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  • Champlin, Richard  (6)
  • Rondon, Gabriela  (6)
  • 1
    In: Blood, American Society of Hematology, Vol. 116, No. 21 ( 2010-11-19), p. 489-489
    Abstract: Abstract 489 Relapse remains the major cause of treatment failure after allogeneic hematopoietic transplantation for AML and MDS. Alloreactive NK cells mediate a potent antileukemic effect and may also enhance engraftment and reduce GVHD. We performed a phase I study infusing “third party” alloreactive NK cells from a haploidentical related donor as a component of the preparative regimen for allotransplantation from a separate HLA identical donor. The goal was to augment the antileukemia cytotoxicity of the preparative regimen by infusion of alloreactive NK cells and improve the overall outcome of hematopoietic transplantation. Patients with advanced AML or high risk MDS in relapse or beyond first remission were eligible. They received the busulfan-fludarabine preparative regimen, followed by infusion of NK cells predicted to be alloreactive by KIR:KIR ligand incompatibility. The NK cell enriched product was produced from a steady state apheresis product by depleting CD3+ cells using the CliniMACS device (Miltenyi Corp). A second step positively selecting CD56+ cells was performed for the first dose level, but discontinued thereafter in order to increase the cell yield. The NK cell product was then cultured in complete media containing rIL-2 for 16 hours and infused intravenously following completion of the busulfan-fludarabine chemotherapy. After 5 days, ATG was administered followed by PBSC infusion from the HLA identical sibling or unrelated donor. Patients received tacrolimus and methotrexate for GVHD prophylaxis. Patients were treated in 4 dose levels of the NK cell enriched product; 1) 106 cells/kg, 2) 5 × 106/kg, 3) 3 × 107/kg, and 4) 3 × 107/kg followed by systemic interleukin-2 0.5 million units/m2 SQ daily for 5 days. CD3+ cells in the NK cell product were required to be 〈 105/kg (median infused 1.1 x104/kg). Median CD56+ cells infused (x106/kg) were from 0.9 (level 1), 1.5 (level 2), and 4.9 (levels 3 and 4). 13 patients were entered. Median age was 51 years (range 2–60). 11 had active disease and 2 were in a second remission. Only mild infusion toxicity occurred with the NK cell infusion. Other toxicities were similar to that experienced with the preparative regimen without NK cells. The 2 patients at dose level 4 tolerated interleukin-2 systemic treatment without fever or increased toxicity. The haploidentical NK cells were transiently detected in the blood by chimerism studies in one patient. Rapid engraftment and hematologic recovery uniformly occurred from the HLA matched PBPC donor in all patients. None had graft failure. Grade 2 acute GVHD developed in 3 patients; all responded to corticosteroid treatment. None developed grade 3 or 4 acute GVHD. 9 of 11 patients with active disease achieved a complete remission. Only one patient died of nonrelapse mortality; she died in remission from infection 2 months post transplant. This trial confirms the feasibility of producing the haploidentical NK cell product, and the lack of major toxicity attributable to the NK cell infusion in combination with an HLA compatible allogeneic transplantation. Infusion of haploidentical alloreactive NK cells was well tolerated and did not interfere with engraftment or increase the rate of GVHD after allogeneic hematopoietic transplantation. This approach merits further phase II and III study designed to reduce relapse and improve the outcome of allogeneic hematopoietic transplantation for AML/MDS. N NK cell doseCells/kg grade 2 GVHD Relapse NRM Alive in CR 4 106 1 4 0 0 4 5 × 106 0 3 0 1 (27+ m) 3 3 × 107 2 1 1 1 (11+ m) 2 3 × 107 + IL-2 0 0 0 2 (9+ m, 4+ m) Disclosures: No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2010
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  • 2
    In: Blood, American Society of Hematology, Vol. 112, No. 11 ( 2008-11-16), p. 58-58
    Abstract: MICA is a highly polymorphic locus located in the Class III region of the HLA system in the short arm of chromosome 6. The products of MICA can elicit humoral allo-recognition and are the ligands of the NKG2-D receptors of natural killer cells. MICA is involved in chronic and possibly acute graft rejection in kidney transplantation. The impact of mismatches in MICA has not been examined systematically in bone marrow transplantation. We hypothesized that donor-recipient MICA mismatches may influence graft-versus-host disease (GVHD) and relapse rates after UD hematopoietic stem cell transplantation (HSCT), and tested this hypothesis in a cohort comprised of all patients with myeloid leukemias transplanted in our institution from January, 2002 to December, 2007 (n=238). Methods: Typing of each of the classical human leukocyte antigen (HLA) and MICA loci was performed by amplification with locus-specific primers of genomic DNA by PCR followed by nucleotide sequencing. For the assignment of MICA alleles, the polymorphisms in exons 2, 3, 4, and 5 were evaluated. Matching grade is described in the GVH direction. Outcomes were acute (a) GVHD incidence, and aGVHD-free survival (time dependent variable, with development of aGVHD as the event), and relapse-free survival (RFS), both estimated by the Kaplan Meier method. Cox proportional hazards regression model was used to estimate the influence of HLA match degree, patient, disease, and transplant-related characteristics on outcomes. Median age was 50 years (range, 18–74; 24% over age 59). Diagnosis were AML/high-risk MDS in 82% (n=195) and CML in 18% (n=43). 42% (n=100) of the patients were in remission (CR) at HSCT. Preparative regimens were ablative in 59% (n=141), and contained ATG, fludarabine and IV busulfan in respectively 96%, 90% and 70% of the HSCT. GVHD prophylaxis was tacrolimus and mini-methotrexate-based in all HSCT. Stem cell source was bone marrow (BM) in 72% (n=172) and peripheral blood (PB) in 28% (n=66) of HSCT. 78 patients have relapsed (34%) and 133 have died (56%). Results. 169 pairs were matched in HLA A, B, C, DRB1, and DQB1 (10/10; 71%); 69 (29%) were & lt;10/10 matches, of which 60 were 9/10 (78% of the mismatches were in HLA class I). One or two HLA-DPB1 mismatches were present in 48% and 25% of the patients, respectively. MICA one or two mismatches were present in 22 pairs (9%). Grade (gd) II–IV and III–IV aGVHD rates for patients with MICA mismatches were 80% and 30%, respectively, versus 40% and 14% for those with no MICA mismatches. Effect of MICA on aGVHD was similar among 10/10 and & lt;10/10 patients. RFS at 52 weeks post HSCT was 0.58 (95%CI 0.5–0.66) versus 0.77 (95%CI 0.59–1) for patients without and with MICA mismatches (P=0.5). Multivariate models are shown in the table. Conclusion: MICA mismatches independently increased the incidence of grade II–IV aGVHD. Multivariate Models for grade II-IV acute GVHD and relapse-free survival (RFS) Variable Incidence Univariate P value Multivariate P, HR and 95% CI Grade II-IV aGVHD Fludarabine-based regimen(yes vs no) 39% vs 72% P=0.0009 0.02; HR 0.51 (0.29–0.9) DPB1 mismatches (0/1 vs 2) 38% vs 52% P=0.06 0.03; HR 0.59 (0.37–0.94) Ablative conditioning (no vs yes) 31% vs 48% P=0.001 0.05; HR 0.65 (0.43–0.99) MICA mismatches (yes vs no) 80% vs 38% P=0.0002 0.002; HR 2.33 (1.37–3.98) RFS Rate at 52 weeks 95%CI Univariate P Multivariate P, HR and 95% CI DPB1 (0 vs 1 vs 2) 0.54(0.42, 0.68) P=0.09 0.03; HR 1.65 (1.051–2.6) 0.55(0.45, 0.67) 0.7(0.58, 0.84) CR at HSCT (yes vs no) 0.78(0.69, 0.87) P & lt;0.0001 & lt;0.0001; HR 0.308 (0.18–0.52) 0.44(0.36, 0.55) (P and HR for aGVHD-free survival) HR: hazard ratio; CI: confidence interval
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2008
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  • 3
    In: Blood, American Society of Hematology, Vol. 108, No. 11 ( 2006-11-16), p. 3125-3125
    Abstract: The HLA class II DP locus encode for both subunits of DPB1 heterodimers, which have low levels of expression on the cell surface of antigen presenting cells. We hypothesized that donor-recipient HLA-DP mismatch would lead to an increased incidence of acute (a) graft-versus-host disease (GVHD), and that 2 mismatches would likely be even more significant. Methods: We studied 84 consecutive patients (pts) with myeloid leukemias in complete remission (CR) transplanted from 01/02 to 02/06. Preparative regimens were ablative IV Busulfan-based (n=58) or Cy/TBI (n=2), and reduced intensity (Fludarabine (Flu)/Bu 130 mg/m2/2 doses plus Gleevec (n=8), and Flu/Melphalan 140 mg/m2 (n=16). Stem cell (SC) source was bone marrow (n=70) or peripheral blood (n=14). ATG was given in 78 cases. GVHD prophylaxis was tacrolimus and mini-methotrexate in all cases, with additional pentostatin in 31 pts. High-resolution typing was sequence-based for HLA-A, B, DRB1; SSP was used for DRB3/4/5, DQB1 and DPB1, and SBT/SSOP for HLA-C. A Cox proportional hazards regression model was used to study aGVHD-free and relapse-free (RFS) survival. Variables with a p-value 〈 0.25 by univariate analysis were included in the multiple regression analysis (MV). Variables were age, gender, weight, conditioning regimen, GVHD prophylaxis, diagnosis, cytogenetics, SC source, ABO group, infused CD34 and CD3 cell dose, and HLA matching. AGVHD-free survival was calculated from transplant date to date of development of grade II–IV GVHD or completion of 100 days of follow-up. Results: Median age was 48 yrs (range, 14–72). Diagnoses were MDS (n=5), AML (n=58), and CML (n=21). 54 pts (64%) were beyond 1st CR; all CML pts were in 〉 1st chronic phase (CP). Sixty-one pts were 10/10 HLA match (A, B, C, DRB1, DQB1), and 23 had one or more mismatches. All but one pt engrafted neutrophils at a median of 13 days. 33 pts (39%) and 13 pts (15%) developed grade II–IV and III–IV aGVHD, respectively. Chronic GVHD incidence was 51%. With a median follow-up of 18 mo. (range,1.3–52) 60 pts are alive; 40 pts have relapsed or died. Median survival has not been reached. Number of DP mismatches and incidence of aGVHD is shown in the table. The following covariates influenced aGVHD-free survival by MV analysis: Flu-based regimen (P=0.005; HR 0.25 (95%CI 0.1–0.66), reduced intensity regimens (p=0.02; HR 0.35 (95%CI 0.15–0.83) and presence of 2 DPB1 mismatches (p=0.02; HR 3.07 (95%CI 1.19–7.95). Presence of 1 DPB1 mismatch was not significantly associated with aGVHD. There was no statistically significant correlation between presence of 2 DP mismatches and RFS (P=0.17;HR 0.3 (95%CI 0.06–1.65);HR 0.75 for 1 mismatch) or with cGVHD. Actuarial 2-yr survival for 10/10 matched pts without DP mismatches (12/12) versus those with DP mismatches is 82% versus 71%(P=0.6). In the 10/10 matched group, GVHD was the cause of death only among recipients of 2 DP mismatches transplants (n=4). Conclusion: Mismatching at HLA-DPB1 may increase the risk of aGVHD following UDT. The role of DP in the development of GVHD and GVL effects merits future study. Incidence of acute GVHD 10 of 10 matches number of DP mismatches grade II–IV grade III–IV 0 8% 0% 1 23% 8% 2 45% 18% 〈 10 of 10 matches number of DP mismatches grade II–IV grade III–IV 0 45% 15% 1 82% 36% 2 80% 40%
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2006
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  • 4
    In: Human Immunology, Elsevier BV, Vol. 71 ( 2010-9), p. S88-
    Type of Medium: Online Resource
    ISSN: 0198-8859
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2010
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  • 5
    In: Blood, American Society of Hematology, Vol. 110, No. 11 ( 2007-11-16), p. 3051-3051
    Abstract: It is unclear how related donor transplants in which the donor-recipient pair is mismatched in one locus (1AgMM) compare to transplants performed using a fully matched unrelated donor (MUD), matching at HLA-A, B, C, DRB1 and DQB1. Here, we performed such a comparison. Methods: We studied 83 consecutive patients (pts) with acute leukemias and myeloproliferative/myelodysplasia syndromes receiving grafts considered to be “1-antigen mismatched related” (1AgMM) and compared their outcomes to that of 134 MUD HSCT performed from 1992 to 2006. Among 83 1AgMM transplants, 40 had prospectively or retrospectively performed high-resolution HLA typing (Hires), of which 6 pts were 8/10 matched, and 43 had only low resolution (Lowres) typing. 8/10 Hires pts were excluded from this analysis. All MUD donor-recipient pairs were typed by Hires. Median age, gender, cytogenetic risk, disease status at transplant and stem cell source were similar. Diagnosis were ALL in 5% of the Hires group, 28% of the Lowres and 0 in the MUD group; AML/MDS cases were 76%, 60% and 87%, respectively; and CML cases were 19%, 12% and 13%, respectively(p 〈 0.001). Preparative regimens were of reduced intensity for 22% of Hires, 21% of Lowres patients and 40% of MUD patients (p=0.02). High-resolution typing was performed for HLA-A, B, DRB1, DQB1 and DPB1, and SBT/SSOP for HLA-C. Cumulative incidence of NRM, acute and chronic GVHD was estimated accounting for death in the absence of event as a competing risk. The Cox’s proportional hazards model was used to compare outcomes beteween groups. Results: With a median follow-up of 15 mo for MUD pts, 27 mo for Hires patients and 33 mo for Lowres pts, OS at 18 mo is 52%, 46% and 22% (p= 0.002 for the comparison of MUD vs Lowres; P=NS for the comparison of MUD vs Hires); NRM is 23%, 26% and 54%(p=0.001 for the comparison of MUD vs Lowres; P=NS for the comparison of MUD vs Hires). Primary graft failure occurred in 1% of MUD, 5% of MOL and 12% of SER (p=0.003). Cumulative incidence of grade II-IV aGVHD is 34% for MUD pts, 48% for Hires and 49% for Lowres(p= 0.07); cGVHD: 34%, 40% and 54%(p=0.01 for the comparison of MUD vs Lowres). We divided the Hires related group according to presence of Class I or Class II mismatches, and compared their outcomes to the MUD group (Table). 23 pts had Class I and 11 pts had Class II mismatches; median follow-up is 26 and 62 mo, respectively. Distribution of gender, diagnosis, cytogenetic risk, dis. status at transplant, ablative/RIC regimens and stem cell source was similar between the 2 groups. Pts with class I mismatches had the worse survival. Conclusion: Cumulative incidence of grade II-IV aGVHD and cGVHD, non-relapse mortality and overall survival of 1AgMM (allele level typing) patients was similar to that observed in a cohort of recipients of molecularly matched, HLA 10/10 MUD transplants. OS at 18 mo HR (95%CI) P C. Incid. gd II-IV aGVHD HR (95% CI) P C. Incid. chronic GVHD HR (95% CI) P C. Incid= cumulative incidence MUD n=134 52% Reference 34% Reference 34% Reference Class I n=23 34% 1.8 (1–3.1) P=0.05 40% 1.3 (0.6–2.9) P=0.4 35% 1.1 (0.5–2.5) P=0.7 Class II n=11 73% 0.6 (0.2–1.9) P=0.4 64% 2.8 (1.3–6.3) P=0.01 48% 2.3 (0.9–5.7) P=0.08
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2007
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 6
    In: Blood, American Society of Hematology, Vol. 110, No. 11 ( 2007-11-16), p. 3050-3050
    Abstract: Little is known of the possible additive effects of mismatches in low expression class II HLA loci (DQB1, DPB1 and DRB3,4,5), and mismatches in MHC class I chain related (MIC-A) on the outcomes of unrelated donor HSCT. We investigated such hypothesis in a group of 139 consecutive patients (pts) with myeloid leukemias transplanted from 01/02 to 02/06 in our institution. All pts received a 8/8 matched graft (HLA-A, -B, -C, -DRB1). Preparative regimens were ablative [IV Busulfan-based (n=93) or Cy/TBI (n=1)], and reduced intensity [(Fludarabine (Flu)/Bu 130 mg/m2/2 doses plus Gleevec (n=7), and Flu/Melphalan 140 mg/m2 (n=38)] . Stem cell (SC) source was bone marrow (n=107) or peripheral blood (n=32). ATG was given in 134 cases. GVHD prophylaxis was tacrolimus and mini-methotrexate in all cases, with additional pentostatin in 38 pts. High-resolution typing was sequence-based for HLA-A, B, DRB1; SSP was used for DRB3/4/5, DQB1 and DPB1, SBT/SSOP for HLA-C and nucleotide-sequencing for MIC-A. A Cox proportional hazards regression model was used to study aGVHD-free and relapse-free survival (RFS). Variables with a p-value 〈 0.25 by univariate analysis were included in the multiple regression analysis (MV). Variables were age, gender, weight, conditioning regimen, GVHD prophylaxis, diagnosis, cytogenetics, SC source, ABO group, infused CD34 and CD3 cell dose, and HLA matching. aGVHD-free survival was calculated from transplant date to date of development of grade II-IV aGVHD or completion of 100 days of follow-up. Results: Median age was 50 yrs (range, 14–75). Diagnoses were MDS (n=19), AML (n=95), CML (n=18) and other MPD diseases (n=6). 61 patients (44%) were in 1st or 2nd CR at transplant; all CML pts were in 〉 1st chronic phase (CP). 133 (96%) patients engrafted neutrophils at a median of 13 days. 49 (35%) and 13 (09%) pts developed grade II-IV and III-IV aGVHD, respectively. Chronic GVHD incidence was 33%. With a median follow-up of 16 months (range, 2–64), 78 pts are alive. Median survival has not been reached. Reduced-intensity conditioning regimen was the only covariate which influenced aGVHD-free survival by MV analysis [p= 0.05; HR 0.53 (95%CI 0.28–1.0)]. Treatment-related mortality was higher among patients who had more than 3 mismatches in the low-expression loci and/or in MIC-A [p=0.01; HR 3.87 (95%CI 1.33–11.26)] . Among the 82 pts who were in CR at transplantation, 10/10 matching status reduced the incidence of grade II-IV aGVHD [p= 0.04; HR 0.35 (95%CI 1.18–0.72)], while the presence of a MIC-A mismatch determined a higher grade II-IV aGVHD incidence [p=0.02; HR 2.7 (95%CI 1.15–6.31)] . Conclusion: Multiple mismatches in low expression loci and in MIC-A increase treatment-related mortality in HLA 8/8 matched donor-recipient pairs in a cumulative manner.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
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    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2007
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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