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1. Agnostic B cell selection approach identifies antibodies against K. pneumoniae that synergistically drive complement activation

Author

van der Lans, Sjors P. A., Bardoel, Bart W., Ruyken, Maartje, de Haas, Carla J. C., Baijens, Stan, Muts, Remy M., Scheepmaker, Lisette M., Aerts, Piet C., van ’t Wout, Marije F. L., Preiner, Johannes, Marijnissen, Renoud J., Schuurman, Janine, Beurskens, Frank J., Kerkman, Priscilla F., and Rooijakkers, Suzan H. M.

Published
2024
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8. Agnostic B cell selection approach identifies antibodies against K. pneumoniae that synergistically drive complement activation

Author

MMB Research line 1, Infection & Immunity, MMB Onderzoek en Onderwijs, CTI, van der Lans, Sjors P A, Bardoel, Bart W, Ruyken, Maartje, de Haas, Carla J C, Baijens, Stan, Muts, Remy M, Scheepmaker, Lisette M, Aerts, Piet C, van 't Wout, Marije F L, Preiner, Johannes, Marijnissen, Renoud J, Schuurman, Janine, Beurskens, Frank J, Kerkman, Priscilla F, Rooijakkers, Suzan H M, MMB Research line 1, Infection & Immunity, MMB Onderzoek en Onderwijs, CTI, van der Lans, Sjors P A, Bardoel, Bart W, Ruyken, Maartje, de Haas, Carla J C, Baijens, Stan, Muts, Remy M, Scheepmaker, Lisette M, Aerts, Piet C, van 't Wout, Marije F L, Preiner, Johannes, Marijnissen, Renoud J, Schuurman, Janine, Beurskens, Frank J, Kerkman, Priscilla F, and Rooijakkers, Suzan H M

Published
2024

9. Klebsiella LPS O1-antigen prevents complement-mediated killing by inhibiting C9 polymerization

Author

MMB Research line 1, Infection & Immunity, Masson, Frerich M, Káradóttir, Salvör, van der Lans, Sjors P A, Doorduijn, Dennis J, de Haas, Carla J C, Rooijakkers, Suzan H M, Bardoel, Bart W, MMB Research line 1, Infection & Immunity, Masson, Frerich M, Káradóttir, Salvör, van der Lans, Sjors P A, Doorduijn, Dennis J, de Haas, Carla J C, Rooijakkers, Suzan H M, and Bardoel, Bart W

Published
2024

12. Human CD45 is an F-component-specific receptor for the staphylococcal toxin Panton–Valentine leukocidin

Author

Tromp, Angelino T., Van Gent, Michiel, Abrial, Pauline, Martin, Amandine, Jansen, Joris P., De Haas, Carla J. C., Van Kessel, Kok P. M., Bardoel, Bart W., Kruse, Elisabeth, Bourdonnay, Emilie, Boettcher, Michael, McManus, Michael T., Day, Christopher J., Jennings, Michael P., Lina, Gérard, Vandenesch, François, Van Strijp, Jos A. G., Lebbink, Robert Jan, Haas, Pieter-Jan A., Henry, Thomas, and Spaan, András N.

Published
2018
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14. Artificial surface labelling of Escherichia coli with StrepTagII antigen to study how monoclonal antibodies drive complement-mediated killing

Author

Muts, Remy M, den Boer, Maurits A, Bardoel, Bart W, Aerts, Piet C, de Haas, Carla J C, Heck, Albert J R, Rooijakkers, Suzan H M, Heesterbeek, Dani A C, Muts, Remy M, den Boer, Maurits A, Bardoel, Bart W, Aerts, Piet C, de Haas, Carla J C, Heck, Albert J R, Rooijakkers, Suzan H M, and Heesterbeek, Dani A C

Abstract

Antibodies play a key role in the immune defence against Gram-negative bacteria. After binding to bacterial surface antigens, IgG and IgM can activate the complement system and trigger formation of lytic membrane attack complex (MAC) pores. Molecular studies to compare functional activity of antibodies on bacteria are hampered by the limited availability of well-defined antibodies against bacterial surface antigens. Therefore, we genetically engineered E. coli by expressing the StrepTagII antigen into outer membrane protein X (OmpX) and validated that these engineered bacteria were recognised by anti-StrepTagII antibodies. We then combined this antigen-antibody system with a purified complement assay to avoid interference of serum components and directly compare MAC-mediated bacterial killing via IgG1 and pentameric IgM. While both IgG1 and IgM could induce MAC-mediated killing, we show that IgM has an increased capacity to induce complement-mediated killing of E. coli compared to IgG1. While Fc mutations that enhance IgG clustering after target binding could not improve MAC formation, mutations that cause formation of pre-assembled IgG hexamers enhanced the complement activating capacity of IgG1. Altogether, we here present a system to study antibody-dependent complement activation on E. coli and show IgM's enhanced capacity over IgG to induce complement-mediated lysis of E. coli.

Published
2023

15. Artificial surface labelling of Escherichia coli with StrepTagII antigen to study how monoclonal antibodies drive complement-mediated killing

Author

Afd Biomol.Mass Spect. and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., Biomolecular Mass Spectrometry and Proteomics, Muts, Remy M, den Boer, Maurits A, Bardoel, Bart W, Aerts, Piet C, de Haas, Carla J C, Heck, Albert J R, Rooijakkers, Suzan H M, Heesterbeek, Dani A C, Afd Biomol.Mass Spect. and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., Biomolecular Mass Spectrometry and Proteomics, Muts, Remy M, den Boer, Maurits A, Bardoel, Bart W, Aerts, Piet C, de Haas, Carla J C, Heck, Albert J R, Rooijakkers, Suzan H M, and Heesterbeek, Dani A C

Published
2023

16. Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye

Author

Egido, Julia E. (author), Toner-Bartelds, Catherine (author), Martins Costa, A.R. (author), Brouns, S.J.J. (author), Rooijakkers, Suzan H.M. (author), Bardoel, Bart W. (author), Haas, Pieter Jan (author), Egido, Julia E. (author), Toner-Bartelds, Catherine (author), Martins Costa, A.R. (author), Brouns, S.J.J. (author), Rooijakkers, Suzan H.M. (author), Bardoel, Bart W. (author), and Haas, Pieter Jan (author)

Abstract

Bacteriophages (phages) are viruses that specifically attack bacteria. Their use as therapeutics, which constitutes a promising alternative to antibiotics, heavily relies on selecting effective lytic phages against the pathogen of interest. Current selection techniques are laborious and do not allow for direct visualization of phage infection dynamics. Here, we present a method that circumvents these limitations. It can be scaled for high-throughput and permits monitoring of the phage infection in real time via a fluorescence signal readout. This is achieved through the use of a membrane-impermeant nucleic acid dye that stains the DNA of damaged or lysed bacteria and new phage progeny. We have tested the method on Pseudomonas aeruginosa and Klebsiella pneumoniae and show that an increase in fluorescence reflects phage-mediated killing. This is confirmed by other techniques including spot tests, colony plating, flow cytometry and metabolic activity measurements. Furthermore, we illustrate how our method may be used to compare the activity of different phages and to screen the susceptibility of clinical isolates to phage. Altogether, we present a fast, reliable way of selecting phages against Gram-negative bacteria, which may be valuable in optimizing the process of selecting phages for therapeutic use., BN/Stan Brouns Lab

Published
2023
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17. Inhibition of cleavage of human complement component C5 and the R885H C5 variant by two distinct high affinity anti-C5 nanobodies

Author

MMB Research line 1, Infection & Immunity, Struijf, Eva M, De la O Becerra, Karla I, Ruyken, Maartje, de Haas, Carla J C, van Oosterom, Fleur, Siere, Danique Y, van Keulen, Joanne E, Heesterbeek, Dani A C, Dolk, Edward, Heukers, Raimond, Bardoel, Bart W, Gros, Piet, Rooijakkers, Suzan H M, MMB Research line 1, Infection & Immunity, Struijf, Eva M, De la O Becerra, Karla I, Ruyken, Maartje, de Haas, Carla J C, van Oosterom, Fleur, Siere, Danique Y, van Keulen, Joanne E, Heesterbeek, Dani A C, Dolk, Edward, Heukers, Raimond, Bardoel, Bart W, Gros, Piet, and Rooijakkers, Suzan H M

Published
2023

18. Colistin resistance mutations in phoQ can sensitize Klebsiella pneumoniae to IgM-mediated complement killing

Author

Infection & Immunity, MMB Research line 1, MMB Research line 2, van der Lans, Sjors P A, Janet-Maitre, Manon, Masson, Frerich M, Walker, Kimberly A, Doorduijn, Dennis J, Janssen, Axel B, van Schaik, Willem, Attrée, Ina, Rooijakkers, Suzan H M, Bardoel, Bart W, Infection & Immunity, MMB Research line 1, MMB Research line 2, van der Lans, Sjors P A, Janet-Maitre, Manon, Masson, Frerich M, Walker, Kimberly A, Doorduijn, Dennis J, Janssen, Axel B, van Schaik, Willem, Attrée, Ina, Rooijakkers, Suzan H M, and Bardoel, Bart W

Published
2023

19. Human Complement Inhibits Myophages against Pseudomonas aeruginosa

Author

MMB Research line 1, Infection & Immunity, MMB Medische Staf, Egido, Julia E, Dekker, Simon O, Toner-Bartelds, Catherine, Lood, Cédric, Rooijakkers, Suzan H M, Bardoel, Bart W, Haas, Pieter-Jan, MMB Research line 1, Infection & Immunity, MMB Medische Staf, Egido, Julia E, Dekker, Simon O, Toner-Bartelds, Catherine, Lood, Cédric, Rooijakkers, Suzan H M, Bardoel, Bart W, and Haas, Pieter-Jan

Published
2023

20. Artificial surface labelling of Escherichia coli with StrepTagII antigen to study how monoclonal antibodies drive complement-mediated killing

Author

MMB Research line 1, Infection & Immunity, Muts, Remy M, den Boer, Maurits A, Bardoel, Bart W, Aerts, Piet C, de Haas, Carla J C, Heck, Albert J R, Rooijakkers, Suzan H M, Heesterbeek, Dani A C, MMB Research line 1, Infection & Immunity, Muts, Remy M, den Boer, Maurits A, Bardoel, Bart W, Aerts, Piet C, de Haas, Carla J C, Heck, Albert J R, Rooijakkers, Suzan H M, and Heesterbeek, Dani A C

Published
2023

21. Genome-wide screen in human plasma identifies multifaceted complement evasion of Pseudomonas aeruginosa

Author

Medical Microbiology, MMB Research line 1, Infection & Immunity, Janet-Maitre, Manon, Pont, Stéphane, Masson, Frerich M., Sleiman, Serena, Trouillon, Julian, Robert-Genthon, Mylène, Gallet, Benoît, Dumestre-Perard, Chantal, Elsen, Sylvie, Moriscot, Christine, Bardoel, Bart W., Rooijakkers, Suzan H.M., Cretin, François, Attrée, Ina, Medical Microbiology, MMB Research line 1, Infection & Immunity, Janet-Maitre, Manon, Pont, Stéphane, Masson, Frerich M., Sleiman, Serena, Trouillon, Julian, Robert-Genthon, Mylène, Gallet, Benoît, Dumestre-Perard, Chantal, Elsen, Sylvie, Moriscot, Christine, Bardoel, Bart W., Rooijakkers, Suzan H.M., Cretin, François, and Attrée, Ina

Published
2023

22. Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye

Author

Medical Microbiology, MMB Research line 1, Infection & Immunity, MMB Medische Staf, Egido, Julia E., Toner-Bartelds, Catherine, Costa, Ana Rita, Brouns, Stan J.J., Rooijakkers, Suzan H.M., Bardoel, Bart W., Haas, Pieter Jan, Medical Microbiology, MMB Research line 1, Infection & Immunity, MMB Medische Staf, Egido, Julia E., Toner-Bartelds, Catherine, Costa, Ana Rita, Brouns, Stan J.J., Rooijakkers, Suzan H.M., Bardoel, Bart W., and Haas, Pieter Jan

Published
2023

23. Inhibition of cleavage of human complement component C5 and the R885H C5 variant by two distinct high affinity anti-C5 nanobodies

Author

Struijf, Eva M., primary, De la O Becerra, Karla I, additional, Ruyken, Maartje, additional, van Oosterom, Fleur, additional, Siere, Danique Y., additional, Heesterbeek, Dani A. C., additional, Dolk, Edward, additional, Heukers, Raimond, additional, Bardoel, Bart W., additional, Gros, Piet, additional, and Rooijakkers, Suzan H.M., additional

Published
2023
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24. Genome-wide screen in human plasma identifies multifaceted complement evasion of Pseudomonas aeruginosa

Author

Janet-Maitre, Manon, primary, Pont, Stéphane, additional, Masson, Frerich M., additional, Sleiman, Serena, additional, Trouillon, Julian, additional, Robert-Genthon, Mylène, additional, Gallet, Benoît, additional, Dumestre-Perard, Chantal, additional, Elsen, Sylvie, additional, Moriscot, Christine, additional, Bardoel, Bart W., additional, Rooijakkers, Suzan H. M., additional, Cretin, François, additional, and Attrée, Ina, additional

Published
2023
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27. Selective Exoenzymatic Labeling of Lipooligosaccharides of Neisseria gonorrhoeae with α2,6-Sialoside Analogues

Author

de Jong, Hanna, Moure, Maria J., Hartman, Jet E. M., Bosman, Gerlof P., Ong, Jun Yang, Bardoel, Bart W., Boons, Geert-Jan, Wosten, Marc M. S. M., Wennekes, Tom, Afd Chemical Biology and Drug Discovery, Sub Chemical Biology and Drug Discovery, Infectiebiologie, Chemical Biology and Drug Discovery, Afd Chemical Biology and Drug Discovery, Sub Chemical Biology and Drug Discovery, Infectiebiologie, and Chemical Biology and Drug Discovery

Subjects
Lipopolysaccharides, lipooligosaccharides, Organic Chemistry, Polysaccharides, Bacterial, Glycosyltransferases, glycoengineering, Biochemistry, N-Acetylneuraminic Acid, Neisseria gonorrhoeae, glycobiology, sialic acid, Cytidine Monophosphate N-Acetylneuraminic Acid, Sialic Acids, Molecular Medicine, Molecular Biology
Abstract

The interactions between bacteria and their host often rely on recognition processes that involve host or bacterial glycans. Glycoengineering techniques make it possible to modify and study the glycans on the host's eukaryotic cells, but only a few are available for the study of bacterial glycans. Here, we have adapted selective exoenzymatic labeling (SEEL), a chemical reporter strategy, to label the lipooligosaccharides of the bacterial pathogen Neisseria gonorrhoeae, using the recombinant glycosyltransferase ST6Gal1, and three synthetic CMP-sialic acid derivatives. We show that SEEL treatment does not affect cell viability and can introduce an α2,6-linked sialic acid with a reporter group on the lipooligosaccharides by Western blot, flow cytometry and fluorescent microscopy. This new bacterial glycoengineering technique allows for the precise modification, here with α2,6-sialoside derivatives, and direct detection of specific surface glycans on live bacteria, which will aid in further unravelling the precise biological functions of bacterial glycans.

Published
2022

30. Soluble MAC is primarily released from MAC-resistant bacteria that potently convert complement component C5

Author

Doorduijn, Dennis J., Lukassen, Marie V., van 't Wout, Marije F.L., Franc, Vojtech, Ruyken, Maartje, Bardoel, Bart W., Heck, Albert J.R., Rooijakkers, Suzan H.M., Afd Biomol.Mass Spect. and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., Biomolecular Mass Spectrometry and Proteomics, Afd Biomol.Mass Spect. and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., and Biomolecular Mass Spectrometry and Proteomics

Subjects
General Immunology and Microbiology, Biochemistry, Genetics and Molecular Biology(all), Neuroscience(all), General Neuroscience, Complement C5, Complement Membrane Attack Complex, General Medicine, Gram-Positive Bacteria, Biochemistry, General Biochemistry, Genetics and Molecular Biology, Immunology and Microbiology(all), Escherichia coli, Humans, Vitronectin, biological phenomena, cell phenomena, and immunity, Complement Activation, Escherichia coli Infections, Genetics and Molecular Biology(all)
Abstract

The Membrane Attack Complex (MAC or C5b-9) is an important effector of the immune system to kill invading microbes. MAC is formed when complement enzymes on the bacterial surface convert complement component C5 into C5b. Although the MAC is a membrane-inserted complex, soluble forms of MAC (sMAC, or terminal complement complex (TCC)) are often detected in sera of patients suffering from infections. Consequently, sMAC has been proposed as a biomarker, but it remains unclear when and how it is formed during infections. Here, we studied mechanisms of MAC formation on bacteria and found that sMAC is primarily formed in human serum by bacteria resistant to MAC-dependent killing. Surprisingly, C5 was converted into C5b more potently by MAC-resistant compared to MAC-sensitiveEscherichia colistrains. Both the increase in C5 conversion and sMAC generation were linked to the expression of lipopolysaccharide (LPS) O-Antigen in the bacterial outer membrane. In addition, we found that MAC precursors are released from the surface of MAC-resistant bacteria during MAC assembly. Release of MAC precursors from bacteria induced lysis of bystander human erythrocytes in the absence of other serum components. However, serum regulators vitronectin (Vn) and clusterin (Clu) can prevent this bystander lysis. Combining size exclusion chromatography with mass spectrometry profiling, we show that sMAC released from bacteria in serum is a heterogeneous mixture of complexes composed of C5b-8, up to 3 copies of C9 and multiple copies of Vn and Clu. Altogether, our data provide molecular insight into how sMAC is generated during bacterial infections. This fundamental knowledge could form the basis for exploring the use of sMAC as biomarker.

Published
2022
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34. Human OTULIN haploinsufficiency impairs cell-intrinsic immunity to staphylococcal α-toxin

Author

Spaan, András N., primary, Neehus, Anna-Lena, additional, Laplantine, Emmanuel, additional, Staels, Frederik, additional, Ogishi, Masato, additional, Seeleuthner, Yoann, additional, Rapaport, Franck, additional, Lacey, Keenan A., additional, Van Nieuwenhove, Erika, additional, Chrabieh, Maya, additional, Hum, David, additional, Migaud, Mélanie, additional, Izmiryan, Araksya, additional, Lorenzo, Lazaro, additional, Kochetkov, Tatiana, additional, Heesterbeek, Dani A. C., additional, Bardoel, Bart W., additional, DuMont, Ashley L., additional, Dobbs, Kerry, additional, Chardonnet, Solenne, additional, Heissel, Søren, additional, Baslan, Timour, additional, Zhang, Peng, additional, Yang, Rui, additional, Bogunovic, Dusan, additional, Wunderink, Herman F., additional, Haas, Pieter-Jan A., additional, Molina, Henrik, additional, Van Buggenhout, Griet, additional, Lyonnet, Stanislas, additional, Notarangelo, Luigi D., additional, Seppänen, Mikko R. J., additional, Weil, Robert, additional, Seminario, Gisela, additional, Gomez-Tello, Héctor, additional, Wouters, Carine, additional, Mesdaghi, Mehrnaz, additional, Shahrooei, Mohammad, additional, Bossuyt, Xavier, additional, Sag, Erdal, additional, Topaloglu, Rezan, additional, Ozen, Seza, additional, Leavis, Helen L., additional, van Eijk, Maarten M. J., additional, Bezrodnik, Liliana, additional, Blancas Galicia, Lizbeth, additional, Hovnanian, Alain, additional, Nassif, Aude, additional, Bader-Meunier, Brigitte, additional, Neven, Bénédicte, additional, Meyts, Isabelle, additional, Schrijvers, Rik, additional, Puel, Anne, additional, Bustamante, Jacinta, additional, Aksentijevich, Ivona, additional, Kastner, Daniel L., additional, Torres, Victor J., additional, Humblet-Baron, Stéphanie, additional, Liston, Adrian, additional, Abel, Laurent, additional, Boisson, Bertrand, additional, and Casanova, Jean-Laurent, additional

Published
2022
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Author

de Jong, Hanna, Moure, Maria J., Hartman, Jet E. M., Bosman, Gerlof P., Ong, Jun Yang, Bardoel, Bart W., Boons, Geert-Jan, Wosten, Marc M. S. M., Wennekes, Tom, de Jong, Hanna, Moure, Maria J., Hartman, Jet E. M., Bosman, Gerlof P., Ong, Jun Yang, Bardoel, Bart W., Boons, Geert-Jan, Wosten, Marc M. S. M., and Wennekes, Tom

Published
2022

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Author

Afd Chemical Biology and Drug Discovery, Sub Chemical Biology and Drug Discovery, Infectiebiologie, Chemical Biology and Drug Discovery, de Jong, Hanna, Moure, Maria J., Hartman, Jet E. M., Bosman, Gerlof P., Ong, Jun Yang, Bardoel, Bart W., Boons, Geert-Jan, Wosten, Marc M. S. M., Wennekes, Tom, Afd Chemical Biology and Drug Discovery, Sub Chemical Biology and Drug Discovery, Infectiebiologie, Chemical Biology and Drug Discovery, de Jong, Hanna, Moure, Maria J., Hartman, Jet E. M., Bosman, Gerlof P., Ong, Jun Yang, Bardoel, Bart W., Boons, Geert-Jan, Wosten, Marc M. S. M., and Wennekes, Tom

Published
2022

37. Novel insights in antimicrobial and immunomodulatory mechanisms of action of PepBiotics CR-163 and CR-172

Author

Sub NMR Spectroscopy, Immunologie, Interne geneeskunde GD, CS_Welfare & emerging diseases, Moleculaire afweer, van Os, Nico, Javed, Ali, Broere, Femke, van Dijk, Albert, Balhuizen, Melanie D, van Eijk, Martin, Rooijakkers, Suzan H M, Bardoel, Bart W, Heesterbeek, Dani A C, Haagsman, Henk P, Veldhuizen, Edwin J A, Sub NMR Spectroscopy, Immunologie, Interne geneeskunde GD, CS_Welfare & emerging diseases, Moleculaire afweer, van Os, Nico, Javed, Ali, Broere, Femke, van Dijk, Albert, Balhuizen, Melanie D, van Eijk, Martin, Rooijakkers, Suzan H M, Bardoel, Bart W, Heesterbeek, Dani A C, Haagsman, Henk P, and Veldhuizen, Edwin J A

Published
2022

38. Soluble MAC is primarily released from MAC-resistant bacteria that potently convert complement component C5

Author

Afd Biomol.Mass Spect. and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., Biomolecular Mass Spectrometry and Proteomics, Doorduijn, Dennis J., Lukassen, Marie V., van 't Wout, Marije F.L., Franc, Vojtech, Ruyken, Maartje, Bardoel, Bart W., Heck, Albert J.R., Rooijakkers, Suzan H.M., Afd Biomol.Mass Spect. and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., Biomolecular Mass Spectrometry and Proteomics, Doorduijn, Dennis J., Lukassen, Marie V., van 't Wout, Marije F.L., Franc, Vojtech, Ruyken, Maartje, Bardoel, Bart W., Heck, Albert J.R., and Rooijakkers, Suzan H.M.

Published
2022

39. Human OTULIN haploinsufficiency impairs cell-intrinsic immunity to staphylococcal a-toxin

Author

MMB, Infection & Immunity, Immuno/reuma patientenzorg, MMB Research line 1, MMB Medische Staf, MS Reumatologie/Immunologie/Infectie, Medische Staf Intensive Care, Spaan, András N., Neehus, Anna Lena, Laplantine, Emmanuel, Staels, Frederik, Ogishi, Masato, Seeleuthner, Yoann, Rapaport, Franck, Lacey, Keenan A., Van Nieuwenhove, Erika, Chrabieh, Maya, Hum, David, Migaud, Mélanie, Izmiryan, Araksya, Lorenzo, Lazaro, Kochetkov, Tatiana, Heesterbeek, Dani A.C., Bardoel, Bart W., DuMont, Ashley L., Dobbs, Kerry, Chardonnet, Solenne, Heissel, Søren, Baslan, Timour, Zhang, Peng, Yang, Rui, Bogunovic, Dusan, Wunderink, Herman F., Haas, Pieter Jan A., Molina, Henrik, Van Buggenhout, Griet, Lyonnet, Stanislas, Notarangelo, Luigi D., Seppänen, Mikko R.J., Weil, Robert, Seminario, Gisela, Gomez-Tello, Héctor, Wouters, Carine, Mesdaghi, Mehrnaz, Shahrooei, Mohammad, Bossuyt, Xavier, Sag, Erdal, Topaloglu, Rezan, Ozen, Seza, Leavis, Helen L., van Eijk, Maarten M.J., Bezrodnik, Liliana, Galicia, Lizbeth Blancas, Hovnanian, Alain, Nassif, Aude, Bader-Meunier, Brigitte, Neven, Bénédicte, Meyts, Isabelle, Schrijvers, Rik, Puel, Anne, Bustamante, Jacinta, Aksentijevich, Ivona, Kastner, Daniel L., Torres, Victor J., Humblet-Baron, Stéphanie, Liston, Adrian, Abel, Laurent, Boisson, Bertrand, Casanova, Jean Laurent, MMB, Infection & Immunity, Immuno/reuma patientenzorg, MMB Research line 1, MMB Medische Staf, MS Reumatologie/Immunologie/Infectie, Medische Staf Intensive Care, Spaan, András N., Neehus, Anna Lena, Laplantine, Emmanuel, Staels, Frederik, Ogishi, Masato, Seeleuthner, Yoann, Rapaport, Franck, Lacey, Keenan A., Van Nieuwenhove, Erika, Chrabieh, Maya, Hum, David, Migaud, Mélanie, Izmiryan, Araksya, Lorenzo, Lazaro, Kochetkov, Tatiana, Heesterbeek, Dani A.C., Bardoel, Bart W., DuMont, Ashley L., Dobbs, Kerry, Chardonnet, Solenne, Heissel, Søren, Baslan, Timour, Zhang, Peng, Yang, Rui, Bogunovic, Dusan, Wunderink, Herman F., Haas, Pieter Jan A., Molina, Henrik, Van Buggenhout, Griet, Lyonnet, Stanislas, Notarangelo, Luigi D., Seppänen, Mikko R.J., Weil, Robert, Seminario, Gisela, Gomez-Tello, Héctor, Wouters, Carine, Mesdaghi, Mehrnaz, Shahrooei, Mohammad, Bossuyt, Xavier, Sag, Erdal, Topaloglu, Rezan, Ozen, Seza, Leavis, Helen L., van Eijk, Maarten M.J., Bezrodnik, Liliana, Galicia, Lizbeth Blancas, Hovnanian, Alain, Nassif, Aude, Bader-Meunier, Brigitte, Neven, Bénédicte, Meyts, Isabelle, Schrijvers, Rik, Puel, Anne, Bustamante, Jacinta, Aksentijevich, Ivona, Kastner, Daniel L., Torres, Victor J., Humblet-Baron, Stéphanie, Liston, Adrian, Abel, Laurent, Boisson, Bertrand, and Casanova, Jean Laurent

Published
2022

40. Novel insights in antimicrobial and immunomodulatory mechanisms of action of PepBiotics CR-163 and CR-172

Author

Longziekten onderzoek 1, MMB Research line 1, Infection & Immunity, van Os, Nico, Javed, Ali, Broere, Femke, van Dijk, Albert, Balhuizen, Melanie D., van Eijk, Martin, Rooijakkers, Suzan H.M., Bardoel, Bart W., Heesterbeek, Dani A.C., Haagsman, Henk P., Veldhuizen, Edwin, Longziekten onderzoek 1, MMB Research line 1, Infection & Immunity, van Os, Nico, Javed, Ali, Broere, Femke, van Dijk, Albert, Balhuizen, Melanie D., van Eijk, Martin, Rooijakkers, Suzan H.M., Bardoel, Bart W., Heesterbeek, Dani A.C., Haagsman, Henk P., and Veldhuizen, Edwin

Published
2022

41. Correction: Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores

Author

MMB Research line 1, Infection & Immunity, MMB, Doorduijn, Dennis J., Heesterbeek, Dani A.C., Ruyken, Maartje, de Haas, Carla J.C., Stapels, Daphne A.C., Aerts, Piet C., Rooijakkers, Suzan H.M., Bardoel, Bart W., MMB Research line 1, Infection & Immunity, MMB, Doorduijn, Dennis J., Heesterbeek, Dani A.C., Ruyken, Maartje, de Haas, Carla J.C., Stapels, Daphne A.C., Aerts, Piet C., Rooijakkers, Suzan H.M., and Bardoel, Bart W.

Published
2022

42. Soluble MAC is primarily released from MAC-resistant bacteria that potently convert complement component C5

Author

MMB Research line 1, Infection & Immunity, MMB, Doorduijn, Dennis J., Lukassen, Marie V., van 't Wout, Marije F.L., Franc, Vojtech, Ruyken, Maartje, Bardoel, Bart W., Heck, Albert J.R., Rooijakkers, Suzan H.M., MMB Research line 1, Infection & Immunity, MMB, Doorduijn, Dennis J., Lukassen, Marie V., van 't Wout, Marije F.L., Franc, Vojtech, Ruyken, Maartje, Bardoel, Bart W., Heck, Albert J.R., and Rooijakkers, Suzan H.M.

Published
2022

44. Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores

Author

Doorduijn, Dennis J., Meri, Seppo, Heesterbeek, Dani A. C., Ruyken, Maartje, Haas, Carla J. C. de, Stapels, Daphne A. C., Aerts, Piet C., Rooijakkers, Suzan H. M., Bardoel, Bart W., and Infectiebiologie

Abstract

Complement proteins can form membrane attack complex (MAC) pores that directly kill Gram-negative bacteria. MAC pores assemble by stepwise binding of C5b, C6, C7, C8 and finally C9, which can polymerize into a transmembrane ring of up to 18 C9 monomers. It is still unclear if the assembly of a polymeric-C9 ring is necessary to sufficiently damage the bacterial cell envelope to kill bacteria. In this paper, polymerization of C9 was prevented without affecting binding of C9 to C5b-8, by locking the first transmembrane helix domain of C9. Using this system, we show that polymerization of C9 strongly enhanced damage to both the bacterial outer and inner membrane, resulting in more rapid killing of several Escherichia coli and Klebsiella strains in serum. By comparing binding of wildtype and ‘locked’ C9 by flow cytometry, we also show that polymerization of C9 is impaired when the amount of available C9 per C5b-8 is limited. This suggests that an excess of C9 is required to efficiently form polymeric-C9. Finally, we show that polymerization of C9 was impaired on complement-resistant E. coli strains that survive killing by MAC pores. This suggests that these bacteria can specifically block polymerization of C9. All tested complement-resistant E. coli expressed LPS O-antigen (O-Ag), compared to only one out of four complement-sensitive E. coli. By restoring O-Ag expression in an O-Ag negative strain, we show that the O-Ag impairs polymerization of C9 and results in complement-resistance. Altogether, these insights are important to understand how MAC pores kill bacteria and how bacterial pathogens can resist MAC-dependent killing.

Published
2021

46. Publisher Correction: Human CD45 is an F-component-specific receptor for the staphylococcal toxin Panton–Valentine leukocidin

Author

Tromp, Angelino T., Van Gent, Michiel, Abrial, Pauline, Martin, Amandine, Jansen, Joris P., De Haas, Carla J. C., Van Kessel, Kok P. M., Bardoel, Bart W., Kruse, Elisabeth, Bourdonnay, Emilie, Boettcher, Michael, McManus, Michael T., Day, Christopher J., Jennings, Michael P., Lina, Gérard, Vandenesch, François, Van Strijp, Jos A. G., Lebbink, Robert Jan, Haas, Pieter-Jan A., Henry, Thomas, and Spaan, András N.

Published
2018
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48. Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores

Author

Meri, Seppo, Doorduijn, Dennis J., Heesterbeek, Dani A. C., Ruyken, Maartje, Haas, Carla J. C. de, Stapels, Daphne A. C., Aerts, Piet C., Rooijakkers, Suzan H. M., Bardoel, Bart W., Meri, Seppo, Doorduijn, Dennis J., Heesterbeek, Dani A. C., Ruyken, Maartje, Haas, Carla J. C. de, Stapels, Daphne A. C., Aerts, Piet C., Rooijakkers, Suzan H. M., and Bardoel, Bart W.

Abstract

Complement proteins can form membrane attack complex (MAC) pores that directly kill Gram-negative bacteria. MAC pores assemble by stepwise binding of C5b, C6, C7, C8 and finally C9, which can polymerize into a transmembrane ring of up to 18 C9 monomers. It is still unclear if the assembly of a polymeric-C9 ring is necessary to sufficiently damage the bacterial cell envelope to kill bacteria. In this paper, polymerization of C9 was prevented without affecting binding of C9 to C5b-8, by locking the first transmembrane helix domain of C9. Using this system, we show that polymerization of C9 strongly enhanced damage to both the bacterial outer and inner membrane, resulting in more rapid killing of several Escherichia coli and Klebsiella strains in serum. By comparing binding of wildtype and ‘locked’ C9 by flow cytometry, we also show that polymerization of C9 is impaired when the amount of available C9 per C5b-8 is limited. This suggests that an excess of C9 is required to efficiently form polymeric-C9. Finally, we show that polymerization of C9 was impaired on complement-resistant E. coli strains that survive killing by MAC pores. This suggests that these bacteria can specifically block polymerization of C9. All tested complement-resistant E. coli expressed LPS O-antigen (O-Ag), compared to only one out of four complement-sensitive E. coli. By restoring O-Ag expression in an O-Ag negative strain, we show that the O-Ag impairs polymerization of C9 and results in complement-resistance. Altogether, these insights are important to understand how MAC pores kill bacteria and how bacterial pathogens can resist MAC-dependent killing.

Published
2021

49. Outer membrane permeabilization by the membrane attack complex sensitizes Gram-negative bacteria to antimicrobial proteins in serum and phagocytes

Author

Heesterbeek, Dani A. C., Muts, Remy M., Hensbergen, Vincent P. van, Aulaire, Pieter de Saint, Wennekes, Tom, Bardoel, Bart W., Sorge, Nina M. van, Rooijakkers, Suzan H.M., Heesterbeek, Dani A. C., Muts, Remy M., Hensbergen, Vincent P. van, Aulaire, Pieter de Saint, Wennekes, Tom, Bardoel, Bart W., Sorge, Nina M. van, and Rooijakkers, Suzan H.M.

Abstract

Infections with Gram-negative bacteria form an increasing risk for human health due to antibiotic resistance. Our immune system contains various antimicrobial proteins that can degrade the bacterial cell envelope. However, many of these proteins do not function on Gram-negative bacteria, because the impermeable outer membrane of these bacteria prevents such components from reaching their targets. Here we show that complement-dependent formation of Membrane Attack Complex (MAC) pores permeabilizes this barrier, allowing antimicrobial proteins to cross the outer membrane and exert their antimicrobial function. Specifically, we demonstrate that MAC-dependent outer membrane damage enables human lysozyme to degrade the cell wall of E. coli. Using flow cytometry and confocal microscopy, we show that the combination of MAC pores and lysozyme triggers effective E. coli cell wall degradation in human serum, thereby altering the bacterial cell morphology from rod-shaped to spherical. Completely assembled MAC pores are required to sensitize E. coli to the antimicrobial actions of lysozyme and other immune factors, such as Human Group IIA-secreted Phospholipase A2. Next to these effects in a serum environment, we observed that the MAC also sensitizes E. coli to more efficient degradation and killing inside human neutrophils. Altogether, this study serves as a proof of principle on how different players of the human immune system can work together to degrade the complex cell envelope of Gram-negative bacteria. This knowledge may facilitate the development of new antimicrobials that could stimulate or work synergistically with the immune system.

Published
2021

50. Outer membrane permeabilization by the membrane attack complex sensitizes Gram-negative bacteria to antimicrobial proteins in serum and phagocytes

Author

Afd Chemical Biology and Drug Discovery, Chemical Biology and Drug Discovery, Heesterbeek, Dani A. C., Muts, Remy M., Hensbergen, Vincent P. van, Aulaire, Pieter de Saint, Wennekes, Tom, Bardoel, Bart W., Sorge, Nina M. van, Rooijakkers, Suzan H.M., Afd Chemical Biology and Drug Discovery, Chemical Biology and Drug Discovery, Heesterbeek, Dani A. C., Muts, Remy M., Hensbergen, Vincent P. van, Aulaire, Pieter de Saint, Wennekes, Tom, Bardoel, Bart W., Sorge, Nina M. van, and Rooijakkers, Suzan H.M.

Published
2021

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