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3. Adenovirus vector produced Zika virus-like particles induce a long-lived neutralising antibody response in mice

Author

Carrera, Julio, primary, Aktepe, Turgut E., additional, Earnest, Linda, additional, Christiansen, Dale, additional, Wheatley, Adam K., additional, Tan, Hyon-Xhi, additional, Chung, Amy W., additional, Collett, Simon, additional, McPherson, Kirsty, additional, Torresi, Joseph, additional, Mackenzie, Jason M., additional, and Simmons, Cameron P., additional

Published
2023
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7. Development of virus-like particles with inbuilt immunostimulatory properties as vaccine candidates.

Author

Collett, Simon, Earnest, Linda, Carrera Montoya, Julio, Edeling, Melissa A., Yap, Ashley, Chinn Yi Wong, Christiansen, Dale, Roberts, Jason, Mumford, Jamie, Lecouturier, Valerie, Pavot, Vincent, Marco, Sergio, Joon Keit Loi, Simmons, Cameron, Gulab, Shivali A., Mackenzie, Jason M., Elbourne, Aaron, Ramsland, Paul A., Cameron, Garth, and Hans, Dhiraj

Subjects
SARS-CoV-2, VIRUS-like particles, PAPILLOMAVIRUSES, HUMAN papillomavirus vaccines, HEPATITIS C virus, ATOMIC force microscopy, PLANT viruses, CYTOSKELETAL proteins
Abstract

The development of virus-like particle (VLP) based vaccines for human papillomavirus, hepatitis B and hepatitis E viruses represented a breakthrough in vaccine development. However, for dengue and COVID-19, technical complications, such as an incomplete understanding of the requirements for protective immunity, but also limitations in processes to manufacture VLP vaccines for enveloped viruses to large scale, have hampered VLP vaccine development. Selecting the right adjuvant is also an important consideration to ensure that a VLP vaccine induces protective antibody and T cell responses. For diseases like COVID-19 and dengue fever caused by RNA viruses that exist as families of viral variants with the potential to escape vaccine-induced immunity, the development of more efficacious vaccines is also necessary. Here, we describe the development and characterisation of novel VLP vaccine candidates using SARS-CoV-2 and dengue virus (DENV), containing the major viral structural proteins, as protypes for a novel approach to produce VLP vaccines. The VLPs were characterised by Western immunoblot, enzyme immunoassay, electron and atomic force microscopy, and in vitro and in vivo immunogenicity studies. Microscopy techniques showed proteins self-assemble to form VLPs authentic to native viruses. The inclusion of the glycolipid adjuvant, α-galactosylceramide (α-GalCer) in the vaccine formulation led to high levels of natural killer T (NKT) cell stimulation in vitro, and strong antibody and memory CD8+ T cell responses in vivo, demonstrated with SARS-CoV-2, hepatitis C virus (HCV) and DEN VLPs. This study shows our unique vaccine formulation presents a promising, and much needed, new vaccine platform in the fight against infections caused by enveloped RNA viruses. [ABSTRACT FROM AUTHOR]

Published
2023
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25. A Hepatitis C Virus DNA Vaccine Encoding a Secreted, Oligomerized Form of Envelope Proteins Is Highly Immunogenic and Elicits Neutralizing Antibodies in Vaccinated Mice

Author

Masavuli, Makutiro Ghislain, primary, Wijesundara, Danushka K., additional, Underwood, Alexander, additional, Christiansen, Dale, additional, Earnest-Silveira, Linda, additional, Bull, Rowena, additional, Torresi, Joseph, additional, Gowans, Eric J., additional, and Grubor-Bauk, Branka, additional

Published
2019
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28. Antigen‐specific CD4+CD25+ T cells induced by locally expressed ICOS‐Ig: the role of Foxp3, Perforin, Granzyme B and IL‐10 ‐ an experimental study.

Author

Christiansen, Dale, Mouhtouris, Effie, Hodgson, Russell, Sutton, Vivien R., Trapani, Joseph A., Ierino, Francesco L., and Sandrin, Mauro S.

Subjects
T cells, GENE flow, KNOCKOUT mice, GENE expression, FLOW cytometry, CYTOTOXIC T cells, GRAFT versus host reaction
Abstract

Summary: We have previously reported that ICOS‐Ig expressed locally by a PIEC xenograft induces a perigraft cellular accumulation of CD4+CD25+Foxp3+ T cells and specific xenograft prolongation. In the present study we isolated and purified CD4+CD25+ T cells from ICOS‐Ig secreting PIEC grafts to examine their phenotype and mechanism of xenograft survival using knockout and mutant mice. CD4+CD25+ T cells isolated from xenografts secreting ICOS‐Ig were analysed by flow cytometry and gene expression by real‐time PCR. Regulatory function was examined by suppression of xenogeneic or allogeneic primed CD4 T cells in vivo. Graft prolongation was shown to be dependent on a pre‐existing Foxp3+ Treg, IL‐10, perforin and granzyme B. CD4+CD25+Foxp3+ T cells isolated from xenografts secreting ICOS‐Ig demonstrated a phenotype consistent with nTreg but with a higher expression of CD275 (ICOSL), expression of CD278 (ICOS) and MHC II and loss of CD73. Moreover, these cells were functional and specifically suppressed xenogeinic but not allogeneic primed T cells in vivo. [ABSTRACT FROM AUTHOR]

Published
2019
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29. Porcine cells express more than one functional ligand for the human lymphocyte activating receptor NKG2D

Author

Tran, Peter D., Christiansen, Dale, Winterhalter, Adam, Brooks, Andrew, Gorrell, Mark, Lilienfeld, Benjamin G., Seebach, Jorg Dieter, Sandrin, Mauro, and Sharland, Alexandra

Subjects
ddc:616, Graft Rejection, Swine, Transplantation, Heterologous/immunology, Kidney Transplantation/immunology, Transplantation, Heterologous, NK Cell Lectin-Like Receptor Subfamily K/*metabolism, hemic and immune systems, chemical and pharmacologic phenomena, Epithelial Cells, Galactosyltransferases/metabolism, Graft Rejection/immunology, Galactosyltransferases, Ligands, Kidney Transplantation, biological factors, Epithelial Cells/metabolism, NK Cell Lectin-Like Receptor Subfamily K, Animals, Humans, Cells, Cultured, Protein Binding
Abstract

BACKGROUND: Xenotransplantation could ameliorate the severe shortage of donor organs. The initial results of transplantation from genetically-modified pig donors to primate recipients suggest that hyperacute rejection can be overcome, but thrombotic microangiopathy and the human anti-pig cellular immune response remain as significant impediments to successful clinical xenotransplantation. NKG2D is an activating immunoreceptor found on human natural killer (HuNK) cells, CD8(+) and gammadelta T cells. Signaling through NKG2D mediates cytotoxicity and cytokine secretion by NK cells and co-stimulation of T cells. METHODS: Chinese hamster ovary P (CHOP) cells were transfected with human NKG2D and used in cell-cell binding studies with porcine epithelial, and endothelial cell lines. Soluble recombinant NKG2D-Fc was used to stain various porcine cells and tissues to indicate ligand expression. Porcine cells were used as targets in cytotoxicity assays with the HuNK cell lines NKL and YT, with and without enzymatic removal of pULBP1 and antibody blockade of NKG2D signaling. RESULTS AND CONCLUSIONS: In this study, we demonstrate the expression of ligands for human NKG2D on porcine cell lines of endothelial and epithelial origin, islet cell clusters and rejecting kidney. HuNK cells were activated to kill pig cells expressing NKG2D ligands, and cytotoxicity was inhibited by antibody blockade of NKG2D. A previous study identified pULBP1 as the principal ligand for human NKG2D on pig aortic endothelial cells. In the current study, renal epithelial and intestinal endothelial cells each expressed high surface levels of pULBP1, but binding of soluble recombinant NKG2D and NKG2D-dependent cytotoxicity against these cells persisted after the enzymatic removal of pULBP1, strongly suggesting the presence of at least one additional functional ligand for human NKG2D in these cell types.

Published
2009

30. A CD46CD[55–46] chimeric receptor, eight short consensus repeats long, acts as an inhibitor of both CD46 (MCP)- and CD150 (SLAM)-mediated cell–cell fusion induced by CD46-using measles virus

Author

Plemper, R., Hammond, A., Fielding, A., Cattaneo, R., Lanteri, Marc, Gerlier, D., De Sousa, Emmanuel, Christiansen, Dale, Kyriakou, Peter, Loveland, Bruce, Wild, Fabian, AstraZeneca, Virologie et pathogenèse virale (VPV), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Immunite et Vaccination, and Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Repetitive Sequences, Amino Acid, viruses, Recombinant Fusion Proteins, Molecular Sequence Data, Immunoglobulins, Receptors, Cell Surface, CHO Cells, Biology, Membrane Fusion, Cell membrane, Measles virus, Membrane Cofactor Protein, Signaling Lymphocytic Activation Molecule Family Member 1, Antigens, CD, Virology, Cricetinae, medicine, Animals, Amino Acid Sequence, Binding site, Receptor, ComputingMilieux_MISCELLANEOUS, Glycoproteins, Membrane Glycoproteins, CD46, biology.organism_classification, female genital diseases and pregnancy complications, Transmembrane protein, medicine.anatomical_structure, Viral replication, Cytoplasm, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology
Abstract

According to their cellular receptor use, measles virus (MV) strains can be separated into two phenotypes, CD46-using and CD46-non-using. A long chimeric receptor, CD46CD[55–46], was generated from the CD46 backbone, encompassing the four short consensus repeat (SCR) domains of CD46 linked via a flexible glycine hinge to SCR1 and SCR2 of CD55, SCR3 and SCR4 of CD46 and the STP, transmembrane and cytoplasmic tail of CD46. This chimeric receptor was proficient for MV binding but deficient in mediating MV-induced cell-to-cell fusion and virus replication, possibly due to the extended distance between the MV haemagglutinin (H) binding site (CD46 SCR1–SCR2) and the cell membrane. When coexpressed with either wild-type CD46 or CD150, this fusion-incompetent receptor exerted a dominant negative effect and inhibited both cell-to-cell fusion and entry of MV with CD46-using, but not CD46-non-using, phenotype. A soluble octameric CD46–C4bpα exhibited similar CD46- and CD150-mediated fusion inhibition properties only against CD46-using MV. This suggests that the long CD46CD[55–46] receptor acts by sequestering incoming MV prior to its binding to the shorter functional CD46 or CD150 receptor.

Published
2002
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31. Prolonged Xenograft Survival Induced by Inducible Costimulator-Ig is Associated With Increased Forkhead Box P3+ Cells

Author

Hodgson, Russell, Christiansen, Dale, Ziolkowski, Andrew, Mouhtouris, Effie, Simeonovic, Charmaine, Ierino, Francesco, Sandrin, Mauro, Hodgson, Russell, Christiansen, Dale, Ziolkowski, Andrew, Mouhtouris, Effie, Simeonovic, Charmaine, Ierino, Francesco, and Sandrin, Mauro

Abstract

Background: Blockade of the inducible costimulator (ICOS) pathway has been shown to prolong allograft survival; however, its utility in xenotransplantation is unknown. We hypothesize that local expression of ICOS-Ig by the xenograft will suppress the T-cell response resulting in significant prolonged graft survival. METHODS.: Pig iliac artery endothelial cells (PIEC) secreting ICOS-Ig were generated and examined for the following: (1) inhibition of allogeneic and xenogeneic proliferation of primed T cells in vitro and (2) prolongation of xenograft survival in vivo. Grafts were examined for Tregs by flow cytometry and cytokine levels determined by quantitative reverse-transcriptase polymerase chain reaction. Results: Soluble ICOS-Ig markedly decreased allogeneic and xenogeneic primed T-cell proliferation in a dose-dependent manner. PIEC-ICOS-Ig grafts were significantly prolonged compared with wild-type grafts (median survival, 34 and 12 days, respectively) with 20% of PIEC-ICOS-Ig grafts surviving more than 170 days. Histological examination showed a perigraft cellular accumulation of Forkhead box P3 (Foxp3) cells in the PIEC-ICOS-Ig grafts, these were also shown to be CD3+CD4+CD25+. Survival of wild-type PIEC grafts in a recipient simultaneously transplanted with PIEC-ICOS-Ig were also prolonged, with a similar accumulation of Foxp3 cells at the periphery of the graft demonstrating ICOS-Ig induces systemic graft prolongation. However, this prolongation was specific for the priming xenograft. Intragraft cytokine analysis showed an increase in interleukin-10 levels, suggesting a potential role in induction/function of CD4+CD25+Foxp3+ cells. Conclusions: This study demonstrates prolonged xenograft survival by local expression of ICOS-Ig, we propose that the accumulation of CD4+CD25+Foxp3+ cells at the periphery of the graft and secretion of interleukin-10 is responsible for this novel observation.

Published
2011

32. Local expression of IDO, either alone or in combination with CD40Ig, IL10 or CTLA4Ig,inhibits indirect xenorejection responses

Author

Mulley, William R, Li, Yu Qin, Wee, Janet L, Dodge, Natalie, Christiansen, Dale, Simeonovic, Charmaine, Ierino, Francesco, Sandrin, Mauro, Mulley, William R, Li, Yu Qin, Wee, Janet L, Dodge, Natalie, Christiansen, Dale, Simeonovic, Charmaine, Ierino, Francesco, and Sandrin, Mauro

Abstract

Background: To overcome cell-mediated xenorejection by transgenic expression of immunomodulatory molecules by a graft, it is likely that expression of multiple molecules will be required. Previous studies support the use of the immunomodulatory agents indoleamine 2,3-dioxygenase (IDO), CD40Ig, interleukin 10 (IL10), and CTLA4Ig for suppression of rejection responses. We examined the effects of local expression of these molecules by a porcine cell line (PIEC) on indirect murine xenorejection responses in vitro and in vivo. Methods: The PIEC stable lines expressing IDO, CD40Ig, and IL10 as single molecules were generated. In addition, PIEC lines expressing IDO with either CD40Ig, IL10 or CTLA4Ig were generated to produce cell lines expressing two molecules. BALB/c mice were primed with wild type PIEC, followed by harvesting splenocytes used as responder cells and PIEC expressing immunomodulatory molecules as stimulators, in proliferation and cytokine assays. In vivo effects of modified PIEC were examined by transplantation of PIEC lines expressing the immunomodulatory molecules under the renal capsule of naïve mice. PIEC grafts were harvested for histological evaluation at days 7 and 14. Results: Proliferation of primed BALB/c splenocytes was inhibited most significantly by IDO compared with control cells (49%, P = 0.02). In addition both Th1 (interferon-gamma) and Th2 (IL4 and IL10) cytokines were markedly inhibited in vitro by IDO expression. IL10 expressing cells did not inhibit proliferation as potently (37%, P = 0.03) whilst CD40Ig lead to an increase in proliferative responses (59%, P = 0.02). Co-expression of CD40Ig, IL10, and CTLA4Ig with IDO resulted in further modest reductions in proliferation compared with IDO expression alone. When transplanted under the renal capsule of BALB/c mice, those grafts expressing IDO demonstrated significantly lower levels of lymphocyte infiltration at days 7 and 14 than control grafts and those expressing CD40Ig, CTLA4Ig or IL

Published
2008

35. Characterization of a CD46 transgenic pig and protection of transgenic kidneys against hyperacute rejection in non-immunosuppressed baboons

Author

Loveland, Bruce E., primary, Milland, Julie, additional, Kyriakou, Peter, additional, Thorley, Bruce R., additional, Christiansen, Dale, additional, Lanteri, Marc B., additional, Regensburg, Mark, additional, Duffield, Maureen, additional, French, Andrew J., additional, Williams, Lindsay, additional, Baker, Louise, additional, Brandon, Malcolm R., additional, Xing, Pei-Xiang, additional, Kahn, Del, additional, and McKenzie, Ian F.C., additional

Published
2009
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