Improved Early Outcomes with T-Cell Replete (TCR) Compared with T-Cell Depleted (TCD) Haploidentical Stem Cell Transplantation (HaploSCT)

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 320-320 ◽  
Author(s):  
Stefan O Ciurea ◽  
Rima M. Saliba ◽  
Ulas D. Bayraktar ◽  
Susan Xie ◽  
Gabriela Rondon ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Conflicts Of Interest ◽  
Conditioning Regimen ◽  
T Cell Subsets ◽  
High Dose ◽  
Cd4 Cells ◽  
Post Transplant ◽  
Cd34 Cells ◽  
Immunologic Reconstitution

Abstract Abstract 320 Background: HaploSCT has been commonly performed with a TCD graft using CD34+ selection; however, this has been limited by a higher non-relapse mortality (NRM) primarily related to infectious complications. An alternative approach using a TCR bone marrow graft and high-dose post-transplant cyclophosphamide (HDPTCy) in the setting of non-myeloablative conditioning has been reported to have lower NRM and acceptable rates of GVHD. Methods: We hypothesized that TCR HaploSCT using HDPTCy is associated with improved immunologic reconstitution, less NRM and better early outcomes compared with TCD HaploSCT, and analyzed 65 consecutive patients (pts) treated at UTMDACC with the same conditioning regimen, fludarabine (40mg/m2/day × 4), melphalan (140mg/m2) and thiotepa (10mg/kg). TCD HaploSCT pts were treated between 2001 and 2009, while TCR patients after 2009. 6 pts in the TCR group >55 years/comorbidities received reduced doses of melphalan (100mg/m2) and thiotepa (5mg/kg). There was no GVHD prophylaxis in the TCD group, while TCR group received HDPTCy (50mg/kg/day × 2) followed by tacrolimus and mycophenolate. Results: The median follow-up was 10 months (range 3.5–25) for the TCR group and 44 (11–79) months for the TCD group. Median age was 45 years (range 20–63) in the TCR group and 36 years (range 18–56) in the TCD group (p=0.02). 28% were > 50 years in the TCR compared with 6% in the TCD group (p=0.02). Diagnoses were: AML/MDS 50% vs. 79%, ALL 13% vs. 12%, CML 16% vs. 6%, lymphoma/CLL 9% vs. 3% in the TCR vs. TCD groups, respectively. Only 13 (41%) and 12 (36%) of pts were in remission at transplant in both groups, respectively (p=0.7). 10/16 (62.5%) pts with AML/MDS in the TCR group had poor risk cytogenetics vs. 13/26 (50%) pts in the TCD group. The donors were 5/10 allele match in 20/32 (63%) and 16/31 (52%) in the two groups, respectively. Median numbers of CD34+ cells infused were 2.5×10e6/kg in the TCR group and 10.5×10e6/kg in the TCD group. All pts in the TCD group had peripheral blood selected CD34+ cells while all but one received bone marrow stem cells in the TCR group. One pt had early death in each group. Primary engraftment was achieved in 94% in the TCR group and 81% in the TCD group (p=0.1). Day-100 NRM for all pts was 9% in the TCR group vs. 21% for the TCD group, and for pts in remission at transplant 0% vs. 42%, respectively (p=0.01). NRM at 1 year for all pts was 16% for the TCR group vs. 42% for the TCD group (p=0.03) (Figure1), while for pts in remission was 0% vs. 67% (p=0.001). The cumulative incidences of grade II-IV aGVHD was 27% vs. 11% (p=0.5) and cGVHD was 8% vs. 18%, in the TCR and TCD group, respectively (p=0.03). OS and PFS at 1 year post-transplant were 66% vs. 30% (p=0.02) and 45% vs. 21% (p=0.03) for the whole group, and 92% vs. 33% (p=0.03) and 80% vs. 25% (p=0.02) for pts in remission at transplant, respectively (Figure1). Improved NRM in the TCR group was related to significantly better immunologic reconstitution of T-cell subsets. On day 30 post transplant there was a significantly better recovery of absolute CD4 cells in the TCR group (median 24 vs. 2, p=0.004) and CD8 cells (median 20.5 vs. 1.5, p=0.036). CD4 cells remained significantly lower in the TCD group until after day 180 when the median CD4 count was 200.5 vs. 64 in the TCR group (p=0.04) while the difference in CD8 counts became non-significantly higher in the TCR after day 90 (median 119 vs. 29, p=0.23). Conclusion: TCR HaploSCT is associated with better immunologic reconstitution and improved early outcomes compared with TCD HaploSCT. Disclosures: No relevant conflicts of interest to declare.

Comparative Analysis on Cellular Components of Bone Marrow Microenvironment in Normal and Aberrant Hematopoiesis

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4808-4808
Author(s):  
Young-Ho Lee ◽  
Young-hee Kwon ◽  
Kyoujung Hwang ◽  
Hyunju Jun ◽  
Byungbae Park ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Conflicts Of Interest ◽  
T Cell Subsets ◽  
Hematopoietic Stem ◽  
Cd34 Cells ◽  
Significant Difference ◽  
Stromal Cell Derived Factor ◽  
And Control ◽  
Cellular Components

Abstract Abstract 4808 Background: It is now evident that hematopoietic stem cells (HSCs) reside preferentially at the endosteal region within the bone marrow (BM) where bone-lining osteoblasts are a key cellular component of the HSC niche that directly regulates HSC fate. We investigated the microenvironmental differences including osteoblastic activities and HSC components in myeloproliferative (chronic myeloid leukemia, CML) and hypogenerative disease (aplastic anemia, AA) as well as normal control (NC). Methods: The immunohistochemistry for osteonectin, osteocalcin, stromal cell derived factor (SDF, CXCL12), T cell, T helper/inducer cell, T suppressor/cytotoxic cell, hematopoietic stem/progenitor (CD34, CD117) and megakaryocytes was performed on BM biopsy specimens from 10 AA patients, 10 CML patients and 10 NC (lymphoma without BM involvement). The positive cells for immunohistochemical stainings except osteocalcin on each slide were calculated on 10 high power fields (HPF, ×400), and then corrected by the cellularity. The positive cells for osteocalcin were counted on the peritrabecular line on each slide, and then corrected by the mean length measured. Results: The CD34+ cells (p=0.012) and megakaryocytes (p<0.0001) were significantly lower in AA than in NC, but CD117+ cells was comparable in AA, CML, and control samples. The osteonectin+ cells (p=0.0003) were lower in CML than in AA and NC, however the osteocalcin+ cells showed wide variation (0-903/2035um) and no significant difference. The SDF+ cells (p<0.0001) was significantly higher in AA and very lower in CML, compared with NC. The counts for T cell and T cell subsets were significantly lower in CML than in NC, and higher in AA than in NC (p<0.0001). Conclusions: Cellular components of BM microenvironment in 2 hematologic diseases representative of myeloproliferation (CML) and hyporegeneration (AA) respectively are quite different. Further studies would be required to explore the role of these components for hematopoiesis and the rationale for therapeutic application. Disclosures: No relevant conflicts of interest to declare.

Donor Antigen Presenting Cell CD80/86 Function Is Required for Eliciting T Cell Resistance Against Progenitor Cell Engraftment in an MHC Matched Allogeneic Non-Myeloablative Transplantation Model.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2137-2137
Author(s):  
Zachary F. Zimmerman ◽  
Robert B. Levy
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Allogeneic Bone Marrow Transplantation ◽  
Allogeneic Bone Marrow ◽  
High Dose ◽  
Allogeneic Bone ◽  
Wild Type ◽  
Marrow Graft ◽  
Post Transplant ◽  
Cell Engraftment

Abstract Immune mediated resistance is a concern in allogeneic bone marrow transplantation (BMT) particularly in the setting of nonmyeloablative conditioning regimens or in situations where prior sensitization to hematopoietic alloantigens has occurred (ie aplastic anemia). This resistance can lead to both delayed reconstitution and rejection. In this investigation, two models of T cell mediated allogeneic bone marrow graft rejection were employed: 1) an MHC disparate, donor antigen sensitized model, ie BALB/c (H-2d) recipients sensitized to B6 (H-2b) hematopoietic antigens transplanted with a high dose (1x107) of B6 BM-TCD following ablative (9.0Gy TBI) conditioning. Resistance in this model was assessed by the presence of donor CFU progenitors early (Day 5) post transplant and 2) a non-myeloablative mHag disparate model, ie C57BL/6 TCD BM à C3H.SW recipients conditioned with 5.5 Gy TBI. In this latter model, BM from B6 wild-type donors at doses <1x107 cells leads to transient donor peripheral chimerism (assessed by monitoring Ly9.1 expression) peaking at day 7–10 post transplant. To study the role of donor APC function in the transplant innoculum in both models, bone marrow was utilized from donors deficient in CD80 and CD86, yielding a “crippled” APC population incapable of co-stimulation through these two primary pathways. In the antigen sensitized model, (ie Tmemory mediated) both lineage committed (CFU-IL3) and multipotential (CFU-HPP) progenitor activity was absent from recipients of both wild-type or CD80/86 −/− BMC. This data suggests that CD80/86 mediated co-stimulation from direct recognition of donor APC is dispensible for eliciting resistance when memory T cells are involved. However, in the unsensitized model, despite the potential for indirect donor antigen presentation, recipients of CD80/86−/− bone marrow exhibited significant engraftment by one month post-transplant (in contrast, recipients of wt bone marrow had universally these rejected grafts by this time point). Significant chimerism in recipients of CD80/86−/− BM was detectable in the B as well as T lymphocyte compartments (>50%). Consistent with the importance of the presence of these co-stimulatory pathways on the donor APC population, 5/7 recipients of mixed (1:1) BM (CD80/86−/− + B6-wt) transplants rejected their grafts. These findings suggest that removal of CD80/86 dependent co-stimulatory pathways in the donor APC population may be insufficient for promoting engraftment in cases where there has been prior donor antigen sensitization, i.e. in the presence of host Tmemory populations. However, absence of CD80/86 co-stimulatory pathways in the donor APC population may be sufficient for overcoming resistance in an MHC matched allo-transplant setting in which prior donor antigen sensitization has not occurred, ie in the presence of host Tnaive populations. These findings have implications in the development of improved non-myeloablative conditioning protocols to facilitate engraftment without global immunosuppressive conditioning.

A Role for Regulatory Cells in Tolerance Induction in Recipients of Haploidentical Combined Non-Myeloablative Bone Marrow and Kidney Transplantation?.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3255-3255
Author(s):  
Giovanna Andreola ◽  
Meredith Chittenden ◽  
Juanita Shaffer ◽  
A. Benedict Cosimi ◽  
Tatsuo Kawai ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Conditioning Regimen ◽  
Tolerance Induction ◽  
In Vitro Assays ◽  
Mixed Chimerism ◽  
Post Transplant ◽  
Hla Dr

Abstract Following an in vivo T cell depleting non-myeloablative conditioning regimen, 5 patients, aged 22–49, received combined kidney and bone marrow transplantation from a haploidentical related donor. Rituximab was included in the conditioning for patients 4 and 5. All patients developed initial mixed chimerism but lost it by day 21; no patient developed GVHD. Four patients discontinued immunosuppression from 240 to 422 days after BMT and have remained off immunosuppression for 9 to 52 months with no evidence of allograft rejection. Flow cytometry was used to assess lymphocyte subsets recovering after transplant. CD3 counts recovered slowly, exceeding 500 cells/μl at days +271, +365, +640 and +450. While memory CD45RO+ cells were most prevalent among CD4+ cells, naïve-type CD4+CD45RA+ cells, presumably arising from the recipient thymus, ranged from 8% to 56% at the time when total CD4 counts recovered to >100 cells/μl (days +165, +21, +352, +240). Notably, a very high proportion of initially recovering T cells were CD3+CD4+ expressing CD25 in all patients as early as day 7 and persisted over 1 year in 2 patients. At approximately day +120 and +365, we further characterized these cells for CD127, FOXP3, CD45RO, CD45RA, HLA-DR and CD62L expression. At Day +120, all 4 patients showed increased frequencies (10.7±4.6%) of CD25+CD127-FOXP3+ regulatory T cells (Treg) within the CD4 population compared to healthy subjects (3.8±0.4%). Expression of CD45RO, CD45RA, CD62L and HLA-DR was variable. By 1 year post-transplant, frequencies of Treg had decreased to levels similar to those in normal subjects. In vitro assays for CD8 and CD4 T cell-mediated alloreactivity (CML/MLR) showed development of long-lasting donor-specific unresponsiveness by 3 months after transplant in Patients 2, 4 and 5, and by 9 months in Patient 1. Responses to 3rd party recovered in all patients after a period of unresponsiveness. In Patient 1, in whom anti-donor CML reactivity declined gradually to become unresponsive by 9 months, depletion of CD4+CD25+ cells revealed a residual anti-donor CML and MLR response at 1year but not at 18 months. In 2 other patients, depletion of CD4+CD25+ cells did not reveal an anti-donor response at time points analyzed from day +122 to 2 years. In patients in whom renal tubular epithelial cells (RTEC) were cultured from the donor kidney, loss of killing activity against donor RTEC was observed post-transplant. The high percentage of Treg recovering early after transplant suggests that they may play a role in initial tolerance induction. This regulatory mechanism may be followed by later deletion of donor-reactive T cells. The variable ability to detect regulation of anti-donor reactivity may reflect the strength of the initial response, as patients with weak pre-transplant anti-donor responses and rapid post-transplant development of donor unresponsiveness did not reveal anti-donor response when Treg were depleted. In addition, infiltration of Treg at the graft site, not revealed by the assays described, might be responsible for tolerance in these patients.

Improved T Cell Recovery After Transplantation of CD3/CD19 depleted Haploidentical Stem Cell Grafts in Pediatric Patients.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4652-4652
Author(s):  
Heiko-Manuel Teltschik ◽  
Tobias Feuchtinger ◽  
Matthias Pfeiffer ◽  
Michael Schumm ◽  
Ingo Mueller ◽  
...  
Keyword(s):  
T Cell ◽  
B Cells ◽  
Conflicts Of Interest ◽  
Negative Impact ◽  
Conditioning Regimen ◽  
T Cell Subsets ◽  
Control Group ◽  
T And B Cells ◽  
Haploidentical Transplantation ◽  
Significant Difference

Abstract Abstract 4652 Profound depletion of T and B cells is a fundamental prerequisite for haploidentical transplantation and allows to minimize GvHD despite HLA incompatibility. However, posttransplant recovery of donor derived T cells is delayed after various graft manipulation procedures and may result in severe infections. Methods to improve this recovery are of great importance. Here we present immune reconstitution data in patients who received CD3/CD19 depleted stem cells in combination with melphalan based or standard conditioning regimens. 32 patients with ALL (n=14), AML/MDS (n=17), CML (n=1) were included. T and B cells were directly depleted using antiCD3/antiCD19 coated magnetic microbeads and the CliniMACSTM device. The patients received either TBI or Bu i.v. and OKT3 (n=9) or a reduced intensity conditioning (“RIC”: Mel 140mg/m2, Flud 160mg/m2, TT 10mg/kg, OKT3, n= 23). Absolute numbers of lymphocyte subsets per microliter on day 90 were compared within these both groups and with a historical control group (patients with leukemias who received CD34 selected grafts and TBI or Bu based standard conditioning regimen in combination with ATG, n=28). CD3+4+, CD3+8+ and total numbers of CD3+ of patients after CD3/CD19 depletion were significantly higher in the RIC-group than in the TBI/Bu-group (mean numbers: 85.83 vs. 38.84; 133.46 vs. 19.69; 270.27 vs. 63.99; p<0.05, unpaired t-test). Comparison with the whole CD34 historical group showed a faster recovery of CD3+4+ in patients with CD3/19 depletion and RIC (104.26 vs. 54.22; p= 0,034) but no significant difference in CD3+8+ and CD3+. Furthermore, subgroups of the CD34 historical population were compared: patients with CD3/19 depletion and RIC had a significantly faster recovery of CD3+4+, CD3+8+ and CD3+ than CD34 patients with TBI (104.26vs. 25.48; 133.46vs. 43.17; 270.27 vs. 65.86; p<0.05) but had no advantage over CD34 patients with non-TBI conditioning. Conclusions: the type of graft manipulation appeared to have an influence on the speed of CD4+ recovery (CD3/19 depletion > CD34 selection). Moreover, the use of TBI had a clear negative impact on all T cell subsets: patients with TBI had a slower recovery than patients with non-TBI conditioning, independent from graft manipulation procedures and probably due to thymic damage. Thus, the use of RIC-protocols in combination with CD3/CD19 depletion may help to speed up the immune recovery after haploidentical transplantation. Further studies are warranted to evaluate the risk of relapse. Disclosures: No relevant conflicts of interest to declare.

Function and Clonality of T-Cells In Myelodysplasia

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4972-4972
Author(s):  
Glenda Davison ◽  
Nicolas Novitzky ◽  
Rygana Abdulla
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Conflicts Of Interest ◽  
Autologous Blood ◽  
Disease Process ◽  
Cell Populations ◽  
Cell Repertoire ◽  
Clonogenic Growth ◽  
Cd34 Cells

Abstract Abstract 4972 Objective: Myelodysplastic (MDS) syndromes are clonal disorders arising from haematopoietic multi-potential stem cells. Research has implied that cells of the immune system play a role in the disease process. The aim of this study was to examine the clonality, response to mitogens, effects of lymphocytes on bone marrow clonogenic progenitors and proliferative capacity of selected marrow progenitors in a well defined cohort of MDS patients. Methodology: Seventeen patients with MDS were accrued. The clonality of the T-cells was studied using the TCR Vβ repertoire kit. In order to assess in vitro function, the lymphocytes were activated using phorbol 12-myristate13 acetate (PMA) and thereafter analysed for expression of the activation antigen CD69 using standard flow cytometry. The results were compared to age matched controls. The effect of the autologous blood mononuclear cell populations on clonogenic growth was studied by culturing Li- selected CD34+ cells with both PMA activated and non-activated autologous lymphocytes using a non-contact double layer culture technique. Results: T-cells did not demonstrate monoclonality, although skewing of the T-cell repertoire was observed. The median percent of CD3+ T-cells expressing CD69 after activation was 30% (9.5 – 95.4%). This was reduced in comparison to age matched controls (p=0.025). The effect of the lymphocytes on clonogenic cell growth was heterogeneous however median values demonstrated a cell dose response in the colony numbers directly related to cultured lymphocyte numbers (p=0.04 compared to no lymphocytes added). In addition, the median CFU-GM scores were higher when cultured with PMA activated lymphocytes (p=0.05). This pattern was not significantly different to normal controls. Background: Our investigations suggest that in this cohort of MDS patients the T-cells were not clonal but had reduced activation capacity when stimulated with the mitogen PMA. However, their ability to stimulate colony growth of autologous bone marrow CD34+ cells was preserved. Furthermore, selected CD34+ MDS cells proliferated well, with clone numbers not different from control. These observations imply that the previously reported abnormal T-cell responses and poor clonogenic growth may be the result of complex interactions between T-cells, the malignant clone and accessory cells in the bone marrow stroma. Further study examining each of these cell populations is required to better understand the mechanism of marrow failure and progression to leukaemia. Disclosures: No relevant conflicts of interest to declare.

Rapid Immune Reconstitution Following Unmanipulated Haploidentical BMT with Post-Transplant High Dose Cyclophosphamide

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3050-3050 ◽  
Author(s):  
Alida Dominietto ◽  
Anna Maria Raiola ◽  
Barbara Bruno ◽  
Maria Teresa van Lint ◽  
Francesco Frassoni ◽  
...  
Keyword(s):  
Immune Reconstitution ◽  
Conflicts Of Interest ◽  
Conditioning Regimen ◽  
High Dose ◽  
Hematopoietic Stem ◽  
Cd4 Counts ◽  
Post Transplant ◽  
Gvhd Prophylaxis ◽  
Donor Type ◽  
Unrelated Cord Blood

Abstract Abstract 3050 Background. Allogeneic hematopoietic stem cell transplantation (HSCT) is the treatment of choice for the majority of hematological malignancies. Early and successful immunologic reconstitution after HSCT reduces morbidity and mortality due to infection complications and improves survival. Aim of the study. We analyzed immune recovery after HSCT in 444 patients according to donor source. Patients and Methods. From January 2005 to June 2011 176 patients were grafted from HLA identical siblings (MSD), 125 from alternative donors (1 antigen mismatched family or unrelated donors) (ALT), 103 from unrelated cord blood grafted intra bone (CBIB) and 40 from haplo-identical mismatched family donors (HAPLO). All patients received unmanipulated bone marrow: 283 after a myeloablative (MA) conditioning regimen (CY-TBI or BU-CY) and 161 after a fludarabine based reduced intensity regimen (RIC). Graft versus host disease (GvHD) prophylaxis was cyclosporin methotrexate (CyA+MTX) for all patients except for CBIB (CyA and mycophenolate, MMF) and for HAPLO transplants which consisted of CyA+MMF and post-transplant high dose cyclophosphamide (HDCY) according to the Baltimore protocol (Lutznik et al BBMT 2008). Anti-thymocyte globulin (ATG) was used only for ALT transplants. Results. We compared immune reconstitution in MA and RIC transplants according to donor type at different time points post BMT. CD3+ absolute median counts/μl in MA conditioning on day+30, +90, +180 were respectively in MSD 477, 565, 700; in ALT donors were 146, 404, 470; in CBIB were 30, 57, 196; for HAPLO transplants they were 195, 182, 499. CD3+ absolute median counts/μl in RIC conditioning on day+30, +90, +180 were respectively in MSD 301, 660, 700; in ALT donors were 506, 186, 721; in CBIB 234, 399, 522; in HAPLO were 178, 276, 1300. CD4+ absolute median counts/μl in MA conditioning on day+30, +90, +180 were respectively in MSD 166, 170, 198; in ALT donors were 36, 86, 111; in CBIB 7, 36, 106; for HAPLO transplants they were 45, 127, 211. CD4+ absolute median counts/μl in RIC conditioning on day+30, +90, +180 were respectively in MSD 89, 189, 274; in ALT donors were 131, 210, 220; in CBIB 52, 110, 130; for HAPLO transplants they were 41, 205, 385. CD8+ absolute median counts/μl in MA conditioning on day+30, +90, +180 were respectively in MSD 280, 389, 500; in ALT donors were 102, 278, 413; in CBIB 42, 16, 51; in HAPLO transplants were 73, 424, 408. CD8+ absolute median counts/μl in RIC conditioning on day+30, +90, +180 were respectively in MSD 196, 432, 300; in ALT donors were 366, 65, 494; in CBIB 71, 167, 199; for HAPLO transplants they were 137, 129, 900. CD3, CD8, and CD4 counts in HAPLO transplants were not statistically different from MSD with the only exception of day +30, both for MA and RIC conditioning. Platelet median counts/μl on day+30, +90, +180 in MA conditioning were in MSD 142, 129, 180, in ALT 75, 101, 147, in CBIB were 19, 77, 128 and for HAPLO transplants were 67, 126, 128; in RIC conditioning platelets counts were in MSD 137, 156, 168, in ALT 33, 134, 142, in HAPLO were 77, 95, 188. Acute GvHD II-IV developed in 29% (MSD) 38% (ALT) 16% (CBIB) and 12% (HAPLO) (p=0.004) in MA conditioning and 40% (MSD) 18% (ALT) 25% (CBIB) and 10% (HAPLO) (p=0.07). Overall Cumulative Incidence of Non-Relapse Mortality (CI-NRM) was respectively 18% (MSD), 35% (ALT), 34% (CBIB), 22% (HAPLO) (p=0.02) in MA conditioning (p=0.02) and was 30% (MSD), 33% (ALT), 45% (CBIB), 0% (HAPLO) (p=0.02) in RIC conditioning (p=0.02). Day+100 CI-NRM was respectively 10% (MSD), 21% (ALT), 19% (CBIB), 12% (HAPLO) in MA conditioning (p=0.01) and 11% (MSD), 19% (ALT), 26% (CBIB), 0% (HAPLO) in RIC conditioning (p=0.02). Death due to infections were respectively 6% (MSD), 26% (ALT), 30% (IBCB), 17% (HAPLO) in MA conditioning and for RIC were 15 (MSD), 36% (ALT), 32% (IBCB), 0% (HAPLO). Conclusions. HAPLO transplant with HDCY post transplant as proposed by the Baltimore group, is associated with (1) rapid immunologic (CD3, CD4, CD8) recovery (2) low infectious death rate, (3) low overall and Day+100 CI-NRM, (4) rapid hematologic recovery. These results are comparable with those achieved with MSD and warrant further studies with HDCY post transplant as a GvHD prophylaxis. Figure: absolute CD4+ counts/μl on day+30, +90, +180, according to donor type in MA conditioning regimen. Disclosures: No relevant conflicts of interest to declare.

211At-Anti-CD45 Radioimmunotherapy Can Replace TBI Prior to Haploidentical Bone Marrow Transplantation and Yield Long-Term Hematopoietic Engraftment

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2417-2417
Author(s):  
Johnnie J. Orozco ◽  
Aimee L. Kenoyer ◽  
Ethan R. Balkin ◽  
Donald K. Hamlin ◽  
D Scott Wilbur ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Transplantation ◽  
Marrow Transplantation ◽  
Conflicts Of Interest ◽  
Conditioning Regimen ◽  
Decay Energy ◽  
Cd8 Cells ◽  
Post Transplant ◽  
Alpha Emitter ◽  
Murine Leukemia

Abstract Haploidentical bone marrow transplantation (haplo-BMT) is both clinically effective and widely available because related-donors can be identified for nearly all recipients. Despite the curative promise of this approach, many patients with hematologic malignancies will relapse after haplo-BMT and more effective preparative regimens are necessary. We have shown that anti-CD45 radioimmunotherapy (RIT) delivers high-doses of radiation to hematolymphoid organs while minimizing the radiation exposure to non-targeted tissues. The efficacy of beta-emitting radionuclides may be limited by their relatively low decay energies (0.66 – 2.3 MeV). We have thus investigated the higher energy alpha-emitter astatine-211 (211At) (average decay energy of 6.8 MeV), for targeted anti-CD45 radioimmunotherapy (RIT) in lieu of total body irradiation (TBI) prior to haploidentical BMT in a murine leukemia model to decrease relapse rates. Groups of five B6SJLF1/J mice (allotype H2-Db) received escalated activities (20, 30 or 40 μCi) of 211At-anti-CD45 antibody [100 μg (0.67 nmol) of B10-30F11] given by tail vein injections on day -2 in place of TBI prior to BMT. Animals received cyclophosphamide (CY; 200 mg/kg/day) on days –3, –2, or –1, and +2 for graft-versus-host disease prophylaxis, either alone, or with fludarabine (FLU; 100 mg/kg/day) for 4 days starting day -6. Transplanted mice received 1.5 × 107 haploidentical bone marrow cells from CB6F1/J mice (allotype H2-Dd) on day 0. Peripheral blood from recipient mice was then assayed monthly by flow cytometry to measure chimerism as the percentage of donor (H2-Dd) circulating CD8+ cells. The highest activity delivered of 40 µCi 211At-anti-CD45 RIT was uniformly lethal without BMT rescue, whereas 60% of transplanted mice at this dose survived to assessment at 1 month. Mice treated with 30 µCi of 211At-anti-CD45 RIT with pre- and post-transplant CY and without TBI or FLU, had high levels of engraftment with an average of 83.7 ± 5.8% donor CD8+ cells 1 month after haploidentical BMT (Table 1). The addition of FLU to 211At-anti-CD45 RIT with CY did not significantly improve chimerism levels, with mean donor CD8+ cells in mice treated with 40 µCi 211At-anti-CD45 RIT of 64.5 ± 41.6% compared to 60.0 ± 13.9% in the absence of FLU (p=0.8668). In addition, mice that received 30 µCi 211At-anti-CD45 RIT and pre-transplant CY on either day –3, –2, or –1 showed mean donor CD8+ cells of 83.7 ± 5.8%, 49.9 ± 29.6% and 55.0 ± 46.2% 1 month after haploidentical-BMT, respectively. Importantly, chimerism levels remained stable 2 months after haploidentical BMT with mean donor CD8+ cells of 80.4 ± 16.6%, 47.0 ± 37.7% and 63.2 ± 10.7% in mice treated with 30 µCi 211At-anti-CD45 RIT and pre-transplant CY on day –3, –2, and –1, respectively. Engraftment using 40 or 30 µCi 211At-anti-CD45 RIT was comparable to using 850 or 1000 cGy TBI (mean donor CD8+ cells of 70.2 ± 18.8% and 60.0 ± 4.6%, respectively) prior to haploidentical BMT. RIT alone without any chemotherapy was insufficient to facilitate clinically relevant rates of donor engraftment, as mice treated with 30 µCi 211At-anti-CD45 RIT and no FLU, CY or TBI had 15.9 ± 7.1% mean donor CD8+ cells 1 month after haploidentical BMT. These results suggest that 211At-anti-CD45 RIT prior to haploidentical BMT with pre– and post–transplant CY can result in high levels of donor hematopoietic cell engraftment in the absence of TBI and FLU. This conditioning regimen may be less toxic and more effective at preventing relapse than TBI-based approaches due to the high linear energy transfer of the alpha emissions, or the high decay energy of targeted 211At deposited over its short effective path-length. On-going studies are assessing the efficacy and toxicity associated with 211At-anti-CD45 RIT compared to a TBI-based haploidentical BMT using a syngeneic murine leukemia model. Abstract 2417. Table 1 Preparative Therapy and CD8+ Donor Chimerism at 1 month Group FLU (100mg/kg/d) pre-BMT CY (200mg/kg) TBI 211 At-anti-CD45 RIT post-BMT CY (200mg/kg) Donor CD8+ % 1 – day –3 – 30 µCi day +2 83.7 ± 5.8 2 – day –2 – 30 µCi day +2 49.9 ± 29.6 3 – day –1 – 30 µCi day +2 55.0 ± 46.2 4 – day –3 – 40 µCi day +2 64.5 ± 41.6 5 d –6 to –3 day –3 – 40 µCi day +2 60.0 ± 13.9 6 – – – 30 µCi day +2 15.9 ± 7.1 7 – day –3 – – day +2 4.4 ± 0.6 8 – – 1000 cGy – day +2 60.0 ± 4.6 Disclosures No relevant conflicts of interest to declare.

T Cells Motility in the Colonic Gvhd Is Influenced By Both Cognate Interaction and Microenvironment

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 3346-3346
Author(s):  
Jieqing Zhu ◽  
Cassandra Secunda ◽  
Michael Parrish ◽  
David Gonzalez ◽  
Ann Haberman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Cell Motility ◽  
Conflicts Of Interest ◽  
Allogeneic Bone Marrow Transplantation ◽  
Allogeneic Bone ◽  
Cd4 Cells ◽  
Cd8 Cells ◽  
Cell Stability

Abstract Graft-versus-host disease (GVHD) accounts for substantial morbidity and mortality after allogeneic bone marrow transplantation. Allograft T cells are stimulated by both host and donor-derived antigen presenting cells (APCs). After differentiating into T effector cells, donor T cells migrate to GVHD target organs where they mediate damage directly and indirectly. However, very little is known about the dynamics of T cells in GVHD target tissues and how their behaviors are affected by the local environments, including by tissue-resident APCs. We utilized two-photon intravital microscopy (2PIM) to analyze T cell locations, motility and interactions with APCs. 129S1/SVLmJ (129) (H-2b) hosts were irradiated and reconstituted with C57Bl/6 (B6) (H-2b) CD11-YFP transgenic mice bone marrow together with B6 dsRed (RFP) CD8 cells and cyan fluorescent protein (CFP) CD4 cells. Mice were imaged from days 25-42 post-transplant. We focused on the colon which is strongly targeted in this model and in humans and which previously has been difficult to image by 2PIM. In syngeneic B6 into B6 recipients, donor CD11c+, CD8+ and CD4+ cells populate the colonic mucosa. However, a greater number of cells accumulated in the allogenic as compared to syngeneic recipients. Based on the analysis of 7-8 mice per group, T cells in allogenic mice were less motile. The mean CD8 speeds in syngeneic and allogeneic mice were 3.38 ± 0.07 µm/min and 2.21 ± 0.03 µm/min, respectively. Mean CD4 speeds in syngeneic and allogeneic mice were 3.11 ± 0.08 µm/min and 2.38 ± 0.04 µm/min, respectively. The majority of T cells were stationary, with few entering or leaving the imaged volume. Perfusion was confirmed by i.v. rhodamine-dextran; therefore, stationary behaviors was not due to tissue viability. Using optical clearing we imaged whole-mount colonic tissue from mucosa to serosa. T cells were aggregated in the sub-mucosa and infiltrated crypts and surprisingly also the muscular layer. Ki67+ T cells were found throughout, especially in the submucosa. Given the 2PIM data which show few new T cells entering the imaged volumes, these cells were likely stimulated to divide locally. Preliminary data with EdU pulsing suggest this to be the case. To determine whether T cell stability is microenvironment- or antigen-driven, we injected OT-1 TCR transgenic effectors which do not recognize any antigen in the recipient and imaged the next day. Most colonic OT-1 cells showed the same stationary behavior as nearby donor-derived CD4 and CD8 cells, suggesting that factors in the GVHD colon microenvironment drive T cell stability. Nonetheless there was also an antigen-driven component as injection of an anti-MHCII antibody (but not isotype control Ab) increased CD4+ T cell motility presumably by disrupting TCR:MHCII interaction. These results show the motility of T cells in the GVHD colon is influence by both TCR:MHC cognate interactions and by the microenvironment. That T cells are dividing and may be activated in situ, suggesting GVHD may be maintained locally. Current studies are focusing on what antigen-independent factors affect T cell motility and on defining the roles played by tissue APCs. Disclosures No relevant conflicts of interest to declare.

scholarly journals EBV-Related Post-Transplant Lymphoproliferative Disease in Allogeneic Hematopoietic Cell Transplantation: Single-Center Experience and Considerations Regarding CNS Involvement

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5706-5706 ◽  
Author(s):  
Anna Vardi ◽  
Ioanna Sakellari ◽  
Andriana Lazaridou ◽  
Apostolia Papalexandri ◽  
Ioannis Batsis ◽  
...  
Keyword(s):  
T Cell ◽  
Research Funding ◽  
Conditioning Regimen ◽  
Lymphoproliferative Disease ◽  
Additional Patient ◽  
Cns Lymphoma ◽  
High Dose ◽  
Cns Involvement ◽  
Post Transplant ◽  
Cell Immunity

Abstract EBV-related post-transplant lymphoproliferative disease (EBV-PTLD) is a serious complication of allo-HCT, for which antithymocyte globulin (ATG) administration has been identified as the most important predisposing factor. Although the development of EBV-PTLD used to be detrimental to patients, use of rituximab has largely changed both the incidence and the outcome of this complication, either administered as preemptive treatment based on peripheral blood (PB) EBV titles, or for the management of the disease itself. We determined the incidence of EBV-PTLD through chart review of 797 consecutive allo-HCT recipients transplanted in our center (7/1990-7/2018) and evaluated factors potentially influencing EBV-PTLD occurrence and outcome. Among 797 allo-HCT recipients (n=465 sibling, n=277 MUD, n=47 haploidentical, n=2 twin and n=6 cord blood), 14 (1.7%) patients developed EBV-PTLD. The diagnosis was confirmed by biopsy in 9/12 cases (7 monomorphic, 1 polymorphic and 1 HL-like PTLD), 1 case developed sole EBV encephalitis, 1 case developed CNS lymphoma, and the remaining 3 cases were clinically diagnosed on the basis of generalized lymphadenopathy coupled with high PB EBV title. The patients suffered from ALL (n=6), AML (n=2), CML (n=2), severe aplastic anemia (n=2), MDS (n=1) and plasmacytic leukemia (n=1). Notably, 6/14 patients had high tumor burden at the time of transplantation [refractory AML (n=1), relapsed refractory ALL (n=2), CML blastic crisis (n=2), refractory MDS RAEB II (n=1)]. EBV-PTLD incidence was significantly higher in MUD versus sibling allo-HCT (3.4% versus 0.6%, p=0.006), in haploidentical versus sibling allo-HCT (6.4% versus 0.6%, p<0.001), and in haploidentical versus conventional allo-HCT (6.4% versus 1.5%, p=0.01). Among the haploidentical allo-HCTs complicated with EBV-PTLD, 2 were T-cell depleted with add-back infusions of transduced donor lymphocytes, and 1 was T-cell replete. These results possibly reflect the standard use of ATG as acute GVHD prophylaxis in MUD and haploidentical allo-HCT. Indeed, 12/14 (85.7%) patients who developed EBV-PTLD had received ATG as part of their conditioning regimen, at doses ranging from 5-10mg/kg (median 5mg/kg), and 1 additional patient had received ATG 20mg/kg as 2nd-line treatment for hyperacute GVHD. The conditioning intensity was not significantly associated with EBV-PTLD development [classic myeloablative (n=8), reduced-toxicity (n=3), reduced-intensity (n=3)]. EBV-PTLD occurred early in the post-transplant period (median: 73, range 41-603 days). In 7/14 cases, it was preceded by the onset of aGVHD (median interval from aGVHD diagnosis: 58 days), in one case it coincided with aGVHD diagnosis +45 days from allo-HCT, and in one case it developed soon (+55 days) after induction of GVHD through DLIs. At EBV-PTLD diagnosis, the median EBV title in PB was 78,400 copies/ml (range 84-2,860,000). Of note, one patient developed sole CNS EBV-PTLD with extremely low PB EBV viral load. This patient had received preemptive rituximab 500mg 7 months prior to CNS EBV-PTLD development, on the basis of elevated PB EBV title (70,900 copies/ml). Since then, PB EBV title was always tested below <500 copies/ml by conventional PCR, and was practically negative at the time of EBV-PTLD diagnosis (84 copies/ml). Ten patients were treated with intravenous rituximab: in six patients the disease resolved, but 3/4 cases with CNS involvement succumbed. The fourth patient with CNS involvement is currently under treatment with high-dose MTX and intravenous rituximab and has achieved partial remission. The remaining four patients were unsuccessfully treated with combinations of bleomycin/vindesine/IFNa/immunoglobulins/DLIs. In conclusion, EBV-PTLD is an early complication of alloHCT, associated mainly with defects in T cell immunity. Selective ATG administration to MUD and haploidentical allo-HCT may explain the higher incidence of EBV-PTLD as compared to sibling allo-HCT, yet we identified higher incidence among haploidentical alloHCTs that cannot be attributed to a particular lymphodepletion scheme. Prompt administration of intravenous rituximab is very effective, except for CNS involvement. Importantly, PB EBV monitoring may be misleading in CNS EBV-PTLD, particularly following preemptive intravenous rituximab administration. Taking into account the dismal outcome, intrathecal rituximab should be considered. Disclosures Vardi: Gilead: Research Funding; Janssen: Honoraria. Gavriilaki:European Hematology Association: Research Funding.

Natalizumab Increases Circulating CD34+ Cells with An Impaired Functional Potential In Patients with Multiple Sclerosis without Affecting the Bone Marrow.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2620-2620
Author(s):  
Christian Saure ◽  
Fabian Zohren ◽  
Thomas Schroeder ◽  
Ingmar Bruns ◽  
Ron Patrick Cadeddu ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Bone Marrow ◽  
Conflicts Of Interest ◽  
T Cell Subsets ◽  
Cell Counts ◽  
Healthy Donors ◽  
Cd34 Cells ◽  
Long Term Treatment ◽  
Significant Difference

Abstract Abstract 2620 Introduction: In our previous report (Zohren et al., Blood 2008) we could show that the blockade of the heterodimer VLA-4 by the monoclonal IgG4 antibody natalizumab leads to a significant increase in circulating CD34+ cells in patients with multiple sclerosis (MS). We now extend our analysis on the influence of natalizumab on CD34+ cells comparing bone marrow (BM) and peripheral blood (PB) derived CD34+ cells of natalizumab patients with those from healthy donors. Methods: A total of 83 patients with MS receiving natalizumab were included. In vitro adhesion, migration and apoptosis assays as well as LTC-IC of immunomagnetically enriched CD34+ cells were conducted. Flow cytometric analyses were performed to assess phenotype and composition of the CD34+ subsets. Results: The median concentration of circulating CD34+ cells was significantly greater compared to normal donors (7.7/μL vs. 1.8/μ L; p= 0.0001) and remained relatively stable during a one year treatment with natalizumab. Leukocyte cell counts, the number of T cell subsets as well as the number of CD19+ B cells and CD56+ natural killer cells were in normal range in PB and BM after short- and long-term treatment with natalizumab. However, we found significantly reduced adhesion and migration abilities of circulating CD34+ cells under natalizumab treatment in comparison to G-CSF mobilized CD34+ cells of healthy donors. Moreover, the self-renewal capacity of these cells was poor. In contrast, no significant difference was seen between the BM of natalizumab patients and the BM of healthy donors with regard to cellularity and proportion of CD34+ cells. In addition, neither co-expression of CD49d nor the adhesion ability of the BM derived CD34+ cells revealed a significant difference between the two collective. Conclusions: Our data indicate that natalizumab mediates an increase in circulating CD34+ cells by impaired homing. These findings argue against the use of natalizumab-exposed PB CD34+ cells for transplantation. Disclosures: No relevant conflicts of interest to declare.

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