Institut Pasteur

Institut National De La Sante Et De La Recherche Medicale

Laleh Majlessi

Roland Brosch

Stewart T Cole

Claude Leclerc

Priscille Brodin

Caroline Demangel

Alexander S Pym

Nadine Honore

Matthias Groschel

Fadel Sayes

Roxane Simeone

Recombinant strains ofbacille Calmette-Guerin (BCG) comprising a heterologous nucleic acid sequence of() are provided. In some embodiments the heterologous nucleic acid sequence ofcomprises a plurality of open reading frames, wherein the plurality of open reading frames comprise open reading frames that encode proteins each at least 95% homologous to the() proteins MMAR5445, MMAR5446, MMAR5447, MMAR5448, MMAR5449, MMAR5450, MMAR5451, MMAR5452, MMAR5453, and MMAR5455. In some embodiments the plurality of open reading frames further comprise an open reading frame that encodes a protein at least 95% homologous to theproteins MMAR5443, MMAR5444, and MMAR5457. In some embodiments the plurality of open reading frames further comprise open reading frames that encode proteins at least 95% homologous to theproteins MMAR5429, MMAR5430, MMAR5431, MMAR5432, MMAR5433, MMAR5434, MMAR5435, MMAR5436, MMAR5437, MMAR5438, MMAR5439, MMAR5440, MMAR5441, MMAR5442, MMAR5454, MMAR5456, MMAR5458, MMAR5459, MMAR5460, and MMAR5461. Pharmaceutical compositions, kits, methods of inducing an immune response againstin a subject, and methods of treating aninfection in a subject are also provided, among other things.

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The present invention relates to a strain ofBCG or, wherein said strain has integrated part or all of the RD1 region responsible for enhanced immunogenicity of the tubercle bacilli, especially the ESAT-6 and CFP-10 genes. These strains will be referred as theBCG::RD1 or::RD1 strains and are useful as a new improved vaccine for preventing tuberculosis and as a therapeutical product enhancing the stimulation of the immune system for the treatment of bladder cancer.

A genetically modified strain oforBCG is provided, wherein the genetically modified strain comprises at least one modified sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, or both, having at least one mutation at T2, Q4, F8, A14, L28, L29, W43, G45, Y51, Q55, Q56, N66, M83, V90, M93, or F94 in SEQ ID NO:1; or at least one mutation at Q3, F7 A13, L27, W42, G44, Y50, Q54, N65, N67, M82, V89, M92, or F93 in SEQ ID NO:2, or a deletion at the terminal end of less than 20 amino acids. In a preferred embodiment, the mutation is at least one of T2H, Q4L, F8I, AI4R, L28A, L29S, W43R, G45T, Q55I, Q56A, N66I, N67A, M83I, V90R, M93T, or F94Q in SEQ ID NO:1, and Q3L, F7I, A13R, L27A, L28S, W42R, Q44T, Q54I, N65I, M82I, V89R, M92T, and F93Q in SEQ ID NO:2. Similarly, the genetically modified strain may also secrete ESAT-6 with a hexa-histidine tag, tetra-cysteine tag, or FLAG-tag, a GFP-fusion, or a short truncation at the C-terminal end of less than 20 amino acids.

The present invention relates to a strain ofBCG or, wherein said strain has integrated part or all of the RD1 region responsible for enhanced immunogenicity of the tubercle bacilli, especially the ESAT-6 and CFP-10 genes. These strains will be referred as theBCG::RDI or::RD1 strains and are useful as a new improved vaccine for preventing tuberculosis and as a therapeutical product enhancing the stimulation of the immune system for the treatment of bladder cancer. These strains are also useful for the expression and presentation of heterologous antigens and molecule that are of therapeutic or prophylactic interest.

Growing community of inventors

Montigny le Bretonneux, France

Laleh Majlessi