Leadd B.v.

Mathieu Hubertus Maria Noteborn

Astrid Adriana Anna Maria Danen-Van Oorschot

Guus Koch

Jennifer Leigh Rohn

Peter Donner

Dominik Mumberg

The invention relates to the field of apoptosis. The invention provides novel therapeutic possibilities, for example novel combinatorial therapies or novel therapeutic compounds that can work alone, sequentially to, or jointly with apoptin, especially in those cases wherein p53 is (partly) non-functional.

Apoptin-associating proteins

The coding information for three putative chicken anemia virus proteins (VP1, VP2, VP3) was inserted into a baculovirus vector and expressed in insect cells. Only lysates of insect cells which have synthesized equivalent amounts of all three recombinant CAV proteins or cells which synthesized mainly VP1 plus VP2 induced neutralizing antibodies directed against CAV in inoculated chickens. Progeny of those chickens were protected against clinical disease after CAV challenge. Inoculation of a mixture of lysates of cells that were separately infected with VP1-, VP2- and VP3-recombinant baculovirus did not induce significant levels of neutralizing antibody directed against CAV and their progeny were not protected against CAV challenge. Expression in the same cell of at least VP1 plus VP2 is required to obtain sufficient protection in chickens. Therefore, recombinant CAV proteins produced by baculovirus vectors can be used as a sub-unit vaccine against CAV infections.

Chicken anemia virus mutants and vaccines and uses based on the viral proteins VP1, VP2 and VP3 or sequences of that virus coding therefor

Phosphorylated Apoptin is described. Apoptin is tumor-specifically phosphorylated and part of the Apoptin apoptotic pathway in tumor cells is elucidated. New therapeutic possibilities, for example, novel therapeutic compounds that can work alone or, sequentially to, or jointly with other known compounds. Also, the use of tumor-specifically phosphorylation of Apoptin for diagnostic purposes is described. Such a diagnostic purpose can, for example, be a method for detecting the presence of cancer cells or cells that are cancer prone or a method to identify a putative cancer inducing agent or a method for the in vitro treatment effect of Apoptin on tumor cells by testing the phosphorylation state of Apoptin. Even more, the invention provides possibilities to further elucidate the apoptotic pathway and to identify for example crucial mediators of phosphorylation in human tumor cells. Interfering with such a mediator could provide new anti-cancer therapies.

Modifications of apoptin

The invention relates to activation of apoptosis by means of interference of the function of Bip-like compounds. Also the invention relates to anti-tumor therapies with compounds, which negatively interfere with Bip-like compounds leading to induction of apoptosis, resulting in the elimination of tumor cells. Also the invention relates to therapies for diseases related to aberrant apoptosis induction, such as auto-immune diseases. Also the invention describes the diagnosis of cells, which are susceptible to apoptin or apoptin-like induced apoptosis.

Methods and means for inducing apoptosis by interfering with Bip-like proteins

The coding information for three putative chicken anemia virus proteins (VP1, VP2, VP3) was inserted into a baculovirus vector and expressed in insect cells. The immunogenic properties of the chicken anemia virus (CAV) proteins produced separately or together in insect-cell cultures were analyzed by inoculating them into chickens. Only lysates of insect cells which have synthesized equivalent amounts of all three recombinant CAV proteins or cells which synthesized mainly VP1 plus VP2 induced neutralizing antibodies directed against CAV in inoculated chickens. Progeny of those chickens were protected against clinical disease after CAV challenge. Inoculation of a mixture of lysates of cells that were separately infected with VP1-, VP2- and VP3-recombinant baculovirus did not induce significant levels of neutralizing antibody directed against CAV and their progeny were not protected against CAV challenge. Our results indicate that expression in the same cell of at least two CAV proteins, VP1 plus VP2, is required to obtain sufficient protection in chickens. Therefore, recombinant CAV proteins produced by baculovirus vectors can be used as a sub-unit vaccine against CAV infections.

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Leiderdorp, Netherlands

Mathieu Hubertus Maria Noteborn