Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Reexamination Certificate
1995-06-07
2001-09-18
Wortman, Donna C. (Department: 1648)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
C435S005000
Reexamination Certificate
active
06291641
ABSTRACT:
FIELD OF THE INVNETION
This invention relates to antigens derived from enterically transmitted nonA
onB hepatitis viral agent, also referred to herein as hepatitis E virus (HEV), and to diagnostic methods, diagnostic assays, vaccine compositions and vaccine methods, which employ such antigens.
REFERENCES
Arankalle, V. A., et al., The Lancet, 550 (Mar. 12, 1988).
Bradley, D., et al., J. Gen. Virol., 69:731 (1988).
Bradley, D. W., et al., Proc. Nat. Acad. Sci., USA, 84:6277 (1987).
Chauhan, et al., Lancet, 341:149 (1993).
Chomczynski, P., et al., Anal. Biochem. 162:156 (1987).
Harlow, E., et al.,
Antibodies: a Laboratory Manual
, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1988).
Huang, C-C., et al., Virology, 1991:550 (1992).
Khuroo, M. S., Am. J. Med., 48:818 (1980).
Khuroo, M. S., et al., Am. J. Med., 70:58 (1981).
Koonin, E. V., et al., Proc. Nat. Acad. Sci. USA, 89:8259 (1992).
Lanford, R. E., et al., In Vitro Cellular and Devel Biol, 25 (2):174 (1989).
Maniatis, T., et al.,
Molecular Cloning: A Laboratory Manual
, Cold Spring Harbor Laboratory (1982).
Reyes, G., et al., Science 247:1335 (1990).
Reyes, G. R., Arch. Virol. Supp. (Review) 7:15 (1993).
Rozanov, M. N., et al., J. Gen. Virol., 73:2129 (1992).
Sambrook, J., et al.,
Molecular Cloning: A Laboratory Manual
, Cold Spring Harbor Laboratory. (1989).
Summers, M. D., et al.,
A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures
, Texas Agricultural Experimental Station Bulletin No. 1555, 1988,
Tam, A., et al., Virology, 185:120 (1991-a).
Tam, A., et al., Hepatitis E virus: CDNA isolation and sequencing, p. 521-524. In Hollinger, F. B. et al. (ed.), Viral Hepatitis and Liver Disease. Williams and Wilkens, Baltimore. (1991-b).
Yarbough, P. O., In: Nishioka, K., Suzuki, H., Mishiro, S., Oda, T. eds.
Viral Hepatitis and Liver Disease
, Tokyo:Springer-Verlag. 367-370 (1994).
Yarbough, P. O., J. Virol., 65(11):5790 (1991).
BACKGROUND OF THE INVENTION
Enterically transmitted non-A
on-B hepatitis viral agent (ET-NANB; also referred to herein as HEV) is the reported cause of hepatitis in several epidemics and sporadic cases in Asia, Africa, Europe, Mexico, and the Indian subcontinent. Infection is usually by water contaminated with feces, although there is some evidence of person to person transmission. The virus does not seem to cause chronic infection. The viral etiology in ET-NANB has been demonstrated by infection of volunteers with pooled fecal isolates; immune electron microscopy (IEM) studies have shown virus particles with 27-34 nm diameters in stools from infected individuals. The virus particles reacted with antibodies in serum from infected individuals from geographically distinct regions, suggesting that a single viral agent or class is responsible for the majority of ET-NANB hepatitis seen worldwide. No antibody reaction was seen in serum from individuals infected with parenterally transmitted NANB virus (also known as hepatitis C virus or HCV), indicating a different specificity between the two NANB types.
In addition to serological differences, the two types of NANB infection show distinct clinical differences. ET-NANB is characteristically an acute infection, often associated with fever and arthralgia, and with portal inflammation and associated bile stasis in liver biopsy specimens (Arankalle, 1988). Symptoms are usually resolved within six weeks. Parenterally transmitted NANB, by contrast, produces a chronic infection in about 50% of the cases. Fever and arthralgia are rarely seen, and inflammation has a predominantly parenchymal distribution (Khuroo, 1980). The course of ET-NANBH is generally uneventful in healthy individuals, and the vast majority of those infected recover without the chronic sequelae seen with HCV. Occasionally the course of disease can be severe, however, as was recently shown by a human volunteer (Chauhan 1993). One peculiar epidemiologic feature of this disease, however, is the markedly high mortality observed in pregnant women; this is reported in numerous studies to be on the order of 10-20% (Khuroo 1981, Reyes 1993). This finding has been seen in a number of epidemiologic studies but at present remains unexplained. Whether this reflects viral pathogenicity, the lethal consequence of the interaction of virus and immune suppressed (pregnant) host, or a reflection of the debilitated prenatal health of a susceptible malnourished population remains to be clarified.
The two viral agents can also be distinguished on the basis of primate host susceptibility. ET-NANB, but not the parenterally transmitted agent, can be transmitted to cynomolgus monkeys. The parenterally transmitted agent is more readily transmitted to chimpanzees than is ET-NANB (Bradley, 1987).
In the earlier-filed parent applications, HEV clones, peptide antigens, and the sequence of the entire HEV genome sequence were disclosed. From HEV ORF-2 expression constructs, recombinant proteins and viral particles were produced.
SUMMARY OF THE INVENTION
In one aspect the invention includes, a nucleic acid sequence encoding a 62K antigen, preferably, having an amino acid sequence selected from the group consisting of SEQ ID NO:3 and SEQ ID NO:4 and homologous sequences therewith.
In a related aspect the invention includes, an expression vector and an expression system for producing a 62K antigen. The expression vector comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:3 and SEQ ID NO:4, wherein the nucleic acid sequence is inserted into the genome of an expression vector such that the nucleic acid sequence is operationally linked to a promoter able to initiate transcription in a cell. The expression system includes a cell transfected with the above expression vector.
In another aspect the invention includes, a 62K antigen wherein the antigen consists essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:15 and SEQ ID NO:16. The 62K HEV antigen may be produced by a process comprising the steps of inserting a DNA sequence encoding a capsid protein of HEV, into a baculovirus expression vector, transfecting an insect cell with the baculovirus expression vector, and culturing the insect cell under conditions sufficient to express the HEV antigen. The 62K antigen may also be produced by the process comprising the steps of, obtaining an HEV capsid derived antigen having at least 549 C-terminal amino acids of an HEV capsid protein and incubating the HEV capsid derived antigen with a baculoviral infected cell lysate under conditions sufficient to cleave the HEV capsid protein to the 62K antigen.
The invention also includes a kit and method for detecting HEV infection in a test individual. In the detecting method, an antigen of the type described is reacted with serum from the test individual, and then examined for the presence of bound antibody. The assay system includes a solid phase system, in which the antigen is carried on a solid support, or a homogeneous system, in which the antigen is associated with a reporter, where antibody binding to the antigen modulates the reporter signal when detected.
The invention further includes a vaccine composition containing in a pharmacologically acceptable carrier, a 62K antigen.
In a related aspect the invention includes a method of inhibiting infection by HEV, by administering to the subject, by parenteral injection, such as intramuscular or intravenous injection, the 62K antigen vaccine composition.
These and other objects and features of the invention will become more fully understood when the following detailed description of the invention is read in conjunction with the accompanying figures.
REFERENCES:
patent: 5686239 (1997-11-01), Reyes et al.
patent: WO 91/15603 (1991-10-01), None
patent: WO 93/14116 (1993-07-01), None
patent: WO 93/14208 (1993-07-01), None
patent: WO 94/06913 (1994-03-01), None
Boswell et al. “Sequence comparison and alignment: the measurement and interpretation of sequence similarity”. Computational Molecular Biology. Arthur Lesk, eds. Oxford University Press, p
Fuerst Thomas R.
McAtee C. Patrick
Yarbough Patrice O.
Zhang Yi-Fan
Dehlinger Peter J.
Fabian Gary
Genelabs Technologies, Inc.
Iota Pi Law Group
Wortman Donna C.
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