title: Clinical-scale selection and viral transduction of human naïve and central memory CD8+ T cells for adoptive cell therapy of cancer patients.
abstract: The adoptive transfer of lymphocytes genetically engineered to express tumor-specific antigen receptors is a potent strategy to treat cancer patients. T lymphocyte subsets, such as naïve or central memory T cells, selected in vitro prior to genetic engineering have been extensively investigated in preclinical mouse models, where they demonstrated improved therapeutic efficacy. However, so far, this is challenging to realize in the clinical setting, since good manufacturing practices (GMP) procedures for complex cell sorting and genetic manipulation are limited. To be able to directly compare the immunological attributes and therapeutic efficacy of naïve (T(N)) and central memory (T(CM)) CD8(+) T cells, we investigated clinical-scale procedures for their parallel selection and in vitro manipulation. We also evaluated currently available GMP-grade reagents for stimulation of T cell subsets, including a new type of anti-CD3/anti-CD28 nanomatrix. An optimized protocol was established for the isolation of both CD8(+) T(N) cells (CD4(-)CD62L(+)CD45RA(+)) and CD8(+) T(CM) (CD4(-)CD62L(+)CD45RA(-)) from a single patient. The highly enriched T cell subsets can be efficiently transduced and expanded to large cell numbers, sufficient for clinical applications and equivalent to or better than current cell and gene therapy approaches with unselected lymphocyte populations. The GMP protocols for selection of T(N) and T(CM) we reported here will be the basis for clinical trials analyzing safety, in vivo persistence and clinical efficacy in cancer patients and will help to generate a more reliable and efficacious cellular product.
authors.affiliation: Research and Development, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Casati
authors.forename: Azam
authors.initials: A
authors.lastname: Varghaei-Nahvi
authors.forename: Steven Alexander
authors.initials: SA
authors.lastname: Feldman
authors.forename: Mario
authors.initials: M
authors.lastname: Assenmacher
authors.forename: Steven Aaron
authors.initials: SA
authors.lastname: Rosenberg
authors.forename: Mark Edward
authors.initials: ME
authors.lastname: Dudley
authors.forename: Alexander
authors.initials: A
authors.lastname: Scheffold
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002454:Cell Differentiation / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008545:Melanoma / Q000276:immunology / Q000628:therapy*; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D014161:Transduction, Genetic
references.citation: N Engl J Med. 2011 Aug 25;365(8):725-33
references.pmid: 21830940
references.citation: Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7739-43
references.pmid: 7644487
references.citation: Nat Rev Immunol. 2006 May;6(5):383-93
references.pmid: 16622476
references.citation: Curr Opin Immunol. 2010 Apr;22(2):251-7
references.pmid: 20171074
references.citation: J Clin Invest. 2008 Jan;118(1):294-305
references.pmid: 18060041
references.citation: J Clin Invest. 2010 Nov;120(11):3953-68
references.pmid: 20978347
references.citation: J Immunol. 2006 Jun 15;176(12):7726-35
references.pmid: 16751420
references.citation: Sci Transl Med. 2012 Mar 28;4(127):127ps8
references.pmid: 22461638
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 17;101(7):1969-74
references.pmid: 14762166
references.citation: Clin Cancer Res. 2011 Jul 1;17(13):4550-7
references.pmid: 21498393
references.citation: J Immunother. 2009 May;32(4):415-23
references.pmid: 19342963
references.citation: Science. 1996 Jun 28;272(5270):1939-43
references.pmid: 8658167
references.citation: N Engl J Med. 2008 Jun 19;358(25):2698-703
references.pmid: 18565862
references.citation: Proc Natl Acad Sci U S A. 2005 Jul 5;102(27):9571-6
references.pmid: 15980149
references.citation: J Clin Invest. 2005 Jun;115(6):1616-26
references.pmid: 15931392
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 20;109(12):4592-7
references.pmid: 22393002
references.citation: Curr Gene Ther. 2011 Aug;11(4):276-87
references.pmid: 21453279
references.citation: J Immunother. 2005 May-Jun;28(3):258-67
references.pmid: 15838383
references.citation: Nat Med. 2008 Nov;14(11):1264-70
references.pmid: 18978797
references.citation: J Clin Oncol. 2011 Mar 1;29(7):917-24
references.pmid: 21282551
references.citation: N Engl J Med. 2000 Oct 5;343(14):1020-34
references.pmid: 11018170
references.citation: Blood. 2003 Jun 1;101(11):4260-6
references.pmid: 12576317
references.citation: Mol Ther. 2011 Mar;19(3):620-6
references.pmid: 21157437
references.citation: Nature. 1999 Oct 14;401(6754):708-12
references.pmid: 10537110
references.citation: Blood. 2009 Jan 29;113(5):1006-15
references.pmid: 18978209
references.citation: Nat Rev Immunol. 2012 Mar 22;12(4):269-81
references.pmid: 22437939
references.citation: Mol Ther. 2010 Apr;18(4):843-51
references.pmid: 20179677
references.citation: J Immunother. 2003 Jul-Aug;26(4):332-42
references.pmid: 12843795
references.citation: J Immunol. 2005 Nov 15;175(10):7046-52
references.pmid: 16272366
references.citation: Int J Cancer. 1992 Jul 30;51(6):973-9
references.pmid: 1386349
references.citation: Blood. 2011 Jan 20;117(3):808-14
references.pmid: 20971955
references.citation: Blood. 2009 Jul 16;114(3):535-46
references.pmid: 19451549
references.citation: Hum Gene Ther. 2011 Jan;22(1):107-15
references.pmid: 20662590
references.citation: Eur J Immunol. 2009 Aug;39(8):2083-7
references.pmid: 19637204
references.citation: Blood. 2011 Feb 10;117(6):1888-98
references.pmid: 21123821
references.citation: Immunity. 2009 Nov 20;31(5):787-98
references.pmid: 19879162
references.citation: J Immunol. 2006 Nov 1;177(9):6548-59
references.pmid: 17056587
references.citation: J Immunol. 2010 Jun 1;184(11):6232-41
references.pmid: 20427762
references.citation: Science. 2006 Oct 6;314(5796):126-9
references.pmid: 16946036
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 13;106(41):17469-74
references.pmid: 19805141
references.citation: Cancer Cell. 2003 May;3(5):431-7
references.pmid: 12781360
references.citation: Blood. 2010 Nov 18;116(20):4099-102
references.pmid: 20668228
references.citation: Blood. 2008 Jun 1;111(11):5326-33
references.pmid: 18276844
references.citation: Blood. 2011 Jan 6;117(1):72-82
references.pmid: 20889925
references.citation: Nat Biotechnol. 2002 Feb;20(2):143-8
references.pmid: 11821859