The interchain disulfide linkage is not a prerequisite but enhances CD28 costimulatory function

Lázár‐Molnár, Eszter; Almo, Steven C.; Nathenson, Stanley G.

doi:10.1016/j.cellimm.2007.02.014

Cited by 23 publications

(21 citation statements)

References 18 publications

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“…We selected CD200 for targeting in part because the natural CD200/ 200R interaction is projected to be sized appropriately to fit within the IS, 12 and we anticipated that entry into the IS would be required for effective CD28 costimulation. The panel of constructs designed varied the fusion regions of CD200R and the CD28 signaling domain to test several factors: the importance of the size of the ectodomain, the contribution of a cysteine bond in the CD28 ectodomain that promotes dimerization, which is known to facilitate CD28 signaling, 28 and the contribution of a glycosylation site in the CD200R ectodomain to a functional conformation. We found, in both murine and human studies, that the receptor that retained a size predicted to permit entry to the IS and that preserved a dimerizing motif was most effective, and we predict that these concepts will inform the design of IFPs using the CD28 costimulatory signal and targeting other ligands.…”

Section: Discussionmentioning

confidence: 99%

“…The next 3 constructs extended the CD28tm into the ec space to incorporate the membrane proximal cysteine (CD28 cys ) that promotes CD28 homodimerization and enhances native CD28 signaling. 28 To compensate for length added by the 9 amino acids (aas) of the ec CD28 domain, the ec region of CD200R in CD200R -9aas -CD28 cys was truncated by 9 aas, whereas the ec domain of CD200R -3aas -CD28 cys was truncated by 3 aas to preserve a potentially structurally important N-linked glycosylation site. Thus, CD200R tm -CD28, CD200R-CD28 tm , and CD200R -9aas -CD28 cys theoretically best approximate the native short spatial distance of CD200R-CD200 (between the T cell and the APC).…”

Section: Cd200r-cd28 Ifps Can Be Expressed By Primary T Cellsmentioning

confidence: 99%

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A CD200R-CD28 fusion protein appropriates an inhibitory signal to enhance T-cell function and therapy of murine leukemia

Oda

Daman

Garcia

et al. 2017

Blood

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Acute myeloid leukemia (AML), the most common adult acute leukemia in the United States, has the poorest survival rate, with 26% of patients surviving 5 years. Adoptive immunotherapy with T cells genetically modified to recognize tumors is a promising and evolving treatment option. However, antitumor activity, particularly in the context of progressive leukemia, can be dampened both by limited costimulation and triggering of immunoregulatory checkpoints that attenuate T-cell responses. Expression of CD200 (OX2), a negative regulator of T-cell function that binds CD200 receptor (CD200R), is commonly increased in leukemia and other malignancies and is associated with poor prognosis in leukemia patients. To appropriate and redirect the inhibitory effects of CD200R signaling on transferred CD8 T cells, we engineered CD200R immunomodulatory fusion proteins (IFPs) with the cytoplasmic tail replaced by the signaling domain of the costimulatory receptor, CD28. An analysis of a panel of CD200R-CD28 IFP constructs revealed that the most effective costimulation was achieved in IFPs containing a dimerizing motif and a predicted tumor-T-cell distance that facilitates localization to the immunological synapse. T cells transduced with the optimized CD200R-CD28 IFPs exhibited enhanced proliferation and effector function in response to CD200 leukemic cells in vitro. In adoptive therapy of disseminated leukemia, CD200R-CD28-transduced leukemia-specific CD8 T cells eradicated otherwise lethal disease more efficiently than wild-type cells and bypassed the requirement for interleukin-2 administration to sustain in vivo activity. The transduction of human primary T cells with the equivalent human IFPs increased proliferation and cytokine production in response to CD200 leukemia cells, supporting clinical translation. This trial was registered at www.clinicaltrials.gov as #NCT01640301.

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Section: Discussionmentioning

confidence: 99%

Section: Cd200r-cd28 Ifps Can Be Expressed By Primary T Cellsmentioning

confidence: 99%

A CD200R-CD28 fusion protein appropriates an inhibitory signal to enhance T-cell function and therapy of murine leukemia

Oda

Daman

Garcia

et al. 2017

Blood

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“…The differences in signaling events initiated by monomeric versus dimeric molecules have not been investigated thoroughly. A monomeric version of CD28 bound CD80 less well and costimulated T cells less efficiently than the wildtype dimer, suggesting that the dimeric nature of CD28 contributes to its function (25). However, we have observed that monomeric or dimeric forms of a chimeric molecule consisting of the murine CD28 extracellular domain fused to the human PD‐1 cytoplasmic tail function equivalently (26).…”

Section: Pd‐1: Cd28 Family On the Outside Sialic Acid Binding Immunomentioning

confidence: 99%

PD‐1 signaling in primary T cells

Riley

2009

Immunological Reviews

674

516

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SummaryProgrammed death-1 (PD-1) is a cell surface molecule that regulates the adaptive immune response. Engagement of PD-1 by its ligands PD-L1 or PD-L2 transduces a signal that inhibits Tcell proliferation, cytokine production, and cytolytic function. While a great deal is known concerning the biological roles PD-1 plays in regulating the primary immune response and in Tcell exhaustion, comparatively little is known regarding how PD-1 ligation alters signaling pathways. PD-1 ligation is known to inhibit membrane-proximal T-cell signaling events, while ligation of the related inhibitory molecule cytotoxic T lymphocyte antigen-4 appears to target more downstream signaling pathways. A major obstacle to an in-depth understanding of PD-1 signaling is the lack of physiologic models in which to study signal transduction. This review focuses on: 1) signaling pathways altered by PD-1 ligation, 2) factors recruited upon PD-1 phosphorylation, and 3) exploring the hypothesis that PD-1 ligation induces distinct signals during various stages of immune-cell differentiation. Lastly, we describe models to dissect the function of the PD-1 cytoplasmic tail using primary cells in the absence of agonist antibodies.

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“…Interestingly, the 28ε cassette does not harbor the extracellular cysteine of CD28 (at position 141), which normally facilitates homodimerization of CD28 at the cell surface. However, this cysteine is not required per se for dimerization and costimulatory function (53). In addition to TCR mispairing and peptide fine specificity, our studies provide the following lines of evidence that TCR:28ε does not hamper safety by constitutively activating T cells: we did not observe activation of NFAT following nonspecific TCR stimulation (Fig.…”

Section: Discussionmentioning

confidence: 54%

TCRs Genetically Linked to CD28 and CD3ε Do Not Mispair with Endogenous TCR Chains and Mediate Enhanced T Cell Persistence and Anti-Melanoma Activity

Govers

Sebestyén

Roszik

et al. 2014

The Journal of Immunology

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Adoptive transfer of T cells that are gene engineered to express a defined TCR represents a feasible and promising therapy for patients with tumors. However, TCR gene therapy is hindered by the transient presence and effectiveness of transferred T cells, which are anticipated to be improved by adequate T cell costimulation. In this article, we report the identification and characterization of a novel two-chain TCR linked to CD28 and CD3ε (i.e., TCR:28ε). This modified TCR demonstrates enhanced binding of peptide–MHC and mediates enhanced T cell function following stimulation with peptide compared with wild-type TCR. Surface expression of TCR:28ε depends on the transmembrane domain of CD28, whereas T cell functions depend on the intracellular domains of both CD28 and CD3ε, with IL-2 production showing dependency on CD28:LCK binding. TCR:28ε, but not wild-type TCR, induces detectable immune synapses in primary human T cells, and such immune synapses show significantly enhanced accumulation of TCR transgenes and markers of early TCR signaling, such as phosphorylated LCK and ERK. Importantly, TCR:28ε does not show signs of off-target recognition, as evidenced by lack of TCR mispairing, as well as preserved specificity. Notably, when testing TCR:28ε in immune-competent mice, we observed a drastic increase in T cell survival, which was accompanied by regression of large melanomas with limited recurrence. Our data argue that TCR transgenes that contain CD28, and, thereby, may provide T cell costimulation in an immune-suppressive environment, represent candidate receptors to treat patients with tumors.

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The interchain disulfide linkage is not a prerequisite but enhances CD28 costimulatory function

Cited by 23 publications

References 18 publications

A CD200R-CD28 fusion protein appropriates an inhibitory signal to enhance T-cell function and therapy of murine leukemia

A CD200R-CD28 fusion protein appropriates an inhibitory signal to enhance T-cell function and therapy of murine leukemia

PD‐1 signaling in primary T cells

TCRs Genetically Linked to CD28 and CD3ε Do Not Mispair with Endogenous TCR Chains and Mediate Enhanced T Cell Persistence and Anti-Melanoma Activity

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