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Article ; Online: Generation of Epstein-Barr Virus Antigen-Specific T Cell Receptors Recognizing Immunodominant Epitopes of LMP1, LMP2A, and EBNA3C for Immunotherapy.

Dudaniec, Krystyna / Westendorf, Kerstin / Nössner, Elfriede / Uckert, Wolfgang

2021 Volume 32, Issue 17-18, Page(s) 919–935

Abstract: Epstein-Barr virus (EBV) infections in healthy individuals are usually cleared by immune cells, wherein ... ...

Abstract	Epstein-Barr virus (EBV) infections in healthy individuals are usually cleared by immune cells, wherein CD8
MeSH term(s)	Epstein-Barr Virus Infections/therapy ; Herpesvirus 4, Human/genetics ; Humans ; Immunodominant Epitopes ; Immunotherapy ; Receptors, Antigen, T-Cell/genetics ; Receptors, Complement 3d ; T-Lymphocytes ; Viral Matrix Proteins/genetics
Chemical Substances	Immunodominant Epitopes ; Receptors, Antigen, T-Cell ; Receptors, Complement 3d ; Viral Matrix Proteins
Language	English
Publishing date	2021-05-14
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1028152-6
ISSN	1557-7422 ; 1043-0342
ISSN (online)	1557-7422
ISSN	1043-0342
DOI	10.1089/hum.2020.283
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Book ; Online ; Thesis: Efficient non-viral T cell engineering for TCR gene therapy by Sleeping Beauty minicircles

Clauß, Julian [Verfasser] / Uckert, Wolfgang [Gutachter] / Matuschewski, Kai [Gutachter] / Pezzutto, Antonio [Gutachter]

2023

Author's details	Julian Clauß ; Gutachter: Wolfgang Uckert, Kai Matuschewski, Antonio Pezzutto
Keywords	Biowissenschaften, Biologie ; Life Science, Biology
Subject code	sg570
Language	English
Publisher	Humboldt-Universität zu Berlin
Publishing place	Berlin
Document type	Book ; Online ; Thesis
Database	Digital theses on the web

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Book ; Online ; Thesis: Generation of Epstein-Barr Virus-specific T Cell Receptorengineered T Cells for Cancer Treatment

Dudaniec, Krystyna [Verfasser] / Matuschewski, Kai [Gutachter] / Meyer, Thomas F. [Gutachter] / Uckert, Wolfgang [Gutachter]

2022

Author's details	Krystyna Dudaniec ; Gutachter: Kai Matuschewski, Thomas F. Meyer, Wolfgang Uckert
Keywords	Biowissenschaften, Biologie ; Life Science, Biology
Subject code	sg570
Language	English
Publisher	Humboldt-Universität zu Berlin
Publishing place	Berlin
Document type	Book ; Online ; Thesis
Database	Digital theses on the web

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Article ; Online: CD8-Specific Designed Ankyrin Repeat Proteins Improve Selective Gene Delivery into Human and Primate T Lymphocytes.

Frank, Annika M / Weidner, Tatjana / Brynza, Julia / Uckert, Wolfgang / Buchholz, Christian J / Hartmann, Jessica

Human gene therapy

2020 Volume 31, Issue 11-12, Page(s) 679–691

Abstract: Adoptive T cell immunotherapy in combination with gene therapy is a promising treatment concept for chronic infections and cancer. Recently, receptor-targeted lentiviral vectors (LVs) were shown to enable selective gene transfer into particular types of ... ...

Abstract	Adoptive T cell immunotherapy in combination with gene therapy is a promising treatment concept for chronic infections and cancer. Recently, receptor-targeted lentiviral vectors (LVs) were shown to enable selective gene transfer into particular types of lymphocytes both
MeSH term(s)	Animals ; Ankyrin Repeat ; CD8-Positive T-Lymphocytes/metabolism ; Cell Line ; Chronic Disease/therapy ; Gene Transfer Techniques ; Genetic Therapy/methods ; Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; Leukocytes, Mononuclear ; Macaca mulatta ; Macaca nemestrina ; Neoplasms/therapy ; Receptors, Antigen, T-Cell/genetics ; Receptors, Antigen, T-Cell/metabolism ; Recombinant Fusion Proteins/genetics ; Single-Chain Antibodies/genetics ; T-Lymphocytes, Cytotoxic/metabolism ; Transduction, Genetic
Chemical Substances	CD8 receptor ; Receptors, Antigen, T-Cell ; Recombinant Fusion Proteins ; Single-Chain Antibodies
Language	English
Publishing date	2020-04-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1028152-6
ISSN	1557-7422 ; 1043-0342
ISSN (online)	1557-7422
ISSN	1043-0342
DOI	10.1089/hum.2019.248
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Article ; Online: Targeting human melanoma neoantigens by T cell receptor gene therapy.

Leisegang, Matthias / Kammertoens, Thomas / Uckert, Wolfgang / Blankenstein, Thomas

The Journal of clinical investigation

2016 Volume 126, Issue 3, Page(s) 854–858

Abstract: In successful cancer immunotherapy, T cell responses appear to be directed toward neoantigens created by somatic mutations; however, direct evidence that neoantigen-specific T cells cause regression of established cancer is lacking. Here, we generated T ... ...

Abstract	In successful cancer immunotherapy, T cell responses appear to be directed toward neoantigens created by somatic mutations; however, direct evidence that neoantigen-specific T cells cause regression of established cancer is lacking. Here, we generated T cells expressing a mutation-specific transgenic T cell receptor (TCR) to target different immunogenic mutations in cyclin-dependent kinase 4 (CDK4) that naturally occur in human melanoma. Two mutant CDK4 isoforms (R24C, R24L) similarly stimulated T cell responses in vitro and were analyzed as therapeutic targets for TCR gene therapy. In a syngeneic HLA-A2-transgenic mouse model of large established tumors, we found that both mutations differed dramatically as targets for TCR-modified T cells in vivo. While T cells expanded efficiently and produced IFN-γ in response to R24L, R24C failed to induce an effective antitumor response. Such differences in neoantigen quality might explain why cancer immunotherapy induces tumor regression in some individuals, while others do not respond, despite similar mutational load. We confirmed the validity of the in vivo model by showing that the melan-A-specific (MART-1-specific) TCR DMF5 induces rejection of tumors expressing analog, but not native, MART-1 epitopes. The described model allows identification of those neoantigens in human cancer that serve as suitable T cell targets and may help to predict clinical efficacy.
MeSH term(s)	Amino Acid Sequence ; Animals ; Base Sequence ; Cell Line, Tumor ; Cyclin-Dependent Kinase 4/genetics ; Cyclin-Dependent Kinase 4/immunology ; DNA Mutational Analysis ; Genetic Therapy ; HLA-A2 Antigen/genetics ; HLA-A2 Antigen/metabolism ; Humans ; MART-1 Antigen/immunology ; Melanoma/genetics ; Melanoma/immunology ; Melanoma/therapy ; Mice, Transgenic ; Protein Binding ; Receptors, Antigen, T-Cell/genetics
Chemical Substances	HLA-A2 Antigen ; MART-1 Antigen ; Receptors, Antigen, T-Cell ; CDK4 protein, human (EC 2.7.11.22) ; Cyclin-Dependent Kinase 4 (EC 2.7.11.22)
Language	English
Publishing date	2016-03-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI83465
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Article ; Online: Antigen receptor-engineered Tregs inhibit CNS autoimmunity in cell therapy using nonredundant immune mechanisms in mice.

Pohar, Jelka / O'Connor, Richard / Manfroi, Benoît / El-Behi, Mohamed / Jouneau, Luc / Boudinot, Pierre / Bunse, Mario / Uckert, Wolfgang / Luka, Marine / Ménager, Mickael / Liblau, Roland / Anderton, Stephen M / Fillatreau, Simon

European journal of immunology

2022 Volume 52, Issue 8, Page(s) 1335–1349

Abstract: ... ...

Abstract	CD4
MeSH term(s)	Animals ; Autoimmunity ; CTLA-4 Antigen ; Cell- and Tissue-Based Therapy ; Forkhead Transcription Factors/genetics ; Immune System Diseases ; Interleukin-10 ; Mice ; Receptors, Antigen ; T-Lymphocytes, Regulatory
Chemical Substances	CTLA-4 Antigen ; Forkhead Transcription Factors ; Receptors, Antigen ; Interleukin-10 (130068-27-8)
Language	English
Publishing date	2022-05-27
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	120108-6
ISSN	1521-4141 ; 0014-2980
ISSN (online)	1521-4141
ISSN	0014-2980
DOI	10.1002/eji.202249845
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Zs.A 489: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Targeting cancer-specific mutations by T cell receptor gene therapy.

Blankenstein, Thomas / Leisegang, Matthias / Uckert, Wolfgang / Schreiber, Hans

Current opinion in immunology

2015 Volume 33, Page(s) 112–119

Abstract: The ease of sequencing the cancer genome, identifying all somatic mutations and grafting mutation-specific T cell receptor (TCR) genes into T cells for adoptive transfer allow, for the first time, a truly tumor-specific and effective therapy. Mutation- ... ...

Abstract	The ease of sequencing the cancer genome, identifying all somatic mutations and grafting mutation-specific T cell receptor (TCR) genes into T cells for adoptive transfer allow, for the first time, a truly tumor-specific and effective therapy. Mutation-specific TCR gene therapy might achieve optimal efficacy with least possible toxicity. Recent clinical data confirm the long-standing evidence from experimental cancer models that antigens encoded by the tumor-specific somatic mutations are potentially the best targets for adoptive T cell therapy. Open questions are, how many somatic mutations create suitable epitopes, whether only individual-specific or also recurrent somatic mutations qualify as suitable epitopes and how neoantigen-specific TCRs are most efficiently obtained. Tumor heterogeneity needs to be considered; therefore, it will be important to identify immunogenic driver mutations that occurred early, are essential for cancer cell survival and present in all cancer cells.
MeSH term(s)	Animals ; Antigens, Neoplasm/genetics ; Antigens, Neoplasm/immunology ; Antigens, Neoplasm/metabolism ; Epitopes, T-Lymphocyte/genetics ; Epitopes, T-Lymphocyte/immunology ; Epitopes, T-Lymphocyte/metabolism ; Histocompatibility Antigens Class I/immunology ; Histocompatibility Antigens Class I/metabolism ; Humans ; Immunotherapy, Adoptive/adverse effects ; Immunotherapy, Adoptive/methods ; Mutation ; Neoplasms/genetics ; Neoplasms/immunology ; Neoplasms/therapy ; Protein Binding ; Receptors, Antigen, T-Cell/genetics ; Receptors, Antigen, T-Cell/metabolism ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; T-Cell Antigen Receptor Specificity/genetics ; T-Cell Antigen Receptor Specificity/immunology ; T-Lymphocyte Subsets/immunology ; T-Lymphocyte Subsets/metabolism ; Treatment Outcome
Chemical Substances	Antigens, Neoplasm ; Epitopes, T-Lymphocyte ; Histocompatibility Antigens Class I ; Receptors, Antigen, T-Cell ; Recombinant Fusion Proteins
Language	English
Publishing date	2015-04
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1035767-1
ISSN	1879-0372 ; 0952-7915
ISSN (online)	1879-0372
ISSN	0952-7915
DOI	10.1016/j.coi.2015.02.005
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Zs.A 2773: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Efficient Non-Viral T-Cell Engineering by Sleeping Beauty Minicircles Diminishing DNA Toxicity and miRNAs Silencing the Endogenous T-Cell Receptors.

Clauss, Julian / Obenaus, Matthias / Miskey, Csaba / Ivics, Zoltán / Izsvák, Zsuzsanna / Uckert, Wolfgang / Bunse, Mario

Human gene therapy

2018 Volume 29, Issue 5, Page(s) 569–584

Abstract: Transposon-based vectors have entered clinical trials as an alternative to viral vectors for genetic engineering of T cells. However, transposon vectors require DNA transfection into T cells, which were found to cause adverse effects. T-cell viability ... ...

Abstract	Transposon-based vectors have entered clinical trials as an alternative to viral vectors for genetic engineering of T cells. However, transposon vectors require DNA transfection into T cells, which were found to cause adverse effects. T-cell viability was decreased in a dose-dependent manner, and DNA-transfected T cells showed a delayed response upon T-cell receptor (TCR) stimulation with regard to blast formation, proliferation, and surface expression of CD25 and CD28. Gene expression analysis demonstrated a DNA-dependent induction of a type I interferon response and interferon-β upregulation. By combining Sleeping Beauty transposon minicircle vectors with SB100X transposase-encoding RNA, it was possible to reduce the amount of total DNA required, and stable expression of therapeutic TCRs was achieved in >50% of human T cells without enrichment. The TCR-engineered T cells mediated effective tumor cell killing and cytokine secretion upon antigen-specific stimulation. Additionally, the Sleeping Beauty transposon system was further improved by miRNAs silencing the endogenous TCR chains. These miRNAs increased the surface expression of the transgenic TCR, diminished mispairing with endogenous TCR chains, and enhanced antigen-specific T-cell functionality. This approach facilitates the rapid non-viral generation of highly functional, engineered T cells for immunotherapy.
MeSH term(s)	CD28 Antigens/genetics ; CD28 Antigens/immunology ; CD28 Antigens/therapeutic use ; Cell Engineering ; Cell Line, Tumor ; Cell Survival/genetics ; Cell Survival/immunology ; DNA Transposable Elements/genetics ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Genetic Vectors/genetics ; Humans ; Immunotherapy, Adoptive/methods ; Interferon Type I/genetics ; Interleukin-2 Receptor alpha Subunit/genetics ; Interleukin-2 Receptor alpha Subunit/immunology ; Interleukin-2 Receptor alpha Subunit/therapeutic use ; Melanoma/genetics ; Melanoma/immunology ; Melanoma/therapy ; MicroRNAs/genetics ; Receptors, Antigen, T-Cell/genetics ; Receptors, Antigen, T-Cell/immunology ; Receptors, Antigen, T-Cell/therapeutic use ; T-Lymphocytes/immunology ; Transposases/genetics
Chemical Substances	CD28 Antigens ; DNA Transposable Elements ; Interferon Type I ; Interleukin-2 Receptor alpha Subunit ; MicroRNAs ; Receptors, Antigen, T-Cell ; Transposases (EC 2.7.7.-)
Language	English
Publishing date	2018-03-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1028152-6
ISSN	1557-7422 ; 1043-0342
ISSN (online)	1557-7422
ISSN	1043-0342
DOI	10.1089/hum.2017.136
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Article ; Online: Immunotherapy with TCR-redirected T cells: comparison of TCR-transduced and TCR-engineered hematopoietic stem cell-derived T cells.

Stärck, Lilian / Popp, Katja / Pircher, Hanspeter / Uckert, Wolfgang

Journal of immunology (Baltimore, Md. : 1950)

2014 Volume 192, Issue 1, Page(s) 206–213

Abstract: Redirecting Ag specificity by transfer of TCR genes into PBLs is an attractive method to generate large numbers of cytotoxic T cells for immunotherapy of cancer and viral diseases. However, transferred TCR chains can pair with endogenous TCR chains, ... ...

Abstract	Redirecting Ag specificity by transfer of TCR genes into PBLs is an attractive method to generate large numbers of cytotoxic T cells for immunotherapy of cancer and viral diseases. However, transferred TCR chains can pair with endogenous TCR chains, resulting in the formation of mispaired TCR dimers and decreased or unspecific reactivity. TCR gene transfer into hematopoietic stem cells (HSCs) is an alternative to create T cells with desired Ag specificity, because in this case expression of endogenous TCR chains is then less likely owing to allelic exclusion. We generated TCR-transduced T cells from peripheral T cells using the lymphocytic choriomeningitis virus-specific P14 TCR. After transfer of the P14 TCR genes into HSCs and subsequent reconstitution of irradiated mice, TCR-engineered HSC-derived T cells were produced. We then compared the Ag-specific T cell populations with P14 TCR-transgenic T cells for their therapeutic efficiency in three in vivo models. In this study, we demonstrate that TCR-transduced T cells and TCR-engineered HSC-derived T cells are comparable in controlling lymphocytic choriomeningitis virus infection in mice and suppress growth of B16 tumor cells expressing the cognate Ag in a comparable manner.
MeSH term(s)	Animals ; Cell Differentiation/genetics ; Cell Differentiation/immunology ; Cytokines/biosynthesis ; Disease Models, Animal ; Gene Expression ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells/cytology ; Hematopoietic Stem Cells/metabolism ; Immunotherapy ; Lymphocytic Choriomeningitis/immunology ; Lymphocytic choriomeningitis virus/immunology ; Melanoma, Experimental/genetics ; Melanoma, Experimental/immunology ; Melanoma, Experimental/mortality ; Melanoma, Experimental/therapy ; Mice ; Mice, Knockout ; Receptors, Antigen, T-Cell/genetics ; Receptors, Antigen, T-Cell/metabolism ; T-Lymphocytes/cytology ; T-Lymphocytes/immunology ; T-Lymphocytes/metabolism ; Transduction, Genetic
Chemical Substances	Cytokines ; Receptors, Antigen, T-Cell
Language	English
Publishing date	2014-01-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3056-9
ISSN	1550-6606 ; 0022-1767 ; 1048-3233 ; 1047-7381
ISSN (online)	1550-6606
ISSN	0022-1767 ; 1048-3233 ; 1047-7381
DOI	10.4049/jimmunol.1202591
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Ud II Zs.37: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Minimal amino acid exchange in human TCR constant regions fosters improved function of TCR gene-modified T cells.

Sommermeyer, Daniel / Uckert, Wolfgang

Journal of immunology (Baltimore, Md. : 1950)

2010 Volume 184, Issue 11, Page(s) 6223–6231

Abstract: TCR gene therapy using adoptive transfer of TCR gene-modified T cells is a new strategy for treatment of cancer. One critical prerequisite for TCR gene therapy is sufficient expression of transferred TCRs. Several strategies to achieve optimal expression ...

Abstract	TCR gene therapy using adoptive transfer of TCR gene-modified T cells is a new strategy for treatment of cancer. One critical prerequisite for TCR gene therapy is sufficient expression of transferred TCRs. Several strategies to achieve optimal expression were developed, including "murinization," which replaces the human TCRalpha and TCRbeta constant regions by their murine counterparts. Using a series of mouse-human hybrid constructs, we have identified nine amino acids responsible for the improved expression of murinized TCRs. Five essential amino acid exchanges were identified in the TCRbeta C region, with exchange of a glutamic acid (human) for a basic lysine (mouse) at position 18 of the C region, being most important. For the TCRalpha C region, an area of four amino acids was sufficient for improved expression. The minimally murinized TCR variants (harboring only nine residues of the mouse sequence) enhanced expression of human TCRs by supporting preferential pairing of transferred TCR chains and a more stable association with the CD3 proteins. Most important, usage of minimally murinized TCR chains improved the function of transduced primary human T cells in comparison with cells transduced with wild-type TCRs. For TCR gene therapy, the utilization of minimally instead of completely murinized constant regions dramatically reduces the number of foreign residues and thereby the risk for immunogenicity of therapeutic TCRs.
MeSH term(s)	Amino Acid Sequence ; Animals ; Blotting, Western ; Cell Separation ; Flow Cytometry ; Gene Expression/genetics ; Gene Expression/immunology ; Gene Expression Regulation/genetics ; Gene Expression Regulation/immunology ; Genes, T-Cell Receptor/genetics ; Genes, T-Cell Receptor/immunology ; Genetic Therapy/methods ; Humans ; Immunoprecipitation ; Mice ; Receptors, Antigen, T-Cell/genetics ; T-Lymphocytes/immunology ; Transduction, Genetic
Chemical Substances	Receptors, Antigen, T-Cell
Language	English
Publishing date	2010-06-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3056-9
ISSN	1550-6606 ; 0022-1767 ; 1048-3233 ; 1047-7381
ISSN (online)	1550-6606
ISSN	0022-1767 ; 1048-3233 ; 1047-7381
DOI	10.4049/jimmunol.0902055
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