Institut Pasteur

National Institutes of Health, a Component of the US Dept. of Health & Human Services

Medical Research Council

Assistance Publique

Other

Institut National De La Sante Et De La Recherche Medicale

Université Pierre Et Marie Curie (Paris Vi)

Institut Pasteur and Centre National De La Recherche Scientifique

Universite De Berne

Universite Paris-vi

Stewart T Cole

Roland Buchrieser-Brosch

Stephen Gordon

Alain Billault

Rolf E Streeck

Pierre Sonigo

Marc Alizon

Simon Wain-Hobson

Jean-Claude Chermann

Solange Chamaret

Douglas B Young

Ying Zhang

Roland Brosch

Beate Heym

Luc Montagnier

Claude Leclerc

Priscille Brodin

Nadine Honore

Laleh Majlessi

Robert C Gallo

Pierre Tiollais

Francoise Barre-Sinoussi

Christine Rouzioux

Mangalasseril G Sarngadharan

Bernard Krust

Mikulas Popovic

Francoise Rey

Willy Rozenbaum

Claudine Axler-Blin

Charles Dauguet

Oliver Danos

Marie-Therese Nugeyre

Olivier Danos

Caroline Demangel

Jacques Gruest, Legal Representative

Francois Brun-Vezinet

Alexander S Pym

Amalio Telenti

Thierry Garnier

Thomas Bodmer

Fran&ccedil;ois Clavel

Francedillaois Clavel

Luc Montangnier

Fran&ccedil;oise Barre-Sinoussi

Stéphen Gordon

Karin Eiglmeier

Francedillaoise Barre-Sinoussi

Glyn Hewinson

Martin Sapp

Wolfgang Philipp

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.

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A method for detectinginvolves contacting a biological sample with the product of expression of at least one ORF contained in a polynucleotide present inand absent inBCG; and detecting whether an immunological complex is formed between the product thereof and antibodies contained in the biological sample, wherein the presence of a complex indicates an infection with

Method for isolating a polynucleotide of interest from the genome of a mycobacterium using a BAC-based DNA library: application to the detection of mycobacteria

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 invention concerns the isolation of nucleotide and peptide sequences in particular for differentiating, in diagnostic terms, an immunization resulting from BCG vaccination of an infection by. Said sequences are eitherBCG/specific orspecific. The invention also concerns a method for detecting said sequences, a method for detecting antibodies generated by the products expressing said sequences as well as kits for implementing said methods. Finally, the invention concerns novel vaccines.

The present invention is the identification of a nucleotide sequence which make it possible in particular to distinguish an infection resulting from the vast majority ofstrains from an infection resulting fromBCG. The subject of the present invention is also a method for detecting the sequences in question by the products of expression of these sequences and the kits for carrying out these methods. Finally, the subject of the present invention is novel vaccines.

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.

The present invention is directed to a method for isolating a polynucleotide of interest that is present or is expressed in a genome of a first mycobacterium strain and that is absent or altered in a genome of a second mycobacterium strain which is different from the first mycobacterium strain using a bacterial artificial chromosome (BAC) vector. The invention further relates to a polynucleotide isolated by this method and recombinant BAC vector used in this method. In addition the present invention comprises method and kit for detecting the presence of a mycobacteria in a biological sample.

The present invention is directed to a method of selection of purified nucleotidic sequences or polynucleotides encoding proteins or part of proteins carrying at least an essential function for the survival or the virulence ofspecies by a comparative genomic analysis of the sequence of the genome ofaligned on the genome sequence ofandandmarker polypeptides of nucleotides encoding the polypeptides, and methods for using the nucleotides and the encoded polypeptides are disclosed.

Comparative mycobacterial geneomics as a tool for identifying targets for the diagnosis, prophylaxis or treatment of mycobacterioses

This invention is in the field of lymphadenopathy virus, which has been designated Human Immunodeficiency Virus Type 1 (HIV-1) to detect the presence of DNA, RNA or antibodies of the lymphadenopathy retrovirus associated with the acquired immune deficiency syndrome or of the lymphadenopathy syndrome by the use of DNA fragments or the peptides encoded by said DNA fragments. The invention further relates tot he DNA fragments, vectors comprising them and the proteins expressed.

DNA sequence of the LTR region of human immunodeficiency virus type 1 (HIV-1)

This invention is in the field of lymphadenopathy virus, which has been designated Human Immunodeficiency Virus Type 1 (HIV-1). This invention relates to a diagnostic means and method to detect the presence of DNA, RNA or antibodies of the lymphadenopathy retrovirus associated with the acquired immune deficiency syndrome or of the lymphadenopathy syndrome by the use of DNA fragments or the peptides encoded by said DNA fragments. The invention further relates to the DNA fragments, vectors comprising them and the proteins expressed.

Cloned DNA sequence related to genomic RNA of human immunodeficiency virus (HIV-1)

A method for isolating a polynucleotide of interest that is present in the genome of a first mycobacterium strain and/or is expressed by the first mycobacterium strain, where the polynucleotide of interest is also absent or altered in the genome of a second mycobacterium strain and/or is not expressed in the second mycobacterium. The method includes (a) contacting the genomic DNA of the first mycobacterium strain under hybridizing conditions with the DNA of a least one clone that belongs to a bacterial artificial chromosome (BAC) genomic DNA library of the second mycobacterium strain, and (b) isolating the polynucleotide of interest that does not form a hybrid with the DNA of the second mycobacterium strain. This invention further pertains to astrain H37Rv genomic DNA library, as well as aBCG strain Pasteur genomic DNA library, and the recombinant BAC vectors that belong to those genomic DNA libraries. This invention also relates to mycobacterial nucleic acids, and methods and kits for using these nucleic acids to detect mycobacteria in a biological sample.

Method for isolating a polynucleotide of interest from the genome of a mycobacterium using a BAC-based DNA library, application to the detection of mycobacteria

A method for isolating a polynucleotide of interest that is present in the genome of a first mycobacterium strain and/or is expressed by the first mycobacterium strain, where the polynucleotide of interest is also absent or altered in the genome of a second mycobacterium strain and/or is not expressed in the second mycobacterium. The method includes (a) contacting the genomic DNA of the first mycobacterium strain under hybridizing conditions with the DNA of a least one clone that belongs to a bacterial artificial chromosome (BAC) genomic DNA library of the second mycobacterium strain, and (b) isolating the polynucleotide of interest that does not form a hybrid with the DNA of the second mycobacterium strain. This invention further pertains to a strain H37Rv genomic DNA library, as well as a BCG strain Pasteur genomic DNA library, and the recombinant BAC vectors that belong to those genomic DNA libraries. This invention also relates to mycobacterial nucleic acids, and methods and kits for using these nucleic acids to detect mycobacteria in a biological sample.

Method for isolating a polynucleotide of interest from the genome of a mycobacterium using a bac-based DNA library: application to the detection of mycobacteria

The molecular cloning and characterization of a novel human retrovirus, designated lymphadenopathy-associated virus, or LAV, is disclosed. LAV was originally isolated from a patient with acquired immune deficiency syndrome (AIDS). A cloned LAV complementary DNA (cDNA) was used to screen a library of recombinant phages constructed from the genomic DNA of LAV-infected T lymphocytes. The nucleotide sequence of an insert obtained from the recombinant phage clone &lgr;J19 was ascertained through M13 shotgun cloning and the dideoxy chain termination sequencing method. The env coding region was identified and various hydrophilic peptides obtained therefrom. These peptides correspond to amino acids 551-577, 594-603, 621-630, 657-679, and 719-758 of the LAV envelope glycoprotein. These peptides should provide suitable diagnostic reagents for the detection LAV-specific antibodies and for the generation of LAV-specific immunological reagents.

Nucleic acids and peptides of human immunodeficiency virus type (HIV-1)

Determined DNA sequenced derived from a papillomavirus genome, their uses for in vitro diagnostic purposes and the production of antigenic compositions

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Clamart, France

Stewart T Cole