Chimeric glycoprotein comprising extracellular domains of PIV-3 F and HN proteins; parainfluenza virus-3F, HN chimeric gene construction and chimeric glycoprotein expression for use in recombinant vaccine

Patent Report Stealth virus-associated diagnosis using permissive cell line; DNA probe construction; virus DNA cloning in Autographa californica polyhedrosis virus vector and expression in Spodopterafrugiperda cell culture for recombinant vaccine Martin WJ World 9220 787; 26 November 1992

Causative agents of the mystery swine disease, vaccine compositions and diagnostic kits; recombinant Lelystad agent production by vector expression in transformant; may be used in recombinant vaccine production Sticht. Cent. Diergeneesh. Inst. World 9221 375; 10 December 1992

Diseases associated with stealth virus (SV) may be diagnosed by a new method in humans or animals by detecting SV in a body sample. Also new are: (1) in vitro culture of SV in a permissive cell line; (2) isolated SV (ATCC VR 2343); (3) SV-infected MRC-5 cells; (4) purified SV-associated toxin; (5) purified antibodies against SV; (6) DNA probes which hybridize specifically with SV nucleic acid; and (7) vaccines containing SV antigens. SV DNA can be isolated by the polymerase chain reaction using retrovirus primers; the amplification product can then be cloned in plasmid pBluescript S K I I + . The amplified product can be used as a probe to screen genomic or cDNA banks. SV DNA can be expressed in standard vector/host systems, especially the Autographa californica nuclear polyhedrosis virus/Spodoptera fruoiperda system. The SV-associated toxin can be isolated e.g. by affinity chromatography using an antibody specific for polyether toxins. This method may be used to diagnose chronic fatigue syndrome, or atypical diseases of the liver, testis, ovary etc. 053-93

An isolated Lelystad Agent(LA) which is the causative agent of Mystery Swine Disease (MSD) is new, the LA corresponding to the isolated LA (CDI-NL-2.91) deposited as I-I102. Also claimd are: i. the recombinant nucleic acid (NA) comprising the nucleotide sequence derived from the genome of LA (sequence specified); ii. the polypeptides encoded by the NA of (i.) (sequence specified); iii. an isolated or synthetic antibody (Ab) which specifically recognizes a part of LA; iv. a vector containing the NA of (i.); v. vaccines for protecting animals against MSD; and vi. diagnostic kits for detecting NA from LA. Knowledge of the NA sequence of LA allows the development of diagnostic tools for LA. LA specific nucleotide sequences can be used as probes or primers in diagnostic techniques such as hybridization, polymerase chain reaction. It is possible to make a vaccine to protect pigs against MSD, in a live, attenuated, killed or recombinant form. 055-93

Chimeric glycoprotein comprising extracellular domains of PIV-3 F and I-IN proteins; parainfluenza virus-3 F, HN chimeric gene construction and chimeric glycoprotein expression for use in recombinant vaccine Upjohn USA 5169 628; 8 December 1992

Vaccine against pneumonic pasteurellosis in cattle; comprising an inactive cytotoxin from Pasteurella haemolytica cultivated in serum-free culture medium Univ. Guelph USA 5165 924; 24 November 1992

A chimeric glycoprotein, consisting of the extracellular domain of parainfluenza virus type-3 (PIV-3) F linked to the extracellular domain of PIV-3 HN protein, is claimed. A vaccine comprising the chimeric glycoprotein is also claimed. Host cells transformed with structural genes encoding the glycoproteins, expression and replication plasmids containing the structural genes, and methods for protecting humans by inoculation with the vaccines, are also disclosed. The vaccine may be combined with vaccines for other diseases to form multi-valent vaccines, and with other medicaments (e.g. antibiotics). The vaccine can be used to produce virus-specific immune responses against PIV-3. In an example, a 1.3kb fragment of a cDNA clone containing the PIV-3 HN gene (plasmid pGPHN1) was ligated into plasmid pGPF2, containing the PIV-3 F glycoprotein gene. The resultant construct was transformed into Escherichia coli HB101. Clones were isolated and selected from the correct orientation of the HN cDNA within the F gene. The junction regions of a properly orientated clone were sequenced. 054-93

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Vaccine, Vol. 11, Issue 7, 1993

A serum-free cell-free vaccine effective againse pneumonic pasteurellosis in cattle is claimed. The vaccine comprises a protective amount of a non-toxic inactive cytotoxin specific for ruminant leukocytes, combined with a serum-free pharmaceutically acceptable carrier. The cytotoxin is isolated with cell-free, endotoxin-free supernatant of a serum-free culture of Pasteurella haemolytiea serotype A1. The supernatant is lyophilized or otherwise concentrated for use. Cattle are vaccinated against pneumonic pasteurellosis by administration of a protective amount of the claimed vaccine. Preparation of the vaccine involves: culturing a P. haemolytiea inoculum, with an O.D. (525 nm) of about 0.18, in serum-free culture medium for 1.5 3 h; periodically measuring the culture broth O.D.; separating the cytotoxin-containing supernatant upon detection of an O.D. (525 nm) of 0.37, which indicates the logarithmic growth phase when an optimum concentration of cytotoxin is produced; and separating solids, including cells, from the supernatant. The vaccine has potential for treatment of other P. haemolytiea infections e.g. mastitis. 056-93