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Approaches to genetic vaccines Britta Wahren Karolinska Institutet Riga 2013.

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Presentation on theme: "Approaches to genetic vaccines Britta Wahren Karolinska Institutet Riga 2013."— Presentation transcript:

1 Approaches to genetic vaccines Britta Wahren Karolinska Institutet Riga 2013

2 Selection of immunogen DNA, RNA or (glyco)protein Whole genes to cover polymorphic MHC in humans All genes to cover structural and regulatory genes Several subtypes for virus variability and polymorphic MHC Vectors like vaccinia or adeno Physical approach Small total vector size Intracellular delivery (skin)- electroporation Delivery devices Total cell targets – repeated injections Divided delivery-avoid immunocompetition Additional adjuvant like cytokine gene, carrier compound

3 Small minicircles were more robust when exposed to shearing forces than the corresponding plasmid. Also as splice correcting vectors, they exhibited a more long-term expression as compared to plasmids in vivo. 3 1 2 3 4 5 Minicircle1x 650 bp Minicircle3x 1 900 bp Plasmid 3 579 bp 1: untreated 2: linearized 3: UV-nicked 4: shot to empty vial 5: shot through mouse hide construct blank shothide shot mc3.12.4 mc3x3.68.6 plasmid3.723.4 % nicked sc nicked

4 Needle injection Dermis Electroporation after needle/jet stream injection Jet stream injection Intracellular delivery

5 Needle injection Dermis Electroporation after needle/jet stream injection Jet stream injection MHC II Secrete d antigens 1. Transfection of somatic and professional antigen presenting cells (APCs) MHC I Somatic cell APC 2. Antigen presentation to and activation of lymphocytes in the draining lymph nodes T cell receptor MHC II Y APC CD8 T cell CD4 T cell B cell MHC I T cell receptor Co- stimmulatory signals 3. Activated lymphocytes

6 Delivery device Biojector im Left arm: Env/rev, 1 inj +/- GMCSF im Right arm: Gag/RT, 1 inj id Left arm: Env/rev, 3 inj +/- GMCSF sc (needle) Right arm: Gag/RT, 2 inj spacer

7 Delivery device electroporator Total length of pulse-train: 0,27 seconds Time Voltage 2x450V 8x110V

8 Repeated injection of DNA, dispersed delivery

9 Volume and concentration Comparison of different immunization protocols using ID EP –BALB/c mice –Imm w 0, 4 and 8 with GagB – ID+EP (15ug) or IM (50ug) –IFNg ELISpot Comparison of different immunization methods – BALB/c mice – 1 imm with GagB – IFNg ELISpot

10 Divided delivery of env and gag genes, subtypes A, B, C Vial 1 Envelope and rev plasmids Vial 2 gag and RT plasmids Vaccine intramuscularly delivered Vial 1 in the left arm, Vial 2 in the right

11 DNA vaccines, licenced and clinical Licenced West Nile virus for horses Hematopoietic necrosis (rhabdovirus) for salmon Melanoma (tyrosine kinase) for dogs, humans? Pig hormonal treatment Clinical small trials HCV for chronic disease, Sällberg et al, SE Prostate heterologous antigen, Pisa et al, SE CEA heterologous and homologous antigen for gi cancer Mellstedt, Wahren et al, SE HIV for healthy individuals, Ho et al, USA and China, HIV for healthy or chronic disease, Wahren et al, SE, IT, TZ, MC; Pantaleo et al, CH, Robinson et al US

12 Acknowledgements: The volunteers SMI/KI Britta Wahren Andreas Bråve Margaret Liu Karl Ljungberg Gunnel Biberfeld Jorma Hinkula Erik Rollman Gunnel Engström Kristian Hallermalm Lindvi Gudmundsdotter Andreas Boberg Susanne Johansson Anne Kjerrström Maria Isaguliants Charlotta Nilsson Katarina Karlén Reinhold Benthin Karolinska/SöS Eric Sandström Bo Hejdeman Lars Eriksson Viveca Holmberg Walter Reed Army Institute of Research Josephine Cox Mary Marovich Nelson Michael Merlin Robb Deborah Birx Richard Stout Pontus Blomberg Jenny Enger Sofia Stenler NIH/NIAID Bernard Moss Patricia Earl Members of the AVIP Consortium University of Munich Volker Erle Georg Gasteiger ISS, Rome Barbara Ensoli Bambino Gesú Paolo Rossi Paolo Palma MUHAS, Dar es Salaam Fred Mhalu Muhammad Bakari Eligius Lyuamuya CytoPulse Richard Walters Anna-Karin Roos Cellectis Nils Carlin Staffan Pauli Bartek Zuber

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