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Host defence peptides of the innate immune system and in the venom of the spider Cupiennius salei

Applicant Nentwig Wolfgang
Number 127500
Funding scheme Project funding (Div. I-III)
Research institution Abteilung Synökologie Institut für Ökologie und Evolution Universität Bern
Institution of higher education University of Berne - BE
Main discipline Zoology
Start/End 01.12.2009 - 31.05.2013
Approved amount 288'000.00
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Keywords (7)

hemolymph; hemocytes; innate immune system; venom gland; antimicrobial peptides; cytolytic peptides; host defence peptides

Lay Summary (German)

Lead
Lay summary
Antimikrobielle Peptide dienen der Verteidigung gegen Mikroorganismen und kommen vermutlich in allen Organismen vor. Man kennt inzwischen tausende solcher Verbindungen nicht nur aus dem Immunsystem eines Organismus, sondern auch aus unterschiedlichen Organen. Bei Spinnen sind bisher nur wenige Arten untersucht worden, vor allem Cupiennius salei, bei der wir in der Vergangenheit einige antimikrobielle Peptide aus dem Gift und aus Hämozyten und Hämolymphe nachweisen konnten. In diesem Forschungsprojekt wollen wir Struktur und Funktion der antimikrobiellen Peptide des Hämolymph-Immunsystems und des Giftes vergleichend untersuchen.Unsere erste Arbeitshypothese nimmt an, dass das Immunsystem von Spinnen konstitutiv ist, d.h. antimikrobiellen Peptide werden kontinuierlich produziert und in Hämozyten gelagert. Wenn eine Infektion erfolgt, werden die antimikrobiellen Peptide in die Hämolymphe freigesetzt. Wir nehmen an, dass die einzelnen Komponenten dieses Systems synergistisch wirken. In Hämozyten von C. salei konnten wir inzwischen 4 Substanzen nachweisen: Ctenidin, C. salei Defensin, Peptid 8 and Mygalin. Wir werden gezielt nach weiteren Substanzen suchen und Interaktionen zwischen ihnen analysieren. Massenspektroskopisch werden wir zudem die Hämolymphe und die Hämozyten von infizierten und nichtinfizierten Spinnen vergleichen, um die Immunantwort dieser Tiere zu analysieren.Unsere zweite Arbeitshypothese betrifft die Giftdrüse von C. salei, in der wir früher bereits eine Reihe von antimikrobiellen Peptiden (Cupiennine) nachweisen konnten. Wir nehmen an, dass die Produktion von Cupienninen auf die Giftdrüsen beschränkt ist, da sich ihre breite membranzerstörende Wirkung in anderen Geweben zu schädlich auswirken würde. Dies könnte bedeuten, dass nach kompletter Entleerung der Giftdrüsen oder bei Spinnenarten ohne Cupiennine eine Verteidigungslücke entsteht. Dies würde die Notwendigkeit eines zweiten Verteidigungssystems unterstreichen, das aus C. salei Defensin oder Ctenidin bestehen könnte. Um unsere Annahmen zu testen, werden wir die Expression von antimikrobiellen Peptiden in der Giftdrüse untersuchen. Wir werden zudem Tiere künstlich infizieren und den zeitlichen Verlauf der Expression von antimikrobiellen Peptiden in verschiedenen Geweben analysieren, um unsere Hypothese der gewebsspezifischen Produktion antimikrobieller Peptide zu testen.Unser Forschungsprojekt wird am Beispiel der Modellspinne C. salei das Immunsystem von Spinnen erforschen. Zudem werden unsere Studien zu antimikrobiellen Peptiden helfen, Möglichkeiten und Grenzen dieser Substanzen aufzuzeigen, welche als mögliche neuen Antibiotika im Zentrum des Interesse stehen.
Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Functional differentiation of spider hemocytes by light and transmission electron microscopy, and MALDI-MS-imaging.
Kuhn-Nentwig L, Kopp LS, Nentwig W, Haenni B, Streitbereger K, Schürch S, Schaller J (2014), Functional differentiation of spider hemocytes by light and transmission electron microscopy, and MALDI-MS-imaging., in Developmental and Comparative Immunology , 43, 59-67.
Main components of spider venoms
Nentwig W, Kuhn-Nentwig L (2013), Main components of spider venoms, in Nentwig W (ed.), Springer, Heidelberg, 191-202.
Spider ecophysiology
Nentwig W (ed) (2013), Spider ecophysiology, Springer, Heidelberg.
Spider venoms potentially lethal to humans
Nentwig W, Kuhn-Nentwig L (2013), Spider venoms potentially lethal to humans, in Nentwig W (ed.), Springer, Heidelberg, 253-264.
The immune system of spiders
Kuhn-Nentwig L, Nentwig W (2013), The immune system of spiders, in Nentwig W (ed.), Springer, Heidelberg, 81-91.
A Venom-derived neurotoxin, CsTx-1, from the spider Cupiennius salei exhibits cytolytic activities
Kuhn-Nentwig L, Fedorova IM, Lüscher BP, Kopp LS, Trachsel C, Schaller J, Streitberger K, Nentwig W, Sigel E, Magazanik LG (2012), A Venom-derived neurotoxin, CsTx-1, from the spider Cupiennius salei exhibits cytolytic activities, in J Biol Chem , 287(30), 25640-25649.
Das Gift der Bananenspinne - Cupiennius salei, eine Kammspinne aus Mittelamerika, macht eine steile wissenschaftliche Karriere
Kuhn-Nentwig L, Nentwig W (2012), Das Gift der Bananenspinne - Cupiennius salei, eine Kammspinne aus Mittelamerika, macht eine steile wissenschaftliche Karriere, in SENCKENBERG natur - forschung - museum, 142(3/4), 114-121.
Multicomponent venom of the spider Cupiennius salei: a bioanalytical investigation applying different strategies
Trachsel C, Siegemund D, Kopp LS, Lüthi C, Cunningham M, Nentwig W, Kuhn-Nentwig L, Schürch S, Schaller J (2012), Multicomponent venom of the spider Cupiennius salei: a bioanalytical investigation applying different strategies, in FEBS J, 279, 2683-2694.
Structural and biochemical characterization of native and recombinant single insulin-like growth factor-binding domain protein (SIBD-1) from the Central American hunting spider Cupiennius salei
Trachsel C, Widmer C, Kämpfer U, Bühr C, Baumann T, Kuhn-Nentwig L, Schürch S, Schaller J, Baumann U (2012), Structural and biochemical characterization of native and recombinant single insulin-like growth factor-binding domain protein (SIBD-1) from the Central American hunting spider Cupiennius salei, in Proteins, 80, 2323-2329.
The cytotoxic mode of action of the venom of Cupiennius salei (Ctenidae).
Kuhn-Nentwig L, Nentwig W (2012), The cytotoxic mode of action of the venom of Cupiennius salei (Ctenidae)., in Nentwig W (ed.), Springer, Heidelberg, 217-228.
Cupiennin 1a exhibits a remarkably broad, non-stereospecific cytolytic activity on bacteria, protozoan parasites, insects, and human cancer cells
Kuhn-Nentwig L, Willems J, Seebeck T, Shalaby T, Kaiser M, Nentwig W (2011), Cupiennin 1a exhibits a remarkably broad, non-stereospecific cytolytic activity on bacteria, protozoan parasites, insects, and human cancer cells, in AMINO ACIDS, 40(1), 69-76.
Purification, cDNA 1 structure and biological significance of a single insulin-like growth factor-binding domain protein (SIBD-1) identified in the hemocytes of the spider Cupiennius salei.
Kuhn-Nentwig L, Largiadèr CR, Streitberger K, Chandru S, Baumann T, Kämpfer U, Schaller J, Schürch S, Nentwig W (2011), Purification, cDNA 1 structure and biological significance of a single insulin-like growth factor-binding domain protein (SIBD-1) identified in the hemocytes of the spider Cupiennius salei., in Insect Biochem Mol Biol, 41, 891-901.
Venom composition and strategies in spiders: is everything possible?
Kuhn-Nentwig L, Stöcklin R, Nentwig W (2011), Venom composition and strategies in spiders: is everything possible?, in Adv Insect Physiol , 40, 1-86.
Ctenidins: antimicrobial glycine-rich peptides from the hemocytes of the spider Cupiennius salei
Baumann T, Kämpfer U, Schürch S, Schaller J, Largiader C, Nentwig W, Kuhn-Nentwig L (2010), Ctenidins: antimicrobial glycine-rich peptides from the hemocytes of the spider Cupiennius salei, in Cell Mol Life Sci, 67(16), 2787-2798.
Expression of defensins in non-infected araneomorph spiders
Baumann T, Kuhn-Nentwig L, Largiader CR, Nentwig W (2010), Expression of defensins in non-infected araneomorph spiders, in Cell Mol Life Sci, 67(15), 2643-2651.
Cytolytic and antimicrobial peptides in the venom of scorpions and spiders.
Kuhn-Nentwig L (2009), Cytolytic and antimicrobial peptides in the venom of scorpions and spiders., in De Lima ME (ed.), University of Minas Gerais Publishing, Belo Horizonte , 249-268.
Spider venom and hemolymph derived cytolytic and antimicrobial peptides
Kuhn-Nentwig L, Trachsel C, Nentwig W (2009), Spider venom and hemolymph derived cytolytic and antimicrobial peptides, in Howl J (ed.), CRC Press, Boca Raton, 447-464.
N-terminal aromatic residues closely impact the cytolytic activity of cupiennin 1a, a major spider venom peptide.
Kuhn-Nentwig L, Sheynis T, Kolusheva S, Nentwig W, Jelinek R, N-terminal aromatic residues closely impact the cytolytic activity of cupiennin 1a, a major spider venom peptide., in Toxicon.

Collaboration

Group / person Country
Types of collaboration
Department of Chemistry, Ben Gurion University of the Negev, Beer-Sheva Israel (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Institute of Clinical Chemistry, Bern University Hospital, Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Laboratory of Toxicology, University of Leuven Belgium (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Atheris Laboratories, Geneva Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Industry/business/other use-inspired collaboration
Swiss Tropical Institute, Basel Switzerland (Europe)
- Research Infrastructure
Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Ac Sciences, St Petersburg, Russia (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Departement Chemie und Biochemie, Universität Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), CCT-La Plata CONICET-UNLP Argentina (South America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Ac. Sciences, Moscow, Russia (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Department of Medicine Laboratory of Biochemistry, Belgium (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Institute of Cell Biology, University of Bern, Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Monash University, Dept. of Pharmacology, Melbourne Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Tagung der Societé Française pour l’étude des Toxines Talk given at a conference The venom of Cupiennius salei 10.12.2012 Paris, France Kuhn-Nentwig Lucia;
27th European Congress of Arachnology Talk given at a conference Barcoding in spiders 02.09.2012 Ljubljana, Slovenia, Slovenia Nentwig Wolfgang;
17th World Congress of the International Society on Toxinology Talk given at a conference Neurotoxins in the venom of Cupiennius salei 08.07.2012 Honolulu, Hawaii, USA, United States of America Nentwig Wolfgang; Baumann Tommy; Kuhn-Nentwig Lucia;
26th European Congress of Arachnology Talk given at a conference Venom components in Cupiennius salei 04.09.2011 Sede Boqer, Israel, Israel Nentwig Wolfgang; Kuhn-Nentwig Lucia;
International Conference on Molecular mechanisms of synaptic transmission Talk given at a conference The venom of Cupiennius salei 19.04.2011 St Petersburg, Russia, Russia Nentwig Wolfgang; Kuhn-Nentwig Lucia;


Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions Das Gift der Bananenspinne macht eine steile wissenschaftliche Karriere International 2012
Media relations: radio, television Berichterstattung über Spinnenforschung SF 1, Einstein German-speaking Switzerland 2010

Associated projects

Number Title Start Funding scheme
113681 Antimicrobial peptides and the dual immune system of the spider Cupiennius salei 01.10.2006 Project funding (Div. I-III)
139078 1.7 mm Micro-Probehead for small volume NMR Spectroscopic Investigations 01.03.2012 R'EQUIP
162564 Functional and phylogenetic importance of antimicrobial peptides and enzymes in spider venoms 01.10.2015 Project funding (Div. I-III)

Abstract

Antimicrobial peptides, cytolytic peptides and comparable substances, recently summarised as host defence peptides, are crucial for the survival of all living organisms. In the last decades more than a thousand such compounds have been isolated not only from different immune-related cells but also from a variety of tissues such as epithelia and glands from a large variety of species from all major taxonomic groups. Due to limited research efforts, host defence peptides have so far only been detected in a few species of spiders. Most records concern cytolytic peptides in spider venom, some records also concern the spider’s immune system (hemocytes and hemolymph). In the here proposed project, which is a continuation of our previous studies on the structure and function of cytolytic and antimicrobial peptides from venom and hemolymph of Cupiennius salei, we want to investigate the similarities and differences of host defence peptides from these two sources.We are focussing our project on two hypotheses. Our first hypothesis states that the innate immune system of spiders is primarily a constitutive system, in which host defence compounds are constitutively produced and stored in hemocytes, until an infection causes their release into the hemolymph. We are assuming that the individual compounds of this immune system act synergistically and we will test this. Meanwhile we know of four substances from hemocytes of C. salei (ctenidin, C. salei defensin, peptide 8, and mygalin). With the aid of a cDNA library we will search for further host defence peptides and we will investigate potential synergisms between these compounds. By means of mass spectrometry, we will also analyse hemolymph and hemocytes of unchallenged and microbially challenged spiders to obtain a deeper understanding of the hemolymph and hemocyte based defence processes.Our second hypothesis focuses on the venom glands of C. salei with their cytolytically acting, a-helical peptides (cupiennins) in the venom. We assume that they are only expressed in venom glands because their broad cytolytic activity is not suitable for other tissues. Since this could indicate a defence gap in the case of empty venom glands or in the case of spider species which do not rely on cytolytical peptides, a second defence system is necessary. This could be represented by host defence peptides, such as C. salei defensin or ctenidin.To verify our assumption, we will investigate the expression of host defence peptides in the venom gland tissue. Moreover, we will cause a bacterial infection of the venom glands and analyse the expression of host defence peptides time-dependently by means of mass spectrometry. A qualitative Real Time PCR assay to investigate the mRNA expression levels of a variety of host defence peptides in different tissues will allow us to test our hypothesis on the tissue-specificity of host defence peptides. Our research proposal is scientifically relevant because (1) our knowledge of the immune system and related aspects of the venom gland of C. salei as the model spider will be signifi-cantly deepened which strengthens basic research. (2) Our recent discovery of one spider defensin meets the actual interest of the pharmaceutical industry into these compounds as promising candidates for the development of new antibiotics. (3) Specific defensins and less specific venom gland derived cytolytic peptides are ideal compounds to investigate the mode of action of host defence peptides, especially with respect to cell membrane specificity. (4) Our systematic screenings for so far unknown host defence compounds and our analyses of synergistic effects between compounds could yield results suitable for fuelling into the development of new antibiotics and new substances against pathogenic microbes.
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