Optimization of individualized graft composition: CD3/CD19 depletion combined with CD34 selection for haploidentical transplantation

Huenecke, Sabine; Bremm, Melanie; Cappel, Claudia; Esser, Ruth; Quaiser, Andrea; Bönig, Halvard; Jarisch, Andrea; Soerensen, Jan; Klingebiel, Thomas; Bader, Peter; Koehl, Ulrike

doi:10.1111/trf.13694

Cited by 13 publications

(17 citation statements)

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“…Depending on the underlying disease, either myeloablative conditioning regimens (MAC) based on total body irradiation (12 Gy TBI) or busulphan, or a reduced-intensity conditioning regimen (RIC) with fludarabine, thiotepa, and melphalan were used. The positive selection of CD34 + PB stem cells with immunomagnetic microbeads and the direct depletion of T and B cells with-coated microbeads under good manufacturing practice (GMP) were performed with-coated microbeads using a CliniMACS (Miltenyi Biotec, Bergisch-Gladbach, Germany) device under GMP as we described previously ( 23 , 25 ). For prophylaxis of GvHD, OKT3 or anti-thymocyte globulin was given.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, T cells need to be depleted prior to the infusion of the haploidentical graft. Different strategies for the purification of haploidentical grafts are available, such as the positive selection of CD34 + stem cells leading to indirect T-cell depletion ( 2 , 15 , 17 ) and the direct depletion of CD3 + T cells or TCR alpha/beta T cells, both in combination with CD19 + B-cell removal ( 20 – 23 ). CD34 + -selected grafts contain highly purified stem cells and can be grafted in the absence of GvHD prophylaxis ( 24 ).…”

Section: Introductionmentioning

confidence: 99%

“…CD34 + -selected grafts contain highly purified stem cells and can be grafted in the absence of GvHD prophylaxis ( 24 ). CD3/CD19 or TCR alpha/beta-depleted grafts contain hematopoietic progenitors, natural killer (NK) cells, and antigen-presenting cells, probably leading to an enhanced graft vs. leukemia effect ( 16 , 20 , 23 ).…”

Section: Introductionmentioning

confidence: 99%

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Joint Modeling of Immune Reconstitution Post Haploidentical Stem Cell Transplantation in Pediatric Patients With Acute Leukemia Comparing CD34+-Selected to CD3/CD19-Depleted Grafts in a Retrospective Multicenter Study

et al. 2018

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Rapid immune reconstitution (IR) following stem cell transplantation (SCT) is essential for a favorable outcome. The optimization of graft composition should not only enable a sufficient IR but also improve graft vs. leukemia/tumor effects, overcome infectious complications and, finally, improve patient survival. Especially in haploidentical SCT, the optimization of graft composition is controversial. Therefore, we analyzed the influence of graft manipulation on IR in 40 patients with acute leukemia in remission. We examined the cell recovery post haploidentical SCT in patients receiving a CD34+-selected or CD3/CD19-depleted graft, considering the applied conditioning regimen. We used joint model analysis for overall survival (OS) and analyzed the dynamics of age-adjusted leukocytes; lymphocytes; monocytes; CD3+, CD3+CD4+, and CD3+CD8+ T cells; natural killer (NK) cells; and B cells over the course of time after SCT. Lymphocytes, NK cells, and B cells expanded more rapidly after SCT with CD34+-selected grafts (P = 0.036, P = 0.002, and P < 0.001, respectively). Contrarily, CD3+CD4+ helper T cells recovered delayer in the CD34 selected group (P = 0.026). Furthermore, reduced intensity conditioning facilitated faster immune recovery of lymphocytes and T cells and their subsets (P < 0.001). However, the immune recovery for NK cells and B cells was comparable for patients who received reduced-intensity or full preparative regimens. Dynamics of all cell types had a significant influence on OS, which did not differ between patients receiving CD34+-selected and those receiving CD3/CD19-depleted grafts. In conclusion, cell reconstitution dynamics showed complex diversity with regard to the graft manufacturing procedure and conditioning regimen.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Joint Modeling of Immune Reconstitution Post Haploidentical Stem Cell Transplantation in Pediatric Patients With Acute Leukemia Comparing CD34+-Selected to CD3/CD19-Depleted Grafts in a Retrospective Multicenter Study

et al. 2018

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“…An effective strategy to prevent the occurrence of lethal GvHD is the in vitro depletion of T cells. Specific depletion techniques are available to remove T and B cells from the grafts (CD3/CD19- or TCRαβ/CD19-depletion) while preserving cells that are beneficial regarding engraftment and GvL/T effect such as NK cells (9, 34, 35). Notably, HLA class I expression is reduced on NB cells which makes them valid targets for NK cell lysis (10, 36).…”

Section: Discussionmentioning

confidence: 99%

“…The removal of potentially alloreactive T cells from the graft by CD3/CD19-depletion allows HLA barriers to be overcome and reduces the induction of harmful graft-versus-host-disease (GvHD). While the risk of EBV post-transplant lymphoproliferative disease (PTLD) is reduced by depletion of CD19 + B cells, CD3/CD19-depleted grafts also seem to facilitate engraftment and to support the graft-versus-leukemia/tumor (GvL/T) effect (9).…”

Section: Introductionmentioning

confidence: 99%

The Synergistic Use of IL-15 and IL-21 for the Generation of NK Cells From CD3/CD19-Depleted Grafts Improves Their ex vivo Expansion and Cytotoxic Potential Against Neuroblastoma: Perspective for Optimized Immunotherapy Post Haploidentical Stem Cell Transplantation

et al. 2019

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Neuroblastoma (NB) is the most common solid extracranial tumor in childhood. Despite therapeutic progress, prognosis in high-risk NB is poor and innovative therapies are urgently needed. Therefore, we addressed the potential cytotoxic capacity of interleukin (IL)-activated natural killer (NK) cells compared to cytokine-induced killer (CIK) cells for the treatment of NB. NK cells were isolated from peripheral blood mononuclear cells (PBMCs) by indirect CD56-enrichment or CD3/CD19-depletion and expanded with different cytokine combinations, such as IL-2, IL-15, and/or IL-21 under feeder-cell free conditions. CIK cells were generated from PBMCs by ex vivo stimulation with interferon-γ, IL-2, OKT-3, and IL-15. Comparative analysis of expansion rate, purity, phenotype and cytotoxicity was performed. CD56-enriched NK cells showed a median expansion rate of 4.3-fold with up to 99% NK cell content. The cell product after CD3/CD19-depletion consisted of a median 43.5% NK cells that expanded significantly faster reaching also 99% of NK cell purity. After 10–12 days of expansion, both NK cell preparations showed a significantly higher median cytotoxic capacity against NB cells relative to CIK cells. Remarkably, these NK cells were also capable of efficiently killing NB spheroidal 3D culture in long-term cytotoxicity assays. Further optimization using a novel NK cell culture medium and a prolonged culturing procedure after CD3/CD19-depletion for up to 15 days enhanced the expansion rate up to 24.4-fold by maintaining the cytotoxic potential. Addition of an IL-21 boost prior to harvesting significantly increased the cytotoxicity. The final cell product consisted for the major part of CD16−, NCR-expressing, poly-functional NK cells with regard to cytokine production, CD107a degranulation and antitumor capacity. In summary, our study revealed that NK cells have a significantly higher cytotoxic potential to combat NB than CIK cell products, especially following the synergistic use of IL-15 and IL-21 for NK cell activation. Therefore, the use of IL-15+IL-21 expanded NK cells generated from CD3/CD19-depleted apheresis products seems to be highly promising as an immunotherapy in combination with haploidentical stem cell transplantation (SCT) for high-risk NB patients.

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Graft Manipulation

Schumm¹,

Lang²,

Handgretinger³

2018

The EBMT Handbook

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Optimization of individualized graft composition: CD3/CD19 depletion combined with CD34 selection for haploidentical transplantation

Abstract: Finally, calculated splitting of the starting product followed by CD3/19 depletion together with CD34+ graft manipulation may enable the composition of optimized grafts with high CD34+-cell and minimal T-cell content.

Cited by 13 publications

References 38 publications

Joint Modeling of Immune Reconstitution Post Haploidentical Stem Cell Transplantation in Pediatric Patients With Acute Leukemia Comparing CD34+-Selected to CD3/CD19-Depleted Grafts in a Retrospective Multicenter Study

Joint Modeling of Immune Reconstitution Post Haploidentical Stem Cell Transplantation in Pediatric Patients With Acute Leukemia Comparing CD34+-Selected to CD3/CD19-Depleted Grafts in a Retrospective Multicenter Study

The Synergistic Use of IL-15 and IL-21 for the Generation of NK Cells From CD3/CD19-Depleted Grafts Improves Their ex vivo Expansion and Cytotoxic Potential Against Neuroblastoma: Perspective for Optimized Immunotherapy Post Haploidentical Stem Cell Transplantation

Graft Manipulation

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