High-level expression from two independent expression cassettes in replication-incompetent adenovirus type 35 vector.
Replication-incompetent adenovirus type 35 (rAd35) represents a potent vaccine carrier that elicits strong, antigen-specific T- and B-cell responses in diverse preclinical models. Moreover, Ad35 is rare in human populations, resulting in the absence of neutralizing antibodies against this carrier, in contrast to the commonly used rAd5. Therefore, rAd35 is being investigated as a vaccine carrier for a number of diseases for which an effective vaccine is needed, including malaria, AIDS and tuberculosis. However, it can be perceived that effective immunization will require insertion of multiple antigens into adenoviral vectors. We therefore wanted to create rAd35 vectors carrying double expression cassettes, to expand within one vector the number of insertion sites for foreign DNA encoding antigenic proteins. We show that it is possible to generate rAd35 vectors carrying two cytomegalovirus promoter-driven expression cassettes, provided that the polyadenylation signals in each expression cassette are not identical. We demonstrate excellent rAd35 vector stability and show that expression of a transgene is not influenced by the presence of a second expression cassette. Moreover, by using two model vaccine antigens, i.e. the human immunodeficiency virus-derived Env-gp120 protein and the Plasmodium falciparum-derived circumsporozoite protein, we demonstrate that potent T- and B-cell responses are induced to both antigens expressed from a single vector. Such rAd35 vectors thus expand the utility of rAd35 vaccine carriers for the development of vaccines against, for example, malaria, AIDS and tuberculosis.
Vogels
Ronald
R
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Zuijdgeest
David
D
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
van Meerendonk
Michelle
M
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Companjen
Arjen
A
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Gillissen
Gert
G
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Sijtsma
Jeroen
J
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Melis
Irene
I
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Holterman
Lennart
L
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Radosevic
Katarina
K
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Goudsmit
Jaap
J
Center of Poverty-Related Communicable Diseases, Academic Medical Center, Amsterdam, The Netherlands.|Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
Havenga
Menzo J E
MJE
Crucell Holland BV, PO Box 2048, 2301 CA Leiden, The Netherlands.
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D003587:Cytomegalovirus / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000276:immunology; D007371:Interferon-gamma / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology; D039104:RNA 3' Polyadenylation Signals / Q000235:genetics; D013154:Spleen / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014614:Vaccines, Synthetic / Q000235:genetics; D014765:Viral Vaccines / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics
D000913:Antibodies, Protozoan; D000914:Antibodies, Viral; D015699:HIV Envelope Protein gp120; D015800:Protozoan Proteins; D014614:Vaccines, Synthetic; D014765:Viral Vaccines; C041595:circumsporozoite protein, Protozoan; D007371:Interferon-gamma