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US 10,240,125 B2Granted
Methods for in vitro memory B cell differentiation and transduction with VSV-G pseudotyped viral vectors
AssigneeImmusoft Corporation
InventorsMei Xu, Matthew Rein Scholz, Eric J. Herbig
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Abstract
The present disclosure relates to the in vitro differentiation of memory B cells to plasmablasts or plasma cells and genetic modification of these cells to express a protein of interest, such as a specific antibody or other protein therapeutic.
Claims14
An in vitro culture method for expressing a nucleic acid of interest in a B cell comprising,
- (a) contacting memory B cells with a composition comprising multimerized soluble CD40L in combination with IL-2, IL-10, IL15 and class P CpG oligodeoxynucleotides (p-ODN) (“CD40L composition”) for about 3 days;
- (b) removing the CD40L composition from the B cells of (a) and contacting the B cells of (a) with IL-2, IL-10, IL-6 and IL-15 under conditions such that the B cells in (b) express CD38;
- (c) transducing the B cells of (b) on day 5 of the in vitro cell culture with a retroviral vector pseudotyped with VSV-G, wherein said retroviral vector comprises the nucleic acid of interest operably linked to a promoter;
- and
- (d) contacting the transduced B cells of (c) with IFN-α, IFN-δ, IL-6 and IL-15;
- thereby expressing the nucleic acid of interest in the B cell.
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Dependencies are inferred from the claim text.