Preparation of antigens of and of vaccines for the virus of myst

Chemistry: molecular biology and microbiology – Virus or bacteriophage – except for viral vector or...

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4242021, 4242091, 4242111, 4242181, C12N 700, A61K 39145, A61K 39155, A61K 39295

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055977219

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BRIEF SUMMARY
CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a US national stage application of PCT/FR93/00026 filed Jan. 13, 1993 and published as WO93/14196 Jul. 22, 1993.


BACKGROUND OF THE INVENTION

The present invention relates to the preparation of antigens of and of vaccines for the virus of Mystery Disease, as well as to the antigens and to the vaccines obtained.
The disease called Mystery Disease (M.D.) or also Porcine Reproductive Respiratory Syndrome (P.R.R.S.) began to acquire an identity of its own in pigs, in 1986 in the United States and in 1990 in Europe. This disease manifests itself essentially in pigs by signs of exhaustion, anorexia and hyperthermia of the order of 40.degree. C., which are conventionally observed in sows in pig farms affected by the disease. These signs are accompanied or followed by reproductive disorders (premature or late farrowing and birth of stillborn, mummified or sickly piglets, and return of the sows to heat). A respiratory syndrome can be observed in piglets with interstitial pneumonia lesions. Older pigs can also be affected by respiratory disorders. All this symptomatology can be accompanied by diseases caused by chance infections conventionally observed in pigs.
In G. Wensvoort et al., Mystery Swine Disease in the Netherlands: the isolation of Lelystad virus, The Veterinary Quarterly, Vol. 13, No. 3, 19 Jul. 1991, the isolation of an agent associated with the disease called Mystery Disease is described, which is characterized as a virus, designated Lelystad virus, and which is presented as the causative agent of the disease. This discovery might constitute a first step for the search for a vaccine against this disease.
A process for the industrial production of this virus or of antigen of this virus was not available, the culture seeming possible only in pig alveolar macrophages.


SUMMARY OF THE INVENTION

Yet, the inventors have succeeded in isolating and in identifying another virus, responsible for this disease. This virus is the Myxovirus type, according to the analysis carried out in the electron microscope, and has the characteristic of not being neutralized by porcine anti-influenza sera H1N1 and H3N2.
A strain of this virus identified under the name P129-294 was deposited in the Collection Nationale de Cultures de Micro-organismes held at the Institut Pasteur under No. 1-1153.
In French Patent Application No. 91 13338 filed on 29 Oct. 1991, the inventors described a process for isolating the virus and its use for the preparation of antigens, a process for the industrial production of this virus, as well as the vaccines produced from the above-mentioned antigens containing an effective vaccinating quantity of antigens, in suitable vehicles.
Yet, parallel to the discovery and use of this other virus, of which a strain has been deposited on Oct. 11, 1991 in the Collection Nationale de Cultures de Microorganismes Instit Pasteur, 25, Rue du Docteur Roux, 75724 Paris Cedex 15, FRANCE under the name P129-294 deposit No. I-7153, they also isolated from the same sow obtained from a German pig farm and identified under the No. 294, a completely different virus, including a strain identified under the name P129-294B. They also isolated another viral strain from organs from a piglet from a German farm, collected on 13 Feb. 1991, this strain being identified under the name P120-117B.
The inventors were able to establish that both viruses, namely the Myxovirus-type virus (for example P129-294), designated below A virus, and the viruses corresponding to the strain P120-117B, called hereinafter B virus, occur associated in the samples collected from sick or infected pigs and in the same organs, and can be considered as joint agents responsible for the disease called Mystery Disease.
The subject of the present invention is therefore a process for isolating the B viruses and its use for the preparation of antigens.
A process isolating a B virus can comprise the passage through pig lung or peritoneal macrophages.
In an improved embodiment of the invention, the iso

REFERENCES:
patent: 4070453 (1978-01-01), Bordt et al.
patent: 5476778 (1995-12-01), Chladek et al.
Wensvoort et al., The Veterinary Quarterly, vol. 13 (No. 3), pp. 121-130, (1991).
Terpstra et al., The Veterinary Quarterly, vol. 13 (No. 3), pp. 131-136, (1991).
Abstract, "Dutch Team Isolates Mystery Pig Disease Agent", Jun. 21, 1991, Animal-Pharm, vol. 230, p. 21 (no author listed).
Baron et al , "Report on the first outbreaks of the porcine . . . ", Ann. Rech. Vet., 1992, vol. 23, pp. 161-166.
Benfield et al, "Characterization of swine infertility . . . ", J. Vet. Diagn. Invest., vol. 4, pp. 127-133, 1992.
Albina et al, "An enzyme linked immunosorbent assay (ELISA) . . . ", Ann. Rech. Vet. vol. 23, pp. 167-176, 1992.
Bonn Symposium Abstract, "Fresh clues to mystery pig disease . . . ", Animal-Pharm, vol. 228, p. 3.

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