bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative Study of Chimerism Dynamics between Allogenic Bone Marrow Transplantation and Nonmyeloablative Stem Cell Transplantation.
Authors: Tang Xiaowen , Wu De Pei , Sun Aining
Published: April 2020

Objective To explore whether the dynamics and outcome of chimerism are different between allogenic bone marrow transplantation (allo-BMT) and nonmyeloablative stem cell transplantation (NST) these two kinds of transplant strategies in order to define factors that significantly influence the early engraftment of donor stem cells.

Methods A retrospective comparative study was performed between 20 patients undergoing BMT and 18 patients receiving NST. There are several differences between NST and standard allo-BMT, such as conditioning regimens, the source and the quantity of hematopoietic stem cells and the protocal of GVHD prophylaxis. But component of two groups patients were similar. Sequential monitoring of chimerism were performed in 38 patients in different periods post transplantation using multiple short tandem repeat(STR) amplification by fluorescence labeling polymerase chain reaction(PCR) combined with a capillary electrophoresis.

Results

The median age of recipient, median quantity of MNC, CD34+ cells and T lymphocyte cells of NST were significantly higher compared with the BMT. The median time for absolute neutrophils recovery of NST(without G-CSF) was similar to that of BMT. While compared to BMT, NST lead to faster engraftment of platelets. The establishment of full donor chimerism (FDC) of NST was much more earlier than that of BMT (1 month VS 3 months, p=0.01). In the early post-transplant period(up to day 30), there was an apparent increase in the incidence of full donor chimerism (FDC) following NST compared with BMT(38.9% VS 20%, p=0.019). Whereas a significantly lower incidence of mixed chimerism(MC) occurred in recipients of NST compare to BMT.(61.2% VS 80%, p=0.02). It suggest that in the early stage after transplantation, the engraftment of donor cells of NST was more rapid than that of BMT. The major relevant factor causing full donor chimerism formation seems to be the high quantity of donor CD34+ cells and T lymphocyte cells. Fludarabine-based reduced-intensity conditioning didn’t result in a significant delay of donor cell engraftment compared to standard conditioning regimen. The incidence of chronic GVHD was much higher in group of NST than that of BMT group(80% VS 50%, p=0.007).

This finding was related to that patients of NST group received the higher doses of CD34+ cells. Conclusion Factors that significantly influence the early kinetics of donor chimerism and donor cell engraftment after transplantation were the quantity of stem cell and T lymphocyte cell of grafts.
Nonmyeloablative Allogeneic Stem Cell Transplantation for a Case of Refractory Systemic Lupus Erythematosus.
Authors: Lu Quanyi , Niu Xiaoqing , Guo Yong , Lu Lulu , Liu Delong
Published: April 2020

Autologous stem cell transplantation (SCT) was effective for severe SLE and may have curative potential in some cases, but recurrence of the disease was still a major problem. Allogeneic SCT for autoimmune disease has been shown to induce long-term remission through graft-versus-autoimmunity effect. Non-myeloablative SCT can induce full-donor Chimerism with much less toxicity. We report a case of 43 year old Chinese female with severe SLE refractory to high dose cyclophosphamide and had lupus nephritis. The patient also had avascular necrosis and was bed-bound. Peripheral blood stem cell transplantation were done from an HLA-matched sibling after non-ablative conditioning using ATG, Fludarabine and busulfan. The patient had good performace status nad had no recurrence at the last follow-up 16 months post-transplantation.
Allogeneic Stem Cell Transplantation for Lymphoid Malignancies.
Authors: Novitzky Nicolas , Thomas Valda , Du Toit Cecile , McDonald Andrew
Published: April 2020

Haematopoietic stem cell transplantation may be curative therapy for selected patients with haematological malignancies. We reviewed our experience in individuals with lymphoid neoplasms (excluding multiple myeloma) who received an allogeneic stem cell transplant from HLA identical sibling donors. Myeloablative conditioning was radiation based (n= 46) or with chemotherapy (n= 12). GvHD prophylaxis was with T-cell depletion (CAMPATH-1 g or H,“ in the bag”). 17 received a BMT while 41 had PBPC (median CD34+ 2.64). Patients receiving BMT had no further immunosuppression, while 12 patients who received PBPC grafts received prednisone 30 mg and another 21 were treated with therapeutic doses of cyclosporine for 90 days. The diagnosis was indolent lymphoma in 14 (median 3 treatment modalities), aggressive variant in 18 (7 with transformed DLBC; 6 with lymphoblastic lymphoma, 2 mantle cell), 5 had chemotherapy responsive (median 4 treatments) recurrent Hodgkin’s lymphoma and 21 had lymphoblastic leukaemia (18 in CR1). Median age was younger in patients with ALL (24 years; range 15–45) than lymphoma (47 years; 29–57; p< 0.001). At a median follow up of 1173 (32–5598) days, 20 have died and 67% survive disease free. Death was from disease recurrence in 15 (10 with ALL), and it was transplant related in 4 while one patient died of AIDS. Survival was significantly worse in patients with lymphoblastic leukaemia (48%; p< 0.001), while 76% of those with lymphomas survive in unmaintained remission (median 1493 days; 32–5598). Survival was significantly associated with diagnosis of lymphoma (p< 0.01), CD34+ cell number above median (p= 0.04) and absence of GvHD (p< 0.01). We conclude that despite T-cell depletion allogeneic stem cell transplantation is very effective therapeutic strategy in patients with lymphoma, as disease recurrence was low (relapse 6/37; p= 0.03), suggesting remaining graft vs. lymphoma effect. To the contrary, this therapy remains less satisfactory for patients with ALL who have poor prognostic factors.
Graft-Versus-Lymphoma Effect in Hodgkin’s Disease: A Possible Role in Maintaining a Disease Control.
Authors: Pastano Rocco , Andreola Giovanna , Gigli Federica , Saronni Luca , Cassatella Maria C. , Peccatori Fedro , Martinelli Giovanni
Published: April 2020

Introduction: non-myeloablative and highly immunosuppressive conditioning regimen instead of conventional bone marrow transplantation have been recently developed. The aim of this treatment is to establish a mixed chimerism and to allow the donor cell-mediated graft versus-tumor effect to occur. This is a well known effect in disease like CML and AML and much less investigated in Hodgkin Lymphoma.

Patients and methods: from September 2003 to November 2004, 5 pts affected by chemorefractory Hodgkin’s lymphoma, underwent a non-myeloablative conditioning regimen followed by allogeneic peripheral blood stem cell transplantation (PBSCT). Median age was 28 ys (range 26–33), 4 M and 1 F, all pts had failed previous conventional treatment including autologous PBSCT. Conditioning regimen were the following: Fludarabine 30 mg/mq, from day −4 to −2 followed by TBI 2Gy on day 0 in 2 pts; TBI 2Gy alone two month after an autologous PBSCT in 1pt; thiotepa 5mg/kg x2, Fludarabine 30mg/mq, Cyclofosfamide 30 mg/kg in 1pt or Rituximab 600 mg/mq on day −5; fludarabine 30 mg/mq + Cyclofosfamide 30 mg/kg from day −4 to day −3 in the other one. All patients received donor peripheral stem cell infusion on day 0. Prophylaxis of aGVHD was performed with Cyclosporine (6.25 mg/kg) po from day −3 to day +36 followed by a rapid tapering until day +56 and Mycophenolate Mofetil (15mg/kg) po from day 0 to +27.

Results: All pts experienced a complete engraftment, obtained a complete response after transplantation and 3/5 them experienced hepatic and/or cutaneous cGvHD. At a median follow-up of 13 months 2/3 pts, who received TBI and/or Fludarabine, are alive after 681 days, 612 days from transplantation and are in CR and SD respectively; both of them had cGvHD; the third one has died at +132 for cerebral hemorrhage. One of the remaining two patients has died at +200 in CR for septic shock after cGvHD, and the last one is still alive in +410 in PD, with no sign of cGvHD.

Conclusions: although limited, our data seem to suggest that even in pts affected by Hodgkin’s lymphoma with a chemoresitant disease, the development of cGvHD after a reduced conditioning allogeneic PBSCT, mostly immunosuppressive, seems to be associated with the presence of a graft-versus lymphoma effect able to mediate a good control of the disease.
Graft vs. Tumor Effects (GVT) in Non Myeloablative Stem Cells Transplant (NST) in Relapse Hodgkin Disease (HD) after Autologous Bone Marrow Transplant (ABMT).
Authors: Milone Jorge H. , Bordone Javier A. , Kusminsky Gustavo , Foncuberta Cecilia , Garcia Juan , Rolon Juliana Martinez , Bullorsky Eduardo , Dengra Cristina , Cerutti Ider
Published: April 2020

The relapse post ABMT is a terminal stage complication in HD. The allogenic transplant contributes to a different therapeutic approach through the immune anti tumor effect of the Graft versus Host Disease (GVHD)

The NST variant of the procedure diminishes the risks of transplant related mortality (TRM) and is associated with induction of mixed hematopoietic chimerism when either HLA-identical sibling or unrelated donor are used, and that powerful GVT effects can be achieved against advanced chemotherapy/resistant hematology disease.

The feasibility, risk and effectiveness of the NST, is analyzed in 13 patients with relapsed HD after ABMT. Seven women and six men with a median age of 26 years old. They had received ≥ of 3 therapeutic lines that included the high doses and the ABMT. The period betwen the diagnosis to NST, had a median time of 36 months. The patients were conditioned with Fludarabine based schemes plus: Melphalan 8, Cyclofosfamide 3, TBI 1, Busulfan 1. The sources of stem cells were, with a HLA-identical sibiling donors in 12 patients, and in 1 with non related donor. 4 patients died due to complications associated with the BMT (TRM 28 %): shock 2, sepsis 1, thrombotic microangiophaty 1. Half of the patients developed some form of GVHD, or it was induced by donor lymphocytes infusion. With an average follow up of 15 months (range: 1–56), 7 patients are alive, 5 of them developed GVHD and are in CR, the other 2 have a short follow up (28 and 75 days). Of 4 patients who progressed only 1 present GVHD. The NST is a feasible procedure in HD relapsed post ABMT. The TRM is 28 %. In this group there is a clear relationship between the control of HD and the appearance of GVHD.
A Successful Syngeneic Bone Marrow Transplant after an Ortotopic Liver Transplantation for Fulminant Hepatitis - Case Report.
Authors: Silla Lucia , Paz Alessandra , Astigarraga Claudia , Zago Alethea , Brum Caroline , Bittencourt Rosane , Fogliatto Laura , Friederich João R. , Leugeur Denise , Silveira Themis R.
Published: April 2020

We report the case of a 12 year old girl admitted to the hospital in August 2004 presenting acute hepatic failure with grade IV encephalopathy without stigmatic of chronic liver disease. Investigation for A, B and C hepatitis was negative as well as the tests for HIV, Toxoplasmosis, EBV, B19 parvovirus and CMV. Diagnosed as fulminant hepatitis she received an ortotopic hepatic transplant 25 days after admission. She had an excellent clinical recovery on tracolimus for immunossupression, except for an asymptomatic pancytopenia unresponsive to drug dose reduction, which progressed to severe bone marrow aplasia in a four months period. A second round of virus screening tests was still negative. She was then submitted to bone marrow transplantation from her twin identical sister utilizing 50mg/kg/day of cyclophosphamide for two days as conditioning regimen. She received 6.6 x 106 CD34+ cells/kg and engrafted at D+22. She was discharged at D+ 33 on tracolimus without any sign of liver or bone marrow rejection. She is presently in complete clinical recovery after 7 months of BMT. This is, from our knowledge, the first report of a successful hepatic transplant followed by an equally successful syngeneic bone marrow transplant.
Prolonged Survival in an Elderly Patient with CD4+ CD56+ Lymphoma of the Skin with Autologous Peripheral Blood Stem Cell Transplantation (PBSCT).
Authors: Nath Rajneesh , Abo Stephen M.
Published: April 2020

CD4+ CD56+ lymphomas are recently described rare hematological neoplasms thought to be originating from plasmacytoid dendritic cells. Clinical features typically include cutaneous nodules associated with lymphadenopathy and/or splenomegaly and bone marrow involvement during the disease course. Complete remission (CR) is frequently obtained (around 80%) with polychemotherapy. However, median time to relapse is about 9 (range 3–18) months and median survival is only 13 months. Age > 60 years is a poor prognostic factor. Long term survival has only been reported following allogeneic SCT (Bone Marrow Transplantation32, 637–646, 2003; Blood99,1556–1563, 2002). We report a case of CD4+ CD56 + lymphoma with prolonged survival after an autologous PBSCT. A 71-year-old male presented in the fall of 2001 with facial erythema that progressed to several purple cutaneous nodules also involving his trunk. Skin biopsy done in June 2002 confirmed the diagnosis of CD4+ CD56+ lymphoma of the skin. Staging studies including CT scan of the chest, abdomen and pelvis, and bone marrow aspiration and biopsy were negative. The patient received chemotherapy with CHOPE (cytoxan 750 mg/m2, adriamycin 50 mg/m2, vincristine 1.4 mg/2, prednisone 100mg x 5 days, etoposide 120 mg/m2) for six cycles starting June 2002, and achieved a CR with disappearance of all skin lesions. CT scan of the chest pretransplant revealed a single 2 cm paratracheal lymph node that was also PET positive. Stem cells were collected using cytoxan 4 gms/m2 followed by 10 mcg/kg/day of G-CSF. The patient underwent autologous stem cell transplantation in February 2003 using BEAM (carmustine 300mg/m2 day −6, etopside 200mg/2 days −5 to −2, cytarabine 400mg/m2 days −5 to −2, melphalan 140 mg/m2 day -1) as a preparative regimen. Neutrophil engraftment occurred on day 10 and platelets engrafted on day 14 post SCT. The paratracheal lymph node was still visible on the PET scan 6 months post SCT. However, restaging studies at one year and beyond have all been negative for disease reoccurrence including a CT chest that revealed a calcified paratracheal lymph node that was negative on PET scan. The patient continues to be in complete remission 37 months from diagnosis and 30 months post SCT. The aggressive nature of CD4+ CD56+ lymphoma is evident by its 9-month median time to relapse and short survival. Therefore, aggressive treatment protocols incorporating stem cell transplant should be utilized in their management and should not be limited to only younger patients. As illustrated by our patient, the use of autologous stem cell transplant in the elderly can be done safely with encouraging results.
Comparison of Hematopoietic Reconstitution after Autologous Transplantation Using Stem Cells after Short- (<12 Months) and Longer-Term Cryostorage.
Authors: Lowenthal Raymond M. , Burbury Kate L. , Ragg Scott J. , Brillant Corinne
Published: April 2020

High-dose chemotherapy with autologous hemopoietic stem cell (HSC) transplantation is an important treatment option for patients with refractory or relapsed non-Hodgkin or Hodgkin lymphoma (NHL or HL). Although it is customary to collect the HSCs at the time the need is apparent, in theory collections carried out at an earlier stage of the disease - preferably when the patient is in remission - may have advantages in that 1) such HSCs will have been less exposed to damaging cytotoxic chemotherapy and 2) early collections may be less likely to be contaminated with malignant cells. However such "prophylactic" or "rainy day" harvests may need to be stored for extended periods before use, raising questions about their functional capacity. Although laboratory studies of the effects of long-term storage on the viability and proliferative capacity of cryopreserved HSCs have demonstrated that the cells remain functional, there have been few studies of clinical outcomes after their reinfusion. Because we have had a long-standing policy of carrying out "rainy day" harvests, we have had the opportunity to retrospectively compare rates of neutrophil and platelet engraftment, and 30-day transplant related mortality (TRM), of patients undergoing HSC transplantation after short-term (<1 year) and longer-term (range 1–9.3 years, median 2.2 years) cryostorage. Of the 133 NHL and HL patients transplanted at our hospital between 1985 and 2005, 98 were in the short-term and 35 in the longer-term group. TRM was 7/98 and 1/35 for the short- and longer-term groups, respectively (P=0.7); no deaths occurred in the 77 patients transplanted after 1998. Hematopoietic reconstitution was achieved in all 125 patients who survived 30 days. Neutrophil recovery (absolute neutrophil count >0.5 x 109/L) was achieved by 12.2 ± 4.2 (SD) days and 14.2 ± 6.5 days post-reinfusion for the short- and longer-term groups respectively (P=0.14). Platelet support was required for a mean 6.6 ± 9.5 and 7.5 ± 7.1 days, respectively (P=0.7). These clinical results demonstrate that HSCs after long-term cryostorage retain engraftment potential similar to that of HSCs stored for lesser periods and that such cells are safe and efficacious for autologous transplantation.
Augmented Preparative Regimens Using Total Body Irradiation or BCNU Prior to Autologous Stem Cell Transplant (AuSCT) Are Well-Tolerated and Yield Substantial Rates of Progression-Free Survival in Patients with Poor Risk Hodgkin Lymphoma (HL) or Diffuse Large B-Cell Lymphoma (DLBCL).
Authors: Ritchie David S. , Seymour John F. , Thompson Sarah , McManus Michael , Prince Miles
Published: April 2020

Introduction: AuSCT using conditioning such as BEAM, for relapsed DLBCL and HL results in progression-free survival (PFS) of 60% of cases in second complete remission (CR). Those who do not achieve a second CR or who have primary refractory disease have significantly reduced benefit from BEAM/AuSCT. Consequently, we have been offering dose intensified conditioning prior to AuSCT with the Stanford regimen using Etoposide 60mg/kg day-4, Cyclophosphamide 100mg/kg day-2 and either BCNU 15mg/kg day-6 or fractionated TBI 12Gy days-8 to -5 to patients with poor risk haematologic malignancy.

Methods: We assessed the outcome of AuSCT in 46 patients with poor risk haematologic disease; DLBCL (n=16), HL (n=16), multiply relapsed indolent lymphoma (n=8) and aggressive “other” lymphomas (n=6) who received conditioning with either the BNCU (n=31) or TBI (n=15) protocols. Of the 32 patients with HL or DLBCL, 28 had previously failed to achieve a CR to salvage chemotherapy; PR (21/32) or induction failure/early progression (7/32).

Results: There were no transplant-related deaths among the 32 HL or DLBCL patients. One case of fatal radiation pneumonitis occurred following consolidative radiotherapy 6 months post AuSCT. The PFS for the combined HL/DLBCL cohort was 65±9% at 12, and 61±9% at 24 months. The 24 month PFS by BCNU or TBI regimen was 63% and 57% respectively (p=0.56). The 24 month PFS for HL or DLBCL treated with either regimen was 54±13% and 68±12% respectively (P=0.34) Additional groups were too heterogeneous to statistically assess. 1 of 6 patients with aggressive “other” lymphoma maintained a durable remission (39 months) and 4/8 patients with indolent lymphoma have ongoing CR at 9 – 59 months follow up.

Conclusions: Stanford TBI or BCNU conditioning prior to AuSCT are well-tolerated and result in substantial rates of PFS in patients with poor risk HL or DLBCL.
Minimal Frequency of Aspirin Resistance after Observed Aspirin Ingestion.
Authors: Schwartz Kenneth A. , Schwartz Dianne E. , Barber Kimberly , Reeves Mathew , DeFranco Anthony C.
Published: April 2020

Poor compliance is an important cause of aspirin resistance. Our goal was to investigate amount of aspirin induced inhibition of platelets independent of compliance. We investigated the net amount of aspirin induced inhibition of platelets by comparing Platelet Prostaglandin Agonist (PPA) stimulated platelet aggregations before, off aspirin, with the platelet inhibition that occurred 2 hours after observed aspirin ingestion. 217 post-myocardial infarction patients who had taken aspirin for at least 30 days were evaluated at two time points. The first time point was 7 days after they had signed informed consent to stop taking aspirin as well as nonsteroidal anti-inflammatory drugs (NSAIDs). The second time point was 2 hrs after they were observed to ingest 325mg of aspirin. Platelets were evaluated using 2 different agonists in a light aggregometer. Arachidonic acid (AA) was used as a qualitative check to see if the patients at the first time point had really refrained from taking aspirin and NSAIDs for 7 days and PPA which mainly activates platelets via the prostaglandin pathway and can detect degrees of aspirin induced platelet inhibition. The slope of the PPA stimulated aggregation curve was used to measure aspirin induced platelet inhibition. The net aspirin response was calculated by determining the paired difference between the PPA slopes at the two time points. Compliance with the directive of not taking aspirin or NSAIDs before the first time point was checked with AA stimulated aggregometry and 45 patients were judged to be noncompliant. We defined aspirin resistance as a minimal change in PPA slope of 15 or less. Of the patients who had complied with the directive not to take aspirin or NSAIDs for 7 days as determined by AA aggregometry only 7 (3.2%) were aspirin resistant. The net aspirin response was directly related to the PPA slope off aspirin (p<0.001). When the 45 patients who were judged by AA aggregometry to be noncompliant with the directive to refrain from taking aspirin or NSAIDs were removed from the analysis, the direct relationship between the slope of the off aspirin PPA aggregation and the net aspirin response remained significant (p<0.001). The amount of platelet inhibition in the compliant group 42±16 was significantly larger than the platelet inhibition in the noncompliant group 36±16 (p=0.02). We conclude that: 1. a small percent of patients, 3.2%, whose net amount of aspirin induced platelet inhibition was less than 15 may be resistant to aspirin because of mechanisms unrelated to compliance and 2. The net amount of aspirin induced inhibition of platelets is directly related to the slope of the “off” aspirin PPA stimulated aggregation curve.
Myeloablative Conditioning of Patients with Myelofibrosis with Myeloid Metaplasia Using i.v. Treosulfan and Autologous Peripheral Blood Progenitor Cell Transplantation (PBPCT) with High Doses of CD34+ Cells Results in Hematologic Responses - 32 Months Follow-Up of Three Patients and Short-Term Results of a New Study.
Authors: Fruehauf Stefan , Buss Eike C. , Toplay Julian , Kreipe Hans H. , Ho Anthony D.
Published: April 2020

Myelofibrosis with myeloid metaplasia (MMM) is a chronic myeloproliferative disorder. Myeloablation with high-dose treosulfan and autologous PBPCT provides a palliative approach. A minimum of 5 x 106 CD34+ cells/kg was collected in all patients after G-CSF-priming (16 μg/kg/d for 4 days). Myeloablation consisted of treosulfan (total dose 42 g/m2). To date we have transplanted 3 patients, all female on an individual basis. We observed a prolonged time to reconstitution of leucocytes > 1/nl post transplantation of 28 days (#1, #2) and 38 days (#3). The observation period post transplantation is 32 months now. Patient #1, who required erythrocyte transfusions twice weekly pretransplant received her last erythrocyte transfusion on day 56; her last Hb-value is 9.5 g/dl. The second patient (#2) recovered to platelet counts higher than pre transplantation (58/nl) at 3 months (143/nl) and had 125/nl at last follow-up. Pat. #3 showed a marked reduction of max. spleen size and a rise in Hb from 9 g/dl to 12 g/dl after 12 months, the last Hb value at 26 months was 7.6 g/dl and she received about 3 erythrocyte transfusions per month again. She then underwent allogeneic transplantation from her sister and is currently alive and well. Three male patients were treated in a new one-armed, multicenter study applying the aforementioned protocol. In two patients we observed again a prolonged reconstitution period of 28 (S1) and 44 (S2) days. In one patient (S1) leukocytosis and thrombocytosis resolved while transfusion dependent anemia persisted. In the second patient (S2) transfusion dependent anemia persisted. He received a backup transplantation but still remains transfusion-dependent. Patient S3 had a critical thrombocytopenia of 16/nl before transplantation and proved to be refractory to thrombocyte transfusions post-transplantation. He developed cerebral hemorrhage and died. The conditioning regimen was well tolerated and despite the prolonged aplasia period neutropenic fever was rare (median 1 day, range 0–6 days). The overall response rate was 60%, the mortality rate was 17% which is both compatible with previous data of autologous transplantation in MMM patients undergoing busulfan conditioning. This multicenter study is continuing to recruit patients (http://leukaemie.krebsinfo.de/kn_home/Studien/studie_101.html).
Phenotypic Correction of von Willebrand Disease Type 3 Blood-Derived Endothelial Cells with Lentiviral Vectors Expressing von Willebrand Factor.
Authors: De Meyer Simon F. , Vanhoorelbeke Karen , Chuah Marinee K. , Pareyn Inge , Gillijns Veerle , Hebbel Robert P. , Deckmyn Hans , Vandendriessche Thierry
Published: April 2020

Von Willebrand disease (VWD) is the most common inherited bleeding disorder, caused by quantitative (type 1 and 3 VWD) or qualitative (type 2 VWD) defects in von Willebrand factor (VWF). Gene therapy is an appealing strategy for treatment of type 3 VWD since it is caused by a single gene defect and since VWF is secreted into the circulation, obviating the need for targeting specific organs or tissues. However, development of gene therapy for VWD has been hampered by the considerable length of the VWF cDNA (8.4 kb) and the inherent complexicity of the VWF protein which requires extensive post-translational processing. The objective of this study consisted of developing a gene-based approach for VWD using lentiviral transduction of blood-outgrowth endothelial cells (BOECs) to express functional VWF. A lentiviral vector encoding the complete human VWF protein was used to transduce BOECs isolated from type 3 VWD dogs resulting in high transduction efficiencies (95.6 ± 2.2 %). These BOECs are completely deficient in VWF due to a point mutation in the VWF gene responsible for impaired VWF synthesis. Transduced VWD BOECs efficiently expressed fully functional vector-encoded VWF (4.6 ± 0.4 U/24hr per 106 cells, figure 1), with normal binding to GPIbα and collagen and normal multimeric pattern resulting in phenotypic correction of these cells, which had not been shown previously. These results indicate that BOECs are attractive target cells for gene therapy of type 3 VWD.
Generation of CMV-Specific T-Lymphocytes for Adoptive Immunotherapy: A Comparison of Artificial Antigen-Presenting Cells and Autologous Dendritic Cells Pulsed with CMV-pp65 Protein-Spanning Pentadecapeptide Pools.
Authors: Kollen Wouter J.W. , Trivedi Deepa , Stephan Matthias , Sadelain Michel , O’Reilly Richard J.
Published: April 2020

Adoptive transfer of virus-specific T-cells constitutes a promising approach for the treatment and prevention of cytomegalovirus (CMV) infections complicating allogeneic HSC transplants. Current strategies employing peptide-pulsed donor-derived dendritic cells (DCs) for antigen presentation are effective, but limited by availability of adequate numbers of DCs and the time required for their generation. We established a series of immediately accessible, replenishable, artificial antigen-presenting cells (AAPCs), consisting of mouse 3T3 cells transduced to express human B7.1, LFA-3, ICAM-1, β2-microglobulin and one HLA class I allele (Papanicolaou et al., Blood 2003). We then compared yields of CMV-peptide-specific T-cells when T-cells from HLA A0201 seropositive donors were sensitized with autologous DCs or HLA A0201 AAPCs loaded with a known HLA A0201-binding immunogenic nonamer of CMV-pp65 (NLVPMVATV) or a pool of pentadecapeptides spanning CMV-pp65, which included the 15-mer No. 123 (AGILARNLVPMVATV) containing this epitope. Peptide loading was performed in the presence or absence of serum in order to distinguish peptide editing mediated by ectopeptidases in the serum or expressed by the different APCs. Results from repeated experiments employing three HLA A0201 CMV-seropositive donors demonstrated that the yields of epitope-specific T-cells following 14–28 day sensitization with AAPCs loaded with the NLVPMVATV nonamer consistently exceeded those generated in response to peptide-loaded autologous DCs, as assessed by enumerating peptide-HLA A0201 tetramer-binding and peptide-specific IFN-γ producing T cells. High and selective yields of NLV-specific T-cells were obtained when the T-cells were sensitized with AAPCs or DCs loaded with the CMV-pp65-spanning pool of 15-mers in the presence or absence of serum suggesting that ectopeptidases expressed on the DCs and AAPCs appropriately clip and edit the 15-mers after binding to HLA A0201. Even higher yields were achieved when the T-cells were sensitized with AAPCs transduced to express the full length CMV-pp65 protein, confirming the potential of the mouse-derived AAPCs to process this protein and load immunogenic epitopes on HLA A0201. T-cells generated in response to peptide-loaded or transduced AAPCs also specifically lysed nonamer-loaded HLA A0201 expressing targets. Taken together, these studies indicate that mouse-derived AAPCs can efficiently process endogenous protein and edit exogenous 15-mer peptides for binding and presentation by the single expressed HLA allele. Use of these AAPCs loaded with overlapping peptides or transduced to express an immunogenic protein may thus provide advantages in terms of immunogenicity, immediate accessibility and broad applicability for rapid production of antigen-specific T-cells for adoptive immunotherapy.
Usefulness of Peripheral Blood Smears in Identifying the Causes of Anemia in Adults.
Authors: McPhedran Peter , Hall Robert B.
Published: April 2020

Anemia is common in hospital patients, being found in about half of the automated complete blood counts (CBCs) done on adults aged 20+ at our hospital. Often the reason for the anemia is immediately apparent (post-operative state, end stage renal disease without erythropoietin treatment), but often it is not. Many tests are available for the differential diagnosis of the causes of the anemia. Morphologic evaluation of a Wright-stained blood smear by a skilled observer is labor intensive, but sometimes useful in the differential diagnosis of anemia. When unexplained anemia is identified in a patient on the Medical Service at our teaching hospital, the ward team may also send an intern or a medical student to check out the smear. The potential diagnostic yield of any of these evaluations (the skilled observer, the intern, the student) is unknown. We did a prospective evaluation of 202 consecutive adults with initially unexplained anemia (Hb <12 in men, <11 in women). Using accepted, pre-established criteria for etiologic diagnosis of the causes of anemia, and available data (plus additional smear evaluations by ourselves, ferritins, free erythrocyte protoporphryns, and a few other tests) we felt we were able to establish the causes of anemia in 86% of the patients. We also referred to standard morphologic criteria for diagnosing specific blood disorders in order to see how much could be learned from the blood smears of these patients, alone. Of 147 patients whose blood smears were of acceptable quality, 30 (21%) of the blood smears were diagnostic (or close to diagnostic), and an additional 46% were supportive of the correct diagnosis. For example, iron deficiency anemia was considered the morphologic diagnosis if the blood smear showed severe hypochromia, “pencil-form” elliptocytes, and thrombocytosis. Hypochromia alone would be considered supportive of the diagnosis of iron deficiency, but require more consideration of alternatives (thalassemia trait, chronic disease, etc). Thus, in evaluating anemia in adults, a good peripheral blood smear, carefully evaluated by a trained observer, is likely to be diagnostic, or very helpful, in 20% or more of the patients.
Risk Reduction of Chronic Graft Versus Host Disease by Anti-CD20 Treatment (Rituximab).
Authors: Van Hoef Marlies E.H.M.
Published: April 2020

Introduction:

Chronic graft versus host disease (cGVHD) occurs in 30–50% of adults transplanted by non-myeloablative (reduced intensity conditioning) allogeneic transplant, whereas it is rare in children. The syndrome complex manifests itselves as auto-immune disease. Dysregulation of B-cell function has also been reported in auto-immune diseases. This is a rationale for B-cell depletion.

We considered that B-cell depletion might have a therapeutic effect in cGVHD and reduce the risks associated with cGVHD. Based on these assumptions we applied risk reduction principles and defined a plan for cure of cGVHD.

Methods:

To identify the risks associated with B-cell depletion in cGVHD we performed a medline search on anti-CD20 or B-cell depletion or rituximab and cGVHD. The publications were analyzed and those applicable to the topic evaluated. Clinical research methods were applied to define a plan for risk reduction of cGVHD.

Results:

The medline search revealed that rituximab was used for B-cell depletion in cGVHD and in auto-immune diseases. In advanced cGVHD complete and partial remissions of a variety of manifestations of cGVHD were reported. Complete remissions could also be induced early during the disease course of single manifestations of cGVHD not responsive to immune suppressive treatment or as initial treatment. The dose of rituximab varied from 1 to 4 weekly courses of rituximab 375 mg/m2, with repetitions of this schedule in advanced disease to induce complete response. During reported follow-up responding manifestations did not recur. Rituximab treatment was well tolerated with appropriate anti-allergic prophylaxis. The results support the concept that anti-CD20 treatment might cure cGVHD manifestations at first diagnosis; it is too early to define whether it also serves as prophylactic for development of new manifestations. The observations were discussed with the company marketing rituximab and a study plan was designed. This study plan is being discussed with study centers for risk reduction of cGVHD by rituximab. The risk reduction plan will be presented
Canadian Variation in Blood Product Demand Is Influenced by Provincial Rates of Coronary Artery Bypass Graft Surgery.
Authors: Pendergrast Jacob M.
Published: April 2020

BACKGROUND: Despite similarities in demography and health care delivery, Canadian provinces exhibit significant variation in population adjusted-rates of blood product usage. A study was conducted to identify factors which underlie this variation.

METHODS: Potential drivers of blood product consumption were identified through literature review, and then mapped to parameters for which provincial data were available from analytic reports published by the Canadian Institute for Health Information. Blood product issues per province for the year 2001 were obtained from annual reports published by Canadian Blood Services and HémaQuébec. All data were controlled for provincial population using data from Statistics Canada. Linear regressions for each parameter were calculated separately for erythrocyte, plasma and platelet issues. Variables that were statistically significant by univariate regression analysis were subjected to multiple regression modeling.

RESULTS: Provincial variation in erythrocyte demand was statistically correlated with the following by univariate regression analysis: population density, coronary artery bypass graft (CABG) procedures and peptic ulcer diagnoses. By multivariate analysis, only CABG procedures continued to have a statistically significant correlation. For plasma issues, univariate analysis revealed average physician age, average specialist age, and CABG procedures as statistically significant drivers. However, by multivariate analysis only CABG procedures retained statistical significance. For platelet issues, average specialist age, cardiovascular procedures, CABG procedures, and number of full-time equivalent general surgeons were statistically significant drivers of provincial variation. However, none of these variables retained statistical significance by multiple regression modeling.

CONCLUSIONS: The population-adjusted rate of coronary artery bypass procedure graft surgery was a statistically significant driver of provincial variation in erythrocyte and plasma issues in the year 2001, a finding which may be of use in demand forecasting.
Characteristics and Survival of 29 Patients with Sezary Syndrome.
Authors: Sampogna Francesca , Frontani Marina , Baliva Giannandrea , Abeni Damiano , Lombardo Giuseppe A. , Monopoli Alessandro , Barbieri Claudio , Benucci Roberto , Marrazza Giuseppe , Alvetreti Gabriele , Didona Biagio , Scala Enrico , Russo Giandomenico
Published: April 2020

We created a relational database of patients with cutaneous T-cell lymphoma (CTCL) to collect in a standardised fashion, anagraphic variables, clinical history, clinical, histological, haematological, and immunological information on CTCL patients hospitalised at IDI-IRCCS, Rome, Italy. At present, there are data on 424 patients, hospitalised from 1983 to July 2005. Active follow-up is performed yearly to ensure a standardised ascertainment of survival time. For deceased patients, the actual date of death (for all causes) is recorded, while surviving patients are censored at the date of last contact.

The database includes 29 patients with Sezary syndrome (SS). Follow-up times ranged from 0 to 105 months. At first hospitalisation the median values of cells/mL were: white blood cells (WBC) 8750, neutrophils 4250, eosinophils 140, basophils 120, lymphocytes 2760, monocytes 500, CD3+ 2780, CD4+ 2431, CD8+ 192, CD19+ 96. Seventeen patients were deceased. We included in the Kaplan-Meier survival analysis only patients who were diagnosed before July 2004 (n=26). Median survival time from diagnosis was 52 months. No significant differences were observed in mortality for WBC (cutoff 9000 cells/uL), neutrophils (cutoff 4500 cells/uL), basophils (cutoff 200 cells/uL), lymphocytes (cutoff 3000 cells/uL), monocytes (cutoff 500 cells/uL), CD3+ (cutoff 2000 cells/uL), CD4+ (cutoff 2000 cells/uL), CD8+ (cutoff 200 cells/uL), CD19+ (cutoff 70 cells/uL). A lower survival was observed for patients with eosinophils <200 cells/uL (p=.08).

Survival in patients with SS does not seem to be influenced by haemathologic parameters. However, patients with long-term survival (>90 months) are observed, and their characteristics should be further investigated.

Survival analysis of 26 patients with Sézary syndrome, Rome, Italy, 1991–2004. Survival analysis of 26 patients with Sézary syndrome, Rome, Italy, 1991–2004.
Economical Analysis of Implementation of External Quality Controls in a General Hospital Hematology Laboratory.
Authors: Recasens Valle , Rubio-Martinez Araceli , Giraldo Pilar , Lucia Jose F. , Dobon Manuela , Montanes Maria A. , Bustamante Eduardo , Garcia-Mata Juan R. , Portillo Juan R. , Giralt Manuel
Published: April 2020

Introduction: The Spanish rule UNE-EN ISO 15189 concerning to Clinical Laboratory establish the realization of external quality controls (EQC) in order to compare results between laboratories. In 2004, external quality controls programs were initiated in our laboratory provided by the AEHH and SEQC.

Aim: To analyze the increase of cost in each determination when we apply these quality measures.

Material and methods: Period of study January 2004– December 2004. Encoded information obtained from ModulabWin informatics system. Determinations provided by AEHH: general hematology (RBC, Hb, HTC, MCV, MHC, HMCC, WBC, reticulocytes, cellular morphology, platelets, protrombine time, partial tromboplastine time, fibrinogen) AT III, F VIII, Hb A2 and F, automated reticulocytes, activated protein C resistance, differential count of WBC: (neutrophiles, monocytes, lymphocytes, eosinophils, basophiles), erythrocyte sedimentation rate (ESR). Determinations provided by SEQC: total proteins, albumin, complement C3 and C4, IgA, IgG, IgM, transferrine, iron serum levels, haptoglobine, ferritine, reactive protein C, b 2-microglobuline. For each ECQ with only one specimen for a unique determination the increase of costs was calculated as follow: EQC cost / total number of determinations. For each EQC with more than one determination the increase of costs was calculated as follow: (EQC cost / total number of parameters)/ total number of determinations.

Results: In ECQ with only one specimen for a unique determination, the increase of cost for each determination was ESR: 0.0033€, AT III: 0,1051€, FVIII: 1.79€, automated reticulocytes: 0,0334€, differential count of WBC 0,0033€. In EQC with more than one determination the increase of cost: were: Albumin and total proteins 0,0030€; C3 and C4: 0,0473€; activated protein C resistance: 0,0059€; IgA, IgG, IgM and b-2 microblobuline: 0,0030€; iron serum levels, Transferrine and Ferritine: 0,0025€; Haptoglobine: 0,0057€; RBC, Hb, HTC MCV, MHC, HMCC, WBC, platelets: 0,0003€ and for protrombine time, partial tromboplastine time and fibrinogen: 0,0007€.

Conclusions: The EQC for F VIII has been the more expensive control related to the number of determinations, however it is justified because our laboratory is reference center for the area. The incorporation of EQC hardly has increase the global costs of different determinations in a General Hematology Laboratory, nevertheless the quality, reproducibility, validity and security of results obtained with this procedure justify the slight increase of costs. This work has been partially supported by a grant from Aragon Health Service.
Study of Treatment Patterns and Consequences of Immune Thrombocytopenic Purpura (ITP).
Authors: Fisher Maxine D. , Yang Grace , Burawski Lauren P. , Grotzinger Kelly M. , Provan Andrew B. , Barron John J.
Published: April 2020

Standard treatment strategies for chronic immune thrombocytopenic purpura (ITP) in adults include oral steroids, high dose IVIG (intravenous immunoglobulin), Anti-D, or splenectomy (in refractory cases). Although guidelines have been developed, optimal and most frequently applied treatment strategies remain uncertain.

In this retrospective administrative claims database study, patient demographics, comorbidities, and treatment characteristics were captured to assist in determining burden of illness on patients with chronic ITP. Patients from five US health plans with medical claims for ITP (ICD-9CM codes 287.3 or 287.5) between 1/1/00 – 2/29/04 were included if they had continuous health plan enrollment 6 months prior to and 12 months after an index diagnosis of ITP. To be considered as chronic, patients also were required to have ≥1 additional drug claims or ITP diagnoses claim(s) six months after the index. Patients with codes for HIV, hepatitis, chemotherapy, or HIT were excluded.

A total of 3,743 patients met the study criteria; 53% were females (n=1,969). Mean age was 57 ± 16.9. Patients had a mean follow-up duration of 976 ± 441 days. Mean Deyo-Charlson Comorbidity Indices were 1.2 ± 1.9 pre-index, 2.0 ± 2.4 post-index of a maximum score of 21. Hypertension (62% pre-index; 51% post-index) and coronary heart disease (38% pre-index; 28% post-index) were common comorbidities. Of concomitant medications, most patients were taking antibiotics pre-index (n=3,008) as well as post-index (n=3,316). Twenty-nine percent of patients were hospitalized during follow-up with ITP recorded during the inpatient stay. The mean number of days to drug treatment was 378 ± 376. Among patients who received pharmacological treatment (n=1,834), the most frequently used first-line and second-line therapies were oral corticosteroids (n=1,742) and cyclosporine (n=75), respectively. Among patients treated with oral corticosteroids with ≥1 year follow up, the average time on treatment was 95 days. Further evaluation of the period between the index ITP diagnosis and drug therapy, and pharmacologic interventions for ITP during hospitalizations are underway.
Dyrk-3 Kinase Is an Erythroid Cell-Intrinsic Attenuator of Stress Erythropoiesis.
Authors: Bogacheva Olga , Bogachev Oleg , Menon Madhu , Valoret Elizabeth , Karur Vinit , Prosser H. , Creasy Caretha , Erickson-Miller Connie , Wojchowski Don
Published: April 2020

During anemia, red cell production depends critically upon erythropoietin and is also supported by Gas-6, oncostatin-M, and BMP4/Madh5 as recently characterized stress erythropoietic factors. Considerably less is understood concerning factors that limit this process. Via gene disruption, and as studied in four functional paradigms, stress erythropoiesis presently is shown to be attenuated by Dyrk-3 kinase. Tissue-wide RT-PCR analyses confirmed Dyrk-3 expression selectively in erythroid progenitor cells, and testes. In Dyrk-3−/− mice, reproductive capacity and steady-state hematopoiesis were normal (with the exception of modest decreases in splenic BFUe). In response to phenylhydrazine- induced anemia, however, reticulocyte production in the absence of Dyrk-3 was sustained, and was elevated to 270% of wild-type controls. Reticulocyte levels (and hematocrits) in Dyrk-3−/− mice also were elevated significantly over controls following 5′-fluorouracil- induced anemia, and a two-fold advantage for Dyrk-3−/− CD71highTer119pos cell formation was observed in short-term bone marrow transplant experiments. These outcomes indicated enhanced rates of Dyrk-3−/− erythroblast development, and using a unique medullary erythroblast expansion system, accelerated development of Dyrk-3−/− Kitpos proerythroblasts to maturing CD71high Ter119pos cells was observed. Given these data, Dyrk-3 is proposed to selectively limit (and possibly apportion) red cell production during stress erythropoiesis.