Polynucleic acids isolated from a porcine reproductive and...

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Virus or component thereof

Reexamination Certificate

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C424S093100, C424S093600, C424S186100, C435S069300, C435S235100, C536S023720

Reexamination Certificate

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06592873

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention concerns DNA isolated from a porcine reproductive and respiratory virus (PRRSV), a protein and/or a polypeptide encoded by the DNA, a vaccine which protects pigs from a PRRSV based on the protein or DNA, a method of protecting a pig from a PRRSV using the vaccine, a method of producing the vaccine, a method of treating a pig infected by or exposed to a PRRSV, and a method of detecting a PRRSV.
2. Discussion of the Background
In recent years, North American and European swine herds have been susceptible to infection by new strains of reproductive and respiratory viruses (see
A.A.S.P.
, September/October 1991, pp. 7-11
; The Veterinary Record
, Feb. 1, 1992, pp. 87-89; Ibid., Nov.30, 1991, pp. 495-496; Ibid., Oct. 26, 1991, p. 370; Ibid., Oct. 19, 1991, pp. 367-368; Ibid., Aug. 3, 1991, pp. 102-103; Ibid., Jul. 6, 1991; Ibid., Jun. 22, 1991, p. 578; Ibid., Jun. 15, 1991, p. 574; Ibid., Jun. 8, 1991, p. 536; Ibid., Jun. 1, 1991, p. 511; Ibid., Mar. 2, 1991, p. 213). Among the first of the new strains to be identified was a virus associated with the so-called Mystery Swine Disease (MSD) or “blue-eared syndrome”, now known as Swine Infertility and Respiratory Syndrome (SIRS) or Porcine Reproductive and Respiratory Syndrome (PRRS).
An MSD consisting of reproductive failure in females and respiratory disease in nursing and weaned pigs appeared in the Midwestern United States in 1987 (Hill et al.,
Am. Assoc. Swine Practitioner Newsletter
4:47 (1992); Hill et al.,
Proceedings Mystery Swine Disease Committee Meeting
, Denver, Colo. 29-31 (1990); Keffaber,
Am. Assoc. Swine Practitioner Newsletter
1:1-9 (1989); Loula,
Agri
-
Practice
12:23-34 (1991)). Reproductive failure was characterized by abortions, stillborn and weak-born pigs. The respiratory disease in nursing and weaned pigs was characterized by fever, labored breathing and pneumonia. A similar disease appeared in Europe in 1990 (Paton et al.,
Vet. Rec.
128:617 (1991); Wensvoort et al.,
Veterinary Quarterly
13:121-130 (1991); Blaha,
Proc. Am. Assoc. Swine Practitioners
, pp. 313-315 (1993)), and has now been recognized worldwide.
This disease has also been called porcine epidemic abortion and respiratory syndrome (PEARS), blue abortion disease, blue ear disease (U.K.), abortus blau (Netherlands), seuchenhafter spatabort der schweine (Germany), Heko-Heko disease, and in the U.S., Wabash syndrome, mystery pig disease (MPD), and swine plague (see the references cited above and Meredith,
Review of Porcine Reproductive and Respiratory Disease Syndrome
, Pig Disease Information Centre, Department of Veterinary Medicine, Madingley Road, Cambridge CB3 OES, U.K. (1992); Wensvoort et al.,
Vet. Res.
24:117-124 (1993); Paul et al.,
J. Clin. Vet. Med.
11:19-28 (1993)). In Europe, the corresponding virus has been termed “Lelystad virus.”
At an international conference in May, 1992, researchers from around the world agreed to call this disease Porcine Reproductive and Respiratory Syndrome (PRRS). The disease originally appeared to be mainly a reproductive disease during its early phases, but has now evolved primarily into a respiratory disease.
Porcine reproductive and respiratory syndrome virus (PRRSV) is a relatively recently recognized swine pathogen associated with porcine reproductive and respiratory syndrome (PRRS). PRRSV is a significant pathogen in the swine industry. PRRSV infections are common in the U.S. swine herds. Outbreaks of PRRS in England have led to cancellation of pig shows.
The symptoms of PRRS include a reluctance to eat (anorexia), a mild fever (pyrexia), cyanosis of the extremities (notably bluish ears), stillbirths, abortion, high mortality in affected litters, weak-born piglets and premature farrowing. The majority of piglets born alive to affected sows die within 48 hours. PRRS clinical signs include mild influenza-like signs, rapid respiration (“thumping”), and a diffuse interstitial pneumonitis. PRRS virus has an incubation period of about 1-2 weeks from contact with a PRRSV-infected animal. The virus appears to be an enveloped RNA arterivirus (
The Veterinary Record
, Feb. 1, 1992). The virus has been grown successfully in pig alveolar macrophages and CL2621 cells (Benfield et al,
J. Vet. Diagn. Invest.,
4:127-133, 1992; Collins et al, Swine Infertility and Respiratory Syndrome/Mystery Swine Disease.
Proc., Minnesota Swine Conference for Veterinarians
, pp. 200-205, 1991), and in MARC-145 cells (Joo, PRRS: Diagnosis,
Proc., Allen D. Leman Swine Conference
, Veterinary Continuing Education and Extension, University of Minnesota (1993), 20:53-55; Kim et al,
Arch. Virol.,
133:477-483 (1993)). A successful culturing of a virus which causes SIRS has also been reported by Wensvoort et al (Mystery Swine Disease in the Netherlands: The Isolation of Lelystad Virus. Vet. Quart. 13:121-130, 1991).
Initially, a number of agents were incriminated in the etiology of this disease (Wensvoort et al.,
Vet. Res.
24:117-124 (1993); Woolen et al.,
J. Am. Vet. Med. Assoc.
197:600-601 (1990)). There is now a consensus that the causative agent of PRRS is an enveloped RNA virus referred to as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), reportedly of approximately 62 nm in diameter (Benfield et al.,
J. Vet. Diagn. Invest.,
4:127-133, 1992).
Virus isolates vary in their ability to replicate in continuous cell lines. Some grow readily, while others require several passages and some grow only in swine alveolar (SAM) cultures (Bautista et al.,
J. Vet. Diagn. Invest.
5:163-165, 1993; see also the Examples hereunder [particularly Table 1]).
PRRSV is a member of an Arterivirus group which includes equine arteritis virus (EAV), lactate dehydrogenase-elevating virus (LDV) and simian hemorrhagic fever virus (SHFV) (Benfield et al., 1992, supra; Plagemann,
Proc. Am. Assoc. Swine Practitioners,
4:8-15 1992; Plagemann and Moennig,
Adv. Virus Res.
41:99-192, 1992; Conzelmann et al.,
Virology,
193:329-339, 1993; Godney et al.,
Virology,
194:585-596, 1993; Meulenberg et al.,
Virology,
192:62-72, 1993). The positive-strand RNA viruses of this Arterivirus group resemble togaviruses morphologically, but are distantly related to coronaviruses and toroviruses on the basis of genome organization and gene expression (Plagemann et al., supra; Spaan et al.,
J. Gen. Virol.
69, 2939-2952 (1988); Strauss et al.,
Annu. Rev. Biochem.
42, 657-683 (1988); Lai,
Annu. Rev. Microbiol.
44, 303-333 (1990); Snijder et al.,
Nucleic Acid Res.
18, 4535-4542 (1990)). The members of this group infect macrophages and contain a nested set of 5 to 7 subgenomic mRNAs in infected cells (Plagemann et al., supra; Meulenberg et al.,
Virology,
192, 62-72 (1993); Conzelmann et al.,
Virology,
193, 329-339 (1993); 15, 16, 17, 18, 19).
The viral genome of European isolates has been shown to be a plus stranded RNA of about 15.1 kb (Conzelmann et al., supra; Meulenberg et al., supra), and appears to be similar in genomic organization to LDV and EAV (Meulenberg et al., supra). However, no serological cross-reaction has been found among PRRSV, LDV and EAV (Goyal et al.,
J. Vet. Diagn. Invest.,
5, 656-664 (1993)).
PRRSV was initially cultivated in swine alveolar macrophage (SAM) cell cultures (Pol et al.,
Veterinary Quarterly,
13:137-143, 1991; Wensvoort et al.,
Veterinary Quarterly,
13:121-130, 1991) and then in continuous cell lines CL2621 (Benfield et al., supra), MA-104, and MARC-145 (Joo,
Proc. Allen D. Leman Swine Conference
, pp. 53-55, 1993). The reproductive and respiratory disease has been reproduced with cell free lung filtrates (Christianson et al.,
Am. J. Vet. Res.,
53:485-488, 1992; Collins et al.,
J. Vet. Diagn. Invest.,
4:117-126, 1992; Halbur et al.,
Proc. Central Veterinary Conference
, pp. 50-59, 1993), and with cell culture-propagated PRRSV (Collins et al., supra, and
Proc. Allen D. Leman Swine Conference
, pp. 47-48, 1993).
Eight open reading frames (also referred to herein as “ORFs” or “genes”) have been identif

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