Institut Pasteur

Centre National De La Recherche Scientifique

Institut National De La Sante Et De La Recherche Medicale

Genticel

Other

Ludwig Institute for Cancer Research Limited

Genkyotex

Inserm

Ladant, Daniel

Daniel Ladant

Claude Leclerc

Agnes Ullmann

Peter Sebo

Benedikt Timmerman

Xavier-Edmond-Edouard Preville

Nicole Guiso

Cecile Bauche

Cécile Bauche

Catherine Fayolle

Nadia Khelef

Mohammed El-Azami El-Idrissi

Pierre Guermonprez

Antoine Danchin

Radim Osicka

Philippe Glaser

Sandra Morel

Véronique Ntsogo Enguene

Octavien Barzu

Gouzel Karimova

Jirina Loucka

Evelyne Krin

Benoît Van Den Eynde

Gilles Dadaglio

Marilyne Davi

Johanna Karst-Spiaczka

Audrey Hessel

Alexandre Chenal

Benoit Van Den Eynde

Xavier Edmond Edouard Preville

The present invention discloses a procedure to produce a monomeric and functional form ofsp, especially, CyaA toxin that can be stably maintained in this functional monodisperse state, even in the absence of any chaotropic agent.

Monomeric and functional adenylate cyclase CyaA toxin

The present invention relates to a method to detect the interaction between a target ligand and a moiety of interest using an adenylate cyclase enzyme (AC) and calmodulin (CaM) as interacting partners, said method comprising: i) expressing in a suitable host cell: (a) a low number of molecules of a first chimeric polypeptide containing AC, and (b) a low number of molecules of a second chimeric polypeptide containing CaM, wherein said AC in said first chimeric polypeptide and/or said CaM in said second chimeric polypeptide has decreased affinity for its interacting partner, wherein said AC in said first chimeric polypeptide is fused to a moiety of interest and said CaM in said second chimeric polypeptide is fused to a target ligand, or conversely, and wherein, when said moiety of interest and said target ligand interact, said AC is activated, and ii) detecting the activation of said AC. The present inventors herein show that only one AC/CaM complex per cell is sufficient to confer a selectable trait to the host cell. Unexpectedly, even less than one AC/CaM complex per cell can be sufficient to confer a selectable trait to the host cell. This surprising result confers a very high sensitivity, that is helpful for screening high affinity interactions, such as antigen-antibody interactions. Moreover, the low expression of the chimeric proteins that is achieved in the present invention allows to characterize toxic moieties, what was not possible before.

In vivo detection of proteins interaction based on adenylate cyclase hybrid system

The invention relates to a novel use of aadenylcyclase toxin in the manufacturing of vectors for targeting in vivo a molecule of interest, specifically to CD11b expressing cells. The invention also relates to an immunogenic composition that primes immune responses, to pharmaceutical compositions and to a new vector for molecule delivery to CD11b expressing cells.

Vectors for molecule delivery to CD11b expressing cells

An immunogenic composition comprising a recombinant protein comprising aCyaA, or a fragment thereof, and a peptide that corresponds to a tumor antigen is provided as a cancer treatment. Methods of treatment with this immunogenic composition are also provided. In an embodiment, the therapeutic composition is a treatment for melanoma, and comprises epitopes from the HLA*0201 epitope. These epitopes include Tyr or GnT-V, and are present in the recombinant proteins CyaA-E5-Tyr and CyaA-E5-GnT-V.

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The invention relates to a recombinant protein comprising one or several polypeptides bearing one or several epitopes of one or several HPV antigens, said polypeptides being inserted in the same or different permissive sites of an adenylate cyclase (CyaA) protein or of a fragment thereof, wherein said CyaA fragment retains the property of said adenylate cyclase protein to target Antigen Presenting Cells. It also concerns polynucleotides encoding the same. The recombinant protein or the polynucleotide can be used for the design of therapeutic means against HPV infection or against its malignant effects.

Recombinant protein carrying human papillomavirus epitopes inserted in an adenylate cyclase protein or fragment thereof therapeutic uses thereof

Recombinant protein carrying human papillomavirus epitopes inserted in an adenylate cyclase protein or fragment thereof. therapeutic uses thereof

The invention relates to modifiedadenylate cyclase toxins which are deficient for CD11b/CD18 binding and to their use in the preparation of pharmaceutical composition for the treatment of whooping cough and/or for the protection againstinfection. The invention also relates to specific fragments ofadenylate cyclase comprising the CD11b/CD18 interaction domain and their use, especially for targeting a molecule of interest to CD11b expressing cells.

A signal amplification system comprises a bacterial multi-hybrid system, and more preferably a two-hybrid system, of at least two chimeric polypeptides containing a first chimeric polypeptide corresponding to a first fragment of an enzyme and a second chimeric polypeptide corresponding to a second fragment of an enzyme or a modulating substance capable of activating said enzyme. The first fragment is fused to a molecule of interest and the second fragment or the modulating substance is fused to a target ligand. The activity of the enzyme is restored by the in vivo interaction between the molecule of interest and the target ligand. Signal amplification is generated and, for example, triggers transcriptional activation. The signal amplification system is useful in a method of selecting a molecule of interest, which is capable of binding to target ligand, wherein the interaction between the molecule of interest and the target ligand is detected with the signal amplification system as a kit therefor. A method of screening for a substance capable of stimulating or inhibiting the interaction between a target ligand and a molecule of interest is also provided.

Bacterial multi-hybrid system and applications thereof

This invention relates to a recombinant plasmid comprising the cyaC and the cyaA genes of Bordetella which directs the expression of Bordetella adenylate cyclase in a transformed host cell. This invention also relates to a recombinat DNA molecule comprising the Bordetella cyaA gene containing at least one insertion of a heterologous DNA sequence at at least one permissive site. This invention further relates to recombinant Bordetella adenylate cyclase comprising a heterologous epitope at a permissive site as well as methods of inducing a specific B cell, helper T cell, and CTL cell immune response.

Recombinant mutants for inducing specific immune responses

A recombinant plasmid comprises the cyaC and the cyaA genes of Bordetella which directs the expression of Bordetella, adenylate cyclase in a transformed host cell. A recombinant DNA molecule can comprise the Bordetella cyaA gene containing at least one insertion of a heterologous DNA sequence at at least one permissive site. In addition, a recombinant Bordetella adenylate cyclase comprises a heterologous epitope at a permissive site. Methods of inducing a specific B cell, helper T cell, and CTL cell immune response are provided.

The adenyl cyclase derivatives are atoxic derivatives in which one or several amino acids are different from those present in the toxic protein, the antibodies formed against these derivatives recognize, however, the toxic protein. These derivatives can be used as vaccines.

Adenyl cyclase derivatives and their biological uses

Growing community of inventors

Cachan, France

Daniel Ladant