CD33/CD10 Ratios in the Blast Region Evaluated by Flow Cytometry Predicts the Response of Chronic Myelogeneous Leukemia to Imatinib.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2595-2595
Author(s):  
Satoko Oka ◽  
Masaki Mori
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Healthy Volunteers ◽  
Peripheral Blood ◽  
Chronic Myelogeneous Leukemia ◽  
Group A ◽  
One Year ◽  
Group B ◽  
Group D

Abstract Abstract 2595 Poster Board II-571 Background: The prediction of the response of chronic myelogeneous leukemia (CML) to imatinib is clinically important. However, no reliable markers are known. Methods: From January 2005 to April 2007, 32 patients with newly diagnosed CML were treated at our hospital. The diagnosis was made based on the existence of the Philadelphia chromosome. A routine bone marrow aspiration was performed to evaluate bone marrow smears, karyotypes, phenotypes, and bcr/abl message levels before the start of imatinib. Phenotypic analysis was conducted using three-color flow cytometry as follows: bone marrow mononuclear cells were stained with fluorescein isothiocyanate-conjugated monoclonal antibody, phycoerythrin-conjugated monoclonal antibody, and peridinin chlorophyll protein-conjugated CD45. A gate was set to identify blasts characterized by intermediate CD45 expression and low side-scatter properties. Phenotypes of the cells in the blast region were analyzed using a flow cytometer (FACSCalibur; BD Biosciences, San Jose, CA, USA). As a control, bone marrow samples were obtained from 8 healthy volunteers and 35 patients with refractory anemia of myelodysplastic syndrome (MDS-RA). Results: All patients were diagnosed as having CML in the chronic phase and given imatinib alone. According to the response after one year of treatment with imatinib, patients showing and not showing a complete cytogenetic response were designated as group A (26 patients) and group B (6 patients), respectively. There were no significant differences between the two groups in terms of the age, peripheral blood cell counts, peripheral blood blast percentages, peripheral blood basophil percentages, bone marrow cell differentials, bone marrow blast percentages, additional chromosomal abnormalities, Sokal's score, Hasford's score, and spleen size before the start of imatinib. The mean imatinib dose per day was higher in group A (349 mg) than in group B (284.4 mg); however, the results were not significant (p=0.051). Group B showed higher percentages of myeloid cells in the blast region than group A, while the former showed lower percentages of B lymphoid cells in the blast region than the latter. CD33/CD10, CD33/CD19, CD13/CD10, and CD13/CD19 ratios in the blast region in group B were significantly higher than in group A: 56.6 vs. 6.0 (p<0.01), 34.5 vs. 6.0 (p<0.01), 33.1 vs. 7.7 (p<0.01), and 23.8 vs. 4.9 (p<0.05), respectively. There were no differences between the two groups regarding the percentage of CD34+ and CD117+ cells. Similar results were obtained, when patients were evaluated after six months of treatment with imatinib. After the six-month imatinib treatment, patients were divided into two groups: one group (group C) showed undetectable bcr/abl messages (17 patients), and the other (group D) showed detectable bcr/abl messages (6 patients). There were no significant differences between the two groups in terms of imatinib dose per day and other clinical data. CD33/CD10, CD33/CD19, CD13/CD10, and CD13/CD19 ratios in the blast region in the group D and in the group C were 11.6 vs. 6.4 (p=0.05), 10.5 vs. 6.1 (p<0.05), 13.9 vs. 8.1 (p<0.05), and 9.4 vs. 5.7 (p<0.05), respectively. CD33/CD10, CD33/CD19, CD13/CD10, and CD13/CD19 ratios in the blast region in the MDS-RA patients and healthy volunteers were 3.2 vs. 0.7, 3.2 vs. 0.5, 2.5 vs. 0.6, and 3.0 vs. 0.6, respectively. Conclusion: An increase in CD33/CD10 ratios in the blast region in CML before the start of imatinib is associated with resistance to the drug. A cut-off value of 30 for CD33/CD10 ratios in the blast region in CML at diagnosis is useful for predicting the response of CML to imatinib after one year of treatment with the drug. Disclosures: No relevant conflicts of interest to declare.

Effects of spaceflight on rat erythroid parameters

1996 ◽  
Vol 81 (1) ◽  
pp. 117-122 ◽  
Author(s):  
Z. Allebban ◽  
L. A. Gibson ◽  
R. D. Lange ◽  
T. L. Jago ◽  
K. M. Strickland ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Life Sciences ◽  
Lower Number ◽  
Ground Control ◽  
Colony Forming Unit ◽  
Human Erythropoietin ◽  
Group A ◽  
Group B ◽  
First Time ◽  
Group D

Hematologic studies were performed on 21 ground control rats and 21 rats flown during the Spacelab Life Sciences-2 14-day mission. Group A (n = 5) was used to collect blood in flight and 9 days postflight, group B (n = 5) was injected with recombinant human erythropoietin (rhEpo), group C (n = 5) received saline as a control, and group D (n = 6) was killed in flight and tissues were collected. Results indicated no significant changes in peripheral blood erythroid elements between flight and ground control rats. The nonadherent bone marrow on flight day 13 showed a lower number of recombinant rat interleukin-3 (rrIL-3)-responsive and rrIL-3 + rhEpo-responsive blast-forming unit erythroid (BFU-e) colonies in flight rats compared with ground control rats. On landing day, a slight increase in the number of rhEpo + rrIL-3-responsive BFU-e colonies of flight animals compared with ground control rats was evident. Nine days postflight, bone marrow from flight rats stimulated with rhEpo alone or with rhEpo + rrIL-3 showed an increase in the number of colony-forming unit erythroid colonies and a decrease in BFU-e colonies compared with ground control rats. This is the first time that animals were injected with rhEpo and subsequently blood and tissues were collected during the spaceflight to study the regulation of erythropoiesis in microgravity.

Concurrent Treatment with Denileukin Diftitox (DD) To Deplete T-Regulatory Cells Enhances Rebound Lymphocytosis and Eosinophilia in Patients Treated with High-Dose IL-2 (HDIL-2) for Metastatic Renal Cell Cancer (MRCC).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1729-1729
Author(s):  
Adi Gidron ◽  
John Eklund ◽  
Brenda Martone ◽  
Alfred W. Rademaker ◽  
Charles Goolsby ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune Response ◽  
Statistical Significance ◽  
Cell Cancer ◽  
Peak Level ◽  
High Dose ◽  
Maximal Increase ◽  
Group A ◽  
Group B ◽  
Group D

Abstract Background: CD4+CD25+hi T cells (Treg) play a suppressive role in immune regulation. DD is an IL-2 receptor specific cytotoxin. We postulated depletion of Treg with DD may enhance immune effector cell populations after HDIL-2 treatment, including rebound lymphocytosis and also eosinophilia which has been reported to be involved in immune response to neoplasm (Mattes J Exp Med 197: 387, 2003). Methods: In this pilot study, 12 pts (8 male, median age 58 yrs) with MRCC were tx with HDIL-2 and DD in different schedules to determine safety and effect on immune response as manifested by changes in Treg, peak lymphocyte, and eosinophil counts. Pts were treated with IL-2 600,000 IU/kg Q8H on days (d) 1–5 and 15–19. Three (group A) and 4 (group B) pts were given 6 and 9ug/kg daily on d8–10 respectively, while 5 (group C) pts received 9ug/kg of DD on d −4 to −2. Nine (group D) pts with metastatic melanoma who received HDIL-2 as above but without DD were included as controls. Flow cytometry was done on days −4, 1,8,10,15,22 for group C and on days 1,8,10,15,22 for groups B, and D. CBC was obtained concurrent or within 24 hours of flow cytometry. Group A pts were evaluated for safety only and were excluded from analysis. Results: Prior to enrollment, all pts had undergone nephrectomy and four patients received interferon-alpha. One pt from group B withdrew from study and was not included in analysis. Administration of DD resulted in a median decline of 25% in Treg number (not significant). DD given before HDIL-2 was associated with a greater increase in Treg post HDIL-2. In Group C there was an increase of rebound median Treg count of 0.88k/ul compared with 0.060k/ul in group B (p=0.025). Absolute lymphocytosis was higher in the combined group getting DD compared to control (median maximal increase of 7.6 vs 4.7 k/ul, respectively) although the difference did not reach statistical significance. However, group C pts had a greater increase in absolute lymphocytosis than did group B pts in which absolute lymphocytosis actually decreased (median increase 10.6 vs. median decrease 0.4 k/ul, p=0.025). A higher peak level of eosinophilia was noted in groups B and C compared with group D (mean increase of 10.5 vs. 4.0 k/ul p=0.2). Group C had a greater peak eosinophilia than group B (11.2 vs 2.2 k/ul p=0.053) Toxicity was manageable and consistent with those seen with HDIL-2. Median HDIL-2 dose given was 21 (range, 14–28). No clinical responses were observed. Of 11 pts included in the analysis 1 pt from group A expired 68 weeks after enrollment. All remaining patients are alive. Survival from enrollment ranges from 11 to 93 weeks. Conclusion: Overall, the combination of DD and HDIL-2 results in a stimulatory effect as manifested by increased rebound lymphocytosis and eosinophilia compared to HDIL-2 alone. Administration of DD in conjunction with HDIL-2 was associated with a rebound in Treg that may be schedule and dose dependent. The results suggest an enhanced immune stimulatory effect as manifested by lymphocytosis and peak eosinophilia in group C. However, this stimulatory effect also extends to Treg that may prove detrimental clinically. Further exploration of these effects in immunotherapy naïve patients would be beneficial.

scholarly journals Impact of Proteomic Profile of Peripheral Blood and Bone Marrow Sera on Molecular Response in Patients with Chronic Myeloid Leukemia: Preliminary Data

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5515-5515
Author(s):  
Nicola Sgherza ◽  
Vito Garrisi ◽  
Giacoma De Tullio ◽  
Simona Serratì ◽  
Angela Iacobazzi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chronic Myeloid Leukemia ◽  
Preliminary Data ◽  
Peripheral Blood ◽  
Myeloid Leukemia ◽  
Differentially Expressed ◽  
Molecular Response ◽  
Proteomic Profile ◽  
Group A ◽  
Group B

Abstract BACKGROUND. Chronic Myeloid Leukemia (CML) is a myeloproliferative neoplasm characterized by an aberrant protein (BCR–ABL) which is a constitutively active tyrosine kinase. According to the latest ELN recommendations for the management of CML, molecular response (MR) is best assessed according to the International Scale (IS) as the ratio of BCR-ABL1 transcripts to ABL1 transcripts, or other internationally recognized control transcripts. It is expressed and reported as BCR-ABL1% on a log scale where 10%, 1%, 0.1%, 0.01%, 0.0032%, and 0.001% correspond to a decrease of respectively 1 (MR1), 2 (MR2), 3 (MR3), 4 (MR4), 4.5 (MR4.5) logs below the standard baseline that was used in the IRIS study. Recent advances in the proteomic field have allowed us to better understand the biology of several cancer types and/or discover new candidate biomarkers, but very few data are available in CML. AIMS. The purpose of this study was to evaluate a possible correlation between depth of MR and proteomic profile in sera samples obtained from the peripheral blood and bone marrow of CML patients. PATIENTS AND METHODS Samples were consecutively and prospectively obtained from 20 CML patients observed between January and June 2014 at the Hematology Unit of the National Cancer Research Centre “Istituto Tumori Giovanni Paolo II” in Bari, Italy. Each individual involved in the study signed an informed consent form authorizing the Institute to utilize their biological tissues for research purposes. All patients at diagnosis displayed the classic t(9;22) Ph chromosome according to standard cytogenetics. The BCR/ABL transcript at RT-PCR was b3a2 in 13 patients and b2a2 in 7 patients. Peripheral blood and bone marrow samples were centrifuged within 30 minutes of sample taking. Serum specimens were immediately collected and frozen at −80°C. Twenty sera from peripheral blood were sampled from 5 patients in MR1 response, four in MR2, eight in MR3, two in MR4 and 1 patient at diagnosis; for eleven patients serum from bone marrow was also available; in particular 2 were sampled from patients in MR1, 3 in MR2, 4 in MR3, 1 in MR4 and 1 at diagnosis. Patients were grouped in two cohorts: the first comprised those with lower molecular response to MR3 (group A: 10 patients) and the second greater than or equal to MR3 (group B: 10 patients). The association of proteomic profile with molecular response was performed using the SELDI ToF Mass Spectrometry platform. Each specimen was spotted on an IMAC30 metal affinity protein-chip, prepared according to the manufacturer's instructions, and analyzed in duplicate. RESULTS Fourteen differentially expressed peaks were highlighted when comparing peripheral sera from group A and group B, but none was statistically significant. When comparing 11 available serum samples from the bone marrow of groups A (6) and B (5), four peaks (m/z 10629, m/z 3889, m/z 7772, m/z 7987) were reported as differentially expressed in a statistically significant way (p<0.05). Focusing the differential expression analysis in peripheral sera only on MR1 patients (including one patient at diagnosis) versus MR4 patients, one peak at m/z 11092 was identified as significantly and differentially expressed (p < 0.05) (Figure 1). Similarly, comparing bone marrow sera only from MR1 and MR4 patients respectively, 32 peaks were differentially expressed. Once again the peak at m/z 11092 resulted under expressed in MR1 patients, and interestingly the single patient at diagnosis had the lowest value. No statistical differences were evidenced when comparing peripheral blood and bone marrow sera obtained from b3a2 and b2a2 patients. CONCLUSIONS These preliminary data suggest that an over-expression of m/z 11092 in serum obtained from peripheral blood and bone marrow could be associated with a deeper molecular response; further investigations are needed on a larger number of patients in order to confirm or refute our results and, to definitively characterize the peak at m/z 11092. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

A Useful Flow Cytometry Panel To Exclude Myelodysplastic Syndrome.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4848-4848
Author(s):  
Ling Zhang ◽  
Jean R. Lopatequi ◽  
Sharron E. Kelly ◽  
Tobi Neer ◽  
Stephen Lee
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Test Group ◽  
Bone Marrow Examination ◽  
Normal Group ◽  
P Value ◽  
Aberrant Expression ◽  
Cd10 Expression ◽  
Group A ◽  
Group B

Abstract Background: Myelodysplastic syndromes (MDS) are a heterogeneous group of hematopoietic disorders, which can be difficult to diagnose based only on morphologic bone marrow examination. Karyotyping can be useful in diagnosing borderline cases. However, only 40% of patients with MDS have an abnormal karyotype. Flow cytometry(FCM) approaches have been described but not clearly defined in MDS due to use of different criteria. We devised a 10-parameter FCM panel, mainly including myeloid and erythroid maturation markers, to differentiate MDS marrows from normal marrows. Design: Bone marrow from 91 patients with cytopenia(s) or anemia were included in the study(10/2005–7/2006). Cases were divided into 3 groups: normal, not morphologically suspicious for MDS, 46 cases, equivocal, morphologically suggestive but not diagnostic of MDS, 20 cases, morphologically diagnostic of MDS, 25 cases. 10 FCM paremeters were performed and scored: hypogranularity,aberrant expression of CD56,lack of CD10 expression,decreased CD64 expression,&lack of CD13 or CD33 expression,&abnormal CD13/CD16 or CD16/CD11b pattern,increase of CD 34 expression gating on all cells excluding erythrocytes,decreased expression of CD71/Glycophorin gating on erythroid precursors. Karyotypings were analyzed. Results: and (see Table 1 and 2). Karyotyping were anlayzed in 86 cases. Cytogenetic abnormalities were found in 2.2% (1/44) of normal group, 5.3% (1/19) of equivocal group and 34.8% (8/43) of MDS group. In MDS group 7 of 8 patients (87.5%) who had both morphologic and cytogenetic diagnosis of MDS were scored &gt;=3 of 8 FCM parameters. Conclusions: Study showed the 8-parameter FCM panel was more predictive of MDS than 10-parameters one. Both CD13/CD16 & CD16/CD11b patterns were considered to be non-specific. In 8-parameter panel, zero score tended to rule out MDS, while score &gt;=3 suggested MDS. The most specific FCM parameters were hypogranularity, CD34, aberrant expression of CD56 or lack of CD10 expression by mature granulocytes. CD71/Glycophorin A might be useful in identifying dysplasia in erythroid lineage. Table 1. Comparison of FCM Parameters in Patients with/without MDS Parameters/Group Normal Group(A)(n=46) Equivocal Group(B)(n=20) MDS Group(C)(n=25) CHITEST(P value)(GRoup A vs C) * These two parameters were deleted in the 8-parameter panel due to their high frequency seen in normal group. Hypogranularity 4 (8.7%) 6 (30%) 18 (72%) 0.0001 CD56 0 (0.0%) 1 (5%) 6 (24%) 0.0005 CD10 7 (15.2%) 7 (35%) 15 (60%) 0.0001 CD64 6 (13.1%) 2 (10%) 6 (24%) 0.2396 CD13 0 (0.0%) 0 (0.0%) 2 (8.0%) 0.0516 CD33 0 (0.0%) 2 (10 %) 0 (0%) 0 CD13/CD16* 23 (50.0%) 7 (35%) 21 (84%) 0.0047 CD34 8 (17.4%) 7 (35%) 13 (52%) 0.0026 CD71 2 (4.3%) 5 (25%) 12 (48%) 0.0001 CD16/CD11b* 27 (58.7%) 8 (40%) 22 (88%) 0.0107 Table 2. Scoring with 8 FCM Parameters in Patients with/without MDS Score/Group Normal Group (A)* (n=46) Equivocal Group (B)* (n=20) MDS Group (C)* (n=25) * Fish Exact T Test: Group A vs C: p&lt;0.0001, A vs B: p=0.0006, B vs C: p=&lt;0.0001 Score=0 21 (45.7%) 3 (15%) 0 (0.0%) Score=1–2 25 (543 %) 14 (70%) 8 (32 %) Score&gt;3 0 (0.0%) 3 (15%) 17 (68%) *Mean score +/− SD 0.65+/−1.06 1.55+/−1.54 2.8+/−1.65

scholarly journals Data Note: COVID-19, social distancing, and pipeline vandalism in Nigeria

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 604
Author(s):  
P. N. Onwuachi-Iheagwara ◽  
B.I Iheagwara
Keyword(s):  
Information System ◽  
Geographical Information System ◽  
Satellite Imagery ◽  
Historical Data ◽  
Geographical Information ◽  
Port Harcourt ◽  
Group A ◽  
One Year ◽  
Group B ◽  
Group D

We present a dataset of the monthly cases of pipeline vandalism in Nigeria from January 2015 to January 2021. Data used in this study were collated from the Monthly Financial and Operations Reports (MFOR) of the Nigeria National Petroleum Corporation (NNPC). Each MFOR provides cases of pipeline vandalism during a 12-month span from five key locations; Mosimi, Kaduna, Port Harcourt, Warri, and Gombe. Recorded incidences of pipeline vandalism from these locations were summed and assembled into five groups; namely: historical data, prior-COVID-19, COVID-19 lockdown, and post-COVID-19 lockdown. The data were grouped based on dates. These dates were January 2015 to July 2019, August 2019 to January 2020, February 2020 to July 2020, and August 2020 to January 2021 respectively. The historical data were further sub-divided into four sub-groups based on the deployment (May 2016) of sophisticated weapons, satellite imagery, and geographical information system into the security apparatus to checkmate pipeline vandalism. The four sub-groups are sub-group A (one-year before deployment), sub-group B (the year of deployment), sub-group C (one-year after deployment), and sub-group D (two-years after deployment). The dates span for each sub-group is May 2015-April 2016, May 2016-April 2017, May 2017-April 2018, and May 2018-April 2019 respectively. After the deployment of GIS devices in May 2016, the accumulated national number of pipeline vandalism cases declined from 400 cases in January 2016 to 293 in February 2016, and 259 cases in March 2016 as opposed to 60, 49, and 94 cases in the same months in 2017; but over the years, 2017 to 2021 these methods have proved less effective, and cases of pipeline vandalism have risen once more. Similar changes in the number of cases and patterns were observed during the COVID-19 movement restrictions. From the dataset, it can be seen that COVID-19 influenced incidences of pipeline vandalism.

Liposomal Nanoparticle-Mediated miR-27b Influences Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells via Peroxisome Proliferator-Activated Receptor Gamma in a Microgravity Environment

2020 ◽  
Vol 12 (11) ◽  
pp. 1301-1308
Author(s):  
Zhiwei He ◽  
Yan Zhu ◽  
Gentao Fan ◽  
Hongbo Qian
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Simulated Microgravity ◽  
Mrna Level ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells ◽  
Group A ◽  
Group B ◽  
Group D

This study was aimed at analyzing the effects of liposomal nanoparticle-based miR-27b on PPARγ and osteogenic differentiation of bone marrow mesenchymal stem cells under microgravity. The rat bone marrow mesenchymal stem cells were set as the research object, and the gyroscope was employed for simulation of microgravity. The cells were randomized into four groups, including the experimental group A (simulated microgravity+liposomal nanoparticle-mediated miR-27b transfection group), as well as the control groups: group B (simulated microgravity+negative control group), group C (simulated microgravity+transfection reagent group) and group D (normal gravity+liposomal nanoparticle-mediatedmiR-27b transfection group). After a two-week osteogenic induction in vitro, staining was performed to assess the lipogenesis rate of the samples. In addition, ALP activity and PPARγ mRNA level was detected. The number of alizarin staining-positive osteogenic nodules and ALP activity (0.21±0.44 King unit) in group A was significantly diminished compared to those in group B, C, and D. Moreover, its lipogenesis rate (9.31±1.02%) and PPARγ mRNA level (1.86±0.39) were significantly higher than those in group B, C, and D (P < 0.05). The number of alizarin staining-positive osteogenic nodules and ALP activity (0.96±0.18 King unit) in group D were significantly reduced in comparison with those in groups B and C, while the lipogenesis rate (4.86±0.77%) and PPARγ mRNA level (0.93±0.34) were significantly higher than those in group B and C (P < 0.05) without difference between group B and group C (P > 0.05). Under a microgravity condition, liposomal nanoparticle-mediated miR-27b can impede the differentiation of BMSCs into osteoblasts via regulating PPARγ signal transduction.

Increased Group B Killer Cell Immunoglobulin-Like Receptor (KIR) Haplotypes with Mismatched MHC Class I and Altered NK Repertoire Distribution in Bone Marrow Failure Syndromes

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2412-2412
Author(s):  
Jeffrey S Painter ◽  
Jong Park ◽  
Michael J Clemente ◽  
Christine L. O'Keefe ◽  
Sheng Wei ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
T Cells ◽  
Nk Cells ◽  
Mhc Class I ◽  
Bone Marrow Failure ◽  
Group A ◽  
Kir Receptors ◽  
Group B ◽  
Kir Genotypes

Abstract Abstract 2412 Background: Killer immunoglobulin like receptors (KIR) are expressed on NK cells and a subset of T cells where they bind HLA-C alleles to discriminate self from non-self during anti-tumor and anti-viral responses. KIR receptors exist in inhibitory and activating forms (KIRDL and KIRDS, respectively) that regulate effector functions. The KIR family consists of 14 genes inherited independently from MHC-class I ligands. An imbalance of KIR receptors tilted in favor of more activating KIR genotypes is associated with improved antiviral responses, but also an increase in autoimmunity. We reported previously that KIR/HLA mismatches occur at a higher frequency in bone marrow failure: Aplastic Anemia (AA), Myelodysplastic Syndrome (MDS), Large Granular Lymphocyte leukemia (LGLL), and Paroxysmal Nocturnal Hemaglobinuria (PNH) (Epling-Burnette et al, Blood (ASH Annual Meeting abstracts 2008 112: abstract 4121). We now present data on a larger cohort of patients along with phenotype analysis and individual KIR genotype analysis. Methods: KIR genotyping by bead array (One Lambda, Inc) was performed on 205 cases and resolved in 199 (97%). Samples from 93 unrelated individuals were used as controls. HLA genotyping was analyzed in 190 and 51 cases and controls, respectively. The diagnosis of MDS (n=72), AA (n=59), LGLL (n=46), and PNH (n=28) were confirmed by pathology review. Flow cytometry for KIR2DS1/DL1 was assessed by staining for CD158a (KIR2DS1/DL1) and CD158b (KIR2DS2/DL2/3) on (CD56+/CD16+) NK and (CD4/CD8/CD3) T cells by multiparameter flow cytometry. Antigens at the HLA-C locus were divided into two groups, the HLA-C1 with alleles encoding Ser-77-Asn80 and HLA-C2 alleles encoding Asn-77-Lys-80 that bind KIR2D2 and KIR2D1, respectively. A mismatch was defined as the presence of the KIR gene in the absence of its cognate ligand. Results: KIR2DL2 and KIR2DS2 gene frequency was increased in the combined BMF group (120/198, 60% DL2 and 117/199, 59%, DS2) compared to controls (43/93, 46% for both DL2 and DS2) (p=0.02 and p=0.058, respectively) in association with a higher frequency of C1 activating and inhibitory receptor mismatches in cases (20 of 190 cases vs. 0 of 51 controls, p=0.009). KIR genotypes can be separated into Group A and B haplotypes with Group A haplotypes containing only one activating KIR (KIR2DS4) and group B haplotypes containing two or more activating KIR. Compared to controls patients with BMF had significantly decreased group A (23%) and increased group B (77%) haplotypes (p0.04). We detected 31 different genotypes in BMF patients and 23 in controls representing both the A and B haplotypes. AA1 was the most common group A genotype in cases (32/35, 91.4%) and controls (49/50, 98%). Specific genotypes that contain KIR2DL2 and DS2 were overrepresented in the BMF cases with AB9 increased in AA (p=0.0054) and AB1 increased in LGLL (p=0.02). KIR phenotyping was performed on NK cells and T cells from 49 BMF patients and 15 controls. KIRs were variably expressed on T cells, and NKG2D had normal expression on NK and T cell populations. CD158a (KIR2DL1/DS1) positive NK cells were significantly reduced in BMF cases compared to controls (17% vs 35%, p0.00008), which was individually significant in AA (18%, p0.003), MDS (20%, p0.01), and LGLL (14%, p=0.0002). The percentage of CD158b (KIR2DS2/DL2/3) positive NK cells, which recognizes the receptors associated with the C1 mismatch, were also significantly reduced in cases compared to controls (30% vs. 43%, p0.02) indicating altered NK repertoire distribution. Conclusions: KIRs and HLA are inherited independently and have a high level of diversity, but previous results suggest that there is an epistatic relationship between these loci because matched KIR/HLA combinations have evolved through selection. In BMF syndromes, increased frequency of KIR2DL2/KIR2DS2 genotypes in the absence of the appropriate HLA C allele may alter NK and T cell effector responses in favor of autoimmunity. Two to 8 different KIR receptors are present on individual NK cells and these are expressed in a stochastic manner. In previous studies, the expression of a receptor did not depend on the presence of the cognate HLA. Our phenotyping data suggests that the genetic disparities or disease characteristics in BMF may influence NK repertoire distribution leading to fewer KIR positive NK cells with possible pathological consequences. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Establishment of Human Acute Monocyte Leukemia Model with Systemic Leukemia in Npg Mice

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 4041-4041
Author(s):  
Jing Xu ◽  
Zhenjiang Li ◽  
Qiong Wu ◽  
Wenfeng He ◽  
Jifu Zheng ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Survival Time ◽  
Peripheral Blood ◽  
Histopathological Examination ◽  
Immunosuppressive Treatment ◽  
Leukemic Cells ◽  
Acute Monocytic Leukemia ◽  
Monocytic Leukemia ◽  
Group A ◽  
Group B

Abstract Although tumor cells are easily to growth in the bodies of immunodeficicent animals such as nude mice and NOD-SCID mice, it's hard for acute leukemia cells to grow in the bone marrow of nude mice or NOD-SCID mice even when mice receive extra immunosuppressive treatment such as splenectomy, cyclophosphamide and irradiation. This study aimed to establish a mice model with systemic leukemia using another highly immunodeficicent NPG mice without immunosuppressive treatment before inoculation. 5-week NPG mice were inoculated with 1x107(Group A) or 5x107 (Group B) SHI-1 cells (a cell line derived from a refractory acute monocytic leukemia patient) via tail vein. One NPG mice in each group was killed by ether randomly at the day 14, 21, 28 after inoculation, other NPG mice were observed the survival time. The leukemic cells engrafted in the NPG mice were detected by the following methods: the blast cells were detected by the blood smear and flow cytometer, the MLL-AF6 fuse gene of SHI-1 cells were detected by PCR amplification, the human CD45 positive cells infiltrated in the organs of NPG mice were detected by histopathological examination and immunohistochemistry. At the day 14 after inoculation with SHI-1 cells, fewer blasts cells were found in the smear of peripheral blood of group B; MLL-AF6 fuse gene could be amplified in the spleen of NPG mice in group A and in spleen and bone marrow in group B (Fig A); histopathological examination had shown that CD45 positive leukemia cell just infiltrated in spleen. At the day 21 after inoculation, more blasts were found in the smear of peripheral blood both in group A and B; MLL-AF6 fuse gene were amplified in the organs of NPG mice such as Spleen, liver, kidney, stomach, lung, heart and bone marrow(Fig A); 5.16% and 0.82% of CD45 and CD33 positive cells were detected in the peripheral blood of NPG mice in group A and B respectively; a green solid neoplasm were found in the kidney of NPG mice in group B, leukemia cells were found in the organ of heart, liver, spleen, stomach, kidney and lung in the NPG mice of both groups by histopathological examination. From the third week, the NPG mice presented anorexia, hunched posture, lethargy and weight loss. For the mice sacrificed in the day 28 after inoculation, the proportion of CD45 and CD33 positive cells in peripheral blood, bone marrow and spleen were 9.60%, 11.4% and 23.20% in group A and were 11.0%,37.80% and 60.5% in group B (Fig B). Green solid tumors were grown in many organs such as kidney, liver, spleen, stomach, heart, lymph node and the soft tissues in the NPG mice killed in day 28 after inoculation and the mice which were dead spontaneous (Fig C); When NPG mice were dead, the weight of spleen in group B is significantly higher than the weight of spleen in group A(P<0.05) (Fig D). The median survival time of NPG in group A and group B is 33 and 30 days respectively. Pathological examination and immunohistochemical staining had shown that leukemic cells could infiltrated to many of the organs of NPG mice and the grade of leukemia infiltration was positively correlated with cells numbers of inoculation and the survival time of NPG mice (Fig E). Altogether, SHI-1 cell could growth in the NPG mice without any pre-immunosuppressive treatment and formed a systemic leukemia in NPG mice as like in acute leukemia patients. This efficient and reproducible model may be a useful tool for the studies of the pathogenesis in acute monocytic leukemia. Figure. Figure. Disclosures No relevant conflicts of interest to declare.

scholarly journals C-Reactive Protein in Peripheral Blood of Patients with Chronic and Aggressive Periodontitis, Gingivitis, and Gingival Recessions

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Stepan Podzimek ◽  
Jaroslav Mysak ◽  
Tatjana Janatova ◽  
Jana Duskova
Keyword(s):  
Periodontal Disease ◽  
Peripheral Blood ◽  
Aggressive Periodontitis ◽  
C Reactive Protein ◽  
Pocket Depth ◽  
Reactive Protein ◽  
Study Results ◽  
Group A ◽  
Group B ◽  
Group D

CRP is a plasma protein that reflects a measure of the acute phase response to inflammation and is one of the markers of choice in monitoring this response. CRP can be used for the prediction and early detection of periodontal disease. The aim of this study was to compare and evaluate the systemic levels of CRP in the peripheral blood samples of patients with chronic and aggressive periodontitis, gingivitis, and gingival recessions and compare them with periodontal clinical parameters. All patients (N=158) were examined prior to the initiation of periodontal treatment. Patients were divided into four groups. Group A consisted of 26 patients with aggressive periodontitis, Group B consisted of 111 patients with chronic periodontitis, Group C consisted of 13 patients with gingivitis, and Group D consisted of 8 patients with gingival recessions. Our study results indicate that CRP levels increase subsequently with the severity of the periodontal disease and that the bleeding on probing index showed much better positive correlation with the CRP levels compared to the pocket depth index in both periodontitis patients groups, especially in aggressive periodontitis patients.

Comparison of one-year prognosis of patients classified as chronic critical lower limb ischaemia according to TASC II or European consensus definition in the COPART cohort

VASA ◽  
2015 ◽  
Vol 44 (3) ◽  
pp. 0220-0228 ◽  
Author(s):  
Marion Vircoulon ◽  
Carine Boulon ◽  
Ileana Desormais ◽  
Philippe Lacroix ◽  
Victor Aboyans ◽  
...  
Keyword(s):  
Medical Treatment ◽  
Critical Limb Ischaemia ◽  
Limb Ischaemia ◽  
Medical Group ◽  
Consensus Definition ◽  
Transcutaneous Oxygen ◽  
European Consensus ◽  
Group A ◽  
One Year ◽  
Group B

Background: We compared one-year amputation and survival rates in patients fulfilling 1991 European consensus critical limb ischaemia (CLI) definition to those clas, sified as CLI by TASC II but not European consensus (EC) definition. Patients and methods: Patients were selected from the COPART cohort of hospitalized patients with peripheral occlusive arterial disease suffering from lower extremity rest pain or ulcer and who completed one-year follow-up. Ankle and toe systolic pressures and transcutaneous oxygen pressure were measured. The patients were classified into two groups: those who could benefit from revascularization and those who could not (medical group). Within these groups, patients were separated into those who had CLI according to the European consensus definition (EC + TASC II: group A if revascularization, group C if medical treatment) and those who had no CLI by the European definition but who had CLI according to the TASC II definition (TASC: group B if revascularization and D if medical treatment). Results: 471 patients were included in the study (236 in the surgical group, 235 in the medical group). There was no difference according to the CLI definition for survival or cardiovascular event-free survival. However, major amputations were more frequent in group A than in group B (25 vs 12 %, p = 0.046) and in group C than in group D (38 vs 20 %, p = 0.004). Conclusions: Major amputation is twice as frequent in patients with CLI according to the historical European consensus definition than in those classified to the TASC II definition but not the EC. Caution is required when comparing results of recent series to historical controls. The TASC II definition of CLI is too wide to compare patients from clinical trials so we suggest separating these patients into two different stages: permanent (TASC II but not EC definition) and critical ischaemia (TASC II and EC definition).

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