Rapid Expansion of Naive CD4+ Cord Blood Lymphocytes Restores Adaptive Immunity within 2 Months After Unrelated Cord Blood Transplantation

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2337-2337
Author(s):  
Robert Chiesa ◽  
Prashant Hiwarkar ◽  
Kanchan Rao ◽  
Waseem Qasim ◽  
Kimberly Gilmour ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Respiratory Tract ◽  
Cord Blood ◽  
Cd4 T Cells ◽  
Viral Infections ◽  
Cord Blood Transplantation ◽  
Cd4 T Cell ◽  
Blood Transplantation ◽  
Unrelated Cord Blood

Abstract Abstract 2337 Viral infections remain a significant problem following haematopoietic cell transplantation (HCT). Recovery of CD4+ T-cells only occurs with return of thymopoiesis 6–12 months after HCT. In an attempt to improve immune reconstitution and reduce viral infections following unrelated cord blood transplantation (CBT), serotherapy was omitted from conditioning. 26 children, median age 19 months (range 1–147) underwent unrelated CBT for immunodeficiency (13) or high risk/relapsed leukemia (13). Pre-transplant co-morbidities included lung aspergillosis (2), adenoviraemia (1), Paraflu3 respiratory tract infection (3). Conditioning was myeloablative in 25 patients. HLA matching was 6/6, 5/6 and 4/6 in 13, 14 and 3 cases respectively (4 double cords). GvHD prophylaxis was mostly cyclosporine/mycophenolate mofetil (24). The median number of nucleated, CD34+ and CD3+ cells infused was 8.12 × 107/kg (range, 4.7–17.72), 3.5 × 105/kg (range, 1.23–11.95) and 5.63 × 106/kg (range, 1.9–24.45). Median time to myeloid and platelet engraftment was 22 (range, 13–38) and 39 (range, 21–110) days. One patient died from conditioning related toxicity and 3 patients died after leukaemia relapse. 22 patients are alive and in remission with a median follow up of 12 months (range 3–66). 18 patients have full donor engraftment while 4 patients are mixed chimeras. Acute GvHD > grade I occurred in 16 patients (grade II n=11; grade III-IV n=5) but in general this responded to steroids and only 1 child developed cGvHD. There was no infection-related mortality, despite early adenoviraemia (4), CMV viraemia (4), HHV6 encephalitis (1) and RSV/Paraflu3 respiratory tract infections (3). The most striking finding was a rapid and sustained CD4+ T-cell expansion with conversion from naive to memory/effector phenotype. The median CD3+, CD4+ and CD8+ counts at 2 months post CBT was 0.85 ×109cells/L (range, 0.44–2.44), 0.55 ×109cells/L (range, 0.06–1.89), and 0.21 ×109cells/L (range, 0.01–1.21) respectively. As a comparison, the median CD4+ T-cell count at 2 months post a HCT from a matched sibling donor was 0.24 ×109cells/L in 23 children evaluated in our institution. In 3 patients who developed adenoviral/CMV reactivation virus-specific T cells were detected by ELISPOT/IFN-g assays as early as 45 days post transplant, and viral infections resolved rapidly. Unrelated CBT without serotherapy results in rapid “peripheral” expansion of naive/memory CD4+ T-cells, and restores cell-mediated immunity in children faster than any other HCT donor source. Disclosures: No relevant conflicts of interest to declare.

Rapid In Vivo Acquisition of Effector Tc1 Phenotype Even before Myeloid Engraftment Predicts Opportunistic Infections (OI) in Unrelated Cord Blood Transplant Recipients.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1415-1415
Author(s):  
Paul Szabolcs ◽  
Young-Ah Lee ◽  
Melissa Reese ◽  
Nelson Chao ◽  
Joanne Kurtzberg
Keyword(s):  
At Risk ◽  
T Cells ◽  
T Cell ◽  
Cord Blood ◽  
Cd4 T Cells ◽  
Viral Infections ◽  
Opportunistic Infections ◽  
Cord Blood Transplantation ◽  
Circulating T Cells ◽  
Unrelated Cord Blood

Abstract Unrelated cord blood transplantation (UCBT) is a viable option for patients who lack HLA-matched sibling donors. However, opportunistic infections (OI), occurring in the first 100 days, remain the major cause of morbidity and mortality. Viral infections are the primary cause of OI death. Despite the large number of T cells in UCB grafts there is no established antiviral immunity in nave UCB lymphocytes available for adoptive transfer. UCB T cells also lack expression of Th1/Tc1 cytokines, Granzymes A, B, and Perforin which are prerequisite cytotoxic effector molecules for controlling viral pathogens. We postulated that following transplant the infused naïve T cells will be critically influenced by the lymphopenic environment and emerging de novo acquired or latent viruses. Phenotypic alterations induced by these pathogens may identify those at risk for OI even before virological/clinical diagnosis is evident. Since our previous analysis of d+ 50 immune profile could not fully separate true risk factors from immunological correlates/consequences of OI we designed a new study. Methods: 70 pediatric recipients of HLA-mismatched UCBT were studied prospectively as soon as the WBC exceeded 0.3/ul, at a median of day+18, typically before myeloid engraftment and prior to the onset of clinical infections. Unprocessed whole blood was stained in “Trucount” (B-D) tubes for absolute T cell and dendritic cell (DC1, DC2) counts, while T cells subsets were similarly analyzed by 4 color FACS from Ficolled specimens for lymphocyte subsets, their activation state, tissue homing molecules. Intracellular (ic) staining identified cytotoxic effectors and inducible cytokine secretion besides characterizing homeostatic proliferation and apoptosis. 2-tailed student’s t-test identified significant variables (p<0.05) between OI + and OI− groups. Results: 39/70 patients developed OI at a median of day +35 (37 viral cases, 1 case of Legionella and 1 Candida), in average 2weeks after immune studies. While only 2/31 non-OI patients died (relapse), 18/39 from the OI group died at a median of 82 days post UCBT, p=0.001. Median f/up of survivors is 28 ± 17 months. Age, malignancy, cell dose had no impact on OI presence. The median values for major leukocytes were; WBC: 0.7×10e3/ul, T cells: 66/ul, NK cells: 46/ul, B cells: 0/ul, CD4+T cells: 42/ul, %CD4+ T cells: 64%. Both the OI+ and OI negative group had comparable WBC, CD3+, CD4+ T cells, NK lymphocytes, or DC1, DC2 subsets. Strikingly, 44% of circulating T cells were in cell cycle (KI-67+) regardless of OI status reflecting vigorous homeostatic expansion with only 16% preserving the CD45RA+/CD62L+ phenotype. Notably, ~9% of the circulating T cells were entering apoptosis (activated Caspase-3+), regardless of OI status. In those who eventually developed OI significantly more CD8+ cells circulated (40% vs 28%, p=0.03), more expressed CCR-5 (82% vs 55%, p=0.009), and more T cells were secreting IFNγ (35% vs 12%, p=0.006). In patients who developed OI significantly more Perforin+/CD8+ T cells circulated (49% vs 26%, p=0.01) and more of them expressed Granzyme B (p=0.05). There was higher BCL-2 expression in T cells from patients developing OI (p=0.04), and greater presence of CD28−/CD57+/CD8+ ‘effector’ CTLs (p=0.014). We conclude that in lymphopenic UCBT recipients subclinical viral infections may skew T cell maturation allowing early identification those at risk for clinical OI.

High Expression of Granzymes and Perforin along with Increased T cell Cycling In Vivo Predicts the Development of Opportunistic Infections (OI) Following Unrelated Cord Blood Transplantation (UCBT).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2238-2238
Author(s):  
Paul Szabolcs ◽  
Young-Ah Lee ◽  
Luciana Marti ◽  
Melissa Reese ◽  
Joanne Kurtzberg
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cord Blood ◽  
Cd8 T Cells ◽  
Viral Infections ◽  
Opportunistic Infections ◽  
Cord Blood Transplantation ◽  
Ki 67 ◽  
Granzyme A ◽  
Blood Transplantation

Abstract Introduction: Unrelated umbilical cord blood transplantation (UUCBT) is a viable option for those who lack HLA-matched sibling donors. However, opportunistic infections (OI) occurring in the first 100 days, remain the major cause of morbidity and mortality. Viral infections are the primary cause of OI death. As previously shown, cord blood T cells have significantly less preformed effector molecules available intracellularly to kill virally infected cells via the perforin-granzyme pathway than adult PB T cells. Since several virally infected patients control their infections in the absence of specific antiviral therapy ( e.g adenoviral enteris, polyoma cystitis) we postulated that the T cell compartment of those UUCBT recipients who experience early viral infections maty upregulate expression of the perforin exocytosis pathway. In parallel the impact of viral infection on T cell turnover would also be appreciable. Here we report on 19 prospectively studied pediatric patients, all full donor chimera, following myeloablative therapy. Methods: On day +50 we determined by 4-color FACS the expression of intracellular Granzyme, A, B, along with perforin. To monitor T cell turnover proliferating cells were identified by monitoring for the KI-67 nuclear antigen. The expression of the antiapoptosis gene BCL-2 was similarly monitored in both CD4+ and CD8+ T cells. We analyzed their association with the development of de novo OI up to day +100 employing Student’s t-test. Results: Mean age of patients was 6.2 years. 10 of 19 patients developed OI (adenovirus x 4, CMV x 7, EBVx1, parainfluenza x 1) with 5/10 patients experiencing more than one viral infections simultaneously) at a median of 29 days after UUCBT. Of those with OI 6/10 died due to their infections while 8/9 without OI are alive at a median of 15.8 months after UUCBT with one death due to leukemic relapse. Table I presents the correlation between the tested parameters with the development of OI. Patients experiencing viral infections had significantly higher % of their T cells in particular CD8+ T cells equipped with effectors of cytotoxicity and were proliferating in higher percentage compared to those with no active infections. However, the anti-apoptotic protein BCL-2 expression was significantly lower in patients experiencing OI that may lead to their shorter life span and overall T cell lymphopenia observed in OI patietns that we have previously detected in a larger cohort of 102 patients (ASBMT 2004 abstract#48). Conclusion: Correlating with active viral infections significant maturation of cord blood T cells is evident as early as 50 days after UUCBT towards acquiring effector molecules of the perforin pathway. Enhanced T cell proliferation is counteracted by reduced expression of BCL-2 that may lead to the lymphopenia in patients with OI. Future strategies aiming to enhance the longevity of antiviral T cells may protect from death due to viral infections. Univariate analysis VARIABLE MEDIAN VALUE FOR PATIENTS WITH OI MEDIAN VALUE FOR PATIENTS WITHOUT OI t-Test p value % Granzyme A+ T cells 52% 9% 0.006 % Granzyme A+ CD8+ T cells 91% 47% <0.001 % Granzyme B+ T cells 36% 6% 0.036 % Granzyme B+ T cells 87% 39% <0.001 % Perforin+ T cells 38% 4% 0.009 % Perforin+ CD8+ T cells 61% 21% <0.001 % Ki-67+ T cells 27% 16% 0.0041 % Ki-67+ CD8+ T cells 35% 16% 0.0037 BCL-2 expression level (MFI) 87 117 0.028

scholarly journals Filgrastim enhances T-cell clearance by antithymocyte globulin exposure after unrelated cord blood transplantation

2018 ◽  
Vol 2 (5) ◽  
pp. 565-574 ◽  
Author(s):  
Coco de Koning ◽  
Julie-Anne Gabelich ◽  
Jurgen Langenhorst ◽  
Rick Admiraal ◽  
Jurgen Kuball ◽  
...  
Keyword(s):  
T Cell ◽  
Cord Blood ◽  
Antithymocyte Globulin ◽  
Cord Blood Transplantation ◽  
Cd4 T Cell ◽  
Blood Transplantation ◽  
T Cell Reconstitution ◽  
Cell Clearance ◽  
Key Points ◽  
Unrelated Cord Blood

Key Points Residual ATG exposure delays CD4+ T-cell reconstitution more severely after CBT than after BMT. Filgrastim (G-CSF), given early after CBT, enhances ATG-mediated T-cell clearance in patients with residual ATG exposure.

scholarly journals Cord blood transplantation recapitulates fetal ontogeny with a distinct molecular signature that supports CD4+ T-cell reconstitution

2017 ◽  
Vol 1 (24) ◽  
pp. 2206-2216 ◽  
Author(s):  
Prashant Hiwarkar ◽  
Mike Hubank ◽  
Waseem Qasim ◽  
Robert Chiesa ◽  
Kimberly C. Gilmour ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cord Blood ◽  
Peripheral Blood ◽  
Cord Blood Transplantation ◽  
Cd4 T Cell ◽  
Molecular Signature ◽  
Blood Transplantation ◽  
T Cell Reconstitution ◽  
Key Points

Key Points Cord blood T cells are ontogenetically distinct from the peripheral blood T cells. Recapitulation of fetal ontogeny after cord blood transplantation results in rapid CD4+ T-cell reconstitution.

A Single Center Experience with Allogeneic Sibling Umbilical-Cord-Blood Transplantation in Children with Malignant and Non-Malignant Disease.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2042-2042
Author(s):  
Yongping Song ◽  
Yanli Zhang ◽  
Baijun Fang ◽  
Xudong Wei ◽  
Lulu Lu ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cord Blood ◽  
Median Time ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Cd4 T Cells ◽  
Cord Blood Transplantation ◽  
Umbilical Cord Blood Transplantation ◽  
Blood Transplantation

Abstract To study the outcome of allogeneic sibling umbilical-cord-blood transplantation (CBT) in children in China, we studied the records of 38 recipients of cord blood from HLA-identical siblings from 1998 through 2004. The mean age at transplant was 10.3+/−5.5 years. Diseases of the patients were leukemia (28), neuroblastoma (3), severe aplastic anaemia (3) and inborn errors of metabolism (four). A volume of 76–208 ml of cord blood was collected from sibling infants soon after delivery. HLA antigens were identical in 32 and one to three antigens mismatched in six. The patients received busulphan and cyclophosphamide as conditioning, and antithymocyte globulin was given to prevent graft rejection. The median number of collected nucleated cells was 3.5 x 107/kg nucleated cells (range, 2.0 to 10.2). T-cell reconstitution was evaluated by using combined approaches of phenotyping analysis of lymphocytes and assessment of ex vivo thymic function by measuring T-cell receptor (TCR) rearrangement excision circles (TRECs). The overall survival is 89% at the median observation time of 72 months. The median time for neutrophil engraftment (absolute neutrophil count >500/mm3) was 16 days (range, 10 to 41days). The median time to become transfusion independent after CBT was 27 days for platelets (range, 15 to 54) and was 28 days for packed red blood cells (range, 19 to 128). Acute GVHD (aGVHD)was observed in 20/38 patients and involved only skin in 16 patients, skin and liver or gut in 2 patients and all 3 organs in another 2 patients. Seventeen of 20 patients had grade 1 to 2 aGVHD toxicity, whereas 3 patients experienced grade 3 to 4 aGVHD. Chronic GVHD (cGVHD) developed in 12 patients. Acute transplant related mortality was 5.5%. Cause of death was persisting non-engraftment till day +100 after transplant. Late mortality occurred in 2 patients: one cGVHD associated haemorrhage 22 months after CBT and one cGVHD associated septicaemia 4 years after CBT. Immunologic reconstitution demonstrated that CBT resulted in consistent and stable T-, B- and natural killer (NK) cell development. The kinetics of development was such that T-cell development occurred between 65 to 180 days. Initial T-cell engraftment consisted predominantly of CD45RO+, CD3+, and CD4+ T cells, and at 12 to 24 months changed to CD45RA+, CD3+, and CD4+ T cells, indicating de novo maturation of T cells. During the first 5 months after transplantation, TCR repertoires were highly abnormal and TREC values low. However, in a longer follow-up (one years or more after transplantation) TREC values and TCR diversity increased and became normal. This data confirms that CBT still should be the first treatment choice if an HLA-identical sibling is available.

Contribution of T-Cell Reconstitution during the Early Phase after Cord Blood Transplantation to Favorable Clinical Outcomes.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2227-2227
Author(s):  
Satoshi Takahashi ◽  
Nobukazu Watanabe ◽  
Jun Ooi ◽  
Akira Tomonari ◽  
Kashiya Takasugi ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cord Blood ◽  
Cd8 T Cells ◽  
Early Phase ◽  
Cord Blood Transplantation ◽  
Interferon Γ ◽  
Cell Recovery ◽  
Memory Phenotype ◽  
Blood Transplantation

Abstract The immaturity of T cells in cord blood is well known in functional assays and phenotypic analyses. During the first several months after cord blood transplantation (CBT), the T cell compartment is recovered by peripheral expansion from those mature and naïve T cells in cord blood grafts and plays an important role in acute graft-versus-host disease (GVHD) and graft-versus-leukemia reaction. Recently, we have reported that adult patients with hematological malignancies receiving CBT from HLA-partially-mismatched unrelated donors (n=68) had a lower risk of severe acute GVHD (> grade II, 7% versus 26%) and transplant-related mortality (9% versus 29% at 1 year) and a higher probability of disease-free survival (74% versus 44% at 2 years) than HLA-matched unrelated bone marrow transplant (BMT) recipients (n=45) in our multivariate analysis (Takahashi et al., Blood, in press). We speculated that the immune reconstitution process over a period of several months after CBT might have contributed to these promising clinical results. Using four-color analysis with CD4, CD8, CD45RA, and CD62L, more than 90% of cord blood CD4+ and CD8+ T cells in the grafts belonged to the naïve fraction. Cytokine expression in cord blood T cells was also suppressed to 0.1% in CD4+ and to 0.9% in CD8+ with positive interferon-γ by intracellular staining, which were significantly lower than those in adult T cells (16.2% in CD4+ and 37.8% in CD8+). Circulating T cell counts normalized after 3 months for CD8+ and 4 months for CD4+ in our CBT recipients, both of which were significantly faster than in previously published studies, which were 9 months for CD8+ and 12 months for CD4+. After T cell recovery, peripheral blood T cells moved from the naïve to the central memory fraction immediately, and then moved to the effector memory fraction. A naïve subset of CD4+ T cells remained (median: 38 cells/μl on day 90, n=12) during the first 3 months, which was significantly higher than in the BMT control (median: 9 cells/μl on day 90, n=5, p=0.015), but showed a low level of CD8+ T cells (median: 14 cells/μl on day 90, n=12), almost the same as in BMT recipients (median: 13 cells/μl on day 90, n=5). Intracellular interferon-γ-producing T cells were detected at 3.4% (0.1–34.2%) in CD4+ and 32.3% (1.1–86.9%) in CD8+ at 1 month post-CBT (n=16), both of which were comparable to post-BMT. To investigate whether these T cells with memory phenotype are functional, we analyzed antigen-specific T-cell recovery using cytomegalovirus (CMV) as a specific antigen. CMV-responsive CD4+ T cells were detected within the first 4 months in all recipients with positive CMV antigenemia (n=13), but CD8+ T cells were detected only in 5 out of 13 cases, probably because of pre-emptive Gancyclovir administration in most antigenemia-positive patients. To conclude, naïve cord blood T cells rapidly increased in number and adopted a memory phenotype showing cytokine-production and antigen-recognition capacity in the early phase after CBT. These data suggest that mature T lymphocytes in cord blood have unique properties and contribute to the favorable clinical outcome of CBT.

scholarly journals Cognate CD4 T-Cell Licensing of Dendritic Cells Heralds Anti-Cytomegalovirus CD8 T-Cell Immunity after Human Allogeneic Umbilical Cord Blood Transplantation

2014 ◽  
Vol 89 (2) ◽  
pp. 1058-1069 ◽  
Author(s):  
T. W. H. Flinsenberg ◽  
L. Spel ◽  
M. Jansen ◽  
D. Koning ◽  
C. de Haar ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cord Blood ◽  
Cord Blood Transplantation ◽  
Survival Rates ◽  
T Cell Immunity ◽  
Cell Responses ◽  
Blood Transplantation ◽  
Cell Immunity ◽  
Cognate Antigen

ABSTRACTReactivation of human cytomegalovirus (CMV) is hazardous to patients undergoing allogeneic cord blood transplantation (CBT), lowering survival rates by approximately 25%. While antiviral treatment ameliorates viremia, complete viral control requires CD8+T-cell-driven immunity. Mouse studies suggest that cognate antigen-specific CD4+T-cell licensing of dendritic cells (DCs) is required to generate effective CD8+T-cell responses. For humans, this was not fully understood. We here show that CD4+T cells are essential for licensing of human DCs to generate effector and memory CD8+T-cell immunity against CMV in CBT patients. First, we show in CBT recipients that clonal expansion of CMV-pp65-specific CD4+T cells precedes the rise in CMV-pp65-specific CD8+T cells. Second, the elicitation of CMV-pp65-specific CD8+T cells from rare naive precursors in cord blood requires DC licensing by cognate CMV-pp65-specific CD4+T cells. Finally, also CD8+T-cell memory responses require CD4+T-cell-mediated licensing of DCs in our system, by secretion of gamma interferon (IFN-γ) by pp65-specific CD4+T cells. Together, these data show that human DCs require licensing by cognate antigen-specific CD4+T cells to elicit effective CD8+T-cell-mediated immunity and fight off viral reactivation in CBT patients.IMPORTANCESurvival rates after stem cell transplantation are lowered by 25% when patients undergo reactivation of cytomegalovirus (CMV) that they harbor. Immune protection against CMV is mostly executed by white blood cells called killer T cells. We here show that for generation of optimally protective killer T-cell responses that respond to CMV, the early elicitation of help from a second branch of CMV-directed T cells, called helper T cells, is required.

In Vitro Assessment of Tolerance and Rejection Occurring After Co-Transplantation of Allogeneic Umbilical Cord Blood and Haploidentical CD34+ Cells as Treatment for Severe Aplastic Anemia

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2352-2352
Author(s):  
Nicole J. Gormley ◽  
Aleah Smith ◽  
Maria Berg ◽  
Lisa Cook ◽  
Catalina Ramos ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cord Blood ◽  
Cord Blood Transplantation ◽  
Neutrophil Recovery ◽  
Post Transplant ◽  
Blood Transplantation ◽  
Cd34 Cells ◽  
Cord Blood Cells

Abstract Abstract 2352 Introduction/Methods: The administration of highly purified haploidentical peripheral blood CD34+ cells combined with an unrelated cord blood transplant results in earlier neutrophil engraftment than is typically seen with a cord blood transplant alone. Chimerism data from pilot trials evaluating this strategy have reported 3 phases of engraftment: 1) early myeloid engraftment from transplanted haplo-CD34+ cells followed by 2) cord blood engraftment resulting in dual chimerism and 3) the subsequent disappearance of haploidentical donor cells with resultant full donor cord chimerism. The mechanism accounting for the disappearance of haploidentical cells has not been defined. Here the clinical results and an in vitro assessment of alloreactivity in three patients that underwent combined haploidentical CD34+ cell and cord blood transplantation for severe aplastic anemia (SAA) are described. The conditioning regimen consisted of cyclophosphamide (60mg/kg/day on days -7 and -6), fludarabine (25mg/m2/day on days -5 to -1), horse ATG (40mg/kg/day on days -5 to -2), and total body irradiation (200cGy on day -1). GVHD prophylaxis consisted of tacrolimus and mycophenolate mofetil. PCR of STRs was used to assess chimerism in T-cell and myeloid lineages and mixed lymphocyte reaction assays(MLR) were performed on peripheral blood samples collected at different time-points post-transplant to assess for alloreactivity against the recipient, the haploidentical donor, or the cord unit. Stimulator cord blood cells for the MLR were obtained from residual cord blood cells remaining in the infusion bag after patient administration and expanded in vitro using anti-CD28/CD3 Dynabeads. Results: Prior to transplantation, all three pts had transfusion dependent SAA associated with severe neutropenia that was refractory to conventional immunosuppressive therapy. Pt 1 had an early transient myeloid recovery (ANC 400 on day+11) from the haploidentical donor followed by engraftment of the cord unit (Cord ANC > 500) on day 21. The patient is currently 2 years post transplant and has 100% cord blood chimerism and is transfusion independent. An MLR assay performed when donor T-cell chimerism was 100% cord, showed evidence for rejection of the haploid cells by cord blood T-cells, with the MLR response to haploidentical donor cells being seven fold higher than the response to fully HLA-mismatched 3rd party cells. In pt 2, neutrophil recovery from the transplanted haploidentical donor occurred on day +10, with chimerism studies showing no evidence for cord engraftment in either myeloid or T-cell lineages at any point post-transplant. The patient is currently 15 months post transplant and is transfusion independent with normal blood counts and sustained “split” chimerism (T-cells recipient in origin with myeloid cells being 100% haploidentical donor). MLR assays showed that the recipient was tolerant to the haploid donor, with no statistically significant difference in the alloreactive response to the haploid donor compared to self. In pt 3, neutrophil recovery from the transplanted haploidentical donor occurred on day +10, with chimerism studies showing split chimerism (T-cell chimerism >90% cord and myeloid chimerism 88–100% haploid donor in origin). MLR assays again showed evidence of rejection of the haploid cells by cord blood T-cells, with a trend towards greater alloreactivity against the haploid donor compared to an HLA mismatched 3rd party on post-transplant day +63. Conclusions: Combined haploidentical CD34+ cell and unrelated cord blood transplantation following highly immunosuppressive conditioning represents a viable treatment option for patients with SAA who lack an HLA-matched donor. Using this approach, 2 of 3 pts had cord blood engraftment associated with early neutrophil recovery from the haploidentical donor. In one pt, the cord unit failed to engraft. Remarkably, sustained engraftment from the haploidentical donor in this pt resulted in transfusion independence. MLR appears to be a useful approach to assess the in vitro alloreactivity of this unique stem cell graft source. In the two pts who had cord engraftment, in vitro MLR assessments established that the disappearance of haploid cells occurred as a consequence of rejection of the haploidentical cells by engrafting cord blood T-cells, rather than from non-immunological haploidentical cell graft failure. Disclosures: No relevant conflicts of interest to declare.

Viral Infection After Unrelated Cord Blood Transplantation In Bordeaux: Incidence and Outcomes. A Single Centre Experience with Eurocord.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4543-4543
Author(s):  
Pierre-Yves Dumas ◽  
Reza Tabrizi ◽  
Stephane Vigouroux ◽  
Marie-Edith Lafon ◽  
Gerald Marit ◽  
...  
Keyword(s):  
Stem Cell ◽  
Viral Infection ◽  
Cord Blood ◽  
Viral Infections ◽  
Hemorrhagic Cystitis ◽  
Cord Blood Transplantation ◽  
Median Number ◽  
Bk Virus ◽  
Blood Transplantation ◽  
Unrelated Cord Blood

Abstract Abstract 4543 Background: Umbilical cord blood is being increasingly used for transplantation, but the ability of neonatal T cells to regulate viral infections seems to be less effective than other stem cell sources. Viral infections remain important cause of morbidity after hematopoietic stem cell transplantation and some virus less monitored could be important pathogens. We performed a retrospective study, using the EUROCORD database, in 45 patients with the aim to analyze the incidence for viral infections in patients transplanted with single or double unrelated cord blood units for hematologic malignancies, during the first two years after transplantation. Methods: Patients received the unrelated umbilical cord blood transplantation at the Bordeaux University Hospital, France, between October 2003 and December 2009. Real-time quantitative polymerase chain reaction was used to measure the EBV and CMV load twice a week and only if presence of symptom for others virus. We included patients with one quantitative positive PCR in blood for EBV, CMV, HHV6 and ADV, one PCR positive for BK virus in urine, and clinical diagnosis for VZV recurrent infections. Herpes virus disease was defined as clinical symptom from the affected organ, combined with the detection of the herpes virus by PCR in organ biopsy or blood. Results: The median age was 41 years (range 8–63) with 5 patients (11.5%) < 15 years old. Among these 45 patients, 75% had acute leukemia or myeloid malignancies and 25% lymphoid malignancies. Administration of acyclovir for prevention of HSV and CMV infections was carried out in all patients. Grafts have been performed with a single unmanipulated cord blood unit in 73% and two unmanipulated units in 27%. Neutrophil and platelet recoveries were defined as the first of 2 consecutive days during which the ANC in the blood was >0.5×109/L and the platelet count was >20×109/L without transfusion support, median time to neutrophil recovery was 24 d (range 5–56 d) and median time to platelet recovery was 34 d (range 0–88 d). The median number of nucleated cells infused in recipients of cord blood was 2.2×107/kg of the recipient's body weight (range, 0.7×107 to 6.63×107/kg). The median number of CD34+ cells in the cord blood grafts was 0.9×105/kg (range, 0.16×105 to 3.4×105/kg). Conditioning varied according to the patient's disease and disease status with 45% myeloablative and 55% reduced intensity. All patients, except one, received a cyclophosphamide containing regimen and 24.4% patients received antithymocyte globulin prior to transplantation. Prophylaxis for acute GVHD consisted of cyclosporine A and MMF in 91%, cyclosporine A alone for 2 patients, with prednisone for 1 patient and with MTX for 1 patient. Among the whole cohort, we observed 77.7% viral infections: 9/45 experienced at least one positive PCR for EBV of whom 1 had lethal PTLD, 10/45 for CMV with one gastrointestinal disease and 25/45 for HHV6 with three encephalitis, two pneumoniae, five skin rash, three gastrointestinal infections manifested as diarrhea or hemorrhagic colitis, and two hepatitis. Furthermore, we observed 6/45 herpes zoster with one disseminated disease, 7/45 hemorrhagic cystitis resulting of BK virus infection and 3 patients who experienced ADV viremia. At 100 days the cumulative incidence (CI) for any viral infection was about 69%, 51% for HHV6 infection, 35% for HHV6 disease, 13% for CMV infection and disease, 18% for EBV infection, 14% for BK virus hemorrhagic cystitis, 5% for ADV infection and cumulative incidence at 200 days about 5% for VZV. Viral infection or disease did not affect significantly the overall survival of the patients. Conclusion: Among the various viral infection/diseases, the high CI of HHV6 infection/disease is of concern. Analysis of risk factors for infection and disease in this specific setting is in progress. Although the morbidity is high, thanks to close monitoring and prompt therapeutic intervention when possible, the mortality directly linked to viral infection remains low. Disclosures: No relevant conflicts of interest to declare.

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