Institut Necker Enfants Malades

Focus

We are interested in the how individual members of the gut microbiota influence the host immune system, how they promote colonization resistance and how they interact with the host during health and disease.

Introduction

The gut microbiota plays an integral part in shaping the host immune system and amongst its many functions, helps to protect the host from pathogens. How individual bacterial species contribute to colonization resistance is still poorly understood. For now, the foremost commensal associated with colonization resistance in the gut, as well as at distal mucosal sites, is the Clostridium-related commensal segmented filamentous bacteria (SFB). SFB is a common commensal in many vertebrates where it colonizes the host at the time of weaning and strongly stimulates secretory IgA responses, innate defenses and a potent Th17 response. SFB is a thereby a key member of the microbiota that potently shapes the immunological milieu of the host and influences immune reactivity. In the lab, we are interested in deciphering the novel interaction of SFB with intestinal epithelial cells at the molecular level, how this interaction fosters physiological inflammation in the host, and how the SFB-mediated immune activation results in enhanced resistance to pathogens in and outside of the gut. Aside a better basic understanding of the unique cross talk between this medically-relevant microbe and the host, our long-term goals is to develop a novel vaccine delivery platform against enteropathogens.

Research objectives

We currently focus our investigations of the host-microbiota interaction on the unique cross talk between the commensal SFB and the host. SFB intimately attaches to intestinal epithelial cells, leading to actin recruitment at the site of contact. This interaction is key for the stimulatory potential of SFB and likely involves the conditioning of the immune cells by SFB-mediated stimulation of epithelial cells. For one, we aim to characterize the tight interaction of SFB with the intestinal epithelial cell using a number of proteomic, molecular and immunological approaches. We also aim to better understand how epithelial cells respond to SFB challenge and to identify and characterize host factors involved in the activation of the host immune system. At the same time, we are interested in the molecular mechanisms, and to what extent, SFB can mediate colonization resistance in and outside of the gut. In addition, we aim to establish genetic manipulation techniques for SFB for mutagenesis studies and to express foreign antigens in order to test SFB as a vaccine delivery platform.

Main publications

Nkamba I, Mulet C, Dubey GP, Gorgette O, Couesnon A, Salles A, Moya-Nilges M, Shima T, Umesaki Y, Nigro G, Krijnse-Locker J, Berard M, Cerf-Bensussan N, Sansonetti PJ, Schnupf P. Intracellular offsprings released from SFB filaments are flagellated. Nature Microbiology. In press.
Schnupf P, Gaboriau-Routhiau V, Sansonetti P, Cerf-Bensussan N. Segmented filamentous bacteria, Th17 inducers and helpers in a hostile world. Current Opinion in Microbiology. 2017, 35: 100-109.
Vonaesch P, Campbell-Valois FX, A Dufour, Sansonetti P, Schnupf P. Shigella flexneri modulates stress granule composition and inhibits stress granule aggregation. Cellular Microbiology. 2016. Jul;18(7):982-97
Schnupf P, Gaboriau-Routhiau V, Gros M, Friedman R, Moya-Nilges M, Nigro G, Cerf-Bensussan N, Sansonetti PJ. Growth and host interaction of mouse Segmented Filamentous Bacterium in vitro. Nature. 2015. Apr 2;520(7545):99-103.
Schnupf P, Gaboriau-Routhiau V, Cerf-Bensussan N. Host-interactions with Segmented Filamentous Bacterium: An unusual trade-off that drives the post-natal maturation of the gut immune system. Seminars in Immunology. 2013. 25(5):342-51.

2019

Nkamba I, Mulet C, Dubey GP, Gorgette O, Couesnon A, Salles A, Moya-Nilges M, Shima T, Umesaki Y, Nigro G, Krijnse-Locker J, Berard M, Cerf-Bensussan N, Sansonetti PJ, Schnupf P^.
Intracellular offsprings released from SFB filaments are flagellated.
Nature Microbiology. In press. ^Corresponding author

2018

Schnupf P^ and Sansonetti PJ^.
Shigella pathogenesis: new insights through advanced methodologies. ASM Publisher, Book chapter in: Cellular and tissue microbiology of intracellular pathogens.
Microbiol Spectr. 2019 Mar;7(2). ^Corresponding authors

Schnupf P, Gaboriau-Routhiau V, Cerf-Bensussan N.
Modulation of the gut microbiota to improve innate resistance.
Current Opinion in Immunology. 2018. 54:137-144.

2017

Schnupf P^, Gaboriau-Routhiau V, Sansonetti P, Cerf-Bensussan N^. Segmented filamentous bacteria, Th17 inducers and helpers in a hostile world. Current Opinion in Microbiology. 2017, 35: 100-109 ^Corresponding authors

Vonaesch P*, Sansonetti P, Schnupf P*,^. Immunofluorescence analysis of stress granule formation after bacterial challenge of mammalian cells. JoVE, (125), e55536, doi:10.3791/55536 (2017). *Co-first authors, ^Corresponding author.

Schnupf P, Gaboriau-Routhiau V, Sansonetti PJ, Cerf-Bensussan N. Chapter 25: Mucosal Interactions with Enteropathogenic Bacteria. Mucosal Immunology, Second edition. Society for Mucosal Immunology. Garland Science. Taylor and Francis Group, LLC. 2017.

Milivojevic M, Dangeard AS, Kasper CA, Tschon T, Emmenlauer M, Pique C, Schnupf P, Guignot J, Arrieumerlou C. ALPK1 controls TIFA/TRAF6-dependent innate immunity against heptose-1,7- bisphosphate of gram-negative bacteria. PLoS Pathogen. 2017;13(2):e1006224.

2016

Vonaesch P*, Campbell-Valois FX, A Dufour, Sansonetti P, Schnupf P*,^. Shigella flexneri modulates stress granule composition and inhibits stress granule aggregation. Cellular Microbiology. 2016. Jul;18(7):982-97. *Co-first authors, ^Corresponding author.

2015

Schnupf P, Gaboriau-Routhiau V, Gros M, Friedman R, Moya-Nilges M, Nigro G, Cerf-Bensussan N, Sansonetti P. Growth and host interaction of mouse Segmented Filamentous Bacterium in vitro. Nature. 2015. Apr 2;520(7545):99-103.

2014

Campbell-Valois FX, Schnupf P, Nigro G, Arena, E, Sachs M, Sansonetti P, Parsot C. Fluorescent transcriptional reporters reveal on∕off switching of the Type Three Secretion Apparatus (TTSA) activity during entry and cell-to-cell spread of Shigella flexneri. Cell Host and Microbe. 2014. 15(2):177-89.

Campbell-Valois FX, Schnupf P, Sansonetti P. Design of a transcription-based secretion activity reporter (TSAR) for the type III secretion apparatus of Shigella flexneri and uses therof. Bio-protocol. 2014. 4(20).

Campbell-Valois FX, Schnupf P, Sansonetti P. Detection of the secreted and cytoplasmic fractions of IpaB, IpaC and IpaD by lysozyme permeabilization. Bio-protocol. 2014. 4(20).

Bolotin A, de Wouters T, Schnupf P, Bouchier C, Loux V, Rhimi M, Jamet A, Dervyn R, Boudebbouze S, Hervé Blottière H, Sorokin A, Snel J, Cerf-Bensussan N, Gaboriau-Routhiau V, van de Guchte M. Genome sequence of Candidatus Arthromitus sp. SFB-mouse-NL, a commensal bacterium with a key role in the postnatal maturation of gut immune functions. GenomeA (ASM). 2014.

2013

Schnupf P, Gaboriau-Routhiau V, Cerf-Bensussan N. Host-interactions with Segmented Filamentous Bacterium: An unusual trade-off that drives the post-natal maturation of the gut immune system. Seminars in Immunology. 2013. 25(5):342-51.

2012

Schnupf P and Sansonetti P. Quantitative RT-PCR profiling of the rabbit immune response: assessment of acute Shigella flexneri infection. PLoS One. 2012. 7(6):e36446.

Teo I, Toms SM, Marteyn B, Barata TS, Simpson P, Johnston KA, Schnupf P, Puhar, A, Bell T, Tang C, Zloh M, Matthews S, Rendle MP, Sansonetti PJ and Shaunak S. Preventing acute gut wall damage in infectious diarrhoeas with glycosylated dendrimers. EMBO Molecular Medicine. 2012. 4(9):866-81.

2010

Schnupf P and Sansonetti P. The gut microbiota and its contribution to homeostasis. Bacterial Virulence: Basic principles, models and global approaches. 2010. WILEY-VCH Verlag GmbH & Co. Pgs 195-214.

Sellge G, Schnupf P and Sansonetti P. Anatomy of the gut barrier and establishment of intestinal homeostasis. Bacterial Virulence: Basic principles, models and global approaches. 2010. WILEY-VCH Verlag GmbH & Co. Pgs 215-250.

2007

Schnupf P, Portnoy DA. Listeriolysin O: a phagosome-specific lysin. 2007. Microbes and Infection. 2007. Aug; 9(10):1176-87.

Schnupf P, Zhou J, Varshavsky A and Portnoy DA. Listeriolysin O secreted by Listeria monocytogenes into the host cell cytosol is degraded by the N-end rule pathway. Infection and Immunity. 2007. Nov; 75(11):5135-47.

2006

Schnupf P, Hofmann, J, Norseen J, Glomski IJ, Schwartzstein H and Decatur AL. Regulated translation of listeriolysin O controls virulence of Listeria monocytogenes. Molecular Microbiology. 2006. Aug; 61(4):999-1012.

Schnupf P, Portnoy DA and Decatur AL. Phosphorylation, ubiquitination, and degradation of listeriolysin O in mammalian cells: Role of the PEST-like sequence. Cellular Microbiology. 2006. Feb; 8(2):353-64.

2005

Schnupf P*, Wei Z*, Poussin MA, Zenewicz LA, Shen H, Portnoy DA, and Goldfine H. Characterization of Listeria monocytogenes expressing anthrolysin O and phosphatidylinositol-specific phospholipase C from Bacillus anthracis. Infection and Immunity. 2005. Oct; 73(10):6639-46. (*contributed equally; listed as Wei et al. in publication).

Brockstedt D, Giedlin MA, Bahjat K, Leong M, Liu W, Luckett W, Gao Y, Schnupf P, Kapadia D, Castro G, Sampson-Johannes A, Herskovits AA, Stassinopoulos A, Bouwer HG, Hearst JE, Portnoy DA, Cook DN, and Dubensky TW Jr. Killed but metabolically active (KBMA) microbes: A new vaccine paradigm for eliciting effector CD8 T cells and protective immunity. Nature Medicine. 2005. Aug; 11(8):853-60.

2004

Punzo C, Plaza S, Seimiya M, Schnupf P, Kurata S, Jaeger J, Gehring WJ. Functional divergence between eyeless and twin of eyeless in Drosophila melanogaster. Development. 2004. Aug; 131(16):3943-53.

2001

Wilson MD, Riemer C, Martindale DW, Schnupf P, Boright AP, Cheung TL, Hardy DM, Schwartz S, Scherer SW, Tsui LC, Miller W, Koop BF. Comparative analysis of the gene-dense ACHE/TFR2 region on human chromosome 7q22 with the orthologous region on mouse chromosome 5. Nucleic Acids Research. 2001. Mar 15; 29(6):1352-65.

2000

Nelson JR, Cooper G, Garner T, Schnupf P. Polymorphic markers for the sea cucumber Parastichopus californicus. Molecular Ecology Notes. 2000. 2(3):233-235.

Pamela obtained her B.Sc. Honors degree in Biology and a Minor in Microbiology from the University of Victoria, British Columbia, Canada in 1998 and her Ph.D. in Microbiology from the University of California at Berkeley in 2005. For her PhD, she investigated the cellular microbiology of the human pathogen Listeria monocytogenes in the laboratory of Dan Portnoy. She then continued in the field of microbial pathogenesis as a postdoctoral fellow with Philippe Sansonetti at the Pasteur Institute in Paris where she explored different aspects of the host-bacterial interaction of the human diarrheal pathogen Shigella flexneri. Pamela then switched to the microbiota field to investigate how commensal bacteria can contribute to the protection of the host from pathogens. For this, she joined Laboratory of Intestinal Immunity of Nadine Cerf-Bensussan the at Imagine Institute where she worked with the immunostimulatory gut commensal segmented filamentous bacteria (SFB) and was the first to set up an in vitro culturing system, starting the field of cellular microbiology for SFB.

Pamela became an INSERM Permanent Researcher in 2016 and joined the Necker Institute for Sick Children (INEM) as a group leader in 2018. Her research is supported through national and international organizations, including a prestigious Bill and Melinda Gates Foundation Grand Challenge grant and a Young Investigator grant from the National Research Agency (ANR).

EDUCATION AND DIPLOMAS
2019 - French accreditation to supervise research (HDR), University Paris Descartes

FUNDING (Research Grants, Fellowships, Scholarships)
2019-2022 - Agence National de la Recherche (ANR) Young Investigator Grant (JCJC). (PI)