Observation on the stability of rat myelosuppression model established by different doses of cyclophosphamide solution

Objective to investigate the stability of different doses of cyclophosphamide solution in establishing rat myelosuppression model. Methods In this study, 30 SD rats were randomly divided into control group (Group C) and model group (M1, M2, M3, M4). The rats in the model groups were injected with cyclophosphamide solution by intraperitoneal injection at doses of 30、40、(30+15) and (40+20) mg/kg respectively to establish a bone marrow suppression rat model. The changes of WBC, lym, Mon, GRA, RBC, Hb and PLT in peripheral blood were observed on the 1st, 5th and 13th day after modeling. Results （1）After modeling, the weight of rats in the model group was lower than that in the control group (P < 0.05). （2）On the 5th day, the levels of WBC, lym, mon and gra in the serum of each model group were lower than those before modeling(P < 0.05). The content of RBC in M1 and M2 groups was lower than that before modeling (P < 0.05), the content of Hb in M1, M2 and M3 groups was lower than that before modeling (P < 0.05), and the content of PLT in M1, M2, m3 and M4 groups was lower than that before modeling (P < 0.05). （3）On the 13th day, the levels of WBC, lym, mon and gra in M1 and M2 groups were higher than those on the 5th day, except for the GRA in M2 group.The levels of WBC and lym in m3 and M4 were lower than those in M1 and M2 (P < 0.05); The contents of RBC and Hb in M1 and M2 groups were lower than those before modeling (P < 0.05); Compared with M1 and M2, the content of Hb in m3 and M4 groups was higher (P < 0.05); The PLT content in the peripheral blood of M3 and M4 rats was higher than that of the 5th day, but lower than that of M1 and M2 groups (P < 0.05). Conclusion The improved method can maintain the stability of rat myelosuppression model.

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Clinical observations of bone marrow transfusion for promoting bone marrow reconstruction after chemotherapy for AIDS-related lymphoma

BMC Immunology ◽

10.1186/s12865-021-00399-8 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Yixuan Liu ◽

Suhong Xie ◽

Lei Li ◽

Yanhui Si ◽

Weiwei Zhang ◽

...

Keyword(s):

Bone Marrow ◽

Immune System ◽

T Lymphocytes ◽

Peripheral Blood ◽

Autologous Bone Marrow ◽

Control Group ◽

Peripheral Vein ◽

Significant Difference ◽

Autologous Bone ◽

Aids Related Lymphoma

Abstract Background This study investigates the effect of autologous bone marrow transfusion (BMT) on the reconstruction of both bone marrow and the immune system in patients with AIDS-related lymphoma (ARL). Methods A total of 32 patients with ARL participated in this study. Among them, 16 participants were treated with conventional surgery and chemotherapy (control group) and the remaining 16 patients were treated with chemotherapy followed by autologous bone marrow transfusion via a mesenteric vein (8 patients, ABM-MVI group) or a peripheral vein (8 patients, ABM-PI group). Subsequently, peripheral blood and lymphocyte data subsets were detected and documented in all patients. Results Before chemotherapy, no significant difference in indicators was observed between three groups of ARL patients. Unexpectedly, 2 weeks after the end of 6 courses of chemotherapy, the ABM-MVI group, and the ABM-PI group yielded an increased level of CD8+T lymphocytes, white blood cells (WBC), and platelet (PLT) in peripheral blood in comparison to the control group. Notably, the number of CD4+T lymphocytes in the ABM-PI group was significantly higher than that in the other two groups. Additionally, no significant difference in haemoglobin levels was observed before and after chemotherapy in both the ABM-MVI and ABM-PI groups, while haemoglobin levels in the control group decreased significantly following chemotherapy. Conclusions Autologous bone marrow transfusion after chemotherapy can promote the reconstruction of both bone marrow and the immune system. There was no significant difference in bone marrow recovery and reconstruction between the mesenteric vein transfusion group and the peripheral vein transfusion group.

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Abstract 239: Stem Cell Therapy Improves Critical Limb Ischemia

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.32.suppl_1.a239 ◽

2012 ◽

Vol 32 (suppl_1) ◽

Author(s):

Alaa Marzouk

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Stem Cell ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Embryonic Stem ◽

Limb Ischemia ◽

Control Group ◽

Chronic Limb Ischemia

Introduction: The journey from single cell to complex being is attributable to stem cells role. Adult stem cells originate during ontogeny & persist in specialized niches within organs. Asymmetric division of each stem cell during differentiation produces : one daughter stem cell & one daughter transit amplifying/intermediate cell having migratory properties. Forced migration of hematopoietic stem/progenitor cells (HSPC) from bone marrow into peripheral blood is called mobilization. Accumulating evidence suggests that attenuation of the chemokine stromal derived factor-1(SDF-1)-CXCR4 axis that plays a pivotal role in retention of HSPC in bone marrow (BM) results in the release of these cells from the BM into peripheral blood. Recently, adult cells have been genetically reprogrammed to an embryonic stem cell like state. Induced pluripotent stem cells (IPSCs) were similar to human embryonic stem cells in morphology, proliferative capacity, expression of cell surface antigens, & gene expression. Treatment of ischemic vascular disease of lower limbs remains a significant challenge. Unfortunately, if medical & surgical salvage procedures fail, amputation is an unavoidable result for those patients. Aim of Work: (Hypothesis) To assess the application of implantation of autologous stem/progenitor cell in the treatment of chronic limb ischemia & to evaluate the safety, efficacy & feasibility of this novel therapeutic approach. Methods: A total of 24 patients with chronic limb ischemia not eligible for arterial reconstruction or endovascular procedures were enrolled & randomized (1:1) to either the implanted group or the control group. Control group: Conventional medical therapy in the form of anti platelet therapy & vasodilators. Implanted group: Subcutaneous injection of 300μ g/day of recombinant human granulocyte colony stimulating factor (G-CSF) for 5 days to mobilize stem/progenitor cells from BM. Total leucocytic count is measured daily to follow up successful mobilization of bone marrow mononuclear cells (BMMNCs). Stem cell Harvesting After 5 days peripheral blood mononuclear cells (PBMNCs) were harvested using a cell separator. Samples from apheresis products are subjected to TLC measurement & immunophenotypic characterization of CD34+ cells by flow cytometry. The collected PBMNCs were implanted by multiple intramuscular injections into ischemic limbs. Results: There was significant increase in pain free walking distance & ankle/brachial index (ABI) & significant decreased rest pain. Effectiveness was documented by : reduced number of amputation, increase ABI & improvement of the quality of life in therapeutic group compared to control group. Conclusion: The novel therapeutic approach of PBMNCs implantation in patients with chronic limb ischemia is safe, feasible & effective in decreasing co-morbidity & rate of amputation. Safety was manifested by absence of complications during G-CSF therapy or during harvesting & injection of the stem cells. Recommendations: 1- Future studies on larger number of patients & longer follow up. 2- Controlled studies using different methods & different cell population (PBMNCs, BMMNCs or MSCs) to compare the outcome of each. 3-Studing the role of endothelial progenitor cell dysfunction in different ischemic diseases to develop successful gene therapy.

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Gengnianchun Recipe Protects Ovarian Reserve of Rats Treated by 4-Vinylcyclohexene Diepoxide via the AKT Pathway

International Journal of Endocrinology ◽

10.1155/2020/9725898 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11

Author(s):

Fangui Zhao ◽

Wenjun Wang

Keyword(s):

Ovarian Reserve ◽

Low Dose ◽

Control Group ◽

High Dose ◽

Model Group ◽

Menopausal Syndrome ◽

Primordial Follicles ◽

Akt Phosphorylation ◽

Vinylcyclohexene Diepoxide ◽

Sd Rats

Diminished ovarian reserve (DOR) refers to a decrease in the number and quality of oocytes. Western treatment of DOR does not improve the ovarian reserve fundamentally, and the effect is limited. Gengnianchun recipe (GNC) is a traditional Chinese medicine formula originally applied to treat menopausal syndrome but is also found to be effective in treating clinical DOR patients. Here we aim to examine the effect of GNC in a DOR rat model induced by 4-vinylcyclohexene diepoxide (VCD), a chemical that selectively destroys ovarian small preantral follicles, and further investigate the possible mechanisms. Female SD rats were randomly divided into four groups: control group (C), model group (M), high-dose GNC group (H), and low-dose GNC group (L). Rats in M, H, and L were administered with VCD and normal saline, high-dose GNC, and low-dose GNC separately. Rat ovaries were harvested either to conduct HE staining for follicle count, immunohistochemistry, or western blot. We found that high dose of GNC significantly increased the ovarian index and sustained the number of primordial follicles and primary follicles in VCD treated rats. Moreover, high dose of GNC significantly increased the ovarian protein expression of mouse vasa homologue (MVH), anti-Müllerian hormone (AMH), follicle-stimulating hormone receptor (FSHR), and estrogen receptor β (ERβ) compared with that in the model group. Besides, high-dose GNC significantly increased ovarian AKT phosphorylation and the expression of downstream forkhead box O3 (FOXO3a). Proapoptosis proteins of Bax, cleaved caspase-3, and poly ADP-ribose polymerase (PARP) were significantly decreased after high-dose GNC treatment compared with those in the model group. Taken together, these findings suggest that high-dose GNC could protect ovarian reserve against VCD-induced toxicity via the activation of the AKT signaling pathway and reduced cell apoptosis in SD Rats. This effect could either be induced by the increased FSHR signaling or by the nontranscriptional activation of ERβ, which requires further investigation.

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Effect of Perceptual Stress Reduction Control Intervention on the Level of Symptoms in Breast Cancer at Different Time Points

Iranian Journal of Public Health ◽

10.18502/ijph.v49i7.3576 ◽

2020 ◽

Author(s):

Zhongru CAO ◽

Yuting LI ◽

Li WANG ◽

Yanhua LIU ◽

Lei ZHANG ◽

...

Keyword(s):

Breast Cancer ◽

Bone Marrow ◽

Health Promotion ◽

Stress Reduction ◽

Intervention Group ◽

Bone Marrow Suppression ◽

Control Group ◽

Time Points ◽

Health Promotion Behavior ◽

Control Intervention

Background: To investigate the effect of perceptual stress reduction control intervention on the level of symptomatic groups at different time points in breast cancer. Methods: A total of 124 breast cancer patients were divided into intervention group and control group, 62 cases in each group. Perioperative nursing and chemotherapy nursing were given to the control group, and the intervention group was given the interventional stress reduction control intervention. The level of symptom clusters of different time points were compared between the two groups. Results: The incidence and severity of myelosuppression in the intervention group were slightly lower than those in the control group. The adverse reactions of bone marrow suppression at T3 were much lower than those in the control group, and the differences were significant (P=0.003, P=0.043). The control group had higher incidence and more severe symptoms of nausea, vomiting and diarrhea than the intervention group (P=0.002, P=0.042). The symptoms of breast pain and swelling at T1 in the intervention group were significantly lower than those in the control group (P=0.000, P=0.000). There was no significant difference in breast symptoms between the two groups at T2 and T3 (p>0.05). At the time of T2 and T3 of chemotherapy, the health promotion behavior scores of the intervention group were higher than the control group, and the difference was statistically significant (PT2=0.000, PT3=0.000). Conclusion: Perceptual stress reduction control intervention can effectively relieve bone marrow suppression, digestive tract discomfort and breast symptoms, and promote health promotion behavior.

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Plasma and Intracellular Concentration of TNF and Its Receptors in Peripheral Blood and Bone Marrow in Different Stages of B-CLL.

Blood ◽

10.1182/blood.v104.11.4781.4781 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4781-4781

Author(s):

Jacek Rolinski ◽

Agnieszka Bojarska-Junak ◽

Iwona Hus ◽

Anna Dmoszynska

Keyword(s):

Bone Marrow ◽

T Cells ◽

B Cells ◽

B Lymphocytes ◽

Peripheral Blood ◽

Biological Effects ◽

Leukemic Cells ◽

Control Group ◽

Stage Of Disease ◽

Positive Correlation

Abstract TNF has been proposed to play a role in the regulation of growth and death of leukemic B-CLL cells. However, the biological effects of TNF on leukemic cells, as well as its role as a prognostic factor need to be further investigated. The aim of the study was to eevaluate the correlation of TNF and its receptors in peripheral blood (PB) and bone marrow (BM) with the stage of B-CLL and some other clinical parameters. PB and BM were taken from 44 newly diagnosed, untreated B-CLL. patients. The control group consisted of 20 healthy subjects. We used flow cytometry technique to assess the capability of T and B lymphocytes to produce TNF and ELISA method to measure plasma levels of TNF and their soluble receptors. We found, that PB and BM plasma TNF concentration in the patients was significantly higher than in the healthy control (2.61 pg/ml. vs 0.62 pg/ml; and 2.91 pg/ml vs 0.75 pg/ml, respectively p<0.001). TNF concentration in PB and BM was significantly higher in Rai stage III–IV than in early stages (p<0.01). There was a correlation between the PB and BM TNF level and lymphocytosis (p<0.005) and the total tumor mass (TTM) (p<0.0001). The PB and BM TNF concentration positively correlated with the percentage of T CD3+ lymphocytes producing intracellular TNF (p<0.01). The percentage of T cells from PB an BM expressing cytoplasmic TNF was significantly higher in patients (PB:39.11±16.97%; BM:40.73±18.19%) than in normal controls (PB:15.74±7.95%; BM:18.80±12.93%) (p< 0.00001; p<0.005, respectively). In PB and BM from B-CLL patients the percentage of CD3+ cells expressing intracellular TNF was significantly higher than the percentage of CD19+/TNF+ cells (p<0.0001). Besides, it was found that the percentage of T cells expressing cytoplasmic TNF positively correlated with the stage of disease (p<0.01). In PB positive correlation were found between the number of T CD3+/TNF+ cells and lymphocytosis (p<0.05) and TTM (p<0.001). The percentage of leukaemic B cells positive for TNF did not correlate with the stage of disease. There was increased expression of TNF-RI and TNF-RII in leukaemic B cells in comparison to normal B-cells was observed (p<0.0001). We found positive correlation between the number of CD5+ B lymphocytes and the levels of soluble TNF-RII (sTNF-RII) (p< 0.05). The sTNF-RII levels in PB and BM significantly correlated with the stage of disease acc. Rai (p<0.0001). Furthermore, the sTNF-RII concentration positively correlated with lymphocytosis and TTM (p<0.0001). These results strongly support the key role TNF in B-CLL pathogenesis. Our results suggest that TNF may function as growth factor for B-CLL cells. CD3+T cells may be the important source of this cytokine in advanced B-CLL. It seems that changes in T cells capability to produce cytoplasmic TNF are associated with disease progression. However, further studies are required to confirm the key role of TNF in B-CLL pathogenesis.

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Parvovirus B19 and Aplastic Anemia: Case Control Study: Preliminary Report from the ITESM Hematology Study Group.

Blood ◽

10.1182/blood.v106.11.3765.3765 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3765-3765

Author(s):

Jose R. Borbolla Escoboza ◽

Marcos E. Garza-Madrid ◽

Luis Villela ◽

Manuel A. Lopez-Hernandez ◽

Jorge Vela-Ojeda

Keyword(s):

Bone Marrow ◽

Aplastic Anemia ◽

Peripheral Blood ◽

Blood Cells ◽

Parvovirus B19 ◽

Bone Marrow Failure ◽

Control Group ◽

Number Of Patients ◽

Two Samples ◽

Sense Primer

Abstract Aplastic anemia (AA) is a classic bone marrow failure syndrome simply defined as peripheral blood pancytopenia and a hypocelular bone marrow, yet the diagnosis must be made by excluding other causes of bone marrow failure. The incidence rate of AA reported by the International Aplastic Anemia and Agranulocytosis Study (IAAAS) in the 1980s was 2 cases per 1 million people. This disease is known to be caused by exposure to radiation, chemotherapy and some viral agents, yet most of the cases are idiopathic. Epstein Barr virus and non-A, non-B or non-C Hepatitis virus have classically been related to the development of some AA cases. Recently there have been some reports of AA following Parvovirus B19 (PvB19) infection. This virus, the only parvoviridae virus capable of infecting humans, attacks erythrocyte precursors attaching to the P antigen in their surface and requiring Beta1 integrin for viral entry. Although PvB19 seems to infect only erytroid precursors, it is widely recognized that the infection with this virus can cause not only anemia, but neutropenia and thrombocytopenia as well, producing aplastic crisis of varying intensity. A correlation has recently been found between PvB19 DNA in peripheral blood and AA in children. We pretend to corroborate this observation and include adult patients in order to improve our understanding of the relationship between PvB19 and AA. So far we have taken peripheral blood samples from 9 AA patients and 9 controls paired by age, sex and community; we plan to include 100 AA patients and their controls from several hospitals around Mexico. DNA was extracted using the PUREGENE DNA extraction kit (Gentra, Minneapolis MN). Nested PCR was performed using the sense primer (P1) 5-AATACACTGTGGTTTTATGGGCCG-3, antisense (P2) 5-CCATTGCTGGTTATAACCACAGGT-3 for the first round and the sense primer (P3) 5-AATGAAAACTTTCCATTTAATGATGTAG-3 and antisense primer (P4) 5-CTAAAATGGCTTTTGCAGCTTCTAC-3for the second round. A DNA sample from a patient with active infectious mononucleosis with positive IgG and IgM against PvB19 in serum was used as positive control. Two samples from the AA group (22%) and 1 from the control group (11%) have turned positive for PvB19 DNA. The reported incidence for the presence of this virusDNA in the peripheral blood of the population is 3%. We expect that, as the number of patients grows, the percentage of positive samples in the control group will decrease, while the percentage of positive samples in the AA group will rise or be sustained. Our partial results point towards a possible relationship between AA and the presence of PvB19 DNA in the peripheral blood cells. It is possible that this virus is one of many factors capable of precipitating the development of AA by limiting the bone marrows capacity to produce blood cells. We are in the process of gathering more samples to prove if a relationship really exists and, if so, future studies will likely shed light upon the mechanism by which PvB19 contributes to the development of AA and other marrow failure syndromes.

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The Immature Platelet Fraction in HIV Patients with Thrombocytopenia.

Blood ◽

10.1182/blood.v110.11.2095.2095 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2095-2095

Author(s):

Brian T. Garibaldi ◽

Rupal B. Malani ◽

Hsin-Chieh Yeh ◽

Deborah Michell ◽

Evan J. Lipson ◽

...

Keyword(s):

Diabetes Mellitus ◽

Bone Marrow ◽

Platelet Count ◽

Peripheral Blood ◽

Reticulocyte Count ◽

Control Group ◽

P Value ◽

Hiv Patients ◽

Immature Platelet Fraction ◽

The Mean

Abstract Thrombocytopenia is a common clinical feature of HIV infection. Given the number of possible etiologies of thrombocytopenia in a patient with known HIV, a peripheral blood test effective in determining the likely pathophysiologic basis of the thrombocytopenia would be a valuable clinical tool. Immature platelets are released early from the bone marrow in response to increased platelet turnover. These platelets contain residual megakaryocyte mRNA and have been termed reticulated platelets. A new assay, the Immature Platelet Fraction (IPF), measures the reticulated platelet count in peripheral blood. Patients with increased destruction of platelets from such conditions as ITP consistently have a higher IPF percent, while patients with decreased platelet production have a low or normal IPF percent. The goal of our study was to determine the performance characteristics of the IPF assay in HIV patients with thrombocytopenia and to see if the IPF percent could be used to help elucidate the etiology of low platelet counts in this group of patients. All adult patients admitted to the Johns Hopkins Hospital with a diagnosis of HIV and a platelet count less than 150,000 were eligible for enrollment. 62 patients were identified from February 2007 to June 2007. 34 control samples were obtained from inpatients with HIV who were not thrombocytopenic. In addition, 81 samples were available from non-HIV historical controls with normal platelet counts. The mean platelet count in the HIV thrombocytopenic group was 92,000 while the mean platelet count in the HIV control group was 254,000 (p value <.001). The mean platelet count in the non-HIV historical control group was 274 (p=.34 when compared to the HIV control group). The mean IPF percent in the HIV thrombocytopenic group was 10.2% as compared to 6.8% in the HIV control group (p=.001). The mean IPF in historical non-HIV controls was 3.1% (p<.001 for both the HIV thrombocytopenic and the HIV control group). Univariate analyses were conducted to identify potential individual predictors of a high IPF percent. Backward selection was then performed using multivariate linear models with a threshold Wald test p-value of 0.05. ITP, diabetes mellitus and cirrhosis were significantly associated with a higher IPF percent with a co-efficient (95% confidence interval) of 6.98 (3.05–10.91), 4.73 (1.39–8.06), and 14.18 (9.7–18.66), respectively. CD4 count, HIV viral load, hepatitis C and reticulocyte count were not correlated with IPF percent. Our results suggest that patients with HIV have increased platelet turnover as compared to patients without HIV. Thrombocytopenic patients with HIV have increased platelet turnover relative to both non-thrombocytopenic HIV patients and to historical non-HIV controls. History of ITP, diabetes mellitus, and cirrhosis are predictive of an elevated IPF percent. Reticulocyte count is not correlated to IPF percent, suggesting that a low reticulocyte count is not a reliable marker for decreased bone marrow production in HIV thrombocytopenia. It is unlikely that the IPF assay alone can be used to determine the pathophysiologic basis of thrombocytopenia in any single patient with HIV. Further work needs to be done to clarify the utility of the IPF assay in this group of patients.

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High Incidence of Peripheral Blood Plasmacytosis In Patients with Dengue Virus Infection: a Prospective Study

Blood ◽

10.1182/blood.v116.21.2772.2772 ◽

2010 ◽

Vol 116 (21) ◽

pp. 2772-2772

Author(s):

Khao T.D. Thai ◽

Josta A. Wismeijer ◽

Catrien M. Zumpolle ◽

Menno D. de Jong ◽

Peter J. Vde ries ◽

...

Keyword(s):

Bone Marrow ◽

Viral Load ◽

Dengue Virus ◽

Peripheral Blood ◽

Extreme Values ◽

Conflicts Of Interest ◽

Dengue Infection ◽

Denv Infection ◽

Bone Marrow Suppression ◽

Median Percentage

Abstract Abstract 2772 Introduction: One of the characteristic features of dengue virus (DENV) infection is the occurrence of leukopenia and thrombocytopenia, probably resulting from virus induced bone marrow suppression. Despite the general bone marrow suppression, polyclonal peripheral blood plasmacytosis has occasionally been described in DENV infected patients. The frequency of peripheral blood (PB) plasmacytosis in patients with dengue infection, the origin of these plasma cells (PCs) and the mechanisms by which they appear in the blood are not known. We initiated this prospective observational study to quantify and describe the kinetics and phenotype of PB plasmacells (PCs) in these patients. Methods: Morphological examination of the peripheral blood smear was performed in 35 sequential returned travelers suspected of DENV infection, with a history of less than 14 days of fever. Flow cytometric (FC) analysis for the characterization and immunophenotyping of lymphocyte subsets and PCs was performed in 31 patients. Follow-up samples were available for 8 patients. Results: Our results show that PB plasmacytosis is a very common hematological finding in DENV infection, with extreme values of up to 36% of total white blood cells in some patients. Depending on the number of days since the onset of fever at presentation, PB plasmacytosis was observed in 64% to 73% of 28 patients with confirmed DENV infection, and in none of 7 patients with other febrile illnesses. PB plasmacytosis was the most pronounced before 7 days after onset of illness and declined rapidly thereafter, to completely disappear after 14 days of illness. The median percentage of PCs at day 7 was 2.5% (range 0–36%; 25–75 interquartile range: 0–8%). The median percentage of PCs was significantly higher in patients with secondary DENV infection than in patients with primary infection (4.5% versus 1.0%; p=0.05). Viral RNA was detectable in 18 of 28 DENV infected patients with a highly variable viral load, but there was no correlation between viral load and percentage of PCs. We found an excellent correlation between percentage of PCs as assessed by morphology and by flow cytometry (r2= 0.85). The majority of CD138+ PCs (89%) had a shared immunophenotype (CD45+/CD19−/CD56−), which differed from normal plasmacells which are generally CD19+. In all cases the PCs were polyclonal. Conclusion: PB plasmacytosis, characterized by a transient presence of polyclonal PCs in the circulation, is a common event in DENV infection and is probably the result of a vigorous humoral immune response to dengue. With an increasing number of travelers to areas where dengue virus is endemic, it is important also for hematologists to recognize this benign cause of sometimes extreme plasmacytosis, for which no invasive procedures such as bone marrow examinations are needed. Disclosures: No relevant conflicts of interest to declare.

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Neutropenia Does Not Delay Lymphopoietic Regeneration in NODSCIDcγ−/− Mice

Blood ◽

10.1182/blood.v118.21.4831.4831 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4831-4831

Author(s):

Stefanie Bugl ◽

Stefan Wirths ◽

R Müller Martin ◽

Märklin Melanie ◽

Tina Wiesner ◽

...

Keyword(s):

Bone Marrow ◽

Peripheral Blood ◽

Nk Cell ◽

Conflicts Of Interest ◽

Control Group ◽

Early Event ◽

Wild Type ◽

Hematopoietic Stem ◽

Lymphoid Progenitors ◽

Fate Decision

Abstract Abstract 4831 Introduction: Previously it was demonstrated that lymphopoiesis is rapidly established after transplantation of wild type stem cells into lymphopenic NODSCIDcγ−/− mice. These data were interpreted as evidence for an “empty” preformed lymphopoietic niche being replenished by lymphoid progenitors. We hypothesized that antibody-induced neutropenia might influence early post transplant fate decision to myeloid rather than lymphoid differentiation resulting in delayed lymphoid reconstitution. Materials and Methods: 25,000 flow sorted CD45.2-expressing wild type Lin-/Sca1+/c-Kit+ (LSK) cells from C57BL/6 mice were transplanted into sublethally irradiated B-/T-/NK-cell deficient NODSCIDcγ−/− mice (CD45.1). Three groups of n = 7 mice received anti-Gr1 or anti-1A8 i.p. every 48 h to induce continuous antibody-mediated neutropenia vs. PBS as control. Blood was harvested at regular intervals to monitor the engraftment. After 16, 22, and 34 days, animals were sacrificed and underwent blood and bone marrow analysis. Results: Hematopoietic regeneration started with the emergence of donor-derived monocytes in all groups as well as neutrophils in the control group as early as 9 days after transplantation. On day 14, B cells were to be detected for the first time, followed by T lymphocytes approximately 20 days after transplantation. Besides the fact that neutrophils were undetectable in the antibody treated groups, the peripheral blood revealed no significant changes between the neutropenic mice and the control group at any point of time. At the bone marrow level, an increase of LSK and granulocyte-macrophage progenitors (GMPs) at the expense of megakaryocyte erythrocyte progenitor cells (MEPs) was found in neutropenic mice. Common lymphoid progenitors (CLPs), however, were not significantly different. Conclusions: The engraftment of wild type donor cells after hematopoietic stem cell transplantation into NODSCIDcγ−/− mice started with the production of monocytes and neutrophils. B-lymphocytes were detectable by day 14 after transplantation. The production of T-cells started around day 20. Continuous antibody-mediated neutropenia did not significantly delay lymphoid regeneration. Although the marrow of neutropenic mice displayed increased proliferation of granulocyte progenitors, CLPs were unchanged. We conclude that the detection of donor-derived lymphocytes in the host peripheral blood is a relatively early event after LSK transplantation. Moreover, antibody induced neutropenia is not sufficient to induce sustainable changes in early hematopoietic fate decisions on the bone marrow level. Disclosures: No relevant conflicts of interest to declare.

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The Novel Use of Hyperbaric Oxygen Therapy to Improve Umbilical Cord Blood Stem Cell Engraftment in Irradiated NOD/SCID Mice: What Did We Learn?

Blood ◽

10.1182/blood.v118.21.4695.4695 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4695-4695

Author(s):

Omar S. Aljitawi ◽

Jeff Radel ◽

Bruce Kimler ◽

Jeff Eskew ◽

Nikhil Parelkar ◽

...

Keyword(s):

Bone Marrow ◽

Stem Cell ◽

Cord Blood ◽

Umbilical Cord Blood ◽

Peripheral Blood ◽

Oxygen Therapy ◽

Photon Emission ◽

Blood Stem Cell ◽

Control Group ◽

Cord Blood Stem Cell

Abstract Abstract 4695 Introduction: UCB is also an attractive stem cell source for transplantation because of the ease of procurement, the absence of donor risks, and reduced risk of transmissible infections. Unfortunately, outcomes of UCB stem cell transplantation are less than optimal because of low cell dose in umbilical cord blood units and defects in homing to bone marrow, which result in both delayed engraftment and delayed immune recovery, both of which increase the mortality rate in UCB transplants. In these experiments, we investigated the use of hyperbaric oxygen therapy (HBO) to improve cord blood stem cell homing and engraftment post irradiation. Method: Six to eight week old NOD/ SCID mice were sublethally irradiated 24 hours prior to cord blood stem cell infusion. The irradiated mice were then divided into two groups (10 mice each). One group of mice served as the control group were mice remained under normoxic conditions. The second group of animals, the HBO group, six hours prior to cord blood stem cell infusion, were exposed to two hours of hyperbaric conditions in a hyperbaric chamber, with 100% oxygen for 2 hours at 2.5 atmospheres absolute (ATA). Erythropoietin (epo) level was checked by ELISA, at time of cord blood stem cell infusion, and was used as a surrogate marker for effective HBO therapy. Prior to CD34+ cord blood stem cell injection, cryopreserved cord blood units obtained from St. Louis Cord Blood Bank were processed for CD34+ selection followed by their transduction with a luciferase lentivirus so that they can be serially imaged, using Xenogen-IVIS imaging system, for engraftment into the bone marrow. Subsequently, cord blood stem cells (7 × 104 / 100 uL) in sterile PBS were administered by tail vein injection. The mice were weighed every other day and were sacrificed at 9–10 weeks post cord blood stem cell infusion. Peripheral blood and spleen tissue were obtained to determine donor engraftment by flow cytometry. Bone tissue was harvested for immunohistochemistry staining for human CD45. Result: Mean epo level in the HBO group was significantly lower than the mean epo level in the control group (226.7 ± 18.02 versus 294.4 ± 14.35 pg/ml (P=.0148)), consistent with effective HBO therapy. The average photon emission, or radiance, measured using the average of ventral and dorsal photon emission values per group, starting week 5, peaked at 7 weeks in the control mice, while peaked at 5 weeks in the HBO group. Since only ventral photon emission values were available during the first 4 weeks, an earlier peak in HBO group could not be excluded. One mouse in each group died, which was thought to be secondary to excessive bleeding related to mandible bleeding. No significant differences in median percentage peripheral blood or spleen engraftment were detected between the two groups. Interestingly, the HBO mice demonstrated a higher mean body weight started day 36 post cord blood stem cell infusion and continued until the end of study. Conclusions: HBO treatment did not result in detectable differences in terms of mean percentage peripheral blood or spleen engraftment at the end of study. However, HBO treated mice, compared to controls, showed an earlier photon emission peak values followed by lower photon emission values overtime. This earlier peak might have corresponded to the significant increase in weight noticed in the HBO mice starting week 5 post cord blood stem cell infusion. *We acknowledge the help we received from St. Louis Cord Blood Bank who provided the cryopreserved cord blood units for this research. Disclosures: No relevant conflicts of interest to declare.

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