Formyl peptides FPR Carbohydrates Peptidoglycan MBL & ficolins

4. C5a Lectin Pathway Alternative Pathway C3a C5b C5b-9 C3bBbC3b C4b2aC3b C4b2a C3bBb C3 C3b FD FB FH FI C5 C3 Properdin H20 Classical Pathway Phagocytosis, Killing Ag presentation Chemotaxis Phagocyte activation Membrane Attack Complex

5. Rolling C3b/iC3b C5a Chemotaxis Activation Diapedesis Firm Adhesion Priming Phagocytosis Killing Adaptive Immunity CP LP AP C3a C5b C5b-9 C3bBbC3b C4b2aC3b C4b2a C3bBb C3 C3b FD FB FH FI C5 C3 fPep Neutrophil Blood vessel Bacteria Properdin H20

6. Innate Immunity In Vitro

7. C5aR FPR CHIPS 121 aa, 14.1 KDa secreted protein pI 9.3 CHemotaxis Inhibitory Protein of Staphylococci C5a C5a fMLP Blocks Migration Blocks Cell Activation Infect Immun 2001 J Exp Med 2004 J Immunol 2004 J Immunol 2004 J Biol Chem 2005 J Mol Biol 2005 Trends Microbiol 2005 J Bacteriol 2006 BMC Immunol 2009 J Biol Chem 2009 Veldkamp De Haas, Veldkamp Postma Haas Postma Haas Rooijakkers Van Wamel Gustafsson Ippel, de Haas

8. FPR FPRL1 Pratt, van Kessel J Immunol 2006 Pratt, van Kessel J Immunol 2009 Stemerding , van Kessel Submitted Hadders, de Haas Submitted FLIPr FLIPr-like Fc  R-I Fc  R-IIa,c Fc  R-IIb CD32 CD64 Fc  R-IIIa Fc  R-IIIb CD16 Inhibits Migration Inhibits Cell Activation Inhibits Phagocytosis

9. Van Wamel J Bacteriol, 2006 Rooijakkers Nature Immunology, 2005 Rooijakkers Cell Microbiol, 2006 Rooijakkers, Milder J Immunol, 2007 Rooijakkers Nature Immunology 2009 Jongerius J Immunol 2009 SCIN-B SCIN-C C4b2a C3bBb C3 C3b SCIN Inhibits Complement Inhibits Phagocytosis

10. Rooijakkers… Microbes Infection, 2005 SAK C3b Bb C3b C4b2a C3b Jongerius J Exp Med, 2007 Jongerius J Biol Chem 2010 C5a C5 Efb Ecb Inhibits Phagocytosis Inhibits Complement Inhibits Cell Activation

11. Bestebroer…de Haas Blood, 2006 Bestebroer…de Haas Blood, 2009 SSL-5 SSL-7 Bestebroer Cell Microbiol 2010 SSL-10 Walenkamp…de Haas, Neoplasia 2009 CXCR4 C5 IgA Inhibits Rolling Inhibits Migration Inhibits Complement Inhibits Cell Activation

12. Mechanism is serum dependent (FH,FI) Serum Serum+Aur C3+Aur 0 5 10 30 60 Time (minutes) C3 α -chain C3 α -chain C3 α -chain AUREOLYSIN CLEAVES C3 Secreted Metalloprotease, 33kD) C3a’ C3 C3b’ Laarman 2010 submitted BlocksComplement Blocks Phagocytosis

13. Rolling C3b/iC3b C5a Chemotaxis Activation Diapedesis Firm Adhesion Priming Phagocytosis Killing Adaptive Immunity CP LP AP C3a C5b C5b-9 C3bBbC3b C4b2aC3b C4b2a C3bBb C3 C3b FD FB FH FI C5 C3 fPep Neutrophil Blood vessel Bacteria Properdin H20

14. Innate Immunity In Vitro

16. SBI SAK SSL-7 Prot A SCIN Efb CHIPS SCIN-B SCIN-C SSL-7 Ecb SCIN SCIN-B SCIN-C Efb SBI SAK CHIPS SSL-6 SSL-5 SSL-5 FLIPr-like FLIPr Eap Efb Ecb Ecb FLIPr-like FLIPr SAK SSL10 Efb Ecb SSL8 Prot A Bestebroer… van Strijp FEMS Microbiol Rev 2009 Serruto…van Strijp, Nature Rev Microbiol 2010 SSL-10 aureolysin SSL-10 SSL-10

17. FORMER LAB MEMBERS Jovanka Bestebroer Ingrid Boer Marleen Huigen Ingrid Koggel Niki Georgiou Andras Vida Frank Coenjaerts Miriam Poppelier Henk van Leeuwen Cristina Pratt Toon van Bommel Bent Postma Miranda Boonstra Willem van Wamel Andreas Peschel Karin Ellen Veldkamp Jörg Köhl Cincinnati, USA Eric Prossnitz Albuquerque Andreas Peschel Tuebingen, Ger Victor Nizet San Diego USA Magnus Hook Houston USA Gerard Lina Lyon Fr Mal Horsburgh Liverpool UK Piet Gros Crystall. UU Rob Liskamp Pharmacy, UU Jan van de Winkel UMCU Corne Pieterse Phytopath, UU Victor Rutten UU Timo Breit UVA Experimental Microbiology COLLABORATORS NICOLE SMITS MAARTEN SCHIJFFELEN MARK JANSSEN SERGEY KONSTANTINOV EDWIN BOEL AD FLUIT SSL’s PIET AERTS MIGNON VUGHTS BAS SUREWAARD ANNEMIEK WALENKAMP CARLA DE HAAS Evasion ERIK HEEZIUS THUY-NGA LE WILLEMIEN VAN ROOIJEN REINDERT NIJLAND PIETER-JAN HAAS CINDY FEVRE support: Mobile Elements & Genomics j.vanstrijp umcutrecht.nl Phagocytosis Evasion ANNETTE STEMERDING ANDRAS SPAAN ANNEMARIE KUIPERS BART BARDOEL FIN MILDER KOK VAN KESSEL MAARTJE RUYKEN ALEX LAARMAN ILSE JONGERIUS SUZAN ROOIJAKKERS Complement Evasion