US 9,121,010 B1 · Continuous porcine kidney cell line constitutively expressing bovine αVβ6 integrin with increased susceptibility to foot and mouth disease virus

Abstract

Foot and out disease virus (FMDV) is worldwide problem. Rapid isolation, serotyping and vaccine matching of FMDV from infected animals is critical to enable the implementation of effective vaccination programs and to stop the spread of infection during outbreaks. Current virus isolation protocols use primary cells, known to be susceptible to FMDV, or baby hamster kidney cells (BHK-21) and other cell lines that are not highly sensitive to some strains of FMDV. The αVβ6 integrin is a principal receptor for FMDV. We therefore transduced the porcine kidney cell line, LFBK, to stably express both the αV and β6 bovine integrin subunits. The LFBK-αVβ6 cell line showed both β6 expression and enhanced susceptibility to FMDV infection for at least 100 cell passages. LFBK-αVβ6 cells are highly sensitive for detecting all serotypes of FMDV from experimentally infected animals, including the porcinophillic strain O/TAW/97 and are thus a sensitive tool for FMDV isolation.

Claims11

Dependent claims

A stably transduced fetal porcine kidney cell line which has been engineered to express a functional bovine αvβ6 integrin receptor, said cell line comprising a host cell transduced with a heterologous nucleic acid molecule comprising a sequence of nucleotides that constitutively express an αv subunit of the bovine αvβ6 integrin receptor and a sequence of nucleotides that constitutively express a β6 subunit of the bovine αvβ6 integrin receptor, wherein said αv subunit and said β6 subunit comprise a sequence of nucleotides which encode the bovine αv subunit and the bovine β6 subunit of the bovine αvβ6 integrin receptor, a functional Foot and Mouth Disease Virus (FMDV) receptor wherein said cell line is the LFBKαvβ6 cell line deposited as ATCC Accession No:

PTA-13047.

A transgenic fetal porcine kidney cell line expressing the bovine αvβ6 integrin receptor, obtained by a method comprising:

a) transducing transformed fetal porcine kidney cells of the transformed LFBK cell line with a vector comprising a sequence that constitutively express an αv subunit of the bovine αvβ6 integrin receptor;
b) contacting the resulting transduced LFBK cells with a selection medium;
and c) obtaining transduced LFBKαv cells, d) transducing said LFBKαv cells with a vector comprising a sequence that constitutively express a β6 subunit of the bovine αvβ6 integrin receptor:
e) selecting transduced LFBKαvβ6 cells to obtain a stable transgenic LFBKαvβ6 cell line, wherein said LFBKαvβ6 cells show consistent expression of αv and β6 subunits as measured by immunohistochemistry and wherein said LFBKαvβ3 cells show increased sensitivity for detection of all seven FMDV subtypes compared to a control LFBK cell line lacking constitutive stable expression of said αv and β6 subunits of said porcine LFBKαvβ6 cells, wherein said cell line is the LFBKαvβ6 cell line deposited as ATCC Accession No:
PTA-13047.

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Dependencies are inferred from the claim text.

Continuous porcine kidney cell line constitutively expressing bovine αVβ6 integrin with increased susceptibility to foot and mouth disease virus

Life of this patent

Continuous porcine kidney cell line constitutively expressing bovine αVβ6 integrin with increased susceptibility to foot and mouth disease virus

Life of this patent

Abstract

Claims11