Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues
Reexamination Certificate
2000-10-12
2003-08-12
Smith, Lynette R. F. (Department: 1645)
Chemistry: natural resins or derivatives; peptides or proteins;
Peptides of 3 to 100 amino acid residues
C424S190100, C424S192100, C424S193100, C424S243100, C424S245100, C424S252100, C530S300000, C530S324000, C530S388200
Reexamination Certificate
active
06605696
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to proteins derived from
Lawsonia intracellularis
and encompasses related proteins, nucleic acids, and immunogenic compositions. The immunogenic compositions are particularly useful in prevention of
L. intracellularis
infections in susceptible animals, such as pigs. The proteins, fragments, and nucleic acids can also be employed as diagnostic agents.
BACKGROUND OF THE INVENTION
Commercially raised pigs are sensitive to a wide spectrum of intestinal diseases or syndromes that are collectively referred to as porcine proliferative enteropathy (PPE). These diseases include intestinal adenomatosis complex (Barker I. K. et al., 1985, In “Pathology of Domestic Animals,” 3
rd
Edition, Vol. 2 p. 1-237, eds. K, V. F. Jubb et al. (Academic Press Orlando)), porcine intestinal adenomatosis (PIA), necrotic enteritis (Rowland A. C. et al., 1976
, Veterinary Record
97:-178-180), proliferative haemorrhagic enteropathy (Love, R. J. et al., 1977
, Veterinary Record
100: 473), regional ileitis (Jonsson, L. et al., 1976, Acta
Veterinaria Scandinavica
17: 223-232), haemorrhagic bowel syndrome (O'Neil, I. P. A., 1970
, Veterinary Record
87:742-747), porcine proliferative enteritis and Campylobacter spp-induced enteritis (Straw, B. E., 1990
, Journal of American Veterinary Medical Association
197: 355-357).
One major type of PPE is non-haemorrhagic and is manifested by porcine intestinal adenomatosis (PIA). This form of PPE frequently causes growth retardation and mild diarrhea. Another important type of PPE is haemorrhagic. It is often fatal, and is manifested by proliferative haemorrhagic enteropathy (PHE) wherein the distal small intestine lumen becomes engorged with blood.
While PPE in pigs is commercially most important, PPE is also a problem in the raising of hamsters (Stills, H. F., 1991
, Infection and Immunology
59: 3227-3236), ferrets (Fox et al. 1989
, Veterinary Pathology
26: 515-517), guinea pigs (Elwell et al., 1981
, Veterinary Pathology
18: 136-139), rabbits (Schodeb et al., 1990
, Veterinary Pathology
27: 73-80) and certain birds (Mason et al, 1998).
The organism that causes PPE is the Campylobacter-like bacterium “
L. intracellularis
” (McOrist S et al, 1995
, International Journal Of Systematic Bacteriology
45: 820-825). This organism is also known as lleal symbiont intracellularis (Stills, 1991, supra). PPE-like diseases in pigs may also be caused by other species of Campylobacter (Gebhart et al., 1983
, American Journal of Veterinary Research
44: 361-367).
L. intracellularis
is located in the cytoplasm of villi and intestinal crypt cells of infected animals, where it causes structural irregularities and enterocyte proliferation. Abscesses form as the villi and intestinal crypts become branched and fill with inflammatory cells.
Current control of PPE relies on the use of antibacterial compounds. There is, however, a need for alternative means of controlling
L. intracellularis
infection.
International Patent Application No. PCT/AU96/00767 describes
L. intracellularis
polypeptides and immunogenic compositions that are useful as vaccines. There is, however, a need for additional compositions that confer resistance to
L. intracellularis
infection.
SUMMARY OF THE INVENTION
The present invention relates to an isolated polynucleotide molecule comprising a nucleotide sequence that is selected from the group consisting of:
a) a nucleotide sequence encoding
L. intracellularis
HtrA, PonA, HypC, LysS, YcfW, ABC1, or Omp100 protein;
b) a nucleotide sequence that is a substantial part of the nucleotide sequence encoding the
L. intracellularis
HtrA, PonA, HypC, LysS, YcfW, ABC1, or Omp100 protein; and
c) a nucleotide sequence that is homologous to the nucleotide sequence of a) or b).
In another aspect, the invention relates to a recombinant vector comprising these polynucleotide molecules, including those encoding a carrier or fusion partner such that expression of the recombinant vector results in a fusion protein comprising the carrier or fusion partner fused to a protein or polypeptide encoded by the nucleotide sequences described above. The invention also encompasses transformed host cells comprising these recombinant vectors and polypeptides produced by such transformed host cells.
In another aspect, the present invention relates to an isolated polypeptide that is selected from the group consisting of:
(a)
L. intracellularis
HtrA, PonA, HypC, LysS, YcfW, ABC1, or Omp100 protein;
(b) a polypeptide having an amino acid sequence that is homologous to that of the
L. intracellularis
HtrA, PonA, HypC, LysS, YcfW, ABC1, or Omp100 protein;
(c) a polypeptide consisting of a substantial portion of the
L. intracellularis
HtrA, PonA, HypC, LysS, YcfW, ABC1, or Omp100 protein or of the polypeptide having an amino acid sequence that is homologous to that of the
L. intracellularis
HtrA, PonA, HypC, LysS, YcfW, ABC1, or Omp100 protein;
(d) a fusion protein comprising the protein or polypeptide of (a), (b) or (c) fused to another protein or polypeptide; and
(e) an analog or derivative of the protein or polypeptide of (a), (b), (c) or (d).
The present invention further provides a polynucleotide molecule comprising a nucleotide sequence of greater than 20 nucleotides having promoter activity and found within SEQ ID NO: 2 from about nt 2691 to about nt 2890.
The present invention further relates to a method of preparing any of these polypeptides, comprising culturing host cells transformed with a recombinant expression vector and recovering the expressed polypeptide from the cell culture. The vector comprises a polynucleotide molecule comprising a nucleotide sequence encoding any of the polypeptides, the nucleotide sequence being in operative association with one or more regulatory elements. Culturing is conducted under conditions conducive to expression of the polypeptide.
In yet another aspect, the invention relates to an isolated antibody that specifically reacts with any of the
L. intracellularis
HtrA, PonA, HypC, LysS, YcfW, ABC1, or Omp100 proteins or polypeptides described above.
The invention also relates to an immunizing composition that comprises an immunologically effective amount of a protein, polypeptide, antibody, or polynucleotide of the invention in combination with a pharmaceutically acceptable carrier. The present invention encompasses a method of immunizing a PPE susceptible animal against
L. intracellularis
infection that comprises administering to the animal the immunizing composition.
The invention also relates to a kit for immunizing a PPE susceptible animal against a disease condition caused or exacerbated by
L. intracellularis
that comprises a container having therein an immunologically effective amount of one of the proteins, polypeptides, antibodies, or polynucleotides described above. The invention also relates to a kit for detecting the presence of
L. intracellularis
, an
L. intracellularis
specific amino acid or nucleotide sequence, or an anti-
L. intracellularis
antibody, comprising a container that has therein a protein, polypeptide, polynucleotide, or antibody of the invention.
REFERENCES:
patent: 5885823 (1999-03-01), Knittel et al.
patent: 843818 (1996-12-01), None
patent: 9639629 (1996-12-01), None
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patent: 0069903 (2000-11-01), None
patent: 0069904 (2000-11-01), None
patent: 0069905 (2000-11-01), None
patent: 0069906 (2000-11-01), None
J.C. Boucher et al.,Journal of Bacteriology, Jan. 1996 pp. 511-523_Two Distinct Loci Affecting Conversion to Mucoidy inPseudomonas aeruginosain Cystic Fibrosis Encode Homologs of the Serine Protease HtrA.
C. Dale et al.Microbiology1998, 144, 2073-2084, Identification and sequencing of the groE operon and flanking genes ofLawsonia intracellularis: use in phylogeny.
Search Report: EP 00 30 9125.
Ford Vanessa L
Ginsburg Paul H.
Kohn & Associates PLLC
Ling Lorraine B.
Pfizer Inc.
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