Metapneumovirus strains and their use in vaccine formulations and as vectors for expression of antigenic sequences

a technology of metapneumovirus and vaccine formulation, which is applied in the field of isolated mammalian negative strand rna virus, can solve the problems of serious respiratory tract disease in infants and children, colorado strain of apv not protecting spf chicks, and the proportion of illnesses observed among mammals cannot be attributed to known pathogens. it reduces the extent of infection and improves the symptoms

Active Publication Date: 2004-01-08
ERASMUS UNIV MEDICAL CENT ROTTERDAM ERASMUS MC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

0050] In certain embodiments, the invention provides a method for identifying a compound useful for the treatment of infections with mammalian MPV, wherein the method comprises: (a) infecting an animal with a mammalian MPV; (b) Administering to the animal a test compound; and (c) determining the effect of the test compound on the infection of the animal, wherein a test compound that reduces the extent of the infection or that ameliorates the symptoms associated with the infection is identified as a compound useful for the treatment of infections with mammalian MPV. In certain embodiments, the invention provides a method for identifying a compound useful for the treatment of infections with mammalian MPV, wherein the method comprises (a) infect

Problems solved by technology

However, a proportion of the illnesses observed among mammals can still not be attributed to known pathogens.
The Colorado strain of APV does not protect SPF chicks against challenge with either a subgroup A or a subgroup B strain of TRT virus.
Parainfluenza viral infection results in serious respiratory tract disease in infants and children.
Treatment options for established RSV disease are limited.
While a vaccine might prevent RSV infection, and/or RSV-related disease, no vaccine is yet licensed for this indication.
A major obstacle to vaccine development is safety.
Finally, primary RSV infection and disease do no

Method used

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  • Metapneumovirus strains and their use in vaccine formulations and as vectors for expression of antigenic sequences
  • Metapneumovirus strains and their use in vaccine formulations and as vectors for expression of antigenic sequences
  • Metapneumovirus strains and their use in vaccine formulations and as vectors for expression of antigenic sequences

Examples

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example 1

6.1 EXAMPLE 1

Specimen Collection, Virus Isolation, Virus Characterization

[0503] Samples of nasopharyngeal aspirates were obtained from hosts to assay for the presence of viruses, and also to characterize those identified. Nasopharyngeal aspirates were collected from children suffering from respiratory tract infection (RTI). In order to determine the identity of the cause of illness, all nasopharyngeal aspirates were tested by direct immmunofluorescence assays (DIF) (See method in Example 9), using fluorescence labeled antibodies against influenza virus types A and B, hRSV, and human parainfluenza virus (hPIV) types 1, 2, and 3. Viruses were also isolated from nasopharyngeal aspirates using rapid shell vial techniques, (Rothbarth et. al., 1999, J of Virol. Methods 78:163-169) on various cell lines, including VERO cells, tertiary cynomolgous monkey kidney (tMK) cells, human endothelial lung (HEL) cells and marbin dock kidney (MDCK) cells. Samples showing cytopathic effects (CPE) after...

example 2

6.2 EXAMPLE 2

Seroprevalence in the Human Population

[0510] To study the seroprevalence of this virus in the human population, sera from humans in different age categories were analyzed by indirect IFA using tMK cells infected with one of the unidentified virus isolates. Studies revealed that antibodies to the virus could be detected in 25% of the children between six and twelve months. Furthermore, by the age of five, nearly 100% of the children were seropositive. In total, 56 sera samples examined by indirect IFA and by VN assay. For 51 of the samples or 91%, the results of the VN assay, i.e., a titer greater than 8, coincided with the results obtained with indirect IFA, i.e., a titer greater than 32. Four samples that were found to be positive by IFA, were negative by the VN assay, i.e., titer less than 8, whereas one serum sample was negative by IFA, i.e., titer less than 32, and was positive by the VN test, i.e., a titer of 16 (FIG. 2).

[0511] IFA conducted on 72 sera samples take...

example 3

6.3 EXAMPLE 3

Genomic Sequence of HMPV Isolate 00-1

[0515] In order to obtain sequence information for the unknown virus isolates, a random PCR amplification strategy known as RAP-PCR (Welsh et. al., 1992, NAR 20:4965-4970) (See Example 19). In short, tMK cells were infected with one of the virus isolates (isolate 00-1) as well as with hPIV-1 that served as a positive control. After both cultures displayed similar levels of CPE, virus in the culture supernatants was purified on continuous 20-60% sucrose gradients. The gradient fractions were inspected for virus-like particles by EM, and RNA was isolated from the fraction that contained approximately 50% sucrose, in which nucleocapsids were observed. Equivalent amounts of RNA isolated from both virus fractions were used for RAP-PCR, after which samples were run side by side on a 3% NuSieve agarose gel. Twenty differentially displayed bands specific for the unidentified virus were subsequently purified from the gel, cloned in plasmid pC...

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Abstract

The present invention provides an isolated mammalian negative strand RNA virus, metapneumovirus (MPV), within the sub-family Pneumoviridae, of the family Paramyxoviridae. The invention also provides isolated mammalian negative strand RNA viruses identifiable as phylogenetically corresponding or relating to the genus Metapneumovirus and components thereof. In particular the invention provides a mammalian MPV, subgroups and variants thereof. The invention relates to genomic nucleotide sequences of different isolates of mammalian metapneumoviruses, in particular human metapneumoviruses. The invention relates to the use of the sequence information of different isolates of mammalian metapneumoviruses for diagnostic and therapeutic methods. The present invention relates to nucleotide sequences encoding the genome of a metapneumovirus or a portion thereof, including both mammalian and avian metapneumovirus. The invention further encompasses chimeric or recombinant viruses encoded by said nucleotide sequences. The invention also relates to chimeric and recombinant mammalian MPV that comprise one or more non-native or heterologous sequences. The invention further relates to vaccine formulations comprising mammalian or avian metapneumovirus, including recombinant and chimeric forms of said viruses. The vaccine preparations of the invention encompass multivalent vaccines, including bivalent and trivalent vaccine preparations.

Description

[0001] This application claims priority to International Application No.: PCT / NL02 / 00040, filed Jan. 18, 2002, which claims priority to European Patent Application 01200213.5, filed Jan. 19, 2001 and European Patent Application 01203985.5, filed Oct. 18, 2001, all of which are incorporated by reference herein in their entireties.[0002] Copending and co-assigned U.S. Patent Application ______, filed on even date herewith, listing Ronaldus Fouchier, Bernadetta van den Hoogen, Albertus Osterhaus, Aurelia Haller, and Roderick Tang as Inventors, entitled "Recombinant Parainfluenza Virus Expression Systems and Vaccines Comprising Heterologous Antigens Derived from Metapneumovirus", is incorporated herein by reference in its entirety.1. INTRODUCTION[0003] The invention relates to an isolated mammalian negative strand RNA virus, metapneumovirus (MPV), within the sub-family Pneumoviridae, of the family Paramyxoviridae. The present invention also relates to isolated mammalian negative strand ...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61K35/64A61K35/74G01N33/53A61K38/00A61K39/00A61K39/155A61K39/395A61K48/00A61P11/00A61P31/14C07KC07K14/11C07K14/115C07K14/135C07K16/10C12N1/15C12N1/19C12N1/21C12N5/10C12N7/00C12N7/01C12N15/09C12N15/86C12Q1/68C12Q1/70G01N33/50G01N33/543G01N33/569
CPCA61K39/155C12N2760/18334A61K2039/5256C07K14/005C07K16/1027C12N7/00C12N15/86C12N2760/18321C12N2760/18322C12N2760/18343C12N2760/18622C12N2760/18722C12N2840/203G01N33/5008G01N33/502G01N33/5091G01N33/56983C12Q1/701A61K2039/5254A61K39/12A61P11/00A61P31/14A61P37/04C12Q2600/158
Inventor FOUCHIER, RONALDUS ADRIANUS MARIAVAN DEN HOOGEN, BERNADETTA GERARDAOSTERHAUS, ALBERTUS DOMINICUS MARCELLINUS ERASMUSHALLER, AURELIATANG, RODERICK
Owner ERASMUS UNIV MEDICAL CENT ROTTERDAM ERASMUS MC
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