The Immunogenicity of Virus-derived 2A Sequences in Immunocompetent Individuals

Overview

Journal Gene Ther

Date 2013 May 24

PMID 23698740

Citations 21

Authors

C Arber

H Abhyankar

H E Heslop

M K Brenner

H Liu

G Dotti

B Savoldo

Affiliations

Soon will be listed here.

Abstract

Genetic engineering of T cells for adoptive immunotherapy in cancer patients has shown significant promise. To ensure optimal antitumor activity and safety, the simultaneous expression of multiple genes is frequently required, and short viral-derived 2A sequences are increasingly preferred for this purpose. Concerns exist, however, that these virus-derived sequences may induce unwanted immune responses, and thus diminish persistence of the gene-modified cells after adoptive transfer. Whereas such responses were absent in immunocompromised recipients, potential immunogenicity in immunocompetent individuals remains a concern. We now address whether ex vivo T cell responses can be elicited against the most widely used 2A sequences (2A-Thosea asigna virus (TAV) or 2A-equine rhinitis virus (ERAV), specifically) in immunocompetent individuals. We used a potent ex vivo culture system previously validated to induce T cell responses even against weakly immunogenic antigens. Of the sixteen donors tested, only five released very low levels of interferon-γ in response to 2A-TAV peptide mixtures (single peptide specificity in three donors, adjacent self-antigen peptide specificity in one donor and nonspecific reactivity in one donor). None of them produced cytotoxic activity or responded to 2A-ERAV. These results suggest that exposure to viral-derived 2A sequences is unlikely to produce unwanted T cell responses in immunocompetent individuals and further supports their continued use for studies of human gene therapy.

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References

Szymczak A, Workman C, Wang Y, Vignali K, Dilioglou S, Vanin E . Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide-based retroviral vector. Nat Biotechnol. 2004; 22(5):589-94. DOI: 10.1038/nbt957. View

Milone M, Fish J, Carpenito C, Carroll R, Binder G, Teachey D . Chimeric receptors containing CD137 signal transduction domains mediate enhanced survival of T cells and increased antileukemic efficacy in vivo. Mol Ther. 2009; 17(8):1453-64. PMC: 2805264. DOI: 10.1038/mt.2009.83. View

Zhang L, Kerkar S, Yu Z, Zheng Z, Yang S, Restifo N . Improving adoptive T cell therapy by targeting and controlling IL-12 expression to the tumor environment. Mol Ther. 2011; 19(4):751-9. PMC: 3070103. DOI: 10.1038/mt.2010.313. View

Johnson L, Morgan R, Dudley M, Cassard L, Yang J, Hughes M . Gene therapy with human and mouse T-cell receptors mediates cancer regression and targets normal tissues expressing cognate antigen. Blood. 2009; 114(3):535-46. PMC: 2929689. DOI: 10.1182/blood-2009-03-211714. View

Hanley P, Cruz C, Savoldo B, Leen A, Stanojevic M, Khalil M . Functionally active virus-specific T cells that target CMV, adenovirus, and EBV can be expanded from naive T-cell populations in cord blood and will target a range of viral epitopes. Blood. 2009; 114(9):1958-67. PMC: 2738578. DOI: 10.1182/blood-2009-03-213256. View

Liu D, Song L, Wei J, Courtney A, Gao X, Marinova E . IL-15 protects NKT cells from inhibition by tumor-associated macrophages and enhances antimetastatic activity. J Clin Invest. 2012; 122(6):2221-33. PMC: 3366399. DOI: 10.1172/JCI59535. View

Craddock J, Lu A, Bear A, Pule M, Brenner M, Rooney C . Enhanced tumor trafficking of GD2 chimeric antigen receptor T cells by expression of the chemokine receptor CCR2b. J Immunother. 2010; 33(8):780-8. PMC: 2998197. DOI: 10.1097/CJI.0b013e3181ee6675. View

Tey S, Dotti G, Rooney C, Heslop H, Brenner M . Inducible caspase 9 suicide gene to improve the safety of allodepleted T cells after haploidentical stem cell transplantation. Biol Blood Marrow Transplant. 2007; 13(8):913-24. PMC: 2040267. DOI: 10.1016/j.bbmt.2007.04.005. View

Di Stasi A, De Angelis B, Rooney C, Zhang L, Mahendravada A, Foster A . T lymphocytes coexpressing CCR4 and a chimeric antigen receptor targeting CD30 have improved homing and antitumor activity in a Hodgkin tumor model. Blood. 2009; 113(25):6392-402. PMC: 2710932. DOI: 10.1182/blood-2009-03-209650. View

10.

Kerkar S, Restifo N . Cellular constituents of immune escape within the tumor microenvironment. Cancer Res. 2012; 72(13):3125-30. PMC: 6327310. DOI: 10.1158/0008-5472.CAN-11-4094. View

11.

Hoyos V, Savoldo B, Dotti G . Genetic modification of human T lymphocytes for the treatment of hematologic malignancies. Haematologica. 2012; 97(11):1622-31. PMC: 3487433. DOI: 10.3324/haematol.2012.064303. View

12.

Ciceri F, Bonini C, Lupo Stanghellini M, Bondanza A, Traversari C, Salomoni M . Infusion of suicide-gene-engineered donor lymphocytes after family haploidentical haemopoietic stem-cell transplantation for leukaemia (the TK007 trial): a non-randomised phase I-II study. Lancet Oncol. 2009; 10(5):489-500. DOI: 10.1016/S1470-2045(09)70074-9. View

13.

Szymczak-Workman A, Vignali K, Vignali D . Design and construction of 2A peptide-linked multicistronic vectors. Cold Spring Harb Protoc. 2012; 2012(2):199-204. DOI: 10.1101/pdb.ip067876. View

14.

Hoyos V, Savoldo B, Quintarelli C, Mahendravada A, Zhang M, Vera J . Engineering CD19-specific T lymphocytes with interleukin-15 and a suicide gene to enhance their anti-lymphoma/leukemia effects and safety. Leukemia. 2010; 24(6):1160-70. PMC: 2888148. DOI: 10.1038/leu.2010.75. View

15.

Restifo N, Dudley M, Rosenberg S . Adoptive immunotherapy for cancer: harnessing the T cell response. Nat Rev Immunol. 2012; 12(4):269-81. PMC: 6292222. DOI: 10.1038/nri3191. View

16.

Quintarelli C, Dotti G, Hasan S, De Angelis B, Hoyos V, Errichiello S . High-avidity cytotoxic T lymphocytes specific for a new PRAME-derived peptide can target leukemic and leukemic-precursor cells. Blood. 2011; 117(12):3353-62. PMC: 3069675. DOI: 10.1182/blood-2010-08-300376. View

17.

Quintarelli C, Vera J, Savoldo B, Giordano Attianese G, Pule M, Foster A . Co-expression of cytokine and suicide genes to enhance the activity and safety of tumor-specific cytotoxic T lymphocytes. Blood. 2007; 110(8):2793-802. PMC: 2018664. DOI: 10.1182/blood-2007-02-072843. View

18.

Quintarelli C, Dotti G, De Angelis B, Hoyos V, Mims M, Luciano L . Cytotoxic T lymphocytes directed to the preferentially expressed antigen of melanoma (PRAME) target chronic myeloid leukemia. Blood. 2008; 112(5):1876-85. PMC: 3401035. DOI: 10.1182/blood-2008-04-150045. View

19.

Di Stasi A, Tey S, Dotti G, Fujita Y, Kennedy-Nasser A, Martinez C . Inducible apoptosis as a safety switch for adoptive cell therapy. N Engl J Med. 2011; 365(18):1673-83. PMC: 3236370. DOI: 10.1056/NEJMoa1106152. View

20.

de Felipe P . Skipping the co-expression problem: the new 2A "CHYSEL" technology. Genet Vaccines Ther. 2004; 2(1):13. PMC: 521497. DOI: 10.1186/1479-0556-2-13. View