Methods for growing African swine fever virus in fetal porcine lung alveolar macrophage cells

Inventors

Zuckermann, Federico A. • Dixon, Linda Kathleen • Portugal, Maria Raquel Seica • Goatley, Lynnette Claire

Assignees

Aptimmune Biologics Inc • University of Illinois System

Abstract

A method for generating progeny of an African swine fever (ASF) virus includes providing an isolated or purified fetal porcine lung alveolar macrophage cell capable of replicating the ASF virus, wherein the cell is cultured for at least 5 passages; exposing the cell to the ASF virus; and allowing the ASF virus to replicate in the cell; thereby generating progeny of the ASF virus.

Core Innovation

Disclosed are methods for generating progeny of an African Swine Fever (ASF) virus by providing an isolated or purified fetal porcine lung alveolar macrophage cell capable of replicating the ASF virus, wherein the cell is cultured for at least 5 passages, exposing the cell to the ASF virus, and allowing the ASF virus to replicate in the cell to thereby generate progeny of the ASF virus. The disclosure includes using fetal porcine lung alveolar macrophage cells identified as ZMAC/ZMAC-1/ZMAC-4, including ATCC PTA-8764, for replication without virus adaptation as described.

Also disclosed are methods for producing an ASF vaccine by providing a modified-live virus (MLV) strain of ASF virus and growing the MLV strain in an isolated or purified fetal porcine lung alveolar macrophage cell capable of replicating the ASF MLV strain. In the disclosed embodiments, the growth of the MLV strain uses culture with the fetal porcine lung alveolar macrophage cell after culturing for at least 5 passages, and optionally includes contacting the cell with a growth factor composition, including macrophage colony stimulating factor (MCSF) or granulocyte-macrophage colony stimulating factor (GMCSF).

The disclosure further describes methods for growing an ASF virus by isolating a fetal porcine lung alveolar macrophage cell from a porcine fetal lung comprising providing a porcine fetal subject, obtaining a cell-containing bronchoalveolar lavage sample from the subject, and separating the macrophage cell from the sample, then culturing the cell and contacting the cell with the ASF virus to allow viral replication and thereby growing the ASF virus. In addition, a method for detecting the presence of an ASF virus in a porcine subject replicates the ASF virus using the isolated or purified fetal porcine lung alveolar macrophage cell cultured for at least 5 passages, contacts the cell with a sample, incubates under suitable conditions, and detects the presence of the ASF virus in the cell.

Claims Coverage

The independent claims cover four inventive areas: generating ASFV progeny, producing an ASF vaccine by growing an ASF modified-live virus in fetal lung alveolar macrophages, growing ASFV by isolating fetal lung alveolar macrophages from bronchoalveolar lavage, and detecting ASFV in a porcine subject by replicating ASFV in fetal lung alveolar macrophages followed by detection.

Across the independent claims, the inventive concept centers on isolated or purified fetal porcine lung alveolar macrophage cells cultured for at least 5 passages to support ASF virus replication, enabling ASFV progeny generation, producing a modified-live ASF vaccine by growing the MLV strain in these cells, growing ASF virus using bronchoalveolar lavage-derived fetal macrophages, and detecting ASF virus in a porcine subject by replicating ASFV in the macrophages and then detecting its presence.

Stated Advantages

Documented Applications