Methods and compositions for genetically modifying and expanding lymphocytes and regulating the activity thereof

A method for genetically modifying a T cell, comprising:

• contacting peripheral blood mononuclear cells (PBMCs) comprising the T cell ex vivo with a replication incompetent recombinant retrovirus to form a transduction reaction mixture, wherein the replication incompetent recombinant retrovirus comprises:
• a) one or more envelope polypeptides on its surface;
• b) one or more activation elements on its surface;
• and
• c) a polynucleotide comprising one or more transcriptional units, wherein the one or more transcriptional units encode a first polypeptide and/or an inhibitory RNA,
• wherein at least one of the one or more activation elements comprises a means for binding CD3, wherein the PBMCs comprising the T cell are contacted ex vivo with the replication incompetent recombinant retrovirus for less than 8 hours before the PBMCs comprising the T cell are washed out of the transduction reaction mixture,
• wherein the method is performed without exposing the PBMCs comprising the T cell to a bovine serum,
• wherein the method is carried out without requiring prior ex vivo stimulation with anti-CD3, anti-CD28, IL-2, or IL-7, and
• wherein said contacting activates the T cell and facilitates membrane fusion of the T cell to the replication incompetent recombinant retrovirus to produce a genetically modified T cell.