Preparation of Stable Single‐Chain Trimers Engineered with Peptide, β2 Microglobulin, and MHC Heavy Chain

Hansen, Ted H.; Yu, Yanbo; Fremont, Daved H.

doi:10.1002/0471142735.im1705s87

Cited by 27 publications

(32 citation statements)

References 29 publications

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“…It has been proven that a particularly powerful strategy to enhance the potency of DNA vaccines involves bypassing the antigen-processing pathway in DNA-transfected antigenpresenting cells (APCs) by expressing an SCT consisting of an immunogenic peptide, β2m and MHC class I heavy chain (22,27,36,37). The SCT localizes to the plasma membrane of APCs and stably displays an antigenic peptide for recognition by CD8 + CTLs (22).…”

Section: Discussionmentioning

confidence: 99%

“…The SCT localizes to the plasma membrane of APCs and stably displays an antigenic peptide for recognition by CD8 + CTLs (22). Therefore, in the present study, we have introduced the SCT platform into our DNA vaccine that consists of KDR2, β2m and H-2D b and examined its ability to induce CTL response to VEGFR2 and its capacity to inhibit tumor-induced angiogenesis and metastasis in mouse models.…”

Section: Discussionmentioning

confidence: 99%

“…It has been demonstrated that the single-chain trimer (SCT) composed of major histocompatibility complex (MHC) class I heavy chain, β2-microglobulin (β2m), and antigen peptide was a particularly powerful strategy used to increase the potency of DNA vaccine against tumor (22)(23)(24)(25)(26) and infection (22,(27)(28)(29)(30)(31). In the present study, we constructed a DNA vaccine encoding an SCT composed of KDR2, β2m and H-2D b [pcDNA3.1(+)-KDR2-β2m-H-2D b , or SCT-KDR2] and tested its ability to induce specific CTL response to VEGFR2 and its capacity to inhibit tumor-induced angiogenesis and metastasis in mouse models.…”

Section: Introductionmentioning

confidence: 99%

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Anti-metastatic effects of DNA vaccine encoding single-chain trimer composed of MHC I and vascular endothelial growth factor receptor 2 peptide

et al. 2015

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Abstract. Vascular endothelial growth factor receptor 2 (VEGFR2)-mediated signaling is the key rate-limiting step in angiogenesis. VEGFR2 serves as the most important target of anti-angiogenic therapy for cancers. Single-chain trimer (SCT) comprising antigen peptide, β2-microglobulin (β2m), and major histocompatibility complex (MHC) class I heavy chain was a particularly powerful strategy involved in the increase of the potency of DNA vaccine against tumors and infections. In the present study, we constructed an SCT-encoding VEGFR2 antigen peptide [aa400-408, also known as kinase insert domain-containing receptor (KDR2)], β2m, and mouse MHC class I heavy chain H-2D b [pcDNA3.1(+)-KDR2-β2m-H-2D b , or SCT-KDR2]. The constructed SCT-KDR2 DNA was efficiently expressed in the human A293 embryonic kidney cell line. Intradermal immunization of C57BL/6 mice with SCT-KDR2 DNA was able to successfully break self-immunological tolerance and induce robust cytotoxic T-lymphocyte (CTL) response to VEGFR2, leading to marked suppression of tumor cell-induced angiogenesis and metastasis in murine models of B16 melanoma and 3LL Lewis lung carcinoma. Taken together, the results showed that VEGFR2-targeted SCT vaccination is an effective modality that can be utilized in anti-angiogenic active immunotherapy for various types of cancer.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Anti-metastatic effects of DNA vaccine encoding single-chain trimer composed of MHC I and vascular endothelial growth factor receptor 2 peptide

et al. 2015

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“…Full-length OVA cDNA was cloned into the pcDNA3.1(+) vector (Invitrogen) using standard techniques and was confirmed by DNA sequencing. Construction and progressive engineering of SCT has been described previously (6,7,9,20). Mutations were introduced to SCT by site-directed mutagenesis (QuikChange II kit; Stratagene).…”

Section: Methodsmentioning

confidence: 99%

Cross-dressed CD8α ⁺ /CD103 ⁺ dendritic cells prime CD8 ⁺ T cells following vaccination

Li¹,

Kim

Herndon³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Activation of naïve cluster of differentiation (CD)8 + cytotoxic T lymphocytes (CTLs) is a tightly regulated process, and specific dendritic cell (DC) subsets are typically required to activate naive CTLs. Potential pathways for antigen presentation leading to CD8 + T-cell priming include direct presentation, cross-presentation, and cross-dressing. To distinguish between these pathways, we designed single-chain trimer (SCT) peptide-MHC class I complexes that can be recognized as intact molecules but cannot deliver antigen to MHC through conventional antigen processing. We demonstrate that cross-dressing is a robust pathway of antigen presentation following vaccination, capable of efficiently activating both naïve and memory CD8 + T cells and requires CD8α + / CD103 + DCs. Significantly, immune responses induced exclusively by cross-dressing were as strong as those induced exclusively through cross-presentation. Thus, cross-dressing is an important pathway of antigen presentation, with important implications for the study of CD8 + T-cell responses to viral infection, tumors, and vaccines. P rofessional antigen-presenting cells (APCs) are typically required to activate naïve cluster of differentiation (CD)8 + T cells, either by direct priming or cross-priming. In direct priming, infected (viral infection) or directly transfected (DNA vaccination) APCs synthesize the foreign antigen and use endogenous MHC class I pathways of antigen presentation to present antigen and prime CD8 + T cells. In cross-priming, APCs are able to capture, process, and present exogenous antigen onto MHC class I molecules through a process known as cross-presentation (1). Cross-priming has been shown to be an essential pathway for immunity to many viral infections and tumors. Although the pathways that lead to cross-presentation remain incompletely understood, increasing evidence suggests that only certain dendritic cell (DC) subsets are efficient in this process.Cross-dressing involves the transfer of intact MHC class I/ peptide complexes between cells without the requirement for further processing, representing an alternative pathway of indirect antigen presentation (2, 3). Although cross-dressed DCs can activate memory CD8 + T cells following viral infection in vivo (4), it remains unclear whether cross-dressing can prime naïve CD8 + T-cell responses, what DC subtypes are required to prime CD8 + T cells by cross-dressing, and how robust this pathway is compared with traditional pathways of indirect antigen presentation. These questions must be addressed before the physiologic relevance of cross-dressing can be evaluated in context.To address these questions, we have taken advantage of Batf3-deficient mice and engineered MHC class I single chain trimer (SCT) constructs. Batf3 −/− mice have a selective loss of CD8α + and CD103 + DCs, without abnormalities in other hematopoietic cell types or architecture (5). DCs from Batf3 −/− mice are deficient in cross-presentation, and cytotoxic T lymphocyte (CTL) responses to viral infection and synge...

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“…80%) used were: HLA-A*0201-binding peptides, NA17-A 1-9 (VLPDVFIRC) (25), and Melan-A [26][27][28][29][30][31][32][33][34][35] (ELAGIGILTV) (26) derived from two human melanomaassociated Ags, HLA-E*0101/03-binding peptides, UL40 [15][16][17][18][19][20][21][22][23] (VMAPRTLLL and VMAPRTLV) derived from human CMV strains (Towne and Toledo) (27,28), and HLA-B*3501-binding self-peptide 37F (LPFDFTPGY) (29). HLA-A*0201/VLPDVFIRC, HLA-A*0201/ELAGIGILTV, HLA-E*0101/ VMAPRTLLL, and HLA-E*0101/VMAPRTLVL monomers were generated by the local recombinant protein facility (SFR Santé, Nantes, France), as previously described (30).…”

Section: Peptides and Recombinant Peptide/hla-e Monomersmentioning

confidence: 99%

Soluble HLA-I/Peptide Monomers Mediate Antigen-Specific CD8 T Cell Activation through Passive Peptide Exchange with Cell-Bound HLA-I Molecules

Allard

Oger

Benlalam

et al. 2014

The Journal of Immunology

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Accumulating evidence that serum levels of soluble class I HLA molecules (sHLA-I) can, under various pathological conditions, correlate with disease stage and/or patient survival, has stimulated interest in defining whether sHLA-I can exert immunological functions. However, despite a mounting number of publications suggesting the ability of sHLA-I to affect immune effectors in vitro, the precise underlying mechanism still remains controversial. In this article, we address potential functions of both classical and nonclassical sHLA-I, using soluble recombinant HLA-I/peptide monomers, and clearly demonstrate their ability to trigger Ag-specific activation of CD8 T cells in vitro. Furthermore, we provide strong evidence that this behavior results from the passive transfer of peptides from monomers to T cell–bound HLA-I molecules, allowing for fratricide representation and activation. Hence, we proposed a unifying model of T cell activation by HLA-I/peptide monomers, reappraising the potential involvement of sHLA-I molecules in the immune response.

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Preparation of Stable Single‐Chain Trimers Engineered with Peptide, β2 Microglobulin, and MHC Heavy Chain

Cited by 27 publications

References 29 publications

Anti-metastatic effects of DNA vaccine encoding single-chain trimer composed of MHC I and vascular endothelial growth factor receptor 2 peptide

Anti-metastatic effects of DNA vaccine encoding single-chain trimer composed of MHC I and vascular endothelial growth factor receptor 2 peptide

Cross-dressed CD8α ⁺ /CD103 ⁺ dendritic cells prime CD8 ⁺ T cells following vaccination

Soluble HLA-I/Peptide Monomers Mediate Antigen-Specific CD8 T Cell Activation through Passive Peptide Exchange with Cell-Bound HLA-I Molecules

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Preparation of Stable Single‐Chain Trimers Engineered with Peptide, β2 Microglobulin, and MHC Heavy Chain

Cited by 27 publications

References 29 publications

Anti-metastatic effects of DNA vaccine encoding single-chain trimer composed of MHC I and vascular endothelial growth factor receptor 2 peptide

Anti-metastatic effects of DNA vaccine encoding single-chain trimer composed of MHC I and vascular endothelial growth factor receptor 2 peptide

Cross-dressed CD8α + /CD103 + dendritic cells prime CD8 + T cells following vaccination

Soluble HLA-I/Peptide Monomers Mediate Antigen-Specific CD8 T Cell Activation through Passive Peptide Exchange with Cell-Bound HLA-I Molecules

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Cross-dressed CD8α ⁺ /CD103 ⁺ dendritic cells prime CD8 ⁺ T cells following vaccination