Title:  Recombinant immunogenic actinomycetale

United States Patent:  6,235,518

Assignee:  Institut Pasteur (Paris, FR)

Filed:  July 26, 1994

PCT NO:  PCT/EP92/01343

102(e) Date:  July 26, 1994

PCT PUB. Date:  December 23, 1993

Foreign Application Priority Data:  Jun 14, 1991[GB] (91401601)

A mycobacteria transformed with an antigen-encoding gene, such as nef, under the control of a Streptomyces stress-responsive promoter, such as the S. albus groES/groEL1 promoter, and preferably associated with a synthetic ribosome binding site. The recombinant mycobacteria can be used as a vaccine against, for example, a pathogen which carries the antigen.

SUMMARY OF THE INVENTION

This invention provides an immunogenic Actinomycetale strain, particularly a strain of mycobacteria, transformed with the following, operably linked, chimaeric DNA sequence, comprising an antigen-encoding gene that: a) is foreign to the Actinomycetale strain; b) encodes a desired antigen, preferably an immunizing antigen; c) is under the control of a promoter foreign to the Actinomycetale strain, preferably a promoter from another strain of Actinomycetale, especially a promoter from another species of Actinomycetale, particularly a Streptomyces promoter, quite particularly a stress-responsive (e.g., heat-shock) promoter; and d) is preferably associated with a ribosome binding site ("RBS") foreign to the Actinomycetale strain, particularly foreign to Actinomycetale and/or synthetic.

This invention also provides the aforementioned chimaeric DNA sequence, especially wherein the antigen-encoding gene codes for an HIV-1 protein (European patent publications ("EP") 0 178 978 and 201,540) or an HIV-2 protein (EP 0 239 425), particularly a Nef protein (e.g., the Nef 1 protein of HIV-1 or Nef 2 protein of HIV-2), or an antigenic fragment or an epitope thereof which can induce an immune response in a mammal, particularly a T or B response, quite particularly wherein the antigen-encoding gene is under the control of the S. albus groES/groEL1 promoter.

This invention further provides a system, such as a plasmid, capable of transforming a Actinomycetale strain. This system comprises a foreign promoter, a foreign ribosome binding site and a DNA fragment coding for an antigen foreign to the Actinomycetale strain, particularly the aforementioned chimaeric DNA sequence, quite particularly pWRIP17.

This invention yet further provides: a) an immunogenic composition, particularly a vaccine, comprising the aforementioned transformed Actinomycetale strain, particularly a strain of mycobacteria, quite particularly BCG, and if necessary, a suitable carrier, as well as, for example, an adjuvant for human use; and b) a method of immunizing a mammalian host, particularly a human, against, for example, a pathogen, comprising the step of administering to the host the aforementioned composition.

This invention also provides: a) a process to produce the desired antigen by culturing the transformed Actinomycetale strain; and b) a process for using the desired antigen or the transformed Actinomycetale strain, for example, as a diagnostic agent and/or an immunogenic agent. An antigenic preparation containing the desired antigen or the transformed Actinomycetale strain can induce the synthesis of specific antibodies or cellular reactions in in vitro and in vivo diagnostic tests and immunogenic treatments.

Claim 1 of 31 Claims

What is claimed is:

1. An immunogenic strain of Actinomycetale comprising a recombinant DNA, wherein said recombinant DNA includes a gene, said gene includes a promoter from a strain different from that of said Actinomycetale strain or from a species different from that of said Actinomycetale strain, and a synthetic ribosome binding site, said gene is capable of being expressed in said immunogenic strain, said gene encodes a protein that is foreign to the species to which said immunogenic strain belongs, and said protein elicits an immune response in a mammal.

____________________________________________
If you want to learn more about this patent, please go directly to the U.S. Patent and Trademark Office Web site to access the full patent.