scholarly journals The Contribution of Toll-like Receptor (TLR) Pathway Hyper-Reactivity to Clonal Selection in Secondary MDS and AML

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4621-4621
Author(s):  
Laura F. Newell ◽  
R. K. Rathbun ◽  
Michael R Garbati ◽  
Kim-Hien T. Dao ◽  
Shernan G. Holtan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Inflammatory Cytokines ◽  
Peripheral Blood ◽  
Clonal Selection ◽  
Cytotoxic Chemotherapy ◽  
Normal Donor ◽  
Growth Responses ◽  
Prior Chemotherapy ◽  
Paradoxical Growth ◽  
Secondary Mds

Abstract Background: Stem cell (HSC) hypersensitivity to inflammatory cytokines and exaggerated TLR-dependent production of such cytokines contribute to bone marrow failure and clonal selection in Fanconi anemia (FA). Clonal neoplasms in FA patients and FA-deficient mice exhibit either resistance or paradoxical proliferative responses to tumor necrosis factor-alpha (TNF) and interferon-gamma (IFN). Because FA MDS/AML shares cytogenetic and clinical features in common with secondary MDS/AML developing after prior MDS or exposure to cytotoxic chemotherapy (sMDS/sAML), we tested the idea that an FA-like TLR/cytokine hypersensitive phenotype might underlie clonal selection in sMDS/sAML. A pilot study of 4 hematologically normal individuals with histories of prior cytotoxic chemotherapy (including alkylating agents) revealed 2 whose committed progenitor cells were hypersensitive to TNF and IFN. Three patients with sMDS/sAML exhibited resistance to IFN and paradoxical growth responses to TNF. To determine the true prevalence of these FA-like phenotypes, we conducted a larger study of patients with sMDS/sAML, quantifying a) progenitor cell growth in response to inflammatory cytokines and b) TLR7/8 or TLR4-dependent cytokine production in peripheral blood monocytes. Methods: Patients with newly diagnosed sMDS or sAML were eligible. All potential subjects with active infections were excluded. Research bone marrow aspirates and peripheral blood (PB) were obtained at the same time. TNF was quantified (ELISA) in the supernatants of PB CD14+ cells cultured in the presence of 1 ng/ml lipopolysaccharide (LPS, a TLR-4 agonist) or 3-5 μM R848 (TLR-7/8 agonist) for 24-hours. BFU-E and CFU-GM were quantified in methylcellulose cultures of low density bone marrow mononuclear cells (LDBMCs) (SCF, IL-3, EPO +/- TNF or IFN). Progenitor responses were classified as resistant/paradoxical if either CFU-GM or BFU-E colonies were >120% of control (normal volunteer bone marrow cells) at TNF 1 ng/ml, or >100% at IFN 0.1 ng/ml; all other responses were categorized as sensitive. TNF production was classified as exaggerated if levels exceeded normal mean values by more than two standard deviations. Results: A total of 22 patients with sMDS/sAML had bone marrow and PB analysis, including 13 patients with complex karyotypes and 9 patients with non-complex karyotypes. An additional 18 patients had PB samples drawn without providing bone marrow samples. Progenitor responses: In colony assays, 18/22 patients demonstrated resistant/paradoxical growth responses to TNF/IFN, a phenomenon seen in patients with both complex and non-complex karyotypes. Interestingly, TNF resistance was observed only in samples that were also IFN resistant. Among patients with a complex karyotype, only 3 demonstrated normal progenitor sensitivity. None of these 3 had received prior chemotherapy (2 with radiation only). Macrophage responses: CD14+ cells from 63% of patients with resistant/paradoxical colony assay responses overproduced TNF in response to either R848 or LPS. Mean TNF levels from CD14+ cells of these patients were higher compared to patients with sensitive colony responses [1756 pg/ml (R848) and 1699 pg/ml (LPS), versus 783 pg/ml (R848) and 959 pg/ml (LPS), for resistant/paradoxical versus sensitive patients respectively]. Among all patients with PB samples (including those with/without marrow results), CD14+ cells from 68% of these patients overproduced TNF. Mean TNF levels in response to R848 or LPS stimulation of patient cells were elevated compared to normal donor controls [1730 pg/ml (R848) and 1795 pg/ml (LPS), versus 719 pg/ml (R848) and 555 pg/ml (LPS), from patient and normal donor CD14+ cells, respectively]. Conclusions: In patients with sMDS/sAML there is a high prevalence of TNF resistance in committed progenitors and TLR-hyper-reactivity in CD14+ cells. In FA the MDS/AML clonal resistance phenotype is selected because it possesses a selective advantage over the unfit pool of non-clonal FA HSC. We suggest that prior chemotherapy can create a fixed phenotype of HSC unfitness (e.g. hypersensitivity to inflammatory cytokines) and that the desultory fitness landscape in that pool permits the selection of cytokine resistant clones. Mechanistic studies should provide opportunities to identify patients at risk of sMDS/sAML and to develop pharmacological approaches to leukemia prevention. Disclosures No relevant conflicts of interest to declare.

Identification of a Novel Hematopoietic Cell Translocation in a Patient with Paroxysmal Nocturnal Hemoglobinuria (PNH).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3717-3717 ◽  
Author(s):  
Gabrielle Meyers ◽  
William H. Babcock ◽  
Robert G. Best ◽  
Charles J. Parker
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Clonal Selection ◽  
Clonal Evolution ◽  
Clonal Expansion ◽  
Structural Rearrangement ◽  
Hematopoietic Cell ◽  
Chromosome 12 ◽  
Gpi Anchor ◽  
Anchor Protein

PNH is an acquired disorder of clonal hematopoiesis, in which the affected hematopoietic stem cells (HSC’s) have a mutation of the X-linked PIG-A gene. PIG-A is required for synthesis of the glycosylphosphatidyl inositol (GPI) moiety that serves as an anchor for a functionally diverse group of membrane proteins. Consequently all GPI-anchored proteins are deficient on progeny of affected HSC’s. GPI-anchor protein deficiency clearly accounts for some of the clinical manifestations of PNH but does not explain either clonal selection or clonal expansion. Further, it is not clear if clonal selection and clonal expansion are independent processes. Observations reported herein suggest that clonal selection and clonal expansion are part of the same process and that PNH is an example of non-malignant clonal evolution. A 31 y/o female presented with complaints of fatigue, fever, numbness in the lower extremity and dark urine. CBC revealed a WBC of 4100/μl, hemoglobin 3.8 gram%, hematocrit of 12.9% and platelet count of 171,000/μl. Laboratory studies were indicative of intravascular hemolysis, and subsequent flow cytometric analysis of the peripheral blood showed that 88% of the PMN were GPI-anchor protein deficient. Bone marrow biopsy showed a normocellular marrow with a relative erythrocytosis but with no evidence of dysplasia. Cytogenetic studies of the bone marrow demonstrated 46XX, ins (12) (p12~13q13q12) in all 20 metaphase cells analyzed. The findings were that of a female with an apparently balanced intrachromosomal inverted insertion at the 400 band level. FISH probes supported the conventional cytogenetics and indicated that the insertion split the TEL locus at 12p13. Subsequent analysis of mitogen stimulated peripheral blood lymphocytes revealed a normal karyotype. These finding show that the rearrangement of chromosome 12 identified in the bone marrow cells is an acquired cytogenetic abnormality and suggest a causal relationship with PNH in this case. TEL, a member of the ETS family of transcription factors is a fusion partner in a number of translocation that result in hematopoietic cell malignancies (e. g., ALL, AML, MDS). While the rearrangement described herein appear to be novel, deletions of 12p12 have been observed in other patients with PNH, and a structural rearrangement of 12q13 (one of the three breakpoints in the insertion in this case) has also been reported in a patient with PNH. Together, these findings suggest the following: (1) non-random cytogenetic abnormalities may be associated with PNH; (2) PNH may be an example of non-malignant clonal evolution in which the PIG-A mutation contributes directly to clonal expansion (in this specific case by acting in concert with a non-transforming proliferative advantage arising from the rearrangement involving TEL); (3) clonal selection and clonal dominance may not be independent process in PNH. Studies designed to characterize the consequences of the structural rearrangement of chromosome 12 in this case are in progress.

scholarly journals Peripheral blood stem cell transplants for multiple myeloma: identification of favorable variables for rapid engraftment in 225 patients

Blood ◽  
1995 ◽  
Vol 85 (2) ◽  
pp. 588-596 ◽  
Author(s):  
G Tricot ◽  
S Jagannath ◽  
D Vesole ◽  
J Nelson ◽  
S Tindle ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Alkylating Agents ◽  
High Dose ◽  
Platelet Recovery ◽  
Prior Chemotherapy ◽  
Prior Therapy ◽  
Cd34 Cells

Abstract Transfusion of autologous peripheral blood stem cells (PBSCs) of good quality ensures fast hematopoietic engraftment after myeloablative therapy with a decrease in procedure-related morbidity and mortality. We have analyzed variables influencing the kinetics of engraftment, and therefore reflecting the quality of PBSC collections, in 225 patients with newly diagnosed or refractory multiple myeloma (MM) who received an autotransplant in support of high dose melphalan (200 mg/m2); 132 of these patients also completed a second transplant. All PBSCs were collected before the first transplant after high-dose cyclophosphamide (6 g/m2) and hematopoietic growth factors, mainly granulocyte- macrophage colony-stimulating factor. PBSCs were administered either alone (91 patients) or with bone marrow (134 patients). A highly significant correlation was observed between the number of CD34+ cells per kilogram infused and prompt recovery of both granulocytes (P = .0001) and platelets (P = .0001). After correction for the proportion of patients with > or = 2 x 10(6)/kg CD34 PBSCs infused and with < or = 12 months of prior therapy, no difference in engraftment kinetics was seen between patients receiving PBSCs only and those also receiving bone marrow. Exposure to chemotherapy, even to < or = 6 months of alkylating agents, significantly delayed hematopoietic recovery posttransplantation. The threshold dose of CD34 cells necessary for prompt engraftment was > or = 2.0 x 10(6)/kg for patients with < or = 24 months of chemotherapy before the first transplant, whereas greater than 5 x 10(6)/kg CD34 cells were required to assure rapid recovery also in those with longer exposure. Such quantities, easily collected in the large majority of patients with shorter exposure (91%), were obtained in only 28% of patients with more than 24 months of prior chemotherapy. Rapid platelet recovery within a narrow range of time (before day 14) was almost invariably seen (94%) when greater than 5 x 10(6)/kg CD34 cells were infused, irrespective of the duration of prior therapy, whereas the range widened progressively when less CD34 cells were infused. In the absence of CD34 measurements, fast recovery of platelets to greater than 50 x 10(9)/L within 14 days after high-dose cyclophosphamide and < or = 12 months of prior chemotherapy were the best predictors of early engraftment. Prudent use of stem cell-damaging agents, such as melphalan and nitrosoureas, is recommended in MM patients who might be candidates for autotransplantation. Alternatively, PBSCs should be collected early after diagnosis.

scholarly journals Cytokine Consistency Between Bone Marrow and Peripheral Blood in Patients With Philadelphia-Negative Myeloproliferative Neoplasms

2021 ◽  
Vol 8 ◽  
Author(s):  
Pu Chen ◽  
Boting Wu ◽  
Lili Ji ◽  
Yanxia Zhan ◽  
Feng Li ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Inflammatory Cytokines ◽  
Peripheral Blood ◽  
Myeloproliferative Neoplasms ◽  
Critical Role ◽  
Primary Myelofibrosis ◽  
Diagnostic Value ◽  
Inflammatory Status ◽  
Bm Niche ◽  
Cytokine Levels

Background: Inflammation might play a critical role in the pathogenesis and progression of Philadelphia-negative myeloproliferative neoplasms (Ph−MPNs) with elevated inflammatory cytokines in peripheral blood (PB). However, the inflammatory status inside the bone marrow (BM), which is the place of malignancy origin and important microenvironment of neoplasm evolution, has not yet been elucidated.Methods: Inflammatory cytokine profiles in PB and BM of 24 Ph-MPNs patients were measured by a multiplex quantitative inflammation array. Cytokines that correlated between PB and BM were selected and then validated by ELISA in a separate cohort of 52 MPN patients. Furthermore, a panel of cytokines was identified and examined for potential application as non-invasive markers for the diagnosis and prediction of fibrosis progress of MPN subtypes.Results: The levels of G-CSF, I-309, IL-1β, IL-1ra, IL-12p40, IL-15, IL-16, M-CSF, MIG, PDGF-BB, and TIMP-1 in BM supernatants were significantly higher than those in PB (all p &lt; 0.05). Linear correlations between BM and PB levels were found in 13 cytokines, including BLC, Eotaxin-2, I-309, sICAM-1, IL-15, M-CSF, MIP-1α, MIP-1δ, RANTES, TIMP-1, TIMP-2, sTNFRI, and sTNFRII (all R &gt; 0.4 and p &lt; 0.05). Levels of BLC, Eotaxin-2, M-CSF, and TIMP-1 in PB were significantly different from those in health controls (all p &lt; 0.05). In PB, levels of TIMP-1 and Eotaxin-2 in essential thrombocythemia (ET) group were significantly lower than those in groups of prefibrotic primary myelofibrosis (pre-PMF) [TIMP-1: 685.2 (322.2–1,229) ng/ml vs. 1,369 (1,175–1,497) ng/ml, p = 0.0221; Eotaxin-2: 531.4 (317.9–756.6) pg/ml vs. 942.4 (699.3–1,474) pg/ml, p = 0.0393] and primary myelofibrosis (PMF) [TIMP-1: 685.2 (322.2–1229) ng/ml vs. 1,365 (1,115–1,681) ng/ml, p = 0.0043; Eotaxin-2: 531.4 (317.9–756.6) pg/ml vs. 1,010 (818–1,556) pg/ml, p = 0.0030]. The level of TIMP-1 in myelofibrosis (MF) &gt;1 group was significantly higher than that in MF ≤ 1 group.Conclusion: Abnormal inflammatory status is present in MPN, especially in its BM microenvironment. Consistency between PB and BM levels was found in multiple inflammatory cytokines. Circulating cytokine levels of BLC, M-CSF, Eotaxin-2, and TIMP-1 reflected inflammation inside BM niche, suggesting potential diagnostic value for MPN subtypes and prognostic value for fibrosis progression.

scholarly journals Reference Values of Human Bone Marrow and Peripheral Blood Levels of 49 Cytokines According to Age and Clonal Hematopoiesis

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 4296-4296
Author(s):  
Noemie Ravalet ◽  
Hélène Guermouche ◽  
Pierre Hirsch ◽  
Frederic Picou ◽  
Nathalie Gallay ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Inflammatory Cytokines ◽  
Reference Values ◽  
Peripheral Blood ◽  
Plasma Concentrations ◽  
Multiple Comparisons ◽  
Clonal Hematopoiesis ◽  
Age Related ◽  
Biological Studies ◽  
Cytokine Levels

Abstract INTRODUCTION Cytokines are involved in many processes, including hematopoiesis and inflammation. Aging is associated with the onset of clonal hematopoiesis (CH) of indeterminate potential, putatively associated with a higher risk of progression to hematological malignancies such as myelodysplastic syndromes or acute myeloid leukemia. Moreover, CH may participate to create a pro-inflammatory environment contributing to the pathogenesis of age-related diseases, such as cardiovascular diseases. This is likely to be driven by or translated in changes in bone marrow (BM) and/or peripheral blood (PB) soluble factors for which reference values still remain unclear, because BM cytokines levels have never been determined in strictly selected healthy people. Indeed, control BM samples classically used in studies are from subjects undergoing surgeries for non-hematologic causes, such as total hip replacement or cardiac surgery, patients suffering from immune thrombocytopenic purpura, brain death patients or allogeneic BM donors. In this study, the BM and PB plasma concentrations of 49 hematopoietic and inflammatory cytokines were measured in a representative panel of 94 healthy adult volunteers and the results were analyzed considering their age and presence of CH. METHOD Ninety-four healthy donors aged from 18.6 to 80.1 years old (yo), including 58 women were recruited for this study (HEALTHOX protocol, CPP Tours, AFSSAPS identifier ID-RCB: 2016-A00571-50 and ClinicalTrials.gov # NCT02789839). The presence or absence of CH (&gt;1% of variant allele frequency) in this cohort is already known (Guermouche H, Ravalet N et al, Blood Adv 2020;4(15):3550-3557). BM samples were obtained through sternal aspiration using a classical procedure in France, and PB sampling was performed at the same time by venipuncture. Samples were collected on sodium heparin or EDTA, centrifuged twice (1200 g, 10 min, 20 C°), aliquoted and stored at -80°C. A 48-plex human cytokine panel assay was used to quantify 48 human cytokines in PB and BM plasmas [beta-NGF, CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CCL27, CLEC11A, CXCL1, CXCL8, CXCL9, CXCL10, CXCL12, FGF-2, G-CSF, GM-CSF, HGF, IFN-alpha-2, IFN-gamma, IL-1 alpha, IL-1 beta, IL-1ra, IL-2, IL-2-RA, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12, IL-12B, IL-13, IL-15, IL-16, IL-17A, IL-18, KITLG, LIF, LT-alpha, M-CSF, MIF, PDGF subunit B, TNF, TNFSF10, VEGF-A]. MIF and FLT3L quantification were performed by ELISA. Regarding CH, the controls were subjects older than 50 yo without CH. Statistical analyses were performed with the R (3.6.3) and Rstudio version 1.2.5042 (www.rstudio.org) software. Comparisons were computed by Wilcoxon and Kruskal-Wallis tests. All pairwise multiple comparisons were performed using Dunn's-test for multiple comparisons of independent samples (PMCMR package). Correlations between cytokine levels in PB and BM were tested with Pearson and Spearman methods. Correlation matrices were plotted using the "corrplot" package. RESULTS CH was detected in 16 volunteers, mostly in individuals over 50 yo. BM and PB plasma samples were studied in 3 age-groups: 18-40, 40-60 and 60-80 yo. With aging, variations were observed for 18 BM cytokine levels, with 7 increasing (FLT3L, CXCL9, HGF, FGF-2, CCL27, IL-16, IL-18) and 8 decreasing (G-CSF, TNF, IL-2, IL-15, IL-17a, IL-4, LT-alpha, IL-1 alpha). In PB, 10 cytokines significantly increased with age (CXCL9, FLT3L, CCL27, CXCL10, HGF, CCL11, IL-16, IL-6, IL-1 beta, CCL2). CH was associated with significantly higher BM levels of MIF and IL-1 beta, lower BM levels of IL-9 and IL-5 and higher PB levels of IL-15, VEGF-A, IL-2, CXCL8, CXCL1 and G-CSF (Table). CONCLUSION In this study and for the first time, we concomitantly analyzed BM and PB concentrations of a panel of hematopoietic and inflammatory cytokines in a cohort of strictly selected healthy volunteers. In addition to the establishment of reference values, useful for various biological studies, and correlations between blood and BM levels, we identify variations in the BM of key cytokines according to age and CH. The differences in these cytokine concentrations, either as causes or as consequences, may shape both the BM microenvironment and hematopoietic processes, eventually leading to the beginning of age-related myeloid malignancies or inflammatory conditions. Figure 1 Figure 1. Disclosures Hirsch: Daiichi Sankyo Oncology: Consultancy; Novartis Pharma: Consultancy. Suner: Sanofi - Genzyme: Consultancy. Delhommeau: Celgene: Consultancy; BMS: Consultancy; Novartis: Consultancy.

Shortened Erythrocyte Life Span and Increased Oxidative Stress In Erythroid Precursors Are Consistent with Normocytic Anemia In Mice with Chronic Inflammation

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3205-3205
Author(s):  
Andrew J. Layman ◽  
Jacqueline M. Langdon ◽  
Qilu Yu ◽  
Olivier D. Prince ◽  
Jeremy D. Walston ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Bone Marrow ◽  
Inflammatory Cytokines ◽  
Peripheral Blood ◽  
Sterile Abscess ◽  
Normocytic Normochromic Anemia ◽  
Erythroid Precursors ◽  
Normochromic Anemia ◽  
Pro Inflammatory Cytokines

Abstract Abstract 3205 The anemia of inflammation is a common co-morbid condition associated with various infections, autoimmune disorders, and other chronic disease states. In general, hemoglobin levels are mildly to moderately decreased. In vitro studies have demonstrated that pro-inflammatory cytokines can inhibit erythropoiesis at various stages of development, but many of these studies are limited to the analysis of early developmental stages and primarily assess proliferation as opposed to maturation or survival. Sufficient data from in vivo studies and whole animal models are lacking. We characterized the anemia in 8–10 week old C57BL/6 mice with turpentine oil-induced sterile abscess. We found that following 3 weeks of sterile abscess, the mice had a normocytic, normochromic anemia. Using in vivo biotin-labeling of peripheral blood, we found the coefficient describing erythrocyte life span significantly decreased from -0.044 ± 0.002 SE in control mice to -0.048 ± 0.002 SE in mice with sterile abscess (p = 0.04). In further support of the mechanism of increased erythrocyte turnover, we found that splenic macrophages isolated from mice with abscess significantly increased expression of Heme-regulated gene 1 (Hrg-1) 2.5 fold, (p < .001) and significantly increased expression of Ferroportin (Fpn) 2.76 fold (p = 0.003). Oxidative stress can lead to increased erythrocyte turnover and can inhibit erythroid maturation. To determine whether erythrocytes from mice with sterile abscess had impaired capacity for survival, we assessed staining of peripheral blood and erythroid precursors with chloromethyl dichlorodihydrofluorescein diacetate, acetyl ester (DCF). DCF fluoresces upon oxidation. We found that mean DCF fluorescence intensity (MFI) in peripheral blood increased from 3.2 ± 1.0 in control mice to 5.5 ± 0.6 in mice with abscess (p=0.004). Similarly, we found a greater percentage of bone marrow derived erythroid precursors from mice with abscess had high DCF staining (16.2 ± 3.9%) when compared to control mice (6.7 ± 1.9%, p=0.003). In conclusion, these data support the hypothesis that increased oxidative stress in erythroid precursors of mice with sterile abscess results in increased erythrocyte turnover and normocytic, normochromic anemia. Future studies will investigate whether pro-inflammatory cytokines inhibit the expression of key erythrocyte anti-oxidant enzymes or whether reactive oxygen species from neighboring granulocytes damage erythroid precursors in the bone marrow. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Peripheral blood stem cell transplants for multiple myeloma: identification of favorable variables for rapid engraftment in 225 patients

Blood ◽  
1995 ◽  
Vol 85 (2) ◽  
pp. 588-596 ◽  
Author(s):  
G Tricot ◽  
S Jagannath ◽  
D Vesole ◽  
J Nelson ◽  
S Tindle ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Alkylating Agents ◽  
High Dose ◽  
Platelet Recovery ◽  
Prior Chemotherapy ◽  
Prior Therapy ◽  
Cd34 Cells

Transfusion of autologous peripheral blood stem cells (PBSCs) of good quality ensures fast hematopoietic engraftment after myeloablative therapy with a decrease in procedure-related morbidity and mortality. We have analyzed variables influencing the kinetics of engraftment, and therefore reflecting the quality of PBSC collections, in 225 patients with newly diagnosed or refractory multiple myeloma (MM) who received an autotransplant in support of high dose melphalan (200 mg/m2); 132 of these patients also completed a second transplant. All PBSCs were collected before the first transplant after high-dose cyclophosphamide (6 g/m2) and hematopoietic growth factors, mainly granulocyte- macrophage colony-stimulating factor. PBSCs were administered either alone (91 patients) or with bone marrow (134 patients). A highly significant correlation was observed between the number of CD34+ cells per kilogram infused and prompt recovery of both granulocytes (P = .0001) and platelets (P = .0001). After correction for the proportion of patients with > or = 2 x 10(6)/kg CD34 PBSCs infused and with < or = 12 months of prior therapy, no difference in engraftment kinetics was seen between patients receiving PBSCs only and those also receiving bone marrow. Exposure to chemotherapy, even to < or = 6 months of alkylating agents, significantly delayed hematopoietic recovery posttransplantation. The threshold dose of CD34 cells necessary for prompt engraftment was > or = 2.0 x 10(6)/kg for patients with < or = 24 months of chemotherapy before the first transplant, whereas greater than 5 x 10(6)/kg CD34 cells were required to assure rapid recovery also in those with longer exposure. Such quantities, easily collected in the large majority of patients with shorter exposure (91%), were obtained in only 28% of patients with more than 24 months of prior chemotherapy. Rapid platelet recovery within a narrow range of time (before day 14) was almost invariably seen (94%) when greater than 5 x 10(6)/kg CD34 cells were infused, irrespective of the duration of prior therapy, whereas the range widened progressively when less CD34 cells were infused. In the absence of CD34 measurements, fast recovery of platelets to greater than 50 x 10(9)/L within 14 days after high-dose cyclophosphamide and < or = 12 months of prior chemotherapy were the best predictors of early engraftment. Prudent use of stem cell-damaging agents, such as melphalan and nitrosoureas, is recommended in MM patients who might be candidates for autotransplantation. Alternatively, PBSCs should be collected early after diagnosis.

▾ Filgrastim – human granulocyte-colony stimulating factor

1993 ◽  
Vol 31 (9) ◽  
pp. 33-36
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Cytotoxic Chemotherapy ◽  
Granulocyte Colony Stimulating Factor ◽  
Risk Of Infection ◽  
Colony Stimulating Factor ◽  
Full Dose ◽  
Stimulating Factor ◽  
Cell Colony

Filgrastim (recombinant human granulocyte-colony stimulating factor) (Neupogen – Amgen-Roche) was the first of several white cell ‘colony stimulating factors’ to be licensed for use in Britain. Filgrastim stimulates specifically the production of neutrophils from progenitor cells in the bone marrow, speeds their maturation and increases their number in the peripheral blood.1,2 Use of filgrastim is intended to reduce the duration and severity of myelosuppression that follows intensive cytotoxic chemotherapy. The manufacturer claims that filgrastim “supports full dose chemotherapy on time”, by moderating neutropenia and reducing the risk of infection that might interfere with the next cycle of treatment. When should filgrastim be given?

Neutrophil pseudoplatelets in subgingival plaque and crevicular fluid samples from periodontal diseased sites

1989 ◽  
Vol 47 ◽  
pp. 862-863 ◽  
Author(s):  
J Hanker ◽  
E.J. Burkes ◽  
G. Greco ◽  
R. Scruggs ◽  
B. Giammara
Keyword(s):  
Electron Microscopy ◽  
Bone Marrow ◽  
Peripheral Blood ◽  
Gingival Crevicular Fluid ◽  
Light And Electron Microscopy ◽  
Subgingival Plaque ◽  
Staining Reaction ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Crevicular Fluid

The mature neutrophil with a segmented nucleus (usually having 3 or 4 lobes) is generally considered to be the end-stage cell of the neutrophil series. It is usually found as such in the bone marrow and peripheral blood where it normally is the most abundant leukocyte. Neutrophils, however, must frequently leave the peripheral blood and migrate into areas of infection to combat microorganisms. It is in such areas that neutrophils were first observed to fragment to form platelet-size particles some of which have a nuclear lobe. These neutrophil pseudoplatelets (NPP) can readily be distinguished from true platelets because they stain for neutrophil myeloperoxidase. True platelets are not positive in this staining reaction because their peroxidase Is inhibited by glutaraldehyde. Neutrophil pseudoplatelets, as well as neutrophils budding to form NPP, could frequently be observed in peripheral blood or bone marrow samples of leukemia patients. They are much more prominent, however, in smears of inflammatory exudates that contain gram-negative bacteria and in gingival crevicular fluid samples from periodontal disease sites. In some of these samples macrophages ingesting, or which contained, pseudoplatelets could be observed. The myeloperoxidase in the ingested pseudoplatelets was frequently active. Despite these earlier observations we did not expect to find many NPP in subgingival plaque smears from diseased sites. They were first seen by light microscopy (Figs. 1, 3-5) in smears on coverslips stained with the PATS reaction, a variation of the PAS reaction which deposits silver for light and electron microscopy. After drying replicate PATS-stained coverslips with hexamethyldisilazane, they were sputter coated with gold and then examined by the SEI and BEI modes of scanning electron microscopy (Fig. 2). Unstained replicate coverslips were fixed, and stained for the demonstration of myeloperoxidase in budding neutrophils and NPP. Neutrophils, activated macrophages and spirochetes as well as other gram-negative bacteria were also prominent in the PATS stained samples. In replicate subgingival plaque smears stained with our procedure for granulocyte peroxidases only neutrophils, budding neutrophils or NPP were readily observed (Fig. 6).

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