Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
2001-04-27
2004-11-23
Housel, James (Department: 1648)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S007210, C435S325000
Reexamination Certificate
active
06821741
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to cell lines which are useful for the rapid detection of enteroviruses. In particular, the invention relates to transgenic buffalo green monkey kidney cell lines which show increased sensitivity to infection by enteroviruses in single-cell type and mixed-cell type cultures, and which are permissive for infection by a broad spectrum of enteroviruses. The invention further relates to transgenic African green monkey kidney cells for detection of enteroviruses.
BACKGROUND OF THE INVENTION
Enteroviruses cause annual epidemics in North America in the period from late summer through early fall. While the majority of the infected individuals are asymptomatic, they are nonetheless capable of transmitting enteroviruses which cause a wide spectrum of diseases, including aseptic meningitis, encephalitis, paralysis, myocarditis, respiratory and gastrointestinal disorders, muscular disability, exanthema and reye's syndrome. In young children, enteroviruses are responsible for aseptic meningitis. Thus, early detection of infection with enteroviruses is critical for disease management.
Thus, to detect and/or isolate enteroviruses from clinical specimens, one approach employed by the prior art has been the use of cell cultures containing a single cell type which is susceptible to infection by enteroviruses, such as buffalo green monkey kidney (BGMK) cells and human lung mucoepidermoid carcinoma cells (NCI-H292, also referred to as H292). However, while BGMK cells are sensitive to some enteroviruses, such as Coxsackie B viruses, their sensitivity is poor to other enteroviruses, such as echoviruses. Similarly, the sensitivity of the H292 cells is variable to different enteroviruses.
Another approach which has been used by the prior art to detect and/or isolate a wider variety of enteroviruses from clinical specimens has employed using a combination of cells [such as primary monkey kidney cells, and cell lines of BGMK cells, human rhabdomyosarcoma (RD) cells, human epidermoid carcinoma (A-549) cells, MRC-5 cells, and others] which are susceptible to enteroviruses. However, even with this multi-cell type approach, and even when incorporating H292 or BGMK cells, the prior art's shell vial cultures and test tube cultures require from about 3 to about 5 day, respectively, to detect enteroviruses.
Thus, what is needed are cells with enhanced sensitivity for enteroviruses to allow rapid detection of enteroviruses, and for cells with a broader spectrum of susceptibility to enteroviruses to allow detection of several types of enteroviruses.
SUMMARY OF THE INVENTION
The invention provides cell lines which are useful for the rapid detection of enteroviruses. In particular, the invention provides transgenic African green monkey kidney cell lines and transgenic buffalo green monkey kidney cell lines. The transgenic buffalo green monkey kidney cell lines have increased sensitivity to infection by enteroviruses in single-cell type and mixed-cell type cultures compared to other cell types which are currently used for enterovirus detection. The transgenic buffalo green monkey kidney cell lines of the invention also are permissive to infection by a larger number of enteroviruses as compared to the cell type from which they were derived.
In particular, the invention provides a transgenic cell line designated BGMK-hDAF.
The invention further provides a cell line established from a transgenic cell line designated BGMK-hDAF, wherein the established cell line has a property selected from the group consisting of (a) increased sensitivity to one or more enteroviruses compared to buffalo green monkey kidney cell line, and (b) permissiveness to echovirus selected from the group consisting of echovirus-6 and echovirus-11. In one embodiment, the cell line has the sensitivity to enterovirus of cell line designated BGMK-hDAF. In another embodiment, the enterovirus is selected from the group consisting of polioviruses, Coxsackie A viruses, Coxsackie B viruses, echoviruses, and enterovirus types 68, 69, 70 and 71.
Also provided by the invention is a transgenic buffalo green monkey kidney cell line expressing human decay accelerating factor, wherein the cell line has a property selected from the group consisting of (a) increased sensitivity to one or more enteroviruses compared to buffalo green monkey kidney cell line, and (b) permissiveness to echovirus selected from the group consisting of echovirus-6 and echovirus-11. In one embodiment, the human decay accelerating factor is encoded by a sequence selected from SEQ ID NO:1 and SEQ ID NO:3. In another embodiment, the transgenic buffalo green monkey kidney cell line has the sensitivity to enterovirus of cell line designated as BGMK-hDAF. In yet another embodiment, the cell line is BGMK-hDAF. In a further embodiment, the enterovirus is selected from the group consisting of polioviruses, Coxsackie A viruses, Coxsackie B viruses, echoviruses, and enterovirus types 68, 69, 70 and 71. In a preferred embodiment, the echovirus is selected from the group consisting of echovirus-4, echovirus-6, echovirus-7, echovirus-9, echovirus-11, echovirus-30. In an alternative embodiment, the Coxsackie virus is selected from the group consisting of Coxsackie virus B1, Coxsackie virus B2, Coxsackie virus B4, Coxsackie virus B5, and Coxsackie virus A9.
The invention additionally provides a composition comprising a transgenic buffalo green monkey kidney cell expressing human decay accelerating factor, wherein the cell has a property selected from the group consisting of (a) increased sensitivity to one or more enterovirus compared to buffalo green monkey kidney cell line, and (b) permissiveness to echovirus selected from the group consisting of echovirus-6 and echovirus-11. In one embodiment, the composition further comprises a cell type other than the transgenic buffalo green monkey kidney cell line, and wherein the transgenic buffalo green monkey kidney cell and the cell type are in mixed-cell type culture. In another embodiment, the cell type is selected from the group consisting of CCD-13 Lu, CCD-8 Lu, CCD-14 Br, CCD-16 Lu, CCD-18 Lu, CCD-19 Lu, Hs888 Lu, MRC-9, CCD-25 Lu, WiDr, DLD-1, COLO205, HCT-15, SW 480, LOVO, SW403, SW48, SW116, SW1463, SW837, SW948, SW1417, FHs74 Int, HCT-8, HCT-116, T84, NCI-H747, NCI-H508, LS123, CaCo-2, HT-29, SK-CO-1, HuTu 80, A253, A704, Hela, Hela, Hela53, L-132, Intestine, BHK-21, Hak, KB, Hep-2, Wish, Detroit 532, FL, Detroit 525, Detroit 529, Detroit 510, WI-38, WI-38 VA13, Citrullinemia, Spik (NBL-10), Detroit 539, Cridu Chat, WI26 VA4, BeWo, SW-13, Detroit 548, Detroit 573, HT-1080, HG 261, C211, Amdur II, CHP 3 (M.W.), CHP 4 (W.W.), RD, HEL 299, Detroit 562, MRC-5, A-549, IMR-90, LS180, LS174T, BGMK, CV-1, and CV-1-hDAF. In a preferred embodiment, the cell type is selected from the group consisting of RD cells, H292 cells, A549 cells, MRC-5 cells, KB cells, and CaCo-2 cells. In a more preferred embodiment, the cell type is CaCo-2 cells.
The invention provides also a composition comprising a transgenic cell designated BGMK-hDAF. In one embodiment, the composition further comprises a cell type other than the BGMK-hDAF cell, and wherein the BGMK-hDAF cell and the cell type are in mixed-cell type culture. In an alternative embodiment, the cell type is selected from the group consisting of CCD-13 Lu, CCD-8 Lu, CCD-14 Br, CCD-16 Lu, CCD-18 Lu, CCD-19 Lu, Hs888 Lu, MRC-9, CCD-25 Lu, WiDr, DLD-1, COLO20S, HCT-15, SW 480, LOVO, SW403, SW48, SW116, SW1463, SW837, SW948, SW1417, FHs74 Int, HCT-8, HCT-116, T84, NCI-H747, NCI-H508, LS123, CaCo-2, HT-29, SK-CO-1, HuTu 80, A253, A704, Hela, Hela, Hela53, L-132, Intestine, BHK-21, Hak, KB, Hep-2, Wish, Detroit 532, FL, Detroit 525, Detroit 529, Detroit 510, WI-38, WI-38 VA13, Citrullinemia, Spik (NBL-10), Detroit 539, Cridu Chat, WI26 VA4, BeWo, SW-13, Detroit 548, Detroit 573, HT-1080, HG 261, C211, Amdur II, CHP 3 (M.W.), CHP 4 (W.W.), RD, HEL 299, Detroit 562, MRC-5, A-549, IMR-90, LS180, LS174T, BGMK, CV-1, and CV-1-hDAF. In another embodiment, the cell type
Foley Shanon A.
Housel James
Medlen & Carroll LLP
University Hospitals of Cleveland
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