Activation of Resting Human Primary T Cells with Chimeric Receptors: Costimulation from CD28, Inducible Costimulator, CD134, and CD137 in Series with Signals from the TCRζ Chain

Abstract

Chimeric receptors that include CD28 signaling in series with TCRζ in the same receptor have been demonstrated to activate prestimulated human primary T cells more efficiently than a receptor providing TCRζ signaling alone. We examined whether this type of receptor can also activate resting human primary T cells, and whether molecules other than CD28 could be included in a single chimeric receptor in series with TCRζ to mediate the activation of resting human primary T cells. Human CD33-specific chimeric receptors were generated with CD28, inducible costimulator, CD134, or CD137 signaling regions in series with TCRζ signaling region and transfected by electroporation into resting human primary T cells. Their ability to mediate Ag-specific activation was analyzed in comparison with a receptor providing TCRζ signaling alone. Inclusion of any of the costimulatory signaling regions in series with TCRζ enhanced the level of specific Ag-induced IL-2, IFN-γ, TNF-α, and GM-CSF cytokine production and enabled resting primary T cells to survive and proliferate in response to Ag in the absence of any exogenous factors. Inclusion of CD28, inducible costimulator, or CD134 enhanced TCRζ-mediated, Ag-specific target cell lysis. Chimeric receptors providing B7 and TNFR family costimulatory signals in series with TCRζ in the same receptor can confer self-sufficient clonal expansion and enhanced effector function to resting human T cells. This type of chimeric receptor may now be used to discover the most potent combination of costimulatory signals that will improve current immunotherapeutic strategies.

Inhibition of p38 MAPK activity in B-NHL Raji cells by treatment with engineered CD20-specific T cells

“…Although engagement of ζ chains is sufficient to induce tumoricidal activity, without adequate co-stimulatory signals, it may not suffice to elicit substantial lymphocyte activation (3). Accumulated evidence has shown that T cells modified with chimeric antigen receptors incorporating a CD28 signaling domain are much more active when tested in vitro and in murine models (6,10,12,14). In the present study, a recombinant anti-CD20scFvFc/ CD28/CD3ζ gene was constructed that provided both primary and costimulatory signals to T cells through the one chimera.…”

To affinity and beyond: Harnessing the T Cell receptor for cancer immunotherapy

“…97 Secondary methods to revert T cell exhaustion in the tumor microenvironment are amplification of co-stimulatory molecules CD28, CD134, CD137. 98 Insertion into third generation of chimeric antigen receptor constructs has been achieved and clinical trials that employ this technology have positive results and show enhanced T cell persistence and function. 80 Technologies that have been developed to deliver high affinity TCR genes to T cells will be effective to modulate enhanced co-stimulatory molecule expression as well.…”

Adoptive cellular immunotherapy for childhood malignancies

“…[101][102][103][104] The genetic manipulation of T cells with implications for tumor therapy is not limited to redirecting specificity, for transgenes can be expressed to improve persistence of the T cells after infusion. For example, to increase the proliferative potential (i) the CAR endodomain can be systematically altered to provide a fully competent activation signal, [105][106][107][108][109][110][111] or (ii) T cells can be modified to enforce expression of stimulatory cytokine transgenes [112][113][114][115][116] or (iii) the CAR can be introduced into T cells which can respond/ proliferate via endogenous TCR to defined (often viral) antigens. [117][118][119] Recognizing that tumor-specific T cells may not traffic to sites of disease, cells can be genetically modified to express desired homing receptors.…”