Affinity Maturation of a T-Cell Receptor-Like Antibody Specific for a Cytomegalovirus pp65-Derived Peptide Presented by HLA-A*02:01

Lee, Se-Young; Ko, Deok-Han; Son, Min-Jeong; Kim, Jeong Ah; Jung, Keunok; Kim, Yong‐Sung

doi:10.3390/ijms22052349

Cited by 6 publications

(5 citation statements)

References 36 publications

(109 reference statements)

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“…For plasmids of peptide-fused Abs, including TEDbodies, DNA encoding the peptide and a G 4 S linker was subcloned in-frame without additional sequences at the C-terminus of the HC of each Ab. Expression plasmids for the anti-EGFR Ab necitumumab [26] and TCRlike Ab C1-17 [27] have been described previously. For the anti-programmed cell death protein 1 (PD1) Ab pembrolizumab (Keytruda ® ) and anti-OX40 (CD134) 1166/1167 Abs, DNA fragments of VH and VL of pembrolizumab (DrugBank Accession No.…”

Section: Expression and Purification Of Tedbodies And Absmentioning

confidence: 99%

“…For the detection of CMV-pMHCI on the cell surface at 37 °C, the indicated cells (1.5 × 10 5 ) were seeded in a 12-well plate in the medium containing 10% FBS and cultured until they were fully attached to the bottom of the plates (~ 12 to 15 h). Next, the cells were treated for 18 h with a synthetic peptide, TEDbody, or control Ab at the indicated concentrations, washed with PBS, and stained with the CMV-pMHCI-specific C1-17 Ab (10 nM) [27] in a blocking solution [PBS (pH 7.4) and 2% FBS] for 1 h at 4 °C. After a wash with the ice-cold blocking solution, the cells were stained with an Alexa Fluor 647-labeled rabbit antimouse IgG Ab (Thermo Fisher Scientific) for 30 min at 4 °C.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…At 37 °C, the 18 h incubation was intended to enable cellular uptake and processing for the surface presentation of CMV-pMHCI. The cell surface presentation of CMV-pMHCI was monitored by flow cytometry using a high-affinity TCR-like Ab (C1-17) specific to CMV-pMHCI [27]. When pulsed with 9-mer CMVp 495-503 or an N-extended 24-mer CMVp 480-503 synthetic peptide, HLA-A*02:01 + MDA-MB-231 and NCI-H889 cells, but not HLA-A*02:01 − LoVo cells, were positively stained with the C1-17 Ab in proportion to the HLA-A*02:01 expression levels at both 4 °C and 37 °C (Fig.…”

Section: Cmv-pmhci Presentation Via a Tedbody Carrying Various Cmvp 4...mentioning

confidence: 99%

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Antibody-mediated delivery of a viral MHC-I epitope into the cytosol of target tumor cells repurposes virus-specific CD8+ T cells for cancer immunotherapy

et al. 2022

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Background Redirecting pre-existing virus-specific cytotoxic CD8+ T lymphocytes (CTLs) to tumors by simulating a viral infection of the tumor cells has great potential for cancer immunotherapy. However, this strategy is limited by lack of amenable method for viral antigen delivery into the cytosol of target tumors. Here, we addressed the limit by developing a CD8+T cell epitope-delivering antibody, termed a TEDbody, which was engineered to deliver a viral MHC-I epitope peptide into the cytosol of target tumor cells by fusion with a tumor-specific cytosol-penetrating antibody. Methods To direct human cytomegalovirus (CMV)-specific CTLs against tumors, we designed a series of TEDbodies carrying various CMV pp65 antigen-derived peptides. CMV-specific CTLs from blood of CMV-seropositive healthy donors were expanded for use in in vitro and in vivo experiments. Comprehensive cellular assays were performed to determine the presentation mechanism of TEDbody-mediated CMV peptide-MHC-I complex (CMV-pMHCI) on the surface of target tumor cells and the recognition and lysis by CMV-specific CTLs. In vivo CMV-pMHCI presentation and antitumor efficacy of TEDbody were evaluated in immunodeficient mice bearing human tumors. Results TEDbody delivered the fused epitope peptides into target tumor cells to be intracellularly processed and surface displayed in the form of CMV-pMHCI, leading to disguise target tumor cells as virally infected cells for recognition and lysis by CMV-specific CTLs. When systemically injected into tumor-bearing immunodeficient mice, TEDbody efficiently marked tumor cells with CMV-pMHCI to augment the proliferation and cytotoxic property of tumor-infiltrated CMV-specific CTLs, resulting in significant inhibition of the in vivo tumor growth by redirecting adoptively transferred CMV-specific CTLs. Further, combination of TEDbody with anti-OX40 agonistic antibody substantially enhanced the in vivo antitumor activity. Conclusion Our study offers an effective technology for MHC-I antigen cytosolic delivery. TEDbody may thus have utility as a therapeutic cancer vaccine to redirect pre-existing anti-viral CTLs arising from previously exposed viral infections to attack tumors.

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Section: Expression and Purification Of Tedbodies And Absmentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Cmv-pmhci Presentation Via a Tedbody Carrying Various Cmvp 4...mentioning

confidence: 99%

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Antibody-mediated delivery of a viral MHC-I epitope into the cytosol of target tumor cells repurposes virus-specific CD8+ T cells for cancer immunotherapy

et al. 2022

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“…In this case, target cells must express the peptide of interest in the context of a specific MHC class I variant, as engineered TCR recognizes both peptide and MHC class I molecules. Accordingly, TCR-based treatments focus on relatively abundant MHC class I alleles, such as HLA-A*02:01 [23]. Tumor targeting TCR sequences can be identified via isolation and deep sequencing strategies of tumor infiltrating lymphocytes (TILs).…”

Section: Targeting Intracellular Proteinsmentioning

confidence: 99%

The Intracellular Proteome as a Source for Novel Targets in CAR-T and T-Cell Engagers-Based Immunotherapy

Arman

Haus-Cohen

Reiter

2022

Cells

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The impressive clinical success of cancer immunotherapy has motivated the continued search for new targets that may serve to guide potent effector functions in an attempt to efficiently kill malignant cells. The intracellular proteome is an interesting source for such new targets, such as neo-antigens and others, with growing interest in their application for cell-based immunotherapies. These intracellular-derived targets are peptides presented by MHC class I molecules on the cell surface of malignant cells. These disease-specific class I HLA–peptide complexes can be targeted by specific TCRs or by antibodies that mimic TCR-specificity, termed TCR-like (TCRL) antibodies. Adoptive cell transfer of TCR engineered T cells and T-cell-receptor-like based CAR-T cells, targeted against a peptide-MHC of interest, are currently tested as cancer therapeutic agents in pre-clinical and clinical trials, along with soluble TCR- and TCRL-based agents, such as immunotoxins and bi-specific T cell engagers. Targeting the intracellular proteome using TCRL- and TCR-based molecules shows promising results in cancer immunotherapy, as exemplified by the success of the anti-gp100/HLA-A2 TCR-based T cell engager, recently approved by the FDA for the treatment of unresectable or metastatic uveal melanoma. This review is focused on the selection and isolation processes of TCR- and TCRL-based targeting moieties, with a spotlight on pre-clinical and clinical studies, examining peptide-MHC targeting agents in cancer immunotherapy.

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“…The approval of this biologic verified TCR-based protein engineering as a valuable therapeutic modality. Lee et al reported a TCR-like antibody specific for HLA-A*02:01 in complex with a peptide derived from human cytomegalovirus (CMV) pp65 protein [ 2 ]. The binding affinity was matured by sequential mutagenesis of complementarity-determining regions using yeast surface display technology for its application to the diagnostics and therapeutics of CMV infection.…”

mentioning

confidence: 99%

Recent Advances in Antibody Therapeutics

Heo

2022

IJMS

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Affinity Maturation of a T-Cell Receptor-Like Antibody Specific for a Cytomegalovirus pp65-Derived Peptide Presented by HLA-A*02:01

Cited by 6 publications

References 36 publications

Antibody-mediated delivery of a viral MHC-I epitope into the cytosol of target tumor cells repurposes virus-specific CD8+ T cells for cancer immunotherapy

Antibody-mediated delivery of a viral MHC-I epitope into the cytosol of target tumor cells repurposes virus-specific CD8+ T cells for cancer immunotherapy

The Intracellular Proteome as a Source for Novel Targets in CAR-T and T-Cell Engagers-Based Immunotherapy

Recent Advances in Antibody Therapeutics

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