Antigen-specific activation and cytokine-facilitated expansion of naive, human CD8+ T cells.
This protocol describes how to evaluate antigen-specific priming of CD8(+) cells 10 d after a single specific stimulation. The assay provides reference conditions, which result in the expansion of a substantial population of antigen-specific T cells from the naive repertoire
Wölfl M, Greenberg PD. Nat Protoc. 2014 Apr;9(4):950-66.

Enhanced-affinity murine T-cell receptors for tumor/self-antigens can be safe in gene therapy despite surpassing the threshold for thymic selection.
Central T-cell tolerance developed in the thymus limits the avidity and anti-tumor efficacy of T cells specific for many promising tumor antigens that are unmodified self-antigens. One approach to overcoming this obstacle is to mutate TCR genes from naturally occurring T cells to enhance the affinity for the target antigen.
Schmitt TM1, Aggen DH, Stromnes IM, Dossett ML, Richman SA, Kranz DM, Greenberg PD. Blood. 2013 Jul 18;122(3):348-56.

Increasing functional avidity of TCR-redirected T cells by removing defined N-glycosylation sites in the TCR constant domain.
This strategy to increase the affinity of TCRs or the functional avidity of TCR-transduced T cells involves selectively removing conserved N-glycoslyation sites in the constant regions of TCR alpha or beta chains to increase functional avidity.
Kuball J, Hauptrock B, Malina V, Antunes E, Voss RH, Wolfl M, Strong R, Theobald M, Greenberg PD. J Exp Med. 2009 Feb 16; 206(2):463-75

Facilitating matched pairing and expression of TCR chains introduced into human T cells.
Cysteines are introduced into the constant region of the engineered alpha and beta chains to promote preferential pairing with each other, increase total surface expression of the introduced TCR chains, and reduce mismatching with endogenous TCR chains.
Kuball J, Dossett ML, Wolfl M, Ho WY, Voss RH, Fowler C, Greenberg PD. Blood. 2007 Mar 15; 109(6):2331-8.

The use of anti-CD3 and anti-CD28 monoclonal antibodies to clone and expand human antigen-specific T cells.
This method uses monoclonal antibodies to T cell activation molecules, CD3 and CD28, as stimulation to efficiently grow CD4+ and CD8+ antigen-specific T cells from single progenitors and expand T cell clones in long-term culture, alleviating the requirement for large amounts of viral or tumor antigens and MHC-compatible APC to sustain the growth of T cell clones.
Riddell SR, Greenberg PD. Immunol Methods. 1990 Apr 17; 128(2):189-201.