Institut De Recherche Pour Le Developpement (ird)

Institut De Recherche Pour Le Developpement

Institut Francais De Recherche Scientifique Pour Le Developpement En Cooperation (orstom)

Institut Pasteur De Tunis

Institut De L'environnement Et De Recherches Agricoles

Institut De Recherche Pour De Developpement (ird)

Virbac Sa

Jean-Loup Lemesre

Philippe Holzmuller

Denis Sereno

Christophe Brugidou

Denis Fargette

Drissa Sereme

Moumouni Konate

Gnissa Konate

Mireille Cavaleyra

Rachel Bras-Gonçalves

Rachel Bras Gonçalves

Amel Garnaoui

Elodie Petitdidier-Lesin

Gérard-Marie Papierok

Rachel Elise Bras Goncalves

Sylvie Gueguen

The invention relates to multi-epitopic peptide compounds obtained from PSA, HwB and LmLRAB proteins ofas well as to pharmaceutical compositions and vaccines for use against one or more leishmaniases.

Multi-epitopic peptide compounds and vaccines against leishmaniasis

The invention provides a composition for the inoculation in a plant of agrobacteria transfected by expression vectors, in order to produce in said plant a protein of interest or a derivative of said protein, by deletion or by mutation, characterized in that it comprises

Means for the transient production in plants of recombinant proteins that can be used in particular in prophylaxis and in therapeutics

The invention relates to nucleic acid constructions, characterized in that they comprise nucleic acids which are isolated in the sense position and which are capable of coding for an immunogenic protein of promastigotes or amastigotes of, said nucleic acids responding to one of the sequences SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 5 et SEQ ID No. 11 and coding for a protein respectively exhibiting a sequence SEQ ID No. 6, SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 et SEQ ID No. 12. The invention can be used for over-expression of the genes ofcoding for an excretion/secretion antigen.

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The invention relates to an expression vector of a gene coding for an antigenic protein ofpromastigote, characterized in that it comprises a PSA gene insert in opposite orientation. The invention can be applied to the development of mutants under-expressing, or no longer expressing, genes coding for an antigenic protein ofpromastigote, and to the therapeutic and/or vaccine-oriented uses thereof.

The process of the invention comprises the implementation of axenic conditions, with use of a liquid single-phase culture medium. For obtaining the amastigote forms, this medium is buffered at a pH of 5.5 to 6.5 and has an osmolarity of at least 400 milliosmoles/kg of liquid, and in particular 400 to 550 milliosmoles/kg of liquid. For obtaining promastigote forms, this medium is buffered at a pH of 7 to 7.5 and has an osmolarity of at least 300 milliosmoles/kg of liquid. This process allows the adaptation and culture in vitro of different stages of tissular parasites, such as leishmanias andor also hematoprotozoa.

Process for the in vitro culture of different stages of tissular parasites

The present invention provides a method for carrying out in vitro the complete developmental sequence culture of tissular parasites, which includes culturing the parasites in a totally defined culture medium which is an axenic . . . monophasic liquid culture medium, free of serum and free of serum-derived or cell-derived macromolecules, proteins and peptides. For obtaining amastigote forms, this medium is buffered at a pH of 5.5 to 6.5 and has an osmolarity of at least 400 milliosmoles/kg of liquid. For obtaining promastigote forms, the medium is buffered at a pH of 7 to 7.5 and has an osmolarity of at least 300 milliosmoles/kg liquid. Application to the in vitro culture of different stages of tissular parasites such as, and hamatoprotozoa is also provided.

The present invention provides a method for carrying out in vitro the complete developmental sequence culture of tissular parasites, which includes culturing the parasites in a totally defined culture medium which is an axenic monophasic liquid culture medium, free of serum and free of serum-derived or cell-derived macromolecules, proteins and peptides. For obtaining amastigote forms, this medium is buffered at a pH of 5.5 to 6.5 and has an osmolarity of at least 400 milliosmoles/kg of liquid. For obtaining promastigote forms, the medium is buffered at a pH of 7 to 7.5 and has an osmolarity of at least 300 milliosmoles/kg liquid. Application to the in vitro culture of different stages of tissular parasites such as Leishmania, and hamatoprotozoa is also provided.

Process for the in vitro culture of different stages of tissue parasites

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Saint Martin de Londres, France

Jean-Loup Lemesre