scholarly journals Coadministration of bone marrow cells and an inhibitor of apoptosis (Z-VAD) promotes bone marrow cell survival and neurological functional recovery after focal cerebral ischemia in adult rat

Stroke ◽  
2001 ◽  
Vol 32 (suppl_1) ◽  
pp. 379-380
Author(s):  
Yi Li ◽  
Jieli Chen ◽  
Michael Chopp
Keyword(s):  
Bone Marrow ◽  
Cerebral Ischemia ◽  
Bone Marrow Cells ◽  
Inhibitor Of Apoptosis ◽  
Apoptotic Cells ◽  
Group 3 ◽  
Group 2 ◽  
Adhesive Removal ◽  
Group 1 ◽  
Marrow Cells

P222 Most bone marrow cells (BMCs) transplanted into the ischemic brain of adult rodents die shortly after they are grafted, similar to the 90%-95% of the fetal cells that die after transplantation into Parkinson s patients. We tested whether coadministration of BMCs and Z-Val-Ala-DL-Asp-fluoromethylketone (Z-VAD), an inhibitor of apoptosis, into the ischemic boundary zone of the striatum and the cortex in rat brain promotes BMC survival. Adult Wistar rats were subjected to transient (2 h) middle cerebral artery occlusion (MCAo). At 1 d after ischemia, saline (n=9, Group 1); BMCs (1x10 6 in 10 :l, n=8, Group 2); or BMCs with Z-VAD (50 nM/ml, n=4, Group 3) were injected into brain. BMCs were harvested from donor adult rats injected with bromodeoxyuridine (BrdU, as a tracer). Rats were subjected to rotarod-motor and adhesive-removal somatosensory functional tests before MCAo and at 1 and 7 d after MCAo. Rats in Group 3 exhibited significant improvement (10.3±2.6 seconds, p<0.05) on the adhesive-removal test at 7 d, compared with those in Group 1 (29.3±9.4 seconds) and Group 2 (21.3±5.5 seconds), respectively. Immunohistochemistry staining was employed to identify BrdU-BMCs, and TUNEL staining was used to identify in situ DNA fragmentation of apoptotic cells. Even though the infarct volume in Group 3 (29.9±9.2%) did not change significantly, compared with Group 1 (34.3±4.0%) and Group 2 (26.6±3.8%); the number of BrdU-BMCs (88,200±7,400, ∼8.8% of 10 6 transplanted cells) increased significantly (p<0.05) in rats in Group 3, compared with that in Group 2 (31,700±9,100, ∼3.2% of 10 6 cells) at 7 d. In the grafting areas, apoptotic cells were less clustered (<90 vs.>240 per region) and apoptotic cells were significantly decreased (12.3±1.7/mm 2 vs. 25.9±2.1/mm 2 , ∼47%, p<0.05). Our data suggest that intracerebral coadministration of bone marrow cells and inhibitors of apoptosis enhance cellular survival of bone marrow cells and improves neurological functional recovery after cerebral ischemia.

scholarly journals Investigation of regenerative and tiss ue-specific activity of tot al RNA of bone marrow cells

2018 ◽  
Vol 20 (3) ◽  
pp. 64-69
Author(s):  
Z. Z. Gonikova ◽  
A. O. Nikolskaya ◽  
L. A. Kirsanova ◽  
M. Yu. Shagidulin ◽  
N. A. Onishchenko ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Specific Activity ◽  
Cell Activity ◽  
Total Rna ◽  
Liver And Kidney ◽  
Group 3 ◽  
Group 2 ◽  
Group 1 ◽  
Marrow Cells

Aim. To establish the ability of the total RNA extracted from the body’s bone marrow cells (BMCs), in which liver tissue was damaged, to serve as a carrier of targeted regenerative signals to this organ.Materials and methods. By method of adoptive transfer in rats (n = 37) the  mitotic and proliferative activity of liver and kidney cells were studied in intact  recipients after intraperitoneal injection: the mononuclear BMCs – 2,5×106;  5,0×106; 3,5×107 cells – group 1 and the total RNA of the same BMCs  (30μg/100g of weight) – group 2 from donors in 12 hours after 70–75% of  hepatectomy; in group 3 (control), a saline solution was injected. RNA from  BMCs was extracted by the method developed by the «Evrogen» firm (Russia) with the reagent Extract RNA.Results. In group 2 in 48 and 72 h. there was the increasing of mitotic and  proliferative cell activity in the liver, but not in the kidneys (control of the  specificity of regenerative signals); in group 1 there was no transfer of  regenerative signals to these organs.Conclusion. The authors believe that the total RNA from BMCs, activated by hepatectomy, accumulates targeted (hepatospecific) regeneration signals, but  they are perceived only when RNA has been obtained by the damaged tissue.

Serum Transferrin Receptor Value as a Universal Indicator of Erythropoietic Activity.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3682-3682
Author(s):  
Young Soo Lee ◽  
Chul Soo Kim ◽  
Jong Weon Choi
Keyword(s):  
Bone Marrow ◽  
Transferrin Receptor ◽  
Bone Marrow Cells ◽  
Reticulocyte Count ◽  
Serum Transferrin Receptor ◽  
Group 4 ◽  
Universal Indicator ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Abstract The serum transferrin receptor (sTfR) is thought a sensitive and quantitative parameter of tissue iron deficiency as well as an indicator of erythropoietic activity. This study was aimed at the verification of a hypothesis that sTfR is a general indicator of erythropoiesis regardless whatever the cause is. A total of 173 patients in heterogeneous diseases who underwent bone marrow study as a workup for anemia were measured for sTfR, reticulocyte maturity index (RMI), erythroid element proportion of bone marrow cells, and other hematologic parameters (hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration, red cell distribution width, absolute reticulocyte count). By immunoenzymometric method sTfR was measured using IDeATMcTfR kids (Orion Diagnostica, Orion, Finland). Reticulocyte count and proportion was measured manually by one expert examiner after standard blood smear and stain. Reticulocyte subpopulation was automatically analyzed by flow cytometry using R-3000 TM (Sysmex, TOA, Japan). RMI was calculated from the equation of (medium fluorescent reticulocyte fraction + high fluorescent reticulocyte fraction) X 100 / low fluorescent reticulocyte fraction. Correlation analysis was done among the variables including sTfR, RMI, erythroid element proportion of bone marrow cells, and other hematologic parameters using SAS 6.12 soft ware. The analysis was carried out for the whole 173 patients to see the general trends and repeated for 4 groups of disease category, arbitrarily divided to group 1 (n=33, iron deficiency or or disease with no predisposition to anemia of chronic disease), group 2 (n=53, hematologic malignancies), group 3 (n=44, solid tumors), and group 4 (n=43, chronic or infectious disease) to see if the trends may be affected by specific diseases. The results showed a solid correlation of sTfR with RMI as well as erythroid precursors in bone marrow, not only in the whole patient population (e.g. sTfR vs RMI, R=0.587, p=0.0001) but also in individual groups (e.g. sTfR vs RMI, R=0.48, p=0.005 in group 1, R=0.69, p=0.0001 in group 2, R=0.58, p=0.0001 in group 3, R=0.81, p=0.0001 in group 4). These findings indicated the significance of sTfR is valid under any clinical setting as a universal indicator of hematopoietic activity. The sTfR can be used as a useful parameter for monitoring of erythropoiesis in a variety disease.

Abelson virus potentiates long-term growth of mature B lymphocytes

1986 ◽  
Vol 6 (1) ◽  
pp. 183-194
Author(s):  
L A Serunian ◽  
N Rosenberg
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
B Cell ◽  
Cell Lines ◽  
Bone Marrow Cells ◽  
Mouse Bone Marrow ◽  
Phenotypic Characteristics ◽  
Group 2 ◽  
Group 1 ◽  
Marrow Cells

Abelson murine leukemia virus (A-MuLV) infection of mouse bone marrow cells usually leads to transformation of pre-B cells. However, when the environment is modified by the continuous presence of lipopolysaccharide (LPS), two novel types of membrane immunoglobulin (mIg)-positive B cell lines are generated. Because the cells which give rise to these cell lines copurify with mIg-positive bone marrow cells, the cell lines arise as a result of A-MuLV interaction with a new type of in vitro target cell. The cell lines generated fall into two groups which differ in several phenotypic characteristics. Group 1 cells are more differentiated than the typical pre-B cell transformant in that they synthesize mIgM and appear to resemble virgin B cells. The group 1 cells do not secrete immunoglobulin and are independent of LPS for growth. In addition, these cell lines synthesize the Abelson P160 protein, contain integrated abl proviral DNA, and are highly tumorigenic in syngeneic animals. The group 2 cell lines differ markedly from both the group 1 cells and from typical, pre-B cell A-MuLV transformants. These cells are mIgG positive and secrete large amounts of immunoglobulin into the culture medium. The cell lines are comprised of both adherent and nonadherent cells and do not synthesize P160 or contain integrated v-abl sequences. The group 2 cells are nontumorigenic in syngeneic animals and require LPS for growth and viability. Both types of cells have remained in culture for over 2 years with no changes in their phenotypic characteristics. This A-MuLV infection system and the novel mIg-positive cell lines may serve as useful models for studying biochemical and molecular properties of mature B cells.

scholarly journals Abelson virus potentiates long-term growth of mature B lymphocytes.

1986 ◽  
Vol 6 (1) ◽  
pp. 183-194 ◽  
Author(s):  
L A Serunian ◽  
N Rosenberg
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
B Cell ◽  
Cell Lines ◽  
Bone Marrow Cells ◽  
Mouse Bone Marrow ◽  
Phenotypic Characteristics ◽  
Group 2 ◽  
Group 1 ◽  
Marrow Cells

Abelson murine leukemia virus (A-MuLV) infection of mouse bone marrow cells usually leads to transformation of pre-B cells. However, when the environment is modified by the continuous presence of lipopolysaccharide (LPS), two novel types of membrane immunoglobulin (mIg)-positive B cell lines are generated. Because the cells which give rise to these cell lines copurify with mIg-positive bone marrow cells, the cell lines arise as a result of A-MuLV interaction with a new type of in vitro target cell. The cell lines generated fall into two groups which differ in several phenotypic characteristics. Group 1 cells are more differentiated than the typical pre-B cell transformant in that they synthesize mIgM and appear to resemble virgin B cells. The group 1 cells do not secrete immunoglobulin and are independent of LPS for growth. In addition, these cell lines synthesize the Abelson P160 protein, contain integrated abl proviral DNA, and are highly tumorigenic in syngeneic animals. The group 2 cell lines differ markedly from both the group 1 cells and from typical, pre-B cell A-MuLV transformants. These cells are mIgG positive and secrete large amounts of immunoglobulin into the culture medium. The cell lines are comprised of both adherent and nonadherent cells and do not synthesize P160 or contain integrated v-abl sequences. The group 2 cells are nontumorigenic in syngeneic animals and require LPS for growth and viability. Both types of cells have remained in culture for over 2 years with no changes in their phenotypic characteristics. This A-MuLV infection system and the novel mIg-positive cell lines may serve as useful models for studying biochemical and molecular properties of mature B cells.

scholarly journals The Positive Impact of Donor Bone Marrow Cells Transplantation into Immunoprivileged Compartments on the Survival of Vascularized Skin Allografts

2021 ◽  
Vol 69 (1) ◽  
Author(s):  
Arkadiusz Jundziłł ◽  
Aleksandra Klimczak ◽  
Erhan Sonmez ◽  
Grzegorz Brzezicki ◽  
Maria Siemionow
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Allogeneic Bone ◽  
Group 4 ◽  
Donor Bone Marrow ◽  
Group 3 ◽  
Group 2 ◽  
Marrow Cells

AbstractUsing the vascularized skin allograft (VSA) model, we compared the tolerogenic effects of different allogeneic bone marrow transplantation (BMT) delivery routes into immunoprivileged compartments under a 7-day protocol immunosuppressive therapy. Twenty-eight fully MHC mismatched VSA transplants were performed between ACI (RT1a) donors and Lewis (RT11) recipients in four groups of seven animals each, under a 7-day protocol of alfa/beta TCRmAb/CsA (alpha/beta-TCR monoclonal antibodies/Cyclosporine A therapy). Donor bone marrow cells (BMC) (100 × 106 cells) were injected into three different immunoprivileged compartments: Group 1: Control, without cellular supportive therapy, Group 2: Intracapsular BMT, Group 3: Intragonadal BMT, Group 4: Intrathecal BMT. In Group 2, BMC were transplanted under the kidney capsule. In Group 3, BMC were transplanted into the right testis between tunica albuginea and seminiferous tubules, and in Group 4, cells were injected intrathecally. The assessment included: skin evaluation for signs and grade of rejection and immunohistochemistry for donor cells engraftment into host lymphoid compartments. Donor-specific chimerism for MHC class I (RT1a) antigens and the presence of CD4+/CD25+ T cells were assessed in the peripheral blood of recipients. The most extended allograft survival, 50–78 days, was observed in Group 4 after intrathecal BMT. The T cells CD4+/CD25+ in the peripheral blood were higher after intrathecal BMC injection than other experimental groups at each post-transplant time point. Transplantation of BMC into immunoprivileged compartments delayed rejection of fully mismatched VSA and induction of robust, donor-specific chimerism.

scholarly journals Donor Recipient Chimeric Cells Induce Chimerism and Extend Survival of Vascularized Composite Allografts

2021 ◽  
Vol 69 (1) ◽  
Author(s):  
Joanna Cwykiel ◽  
Arkadiusz Jundzill ◽  
Aleksandra Klimczak ◽  
Maria Madajka-Niemeyer ◽  
Maria Siemionow
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Fluorescent Microscopy ◽  
Study Endpoint ◽  
Solid Organ ◽  
Post Transplant ◽  
Group 4 ◽  
Cell Based Therapy ◽  
Group 3 ◽  
Group 1

AbstractThis study evaluated the efficacy of donor recipient chimeric cell (DRCC) therapy created by fusion of donor and recipient derived bone marrow cells (BMC) in chimerism and tolerance induction in a rat vascularized composite allograft (VCA) model. Twenty-four VCA (groin flaps) from MHC-mismatched ACI (RT1a) donors were transplanted to Lewis (RT1l) recipients. Rats were randomly divided into (n = 6/group): Group 1—untreated controls, Groups 2—7-day immunosuppression controls, Group 3—DRCC, and Group 4—DRCC with 7-day anti-αβTCR monoclonal antibody and cyclosporine A protocol. DRCC created by polyethylene glycol-mediated fusion of ACI and Lewis BMC were cultured and transplanted (2–4 × 106) to VCA recipients via intraosseous delivery route. Flow cytometry assessed peripheral blood chimerism while fluorescent microscopy and PCR tested the presence of DRCC in the recipient’s blood, bone marrow (BM), and lymphoid organs at the study endpoint (VCA rejection). No complications were observed after DRCC intraosseous delivery. Group 4 presented the longest average VCA survival (79.3 ± 30.9 days) followed by Group 2 (53.3 ± 13.6 days), Group 3 (18 ± 7.5 days), and Group 1 (8.5 ± 1 days). The highest chimerism level was detected in Group 4 (57.9 ± 6.2%) at day 7 post-transplant. The chimerism declined at day 21 post-transplant and remained at 10% level during the entire follow-up period. Single dose of DRCC therapy induced long-term multilineage chimerism and extended VCA survival. DRCC introduces a novel concept of customized donor-recipient cell-based therapy supporting solid organ and VCA transplants.

Mobilization of Stem Cells from the Bone Marrow Is Required for Neovascularization of Ischemic Limbs in Mice.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4233-4233
Author(s):  
Jeong-A Kim ◽  
Chang -Hoon Lee ◽  
Jin-A. Yoon ◽  
Woo-Sung Min ◽  
Chun-Choo Kim
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hind Limb ◽  
Limb Ischemia ◽  
Hind Limb Ischemia ◽  
Ischemic Limb ◽  
Group 3 ◽  
Group 2 ◽  
Surgically Induced ◽  
Group 1

Abstract We examined whether the injection of bone marrow mononuclear cells (BM-MNCs) or mesenchymal stem cells (MSCs) might augment angiogenesis and collateral vessel formation in a mouse model of hind limb ischemia. C57BL/6 BM-MNCs were isolated by centrifugation through a Histopaque density gradient and MSCs were obtained from C57BL/6 bone marrow and cultured in low-glucose DMEM media. Unilateral hind limb ischemia was surgically induced in C57BL/6 mice (control; n=4), and autologous BM-MNCs (Group 1; n=4, 1.8±0.2 x107/animal) or MSCs (Group 2; n=4, 1.0±0.14 x106/animal) or BM-MNCs and MSCs (Group 3; n=4, 2.3±0.1 x107 and 1.1±0.21 x106/animal) were transplanted into the ischemic tissue. Six weeks after transplantation, the group 1, group 2 and group 3 had a higher capillary/muscle ratio (0.82±0.12 vs 0.85±0.08 vs 0.97 ±0.03) than control (0.46±0.12, p&lt;0.05) (Fig. 1). This result suggested that direct local transplantation of autologous BM-MNCs or MSCs seems to be a useful strategy for therapeutic neovascularization in ischemic tissues. Next, we evaluated whether bone marrow derived stem cells were participated in the process of local injected stem cells forming new vessels. In general, mobilizing stem cells from bone marrow to local site, MMP-9 has been known as an important molecule. So we used the MMP-9 deficient KO mice and wild type, 129SvEv mice were used in the experiments. Autologous BM-MNCs and MSCs were transplanted into the ischemic limb in MMP-9 (−/−) (n=4) after unilateral hind limb ischemia was surgically induced and then the same experiments was done in MMP-9 (+/+) mice (n=4). The number of the injected BM-MNCs and MSCs was 2.2±0.05 x107 and 0.87±0.17 x106/animal in MMP-9 (−/−). And the number of the injected BM-MNCs and MSCs was 2.1±0.17 x107 and 0.98±0.09 x106/animal in MMP-9 (+/+). No difference was seen in the BM-MNCs and MSCs were injected or not (0.52±0.07 vs 0.49±0.03,) in MMP-9 (−/−). But, in the case that BM-MNCs and MSCs were injected, the higher capillary/muscle ratio was seen in MMP-9 (+/+) compared to control (0.86 ±0.09 vs 0.49±0.03, P&lt;0.05) (Fig 2). This data indicated that the mobilization of bone marrow derived stem cells would have an important role in the neovasculrization although the stem cells were injected directly into the muscle of ischemic limb. Figure Figure Figure Figure

Phosphorus-31 Magnetic Resonance Spectroscopy of Cerebral Ischemia in Cats

Neurosurgery ◽  
1990 ◽  
Vol 27 (2) ◽  
pp. 240-246 ◽  
Author(s):  
Hidenori Kobayashi ◽  
Minoru Hayashi ◽  
Hirokazu Kawano ◽  
Yuji Handa ◽  
Masanori Kabuto ◽  
...  
Keyword(s):  
Cerebral Ischemia ◽  
Magnetic Resonance ◽  
Magnetic Resonance Spectroscopy ◽  
Inorganic Phosphate ◽  
Left Subclavian Artery ◽  
Resonance Spectroscopy ◽  
Brachiocephalic Trunk ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Abstract The energy metabolism of the brain was measured in three types of ischemic models in the cat using phosphorus-31 magnetic resonance spectroscopy. The cerebral ischemia was produced as follows. In Group 1, two balloons were inflated in the left subclavian artery and the brachiocephalic trunk. In Group 2, the left middle cerebral artery was occluded through a transorbital approach. A combination of the two was employed in Group 3. Phosphorus-31 magnetic resonance spectra were obtained serially during 2 hours of ischemia. Immediately after occlusion, peaks of phospho-creatine and adenosine triphosphate decreased, whereas the peak of inorganic phosphate increased and split in two. Intracellular pH determined by chemical shift of the inorganic phosphate peak decreased. These changes were more pronounced in Group 3 when compared with the other groups. Histological study showed no infarction in Group 1 and infarcted areas in Groups 2 and 3. The size of the infarcted area in Group 3 was larger than that in Group 2. These results suggest that the model of middle cerebral artery occlusion potentiated with the occlusion of the brachiocephalic trunk and the left subclavian artery by balloon catheters is a reliable stroke model and that phosphorus-31 magnetic resonance spectroscopy is useful to understand the pathophysiological state of cerebral ischemia in vivo.

scholarly journals Feline Leukemia Virus-associated Enteritis—A Condition with Features of Feline Panleukopenia

1987 ◽  
Vol 24 (1) ◽  
pp. 1-4 ◽  
Author(s):  
M. Reinacher
Keyword(s):  
Bone Marrow ◽  
Leukemia Virus ◽  
Feline Leukemia Virus ◽  
Lymphoid Tissues ◽  
Post Mortem ◽  
The Third ◽  
Histopathological Alterations ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Infection with feline leukemia virus (FeLV) was demonstrated immunohistologically in 218 necropsied cats suffering from enteritis. The animals were divided into three groups according to histopathological criteria. The first group exhibited the signs of feline panleukopenia in intestine, lymphoid tissues, and bone marrow. Only 1.6% of these animals were FeLV-infected. The animals of the second group had histopathological alterations as seen in cats suffering from feline panleukopenia, but these were found only in the intestine and not in lymphoid tissues or bone marrow. Of these 71.9% were infected with FeLV. The third group consisted of all other cats suffering from enteritis of which 6.3% were FeLV-positive. The association between FeLV infection and the lesions seen in the animals of group 1 (feline panleukopenia) and group 3 (other types of enteritis) is statistically not significant whereas the alterations exhibited by the cats of group 2 are significantly FeLV-associated. Cats with FeLV-associated enteritis (group 2) are of a mean age of about 2.5 years and are significantly older than animals with feline panleukopenia which are of a mean age of about half a year. Thus a FeLV-associated enteritis exists as a histopathologically recognizable condition which sometimes might be mistaken for feline panleukopenia in routine post-mortem investigations.

WT1 Levels At Diagnosis and POST-Induction Provide Prognostic Information in Adult De Novo AML. Results From the Spanish Cetlam Group.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2524-2524
Author(s):  
Josep F Nomdedeu ◽  
Montserrat Hoyos ◽  
Maite Carricondo ◽  
Elena Bussaglia ◽  
Camino Estivill ◽  
...  
Keyword(s):  
Bone Marrow ◽  
De Novo ◽  
High Dose ◽  
Prognostic Impact ◽  
Post Induction ◽  
Group 3 ◽  
Group 2 ◽  
De Novo Aml ◽  
Intermediate Dose ◽  
Group 1

Abstract Abstract 2524 WT1 monitoring is an almost universal target to follow de novo AML. Its exppression in myeloid malignancies is upregulated in parallel to the blast percentage. Recently, WT1 determination has been standardized as result of an European Leukemia Net initiative. Early reports have demonstrated that the best results are obtained when peripheral blood is used to establish clinical predictions. Pediatric studies in AML have shown that raised WT1 levels after induction associate with unfavourable outcome. Despite all the mentioned, WT1 quantitation has not yet gained widespread use, in part because some AML show normal WT1 levels at diagnosis. To investigate the prognostic impact of the normalized bone marrow WT1 levels at diagnosis and post-induction in a consecutive series of de novo AML patients enrolled in the CETLAM group trials. Available bone marrow samples at diagnosis (586 cases) and post induction (367 cases) were obtained in each participating center and sent to the CETLAM repository center at the Hospital de la Santa Creu i Sant Pau for complete immunophenotype and molecular analyses. One μg of RNA was reverse transcribed to cDNA in a total reaction volume of 20μl containing Cl2Mg 5mM, 10× Buffer, DTT 10mM, dNTP's 10mM each, random hexamers 15μM, RNAsin 20 units (Promega) and 200 units of MMLV enzyme. WT1 expression levels were determined by real-time quantitative polymerase chain reaction (RQ-PCR) in an ABI PRISM 7700® Genetic Analyzer (Applied Biosystems, Foster City, CA) using the primers and conditions described by the ELN group (Cilloni et al J. Clin. Oncol 2009;27:5195-201). For WT1 copy number titration, the IPSOGEN® (Marseille, France) plasmid was employed. Results were expressed as copies and four normal bone marrow samples were used as test controls. Patients were treated between 2004 and 2011 according to the CETLAM03 protocol. Adults up to 70 years of age received induction chemotherapy with idarubicin, intermediate-dose cytarabine and etoposide, followed by consolidation with mitoxantrone and intermediate-dose ara-C. Subsequently, patients with favourable cytogenetics at diagnosis received one cycle of high-dose cytarabine.G-CSF priming during induction and consolidation was used. Patients with favorable cytogenetics and high leukocyte counts at diagnosis were treated with autologous transplantation instead of high-dose cytarabine. Furthermore, patients with a normal karyotype but an adverse molecular profile (FLT3 mutations or MLL rearrangements) were allocated to the treatment for unfavorable cases; this included allogeneic transplantation from an HLA-identical donor. Overall survival (OS) was measured from the date of enrolment until the date of death. Leukemia-free survival (LFS) for patients who achieved a CR was calculated from the date of CR to relapse or death. OS and LFS were plotted by the Kaplan-Meier method; differences between curves were analyzed by the log-rank test. The probability of relapse was calculated using cumulative incidence estimates and taking into account the competing risk of death in remission. A WT1 cut-off value of 5065.2 copies at diagnosis was obtained. Two hundred and four samples had WT1 levels greater than this value, whereas 382 samples showed levels below this cut-off. These groups had statistically different OS 55±3 vs 33±5 p<0.001, LFS 52±3 vs 30±6 p:0.004 and CIR 34±3 vs 56±6 p<0.001. As regards the post-induction results, four groups were established: Group 0 (135 patients) with WT1 levels between 0 and 17.5 copies, Group 1 (107 patients) with WT1 values ranging from 17.6 to 76 copies, Group 2 (54 patients) with WT1 between 76.1 and 170.5 copies and Group 3 (71 patients) with WT1 levels after induction greater than>170.6 copies. These groups showed statistically significant differences(p<0.001) in terms of OS: Group 0 59±4 months, Group 1 50±5 months, Group 2 45±7 months and Group 3 23±6 months. LFS was also statiscally different: Group 0: 58±4, Group 1: 46±5, Group 2: 39±8 and Group 3:19±8 (all p<0.001). Lastlly, CIR was markedly different between the four groups: Group 0:25±4, Group 1: 44±5, Group 2: 46±8 and Group 3: 68±8(p<0.001) . WT1 quantitation at diagnosis and post-induction provide a simple and well standardized measurement of the prognostic risk of adult AML patiens. Larger series need to be analyzed to ascertain whether this determination could be incorporated to initial AML risk stratification. Disclosures: No relevant conflicts of interest to declare.

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