title: Purification of CD4+ T cells for adoptive immunotherapy after allogeneic hematopoietic stem cell transplantation.
abstract: Donor lymphocyte infusions (DLIs) are used for adoptive immunotherapy to prevent or treat relapse and infectious complications after allogeneic hematopoietic stem cell transplantation (HSCT). Unmanipulated DLIs are associated with a risk of graft-versus-host disease (GVHD), probably related to CD8(+) T cell activity. We investigated an automated clinical-scale human-CD4(+)-cell purification method to deplete CD8(+) cells. Twenty-four stem cell recipients received a total of 24 leukapheresis products being enriched for CD4(+) cells using magnetic associated cell sorting (MACS) with an automated device (CliniMACS(®)) before DLIs. MACS resulted in a mean CD4(+) cell count of 16 × 10(6)/kg bw corresponding to 3.4-fold CD4(+) cell enrichment. Mean yield and purity of CD45(+)CD3(+)CD4(+)CD14(-)7AAD(-) were 74% ± 23% and 82% ± 11%, respectively. Median initial dose of DLIs was 1.1 × 10(6) CD4(+)/kg. During a median follow-up of 25 months, 7 (30%) patients experienced GVHD (acute II-IV: n = 4, 17%; acute III-IV: n = 2, 8%; chronic limited: n = 2, 8%; chronic extensive: n = 1, 4%). Thirteen of 21 further evaluable patients (62%) showed measurable clinical response, 2 patients with therapy refractory infectious complications (HSV) showed remarkable immunologic improvement. Automated enrichment of CD4(+) by magnetic cell sorting provides an efficient and rapid method for processing donor lymphocytes. Additional studies should further investigate this approach in terms of efficacy and the risk of GVHD.
authors.affiliation: Department of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Klyuchnikov
authors.forename: Andreas
authors.initials: A
authors.lastname: Sputtek
authors.forename: Olga
authors.initials: O
authors.lastname: Slesarchuk
authors.forename: Michael
authors.initials: M
authors.lastname: Lioznov
authors.forename: Thomas
authors.initials: T
authors.lastname: Stübig
authors.forename: Ulrike
authors.initials: U
authors.lastname: Bacher
authors.forename: Gitta
authors.initials: G
authors.lastname: Amtsfeld
authors.forename: Edeltraut
authors.initials: E
authors.lastname: Merle
authors.forename: Marie-Luise
authors.initials: ML
authors.lastname: Reckhaus
authors.forename: Boris
authors.initials: B
authors.lastname: Fehse
authors.forename: Christine
authors.initials: C
authors.lastname: Wolschke
authors.forename: Raissa
authors.initials: R
authors.lastname: Adjallé
authors.forename: Francis
authors.initials: F
authors.lastname: Ayuk
authors.forename: Axel
authors.initials: A
authors.lastname: Zander
authors.forename: Nicolaus
authors.initials: N
authors.lastname: Kröger
mesh_terms: D000328:Adult; D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D015331:Cohort Studies; D005260:Female; D005500:Follow-Up Studies; D006086:Graft vs Host Disease / Q000276:immunology / Q000517:prevention & control; D019337:Hematologic Neoplasms / Q000276:immunology / Q000628:therapy*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation / Q000379:methods*; D016219:Immunotherapy, Adoptive / Q000009:adverse effects / Q000379:methods*; D008297:Male; D008875:Middle Aged; D009101:Multiple Myeloma / Q000276:immunology / Q000628:therapy; D009894:Opportunistic Infections / Q000517:prevention & control / Q000628:therapy; D012189:Retrospective Studies; D055502:Secondary Prevention; D016019:Survival Analysis; D055815:Young Adult