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AssigneeWashington State University Research Foundation
InventorsSubramaniam Srikumaran, Weiguo Liu, Sudarvili Shanthalingam
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Particular aspects provide novel recombinant cells and cell lines (e.g., macrophage cell lines) that are permissive for propagation of porcine reproductive and respiratory syndrome virus (PRRSV) propagation in vitro or in vivo. In certain aspects, novel nucleic acid sequences encoding porcine sialoadhesin were transfected into existing macrophage cell-lines from other species, rendering them permissive to PRRSV infection, and suitable for propagation of PRRSV. Particular aspects provide exemplary recombinant cloned cell lines that support the replication of PRRSV, with an obtainable PRRSV titre of between 2×105/ml and 2×106/ml. Additional aspects provide novel nucleic acid sequences and polymorphisms thereof that encode for porcine sialoadhesin. Further aspects provide PRRSV propagation and preparation methods using the novel recombinant cell lines, and methods for PRRSV antigen and vaccine production using same. Yet further aspect provide transgenic, chimerical or engrafted animals having cell comprising nucleic acid sequences encoding porcine sialoadhesin.
A recombinant macrophage cell comprising a transfected nucleic acid that encodes a cell-surface porcine sialoadhesin receptor or portion thereof, the cell-surface receptor or portion thereof sufficient to provide for porcine reproductive and respiratory syndrome virus (PRRSV) binding, endocytosis, or susceptibility to PRRSV infection.
A recombinant cell comprising a transfected nucleic acid that encodes a polypeptide comprising a sequence selected from the group consisting of SEQ ID NO:2, a sequence having greater than 99% sequence identity with SEQ ID NO:2, and PRRSV-binding portions thereof.
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A method for propagating porcine reproductive and respiratory syndrome virus (PRRSV), comprising:
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An isolated nucleic acid comprising a sequence that encodes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, a sequence having greater than 99% sequence identity with SEQ ID NO:2, and PRRSV-binding portions thereof.
A recombinant expression system comprising an expression vector into which is inserted a nucleic acid that encodes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, a sequence having greater than 99% sequence identity with SEQ ID NO:2, and PRRSV-binding portions thereof.
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A non-human transgenic or chimeric animal, harboring a nucleic acid transgene that encodes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NOS:2 or 4, and PRRSV-binding portions thereof.
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Dependencies are inferred from the claim text.