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The cross-talk between chemotherapy and immunity in murine and ovine neosporosis disease

English title The cross-talk between chemotherapy and immunity in murine and ovine neosporosis disease
Applicant Hemphill Andrew
Number 165782
Funding scheme Project funding (Div. I-III)
Research institution Institut für Parasitologie Universität Bern
Institution of higher education University of Berne - BE
Main discipline Immunology, Immunopathology
Start/End 01.04.2016 - 31.03.2019
Approved amount 678'000.00
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All Disciplines (2)

Discipline
Immunology, Immunopathology
Veterinary Medicine

Keywords (8)

Neospora caninum; recombinant antigens; cytotoxicity; mouse model; calcium dependent kinase inhibitors; Apicomplexa; sheep model; immunity

Lay Summary (German)

Lead
Neospora caninum, ein weltweit verbreiteter Aborterreger und Verursacher von neuromuskulären Störungen bei Rindern und Hunden, lebt unbemerkt in chronisch infizierten Tieren und wird während der Trächtigkeit transplazental auf die Nachkommen übertragen. Das Mausmodell ist ein häufig verwendetes Infektionsmodell, aber aufgrund der immunologischen Unterschiede zwischen Rindern und Mäusen ist dessen Relevanz grundsätzlich in Frage zu stellen. Als alternatives Infektionsmodell wurde ein experimentelles Infektionsmodel in Schafen etabliert.
Lay summary

Ziel des Projektes ist, neue Massnahmen zur Prävention und Therapie der Neosporose zu entwickeln. Die Evaluation von Inhibitoren von Ca2+-abhängigen Protein Kinasen (bumped kinase inhibitors oder BKIs) in vitro und im Mausmodell hat gezeigt, dass diese dazu geeignet wären, die transplazentäre Infektion von Nachkommen in Neospora caninum infizierten Tieren zu hemmen. Diese Wirkstoffe töten die Parasiten in vitro erst nach langer Behandlung ab, sind also nicht per se parasitozid, aber induzieren die Bildung von multizelluären Parasitenstadien, die eine deregulierte Antigenexpression aufweisen. Im Mausmodell konnte aber gezeigt werden, dass eine kurze Behandlungsdauer die Infektion eliminiert. Wir vertreten die Hypothese, dass diese multizelluären Stadien, die durch diese Wirkstoffe induziert werden, Antigene exprimieren, die es dem Wirt ermöglichen eine sehr wirksame Immunität aufzubauen, und so den Parasiten abzutöten. Dies soll in diesem Projekt genauer untersucht werden. Dabei wird die Immunantwort von behandelten und unbehandelten N. caninum infizierten Mäusen und Schafen genauer untersucht, und parallel dazu wird das Proteom dieser BKI-induzierten multizellulären Komplexe bestimmt. Diejenigen Antigene welche CD4 und CD8 T Zellen aktivieren, werden dann rekombinant exprimiert und als Vakzinkandidaten weiter validiert. Desweitern wird die Wirksamkeit eines dieser BKIs, BKI1294, im trächtigen Schafmodell weiter evaluiert, dies im Hinblick auf spätere Versuche in Rindern.

Direct link to Lay Summary Last update: 31.03.2016

Responsible applicant and co-applicants

Employees

Publications

Publication
Targeting of the mitochondrion by dinuclear thiolato-bridged arene ruthenium complexes in cancer cells and in the apicomplexan parasite Neospora caninum
Basto Afonso P., Anghel Nicoleta, Rubbiani Riccardo, Müller Joachim, Stibal David, Giannini Federico, Süss-Fink Georg, Balmer Vreni, Gasser Gilles, Furrer Julien, Hemphill Andrew (2019), Targeting of the mitochondrion by dinuclear thiolato-bridged arene ruthenium complexes in cancer cells and in the apicomplexan parasite Neospora caninum, in Metallomics, 11(2), 462-474.
Development and characterization of monoclonal antibodies against Besnoitia besnoiti tachyzoites
García-Lunar P., Schares G., Sanz-Fernández A., Jiménez-Meléndez A., García-Soto I., Regidor-Cerrillo J., Pastor-Fernández I., Hemphill A., Fernández-Álvarez M., Ortega-Mora L. M., Álvarez-García G. (2019), Development and characterization of monoclonal antibodies against Besnoitia besnoiti tachyzoites, in Parasitology, 146(02), 187-196.
In vitro treatment of Besnoitia besnoiti with the naphto-quinone buparvaquone results in marked inhibition of tachyzoite proliferation, mitochondrial alterations and rapid adaptation of tachyzoites to increased drug concentrations
Müller Joachim, Manser Vera, Hemphill Andrew (2019), In vitro treatment of Besnoitia besnoiti with the naphto-quinone buparvaquone results in marked inhibition of tachyzoite proliferation, mitochondrial alterations and rapid adaptation of tachyzoites to increased drug concentrations, in Parasitology, 146(1), 112-120.
Virulence in Mice of a Toxoplasma gondii Type II Isolate Does Not Correlate With the Outcome of Experimental Infection in Pregnant Sheep
Sánchez-Sánchez Roberto, Ferre Ignacio, Regidor-Cerrillo Javier, Gutiérrez-Expósito Daniel, Ferrer Luis Miguel, Arteche-Villasol Noive, Moreno-Gonzalo Javier, Müller Joachim, Aguado-Martínez Adriana, Pérez Valentín, Hemphill Andrew, Ortega-Mora Luis Miguel, Benavides Julio (2019), Virulence in Mice of a Toxoplasma gondii Type II Isolate Does Not Correlate With the Outcome of Experimental Infection in Pregnant Sheep, in Frontiers in Cellular and Infection Microbiology, 8, 1-18.
Immunization with a cocktail of antigens fused with OprI reduces Neospora caninum vertical transmission and postnatal mortality in mice
Aguado-Martínez Adriana, Basto Afonso P., Tanaka Shun, Ryser Lorenz T., Nunes Telmo P., Ortega-Mora Luis-Miguel, Arranz-Solís David, Leitão Alexandre, Hemphill Andrew (2019), Immunization with a cocktail of antigens fused with OprI reduces Neospora caninum vertical transmission and postnatal mortality in mice, in Vaccine, 37(3), 473-483.
Accessible and distinct decoquinate derivatives active against Mycobacterium tuberculosis and apicomplexan parasites
Beteck Richard M., Seldon Ronnett, Coertzen Dina, van der Watt Mariëtte E., Reader Janette, Mackenzie Jared S., Lamprecht Dirk A., Abraham Matthew, Eribez Korina, Müller Joachim, Rui Feng, Zhu Guang, de Grano Ruel Valerio, Williams Ian D., Smit Frans J., Steyn Adrie J. C., Winzeler Elizabeth A., Hemphill Andrew, Birkholtz Lyn-Marie, Warner Digby F., N’Da David D., Haynes Richard K. (2018), Accessible and distinct decoquinate derivatives active against Mycobacterium tuberculosis and apicomplexan parasites, in Communications Chemistry, 1(1), 62-62.
Endochin-Like Quinolones Exhibit Promising Efficacy Against Neospora Caninum in vitro and in Experimentally Infected Pregnant Mice
Anghel Nicoleta, Balmer Vreni, Müller Joachim, Winzer Pablo, Aguado-Martinez Adriana, Roozbehani Mona, Pou Sovitj, Nilsen Aaron, Riscoe Michael, Doggett J. Stone, Hemphill Andrew (2018), Endochin-Like Quinolones Exhibit Promising Efficacy Against Neospora Caninum in vitro and in Experimentally Infected Pregnant Mice, in Frontiers in Veterinary Science, 5, 1-15.
Characterization of a multi-epitope peptide with selective MHC-binding capabilities encapsulated in PLGA nanoparticles as a novel vaccine candidate against Toxoplasma gondii infection
Roozbehani Mona, Falak Reza, Mohammadi Mohsen, Hemphill Andrew, Razmjou Elham, Meamar Ahmad reza, Masoori Leila, Khoshmirsafa Majid, Moradi Maryam, Gharavi Mohammad Javad (2018), Characterization of a multi-epitope peptide with selective MHC-binding capabilities encapsulated in PLGA nanoparticles as a novel vaccine candidate against Toxoplasma gondii infection, in Vaccine, 36(41), 6124-6132.
Repurposing of commercially available anti-coccidials identifies diclazuril and decoquinate as potential therapeutic candidates against Besnoitia besnoiti infection
Jiménez-Meléndez Alejandro, Rico-San Román Laura, Hemphill Andrew, Balmer Vreni, Ortega-Mora Luis Miguel, Álvarez-García Gema (2018), Repurposing of commercially available anti-coccidials identifies diclazuril and decoquinate as potential therapeutic candidates against Besnoitia besnoiti infection, in Veterinary Parasitology, 261, 77-85.
TbLpn, a key enzyme in lipid droplet formation and phospholipid metabolism, is essential for mitochondrial integrity and growth of Trypanosoma brucei Role of lipin in T. brucei growth
Dawoody Nejad Ladan, Serricchio Mauro, Jelk Jennifer, Hemphill Andrew, Bütikofer Peter (2018), TbLpn, a key enzyme in lipid droplet formation and phospholipid metabolism, is essential for mitochondrial integrity and growth of Trypanosoma brucei Role of lipin in T. brucei growth, in Molecular Microbiology, 109(1), 105-120.
The significance of cryptosporidiosis for the health of calves in SwitzerlandDie Bedeutung der Cryptosporidiose für die Kälbergesundheit in der Schweiz
Olias P, Dettwiler I, Hemphill A, Deplazes P, Steiner A, Meylan M (2018), The significance of cryptosporidiosis for the health of calves in SwitzerlandDie Bedeutung der Cryptosporidiose für die Kälbergesundheit in der Schweiz, in Schweiz Arch Tierheilkd, 160(6), 363-374.
Modulation of cis- and trans- Golgi and the Rab9A-GTPase during infection by Besnoitia besnoiti , Toxoplasma gondii and Neospora caninum
Cardoso Rita, Wang Junhua, Müller Joachim, Rupp Sebastian, Leitão Alexandre, Hemphill Andrew (2018), Modulation of cis- and trans- Golgi and the Rab9A-GTPase during infection by Besnoitia besnoiti , Toxoplasma gondii and Neospora caninum, in Experimental Parasitology, 187, 75-85.
Safety and efficacy of the bumped kinase inhibitor BKI-1553 in pregnant sheep experimentally infected with Neospora caninum tachyzoites
Sánchez-Sánchez Roberto, Ferre Ignacio, Re Michela, Vázquez Patricia, Ferrer Luis Miguel, Blanco-Murcia Javier, Regidor-Cerrillo Javier, Pizarro Díaz Manuel, González-Huecas Marta, Tabanera Enrique, García-Lunar Paula, Benavides Julio, Castaño Pablo, Hemphill Andrew, Hulverson Matthew A., Whitman Grant R., Rivas Kasey L., Choi Ryan, Ojo Kayode K., Barrett Lynn K., Van Voorhis Wesley C., Ortega-Mora Luis Miguel (2018), Safety and efficacy of the bumped kinase inhibitor BKI-1553 in pregnant sheep experimentally infected with Neospora caninum tachyzoites, in International Journal for Parasitology: Drugs and Drug Resistance, 8(1), 112-124.
Activities of 11-Azaartemisinin and N -Sulfonyl Derivatives against Neospora caninum and Comparative Cytotoxicities
Harmse Rozanne, Wong Ho Ning, Smit Frans J., Müller Joachim, Hemphill Andrew, N'Da David D., Haynes Richard K. (2017), Activities of 11-Azaartemisinin and N -Sulfonyl Derivatives against Neospora caninum and Comparative Cytotoxicities, in ChemMedChem, 12(24), 2094-2098.
Advances in bumped kinase inhibitors for human and animal therapy for cryptosporidiosis
Hulverson Matthew A., Choi Ryan, Arnold Samuel L.M., Schaefer Deborah A., Hemphill Andrew, McCloskey Molly C., Betzer Dana P., Müller Joachim, Vidadala Rama S.R., Whitman Grant R., Rivas Kasey L., Barrett Lynn K., Hackman Robert C., Love Melissa S., McNamara Case W., Shaughnessy Thomas K., Kondratiuk Alison, Kurnick Matthew, Banfor Patricia N., Lynch James J., Freiberg Gail M., Kempf Dale J., Maly Dustin J., Riggs Michael W., et al. (2017), Advances in bumped kinase inhibitors for human and animal therapy for cryptosporidiosis, in International Journal for Parasitology, 47(12), 753-763.
ApiCOWplexa 2017 – 4th International Meeting on Apicomplexan Parasites in Farm Animals
Hemphill Andrew, Leitão Alexandre, Ortega-Mora Luis-Miguel, Cooke Brian M. (2017), ApiCOWplexa 2017 – 4th International Meeting on Apicomplexan Parasites in Farm Animals, in International Journal for Parasitology, 47(12), 697-699.
In vitro efficacy of bumped kinase inhibitors against Besnoitia besnoiti tachyzoites
Jiménez-Meléndez Alejandro, Ojo Kayode K., Wallace Alexandra M., Smith Tess R., Hemphill Andrew, Balmer Vreni, Regidor-Cerrillo Javier, Ortega-Mora Luis M., Hehl Adrian B., Fan Erkang, Maly Dustin J., Van Voorhis Wesley C., Álvarez-García Gema (2017), In vitro efficacy of bumped kinase inhibitors against Besnoitia besnoiti tachyzoites, in International Journal for Parasitology, 47(12), 811-821.
In vitro screening of the open source Pathogen Box identifies novel compounds with profound activities against Neospora caninum
Müller Joachim, Aguado Adriana, Laleu Benoît, Balmer Vreni, Ritler Dominic, Hemphill Andrew (2017), In vitro screening of the open source Pathogen Box identifies novel compounds with profound activities against Neospora caninum, in International Journal for Parasitology, 47(12), 801-809.
Neospora caninum in non-pregnant and pregnant mouse models: cross-talk between infection and immunity
Aguado-Martínez Adriana, Basto Afonso P., Leitão Alexandre, Hemphill Andrew (2017), Neospora caninum in non-pregnant and pregnant mouse models: cross-talk between infection and immunity, in International Journal for Parasitology, 47(12), 723-735.
The single cyclic nucleotide-specific phosphodiesterase of the intestinal parasite Giardia lamblia represents a potential drug target
Kunz Stefan, Balmer Vreni, Sterk Geert Jan, Pollastri Michael P., Leurs Rob, Müller Norbert, Hemphill Andrew, Spycher Cornelia (2017), The single cyclic nucleotide-specific phosphodiesterase of the intestinal parasite Giardia lamblia represents a potential drug target, in PLOS Neglected Tropical Diseases, 11(9), e0005891-e0005891.
Extended-spectrum antiprotozoal bumped kinase inhibitors: A review
Van Voorhis Wesley C., Doggett J. Stone, Parsons Marilyn, Hulverson Matthew A., Choi Ryan, Arnold Samuel L.M., Riggs Michael W., Hemphill Andrew, Howe Daniel K., Mealey Robert H., Lau Audrey O.T., Merritt Ethan A., Maly Dustin J., Fan Erkang, Ojo Kayode K. (2017), Extended-spectrum antiprotozoal bumped kinase inhibitors: A review, in Experimental Parasitology, 180, 71-83.
Characterization of the Activities of Dinuclear Thiolato-Bridged Arene Ruthenium Complexes against Toxoplasma gondii
Basto Afonso P., Müller Joachim, Rubbiani Riccardo, Stibal David, Giannini Federico, Süss-Fink Georg, Balmer Vreni, Hemphill Andrew, Gasser Gilles, Furrer Julien (2017), Characterization of the Activities of Dinuclear Thiolato-Bridged Arene Ruthenium Complexes against Toxoplasma gondii, in Antimicrobial Agents and Chemotherapy, 61(9), 1-10.
Development of a murine vertical transmission model for Toxoplasma gondii oocyst infection and studies on the efficacy of bumped kinase inhibitor (BKI)-1294 and the naphthoquinone buparvaquone against congenital toxoplasmosis
Müller Joachim, Aguado-Martínez Adriana, Ortega-Mora Luis-Miguel, Moreno-Gonzalo Javier, Ferre Ignacio, Hulverson Matthew A., Choi Ryan, McCloskey Molly C., Barrett Lynn K., Maly Dustin J., Ojo Kayode K., Van Voorhis Wes, Hemphill Andrew (2017), Development of a murine vertical transmission model for Toxoplasma gondii oocyst infection and studies on the efficacy of bumped kinase inhibitor (BKI)-1294 and the naphthoquinone buparvaquone against congenital toxoplasmosis, in Journal of Antimicrobial Chemotherapy, 72(8), 2334-2341.
Two Novel Calcium-Dependent Protein Kinase 1 Inhibitors Interfere with Vertical Transmission in Mice Infected with Neospora caninum Tachyzoites
Müller Joachim, Aguado-Martínez Adriana, Balmer Vreni, Maly Dustin J., Fan Erkang, Ortega-Mora Luis-Miguel, Ojo Kayode K., Van Voorhis Wesley C., Hemphill Andrew (2017), Two Novel Calcium-Dependent Protein Kinase 1 Inhibitors Interfere with Vertical Transmission in Mice Infected with Neospora caninum Tachyzoites, in Antimicrobial Agents and Chemotherapy, 61(4), 1-12.
First Characterization of the Neospora caninum Dense Granule Protein GRA9
Leineweber Margret, Spekker-Bosker Katrin, Ince Vanessa, Schares Gereon, Hemphill Andrew, Eller Silvia K., Däubener Walter (2017), First Characterization of the Neospora caninum Dense Granule Protein GRA9, in BioMed Research International, 2017, 1-15.
Neosporosis in animals
DubeyJitender, HemphillAndrew, Calero-BernalRafael, ScharesGereon (2017), Neosporosis in animals, CRC Press, Taylor & Francis Group, Boca Raton.
Vaccines for bovine neosporosis: current status and key aspects for development
Horcajo P., Regidor-Cerrillo J., Aguado-Martínez A., Hemphill A., Ortega-Mora L. M. (2016), Vaccines for bovine neosporosis: current status and key aspects for development, in Parasite Immunology, 38(12), 709-723.
Open Source Drug Discovery with the Malaria Box Compound Collection for Neglected Diseases and Beyond
Van Voorhis Wesley C., Adams John H., Adelfio Roberto, Ahyong Vida, Akabas Myles H., Alano Pietro, Alday Aintzane, Alemán Resto Yesmalie, Alsibaee Aishah, Alzualde Ainhoa, Andrews Katherine T., Avery Simon V., Avery Vicky M., Ayong Lawrence, Baker Mark, Baker Stephen, Ben Mamoun Choukri, Bhatia Sangeeta, Bickle Quentin, Bounaadja Lotfi, Bowling Tana, Bosch Jürgen, Boucher Lauren E., Boyom Fabrice F., et al. (2016), Open Source Drug Discovery with the Malaria Box Compound Collection for Neglected Diseases and Beyond, in PLOS Pathogens, 12(7), e1005763-e1005763.
Apicomplexans pulling the strings: manipulation of the host cell cytoskeleton dynamics
CARDOSO RITA, SOARES HELENA, HEMPHILL ANDREW, LEITÃO ALEXANDRE (2016), Apicomplexans pulling the strings: manipulation of the host cell cytoskeleton dynamics, in Parasitology, 143(08), 957-970.
Drug target identification in protozoan parasites
Müller Joachim, Hemphill Andrew (2016), Drug target identification in protozoan parasites, in Expert Opinion on Drug Discovery, 11(8), 815-824.
Flagellar membrane fusion and protein exchange in trypanosomes; a new form of cell-cell communication?
Imhof Simon, Fragoso Cristina, Hemphill Andrew, von Schubert Conrad, Li Dong, Legant Wesley, Betzig Eric, Roditi Isabel (2016), Flagellar membrane fusion and protein exchange in trypanosomes; a new form of cell-cell communication?, in F1000Research, 5, 682-682.

Collaboration

Group / person Country
Types of collaboration
Faculty of Health Sciences, North-West University, Potchefstroom 2520 South Africa (Africa)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Institute of Parasitology, University of Bern Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Institute for Virology and Immunology, Vetsuisse Faculty, Bern Switzerland (Europe)
- Research Infrastructure
Institute of Animal Pathology, Vetsuisse Faculty, Bern Switzerland (Europe)
- Research Infrastructure
Institute of Anatomy, Medical Faculty, University of Bern Switzerland (Europe)
- Research Infrastructure
University of Washington, Seattle, USA, Department of Medicine Division of Allergy and Infectious Di United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Industry/business/other use-inspired collaboration
Institute of Infectious Diseases, University of Bern Switzerland (Europe)
- Research Infrastructure
Proteomics & mass spectrometry core facility at the Department of Clinical Research, Bern Switzerland (Europe)
- Research Infrastructure
Veterinary Faculty, University of Lisbon Portugal (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Institute of Parasitology, Vetsuisse Faculty Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
VA Portland Health Care System Research and Development Service, Portland, OR United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Department of Parasitology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran Iran (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel
Universidad Complutense Madrid, Veterinary Faculty Spain (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
PhD mid-term evaluation, University of Bern Individual talk Molecular changes of Neosporacaninum treated with Calcium dependent protein kinase 1 inhibitor 1294 and implications on the immune response. 12.02.2019 Bern, Switzerland Winzer Pablo Arnold;
Vetsuisse Veterinary Public Health Conference, Zürich, Switzerland, Talk given at a conference Comparative study between pregnant mouse interference test and zebrafish embryo development test: a possible replacement in anti-parasitic chemotherapy trials? 29.11.2018 Zürich, Switzerland de Amorim Cardoso Rita; Winzer Pablo Arnold;
CIISA congress: exploring the boundaries of animal, veterinary and biomedical sciences Talk given at a conference Novel drugs and drug targets for the treatment of Toxoplasma gondii infections 16.11.2018 Lisbon, Portugal Hemphill Andrew;
Swiss Society of Tropical Medicine and Parasitology PhD/Doctoral meting, Sigriswill, Switzerland Talk given at a conference Chemotherapy agaoinst neosporosis 01.11.2018 Sigriswil, Switzerland Winzer Pablo Arnold;
European Coccidiosis Discussion Group, Royal Veterinary College London Talk given at a conference Drugs and drug targets in pregnant models for toxoplasmosis and neosporosis 17.05.2018 London, Great Britain and Northern Ireland Hemphill Andrew;
Invited seminar, Royal Veterinary College, London Individual talk Vaccination against Neospora caninum infection: studies in the mouse model 16.05.2018 London, Great Britain and Northern Ireland Hemphill Andrew;
Host Pathogen Interactions Symposium, Vetsuisse Bern, Haus der Universität Talk given at a conference Comparative study between pregnant mouse interference test and zebrafish embryo development test 04.05.2018 Bern, Switzerland de Amorim Cardoso Rita; Hemphill Andrew; Winzer Pablo Arnold;
Seminarios investigación platesa, Universidad Completense de Madrid Individual talk Studies on novel preventive and therapeutic tools to tackle diseases caused by apicomplexan parasites 24.04.2018 Madrid, Spain Hemphill Andrew;
Fishclub Bern Seminar Series Individual talk Comparative study between pregnant mouse interference test and zebrafish embryo development test: a possible replacement in anti-parasitic chemotherapy trials? 29.03.2018 Bern, Switzerland Winzer Pablo Arnold; Hemphill Andrew;
Deutsche Gesellschaft für Parasitologie Annual Meeting Talk given at a conference Bumped kinase inhibitor 1294 induces the formation of multi-nucleated complexes and alters protein expression in Neospora caninum 21.03.2018 berlin, Germany Hemphill Andrew; Winzer Pablo Arnold; Aguado-Martinez Adriana;
Graduate School of Life Sciences Symposium 2018 Poster Bumped kinase inhibitor 1294 induced formation of multi-nucleated complexes in Neospora caninumand transformation of protein expression. 01.02.2018 Bern, Switzerland de Amorim Cardoso Rita; Hemphill Andrew; Winzer Pablo Arnold;
SSTMP student meeting Talk given at a conference Neospora caninum- Multinucleated complexes induced by BKI-1294. 02.11.2017 Schwarzenberg, Switzerland Winzer Pablo Arnold;
Apicowplexa 2017: 4th international meeting on apicomplexan parasites in farm animals Talk given at a conference Bumped kinase inhibitors and their effects on the host-parasite relationship during experimental N. caninuminfection in vitro and in mice 11.10.2017 Madrid, Spain Hemphill Andrew; Aguado-Martinez Adriana; Winzer Pablo Arnold;
Apicowplexa 2017: 4th international meeting on apicomplexan parasites in farm animals Poster Significant reduction of Neospora caninum vertical transmission and postnatal mortality by a toll-like-receptor 2-targeting vaccine formulation in the pregnant mouse model of neosporosis 11.10.2017 Madrid, Spain Aguado-Martinez Adriana; Hemphill Andrew;
Apicowplexa 2017: 4th international meeting on apicomplexan parasites in farm animals Poster Comparative study between pregnant mouse interference test and zebrafish embryo development test: a possible replacement in anti-parasitic chemotherapy trials? 11.10.2017 Madrid, Spain Winzer Pablo Arnold; de Amorim Cardoso Rita; Hemphill Andrew;
Apicowplexa 2017: 4th international meeting on apicomplexan parasites in farm animals Talk given at a conference Drugs and drug targets in pregnant models for Toxoplasma and Neospora infection. Apicowplexa 4 11.10.2017 Madrid, Spain Aguado-Martinez Adriana; de Amorim Cardoso Rita; Hemphill Andrew; Winzer Pablo Arnold;
Annulal meeting of the Swiss Society of Tropical Medicine and Parasitology Talk given at a conference Bumped kinase inhibitors and their effects during experimental infection with the apicomplexan parasites Neospora caninum & Toxoplasma gondii: Proof of concept in pregnant mouse models 30.08.2017 Basel, Switzerland Hemphill Andrew;
Invited seminar, Friedrich Löffler Institute, Insel Riems Individual talk Of mice and sheep: vaccination and drug treatment of Neospora caninum infections 05.05.2017 Insel Riems, Germany Hemphill Andrew;
Institute Seminar, Institute of Parasitology, University of Zürich Individual talk Of mice and sheep: prevention and treatment of Neospora caninum infections 18.10.2016 Zürich, Switzerland Hemphill Andrew;
Annual Meeting of the Swiss Society of Tropical Medicine and Parasitology Poster Immunization with Neospora caninum antigens fused to a TLR2 ligand increases pup survival in a nesporosis pregnant mouse model 01.09.2016 Montreux, Switzerland Hemphill Andrew; Aguado-Martinez Adriana;


Knowledge transfer events

Active participation

Title Type of contribution Date Place Persons involved
Toxoplasma. Postgraduate course in Parasitology, Ministre de l’Enseignement Sup.eieur et de la Recherche Scientifique, Monastir University Talk 06.12.2017 Monastir, Switzerland Hemphill Andrew;
Drugs and targets in parasitic diseases: general considerations. Postgraduate course in Parasitology, Ministre de l’Enseignement Superieur et de la Recherche Scientifique, Monastir University, Talk 06.12.2017 Monastir, Tunisia Hemphill Andrew;
Scientific writing Postgraduate course in Parasitology, Ministre de l’Enseignement Superieur et de la Recherche Scientifique, Monastir University Workshop 06.12.2017 Monastir, Tunisia Hemphill Andrew;


Associated projects

Number Title Start Funding scheme
173718 Ruthenium Complexes for the Treatment of Protozoan Diseases of Medical and Veterinary Importance 01.09.2017 Sinergia
146162 Of mice and sheep: prevention and treatment of Neospora caninum infections 01.04.2013 Project funding (Div. I-III)
184662 Effects of a double-edged sword: exploiting the interaction between immunity and chemotherapy in murine and ovine models of congenital neosporosis and toxoplasmosis 01.04.2019 Project funding (Div. I-III)

Abstract

Background: The apicomplexan Neospora caninum represents one of the major infectious causes of bovine abortion worldwide, and also causes fetal loss in sheep and other ruminants. Parasite proteins with invasive functions, released from rhoptries, micronemes and dense granules, are highly interesting vaccine targets, but information on parasite antigens specifically targeted by T cells is sparse. The development of subunit vaccines against neosporosis has been hampered by the fact that an immune response that is induced in a non-pregnant animal prior to pregnancy is generally not maintained, or when maintained, not appropriately, for pregnancy. Thus, alternative approaches for defining vaccine candidates are needed. Experience from other disease models shows that T cell antigens could constitute prime vaccine candidates. In addition, chemotherapy could offer an alternative and/or addition to vaccination. In this respect, inhibitors of calcium-dependent protein kinase 1, such as the bumped kinase inhibitor BKI-1294, exhibit outstanding efficacy in the non-pregnant and pregnant neosporosis mouse models, and BKI-1294 has emerged as a prime candidate to be further evaluated in a ruminant model. BKI-1294 does not kill the parasites instantly but interferes in the separation of daughter zoites. This leads to the formation of large intracellular multinucleated complexes, which remain viable for several days to weeks in vitro. Preliminary studies indicate that this could also impact on the immune response against the parasite in vivo, and this cross-talk between chemotherapy and immunity aspect will be further elaborated in this project. Working hypotheses: (1.) BKI-1294 treatment in mice enhances the immune response against infection, which in turn supports treatment efficacy.(2.) Antigens targeted by N. caninum-specific CD4+ and CD8+ T cells represent potential vaccine candidates. (3.) BKI-1294 limits neosporosis and also impacts on the immune response in sheep. Specific aims:(1.) Characterization of (i) the proteome of multinucleated complexes in N. caninum-infected host cells treated with BKI-1294 in vitro, and of (ii) the immune response in BKI-1294-treated and non-treated infected T cell deficient and wildtype mice. (2.) Identification and characterization of Neospora T-cell target antigens, assessment of their vaccine potential in pregnant mice, and preliminary evaluation of their antigenicity in N. caninum infected pregnant sheep.(3.) Assessment of BKI-1294 therapy in the pregnant neosporosis sheep model, and its impact on the ovine immune response.Experimental design and/or methods: (1.) BKI-1294-induced proteome changes will be analyzed in a time-course experiment by immunoblotting followed by LC-MS. Overexpressed proteins that are potentially targeted by the immune response will be identified by immunoblotting and recombinantly expressed. Innate and acquired immune responses in BKI-treated or non-treated Neospora-infected pregnant or non-pregnant mice are comparatively analyzed, and the effects of BKI-treatment on immunity assessed by re-challenge experiments. Experiments in athymic nude mice will be performed to study the role of T cells in compound efficacy.(2.) Generation of N. caninum-specific CD4+ and CD8+ T cell lines (NcTcs) from pregnant and non-pregnant mice with acute or chronic infection, which will be studied with regard to proliferation, phenotype (CD4+ and CD8+ populations), polarization and cytotoxicity. We will identify proteins targeted by T cells through screening antigens previously studied in the group as vaccine candidates, antigens shown to be upregulated in virulent isolates, and antigens upregulated in multinucleated complexes. The targeted antigens are then assessed for their immunoprotective potential in the pregnant neosporosis mouse model.(3.) BKI-1294 efficacy will be studied in the N. caninum-infected pregnant sheep model. Parameters in dams (clinical signs such as fever and abortion) and offspring (fetal death, stillbirth, birth of weak offspring, neurological disorders, congenital transmission in healthy lambs) will be assessed. Tissue samples of aborted offspring and/or newborn lambs will be analyzed by (immuno)histology and real time PCR for measuring parasite and lesion presence and infection intensity. Cellular and humoral peripheral responses and comparative detection of cytokines in placental tissues will be analyzed by real time PCR, ELISA and IHC to study the role of immunity during BKI-1294 treatment in sheep. Expected value of the project: This project aims to develop novel tools to prospectively limit economic losses due to neosporosis, but has a wider impact in that CDPK1 is an important drug target in other apicomplexan parasites such as Toxoplasma, Cryptosporidium, and Besnoitia among others. For the development of preventive or therapeutical measures against neosporosis, it is important to show whether and how BKI-1294 affects the immunogenicity of the parasite, and whether this change could be a relevant factor for improving protective immunity and contribute to compound efficacy. As the role of T cell immunity in neosporosis has not been widely explored, we aim to identify novel T-cell targets, and these can constitute novel vaccine candidates. This project aims to shed light on the relationship between immunity and BKI-1294 therapy, and provides important information on how vaccination could be coupled to a drug treatment strategy, both in mice but also in a ruminant model that largely mimics infection in cattle. Prospectively, a combined immune-chemotherapeutical approach could represent an interesting option for future development of effective measures against neosporosis, and possibly also related apicomplexans such as Toxoplasma, Besnoitia among others.
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