Pig dysentery vaccine production by culturing Treponema hyodysenteriae in a culture medium containing cattle cholesterol fraction

Patent Report DNA sequence comprising a portion of g a g region Z of LAV encoding protein immunologically reactive with antibodies to LAV/HTLV-III Genetic Systems World 8606 099; 23 October 1986 A new D N A sequence (I) comprises a portion of the gag region of the L A V genome: the portion coding for a protein which is immunologically reactive with antibodies to LAV/ HTLV-III. A recombinant plasmid capable of replication in bacteria is also described which can be introduced into a bacterial host to produce proteins which are immunologically reactive with anitbodies to LAV/HTLV-III. The recombinant proteins can be used as immunogens and antigens in immunoassays for determining the presence of antibodies to LAV/ HTLV-III. The proteins can also be used as vaccines for human use against infection caused by the virus responsible for AIDS. D N A is isolated from LAV-infected T-cells of a healthy donor, partially digested with restriction endonuclease, and fragments the size of the L A V genome (9.2 kb) are ligated with a suitable vector, then packaged into phage particles. These are then used to transform bacteria and clones are screened for L A V inserts. The desired region of L A V may then be subcloned. 046-87 Particles comprising the major polypeptide of hepatitis B virus surface antigen modified by insertion of foreign amino acid sequence, secreted into the supernatant of transformed cells; use in vaccine production Inst. Pasteur Eur. 201 416; 12 November 1986 Particles contain a sufficient quantity of the amino acid sequences characteristic of the major polypeptide (I) of hepatitis B virus surface antigen to preserve the structure of this antigen, but include in (I) at least one foreign amino acid sequence (II) of 6-13 residues, preferably itself having an antigenic site. (II) Is inserted in a hydrophilic region normally exposed at the particle surface. Alternatively, 1 or more of the amino acids in these regions are replaced by (II). Also new are: (1) recombinant D N A coding for the S region of the viral genome, modified by a sequence coding for (II) inserted into the sequences coding for the hydrophilic regions, under the control of the SV40 promoter, allowing expression in human, animal or yeast cells; and (2) cell lines transformed with these D N A sequences. The particles are useful for vaccine production. They can be isolated in a CsC1 density gradient free of Danes particles or other antigens. When (II) contains an antigenic determinant, the particles may be used for preparation of mixed vaccines. 047-87 New DNA molecules encoding Plasmodium falciparum antigens, SHARP ARP and MESA antigen isolation and sequence analysis; application to polypeptide production and malaria vaccine preparation Walter and Eliza Hall Inst. Med. Res. World 8606 075; 23 October 1986 A new D N A molecule (I) comprises a nucleotide sequence corresponding to all or a portion of the base sequence coding for an antigen of Plasmodium falciparum, i.e. the small histidine-alanine rich protein (SHARP), the asparagine rich protein (ARP), the mature-parasite-infected erythrocyte surface antigen (MESA) or other antigens of P. falciparum. (I) Is capable of being expressed as a polypeptide (II) displaying the above antigenicity in e.g. Escherichia coli. A fused polypeptide with an additional polypeptide as the N-terminal sequence fused to (II) is also described. (II) May be used in vaccine compositions to stimulate an immune response against P. falciparum, the causative agent of human malaria. The invention describes cloned D N A molecules from P. falciparum isolate FCQ27/PNG(FC27) in X-gt/ll-Amp3 (Amp3), and the affinity purification of human antibodies on lysates of ~-Amp3 antigen-positive clones. The isolation of SHARP, A R P and MESA from expression libraries and their nucleotide sequence analysis is described. 048-87 154

Vaccine, Vol. 5, June 1987

New DNA sequence comprising part of e n v region of LAV genome potentially useful for AIDS vaccine development and corresponding transformational protein product; cloning and expression in Escherichia coli Genetic Systems Eur. 201 716; 20 November 1986 A D N A sequence (I) comprises a portion of the env region of the L A V virus genome encoding a protein immunologically reactive with antibodies to LAV. A recombinant plasmid is described which contains (I), and hosts transformed with the plasmid produce a protein (II) which reacts with antibodies to LAV. The (II) sequence starts with isoleucine (1) and ends with threonine (173). It can be used in an AIDS vaccine composition or in an assay to detect L A V antibodies in biological fluids. An example describes the subcloning of a L A V genomic clone, X-J19, in the plasmid vector pUC18 to give pBT-1, which was further subcloned to give pRS-3, containing env, F and LTR region sequences. The env and part of the F sequences were subcloned into M13mp18 and regions of the env sequence transferred into the trpE inducible expression vector. The env D N A was inserted in-frame downstream of the trpE gene, resulting in expression of a trpE-env fusion protein in Escherichia coil transformed with the construct. 049-87 Pig dysentery vaccine production by culturing Treponema hyodysenteriae in a culture medium containing cattle cholesterol fraction

Mobay Eur. 201 796; 20 November 1986 The production of vaccines against pig dysentery is effected by culturing Treponema hyodysenteriae in a culture medium containing a cholesterol-rich cattle fraction (1) and using the resulting cells to prepare a vaccine. The vaccines produce significant resistance to pig dysentery after only 2 i.m. injections, unlike known vaccines which require several i.v. injections. (I) Is obtained by contacting a liquid cholesterolcontaining cattle plasma or serum with a silica adsorbent, separating the adsorbent and subjecting it to freeze-thaw treatment, eluting the cholesterol at pH 9-11.5, concentrating the eluate by ultrafiltration, adjusting the pH to 11.0-11.4, dialysing against Na2CO3 and water, concentrating to a cholesterol level of 50-3000 mg d1-1 by ultrafiltration, adjusting the pH to 7-11, and heating at 50-100°C for 0.5-24 h. The culture medium contains 0.1-5 (especially 1-2.5) vol.% (I). The cultured cells are killed with formaldehyde etc., harvested, suspended in a vehicle, and mixed with an adjuvant to give a vaccme. 050-87

New avirulent heat-sensitive double mutant of CVS strain rabies virus with high stability against reversion; useful for production of vaccine CNRS Eur. 202 142; 20 November 1986 The avirulent, heat-sensitive double mutant of the CVS strain of rabies virus deposited at the Pasteur Institute as 1-433 is described, together with antirabies vaccines which contain the mutant. The double mutant is stable in contrast to those mutants having a single mutation, and can be safely manipulated on a large scale. The CVS parent strain has an amino acid sequence differing from that previously described. It has Ile-36 in place of Thr and Leu-122 in place of Val. The heatsensitive mutation is replacement of Leu-132 by Phe; this results in a total loss of pathogenicity when given by i.m. injection, but is not stable. The second mutation is replacement of Arg-33 by Gln. The mutant is isolated from strains containing the heat-sensitive mutation by using monoclonal antibodies. The mutation is not stable when present alone. The new double mutant may be passaged many times in mouse brain tissue without reversions and can be propagated