Platelets: bridging hemostasis, inflammation, and immunity Tập 35 Số 3 - Trang 254-261 - 2013
Craig N. Jenne, Rossana Urrutia, Paul Kubes
SummaryAlthough the function of platelets in the maintenance of hemostasis has been studied in great detail, more recent evidence has highlighted a central role for platelets in the host inflammatory and immune responses. Platelets by virtue of their large numbers and their ability to rapidly release a broad spectrum of immunomodulatory cytokines, chemokines, and other mediators act as circulating sentinels. Upon detection of a pathogen, platelets quickly activate and begin to drive the ensuing inflammatory response. Platelets have the ability to directly modulate the activity of neutrophils (phagocytosis, oxidative burst), endothelium (adhesion molecule and chemokine expression), and lymphocytes. Due to their diverse array of adhesion molecules and preformed chemokines, platelets are able to adhere to leukocytes and facilitate their recruitment to sites of tissue damage or infection. Furthermore, platelets directly participate in the capture and sequestration of pathogens within the vasculature. Platelet–neutrophil interactions are known to induce the release of neutrophil extracellular traps (NETs) in response to either bacterial or viral infection, and platelets have been shown to internalize pathogens, sequestering them in engulfment vacuoles. Finally, emerging data indicate that platelets also participate in the host immune response by directly killing infected cells. This review will highlight the central role platelets play in the initiation and modulation of the host inflammatory and immune responses.
Evaluation of the diagnostic reliability of different RBC indices and formulas in the differentiation of the β‐thalassaemia minor from iron deficiency in Palestinian population Tập 30 Số 4 - Trang 324-330 - 2008
Mahmoud M. Sirdah, Issa S. Tarazi, Esmat Najjar, Rula Haddad
Summaryβ‐thalassaemia minor and iron deficiency are the most common causes of microcytosis and/or hypochromasia. The present study evaluates the diagnostic reliability of different RBC indices and formulas, as well as our proposed formula, in the differentiation of the β‐thalassaemia minor from iron deficiency in Palestinian population. Complete blood count (CBC) parameters of 2196 certainly diagnosed (1272 β‐thalassaemia minor and 924 iron deficiency) samples were used to evaluate the following indices and formulas: Bessman index (RDW), Mentzer formula (MCV/RBC), England and Fraser formula (MCV − RBC − 5 × Hb − 3.4), Shine and Lal formula (MCV2 × MCH/100), Ehsani formula (MCV − 10 × RBC), Srivastava formula (MCH/RBC), Green and King formula (MCV2 × RDW/Hb × 100), red distribution width index RDWI (RDW × MCV/RBC), RDW/RBC, as well as our formula (MCV − RBC − 3 × Hb). For each index and formula, the receiver operative characteristic (ROC) curve was constructed to calculate the area under the curve (AUC), in addition, sensitivity, specificity, and likelihood ratios were calculated. No significant differences were reported between our formula, Green‐King formula and the RDWI (P > 0.05) in discriminating β‐thalassaemia minor from iron deficiency (AUC = 0.914, 0.909 and 0.907 respectively). However, the three indices and formula showed the highest efficiencies and they were significantly (P < 0.05) better than the others in the discrimination efficiency .It was concluded that our formula, Green–King formula and the RDWI provided the highest reliabilities in differentiating β‐thalassaemia minor from iron deficiency in Palestinian population while Bessman index was poor and ineffective for that purpose.
Selected Stro‐1‐enriched bone marrow stromal cells display a major suppressive effect on lymphocyte proliferation Tập 31 Số 1 - Trang 9-19 - 2009
Aisha Nasef, Y. Z. ZHANG, Christelle Mazurier, Sandrine Bouchet, Morad Bensidhoum, Sabine François, N C Gorin, M López, Dominique Thierry, L Fouillard, Alain Chapel
SummaryMesenchymal stem cells (MSCs) have an immunosuppressive effect and can inhibit the proliferation of alloreactive T cells in vitro and in vivo. Cotransplantation of MSCs and hematopoietic stem cells (HSCs) from HLA‐identical siblings has been shown to reduce the incidence of acute graft‐vs.‐host disease. MSCs are heterogeneous and data on the inhibitory effects of different MSC subsets are lacking. The antigen Stro1 is a marker for a pure primitive MSC subset. We investigated whether Stro‐1‐enriched induce a more significant suppressive effect on lymphocytes in a mixed lymphocyte reaction (MLR), and whether this action is related to a specific gene expression profile in Stro‐1‐enriched compared to other MSCs. We demonstrated that the Stro‐1‐enriched population elicits a significantly more profound dose‐dependent inhibition of lymphocyte proliferation in a MLR than MSCs. One thousand expanded Stro‐1‐enriched induced an inhibitory effect comparable to that of 10 times as many MSCs. Inhibition by Stro‐1‐enriched was more significant in contact‐dependent cultures than in noncontact‐dependant cultures at higher ratio. The Stro‐1‐enriched inhibitory effect in both culture types was linked to increased gene expression for soluble inhibitory factors such as interleukin‐8 (IL‐8), leukemia inhibitory factor (LIF), indoleamine oxidase (IDO), human leukocyte antigen‐G (HLA‐G), and vascular cell adhesion molecule (VCAM1). However, tumor growth factor‐β1 (TGF‐β) and IL‐10 were only up‐regulated in contact‐dependant cultures. These results may support using a purified Stro‐1‐enriched population to augment the suppressive effect in allogeneic transplantation.
Technical issues: flow cytometry and rare event analysis Tập 35 Số 3 - Trang 344-350 - 2013
Benjamin D. Hedley, Michael Keeney
SummaryFlow cytometry has become an essential tool for identification and characterization of hematological cancers and now, due to technological improvements, allows the identification and rapid enumeration of small tumor populations that may be present after induction therapy (minimal residual disease, MRD). The quantitation of MRD has been shown to correlate with relapse and survival rates in numerous diseases and in certain cases, and evidence of MRD is used to alter treatment protocols. Recent improvements in hardware allow for high data rate collection. Improved fluorochromes take advantage of violet laser excitation and maximize signal‐to‐noise ratio allowing the population of interest to be isolated in multiparameter space. This isolation, together with a low background rate, permits for detection of residual tumor populations in a background of normal cells. When counting such rare events, the distribution is governed by Poisson statistics, with precision increasing with higher numbers of cells collected. In several hematological malignancies, identification of populations at frequencies of 0.01% and lower has been attained. The choice of antibodies used in MRD detection facilitates the definition of a fingerprint to identify abnormal populations throughout treatment. Tumor populations can change phenotype, and an approach that relies on ‘different from normal’ has proven useful, particularly in the acute leukemias. Flow cytometry can and is used for detection of MRD in many hematological diseases; however, standardized approaches for specific diseases must be developed to ensure precise identification and enumeration that may alter the course of patient treatment.
Comparison of the MagNA pure LC automated system and the RiboPure‐Blood RNA manual method for RNA extraction from multiple myeloma bone marrow samples conserved in an RNA stabilizer Tập 29 Số 2 - Trang 139-144 - 2007
Guillermo García‐Effrón, Soledad Gamarra, Almudena Crooke, Pilar Martínez‐Sánchez, Juan José Lahuerta, Joaquín Martínez‐López
SummaryA total of 62 frozen bone marrow specimens conserved in RNA later® (Ambion) were processed using two different extraction methods, the MagNA Pure LC system (MAG; Roche) and the manual RiboPure‐Blood RNA method (RIBO; Ambion); Beta glucoronidase RNA (GUS) was amplified by LightCycler PCR to evaluate the quality of both extraction procedures. Less than 1000 GUS copies/ml was detected in 26 of 62 specimens (41.94%) processed by MAG and in five of 62 specimens (8.06%) processed by RIBO. Moreover, RNA recovery from the 62 specimens by MAG is, on average, 2.91 cycle threshold‐fold higher than RIBO (P = 0.0008). Furthermore, we compared the extraction times and reagent costs of both methods. In conclusion, RNA extraction using MAG is faster to process 32 samples and less expensive than RIBO but it is not sensitive enough to be employed for research purpose in our laboratory.
GATA1 mutation analysis and molecular landscape characterization in acute myeloid leukemia with trisomy 21 in pediatric patients Tập 43 Số 4 - Trang 713-723 - 2021
А. В. Панферова, Marina Gaskova, Eugenyi Nikitin, Pavel Baryshev, Natalia Timofeeva, Anna Kazakova, V. E. Matveev, Ekaterina Mikhailova, А. М. Попов, И. И. Калинина, Lili Hachatrian, Aleksey Maschan, Michael Maschan, Galina Novichkova, Yulia Olshanskaya
AbstractIntroductionAccurate detection of GATA1 mutation is highly significant in patients with acute myeloid leukemia (AML) and trisomy 21 as it allows optimization of clinical protocol. This study was aimed at (a) enhanced search for GATA1 mutations; and (b) characterization of molecular landscapes for such conditions.
MethodsThe DNA samples from 44 patients with newly diagnosed de novo AML with trisomy 21 were examined by fragment analysis and Sanger sequencing of the GATA1 exon 2, complemented by targeted high‐throughput sequencing (HTS).
ResultsAcquired GATA1 mutations were identified in 43 cases (98%). Additional mutations in the genes of JAK/STAT signaling, cohesin complex, and RAS pathway activation were revealed by HTS in 48%, 36%, and 16% of the cases, respectively.
ConclusionsThe GATA1 mutations were reliably determined by fragment analysis and/or Sanger sequencing in a single PCR amplicon manner. For patients with extremely low blast counts and/or rare variants, the rapid screening with simple molecular approaches must be complemented with HTS. The JAK/STAT and RAS pathway‐activating mutations may represent an extra option of targeted therapy with kinase inhibitors.
Proteomics analysis reveals alterations of NK cells in patients with severe aplastic anemia Tập 42 Số 3 - Trang 308-315 - 2020
Hui Liu, Tian Zhang, Yingying Chen, Chunyan Liu, Weiwei Qi, Yuanyuan Shao, Hong Yu, Tong Chen, Shaoxue Ding, Yang Li, Ting Wang, Zonghong Shao, Rong Fu
AbstractIntroductionSevere aplastic anemia (SAA) is a disease characterized by severe pancytopenia and hematopoietic failure of bone marrow. Natural killer (NK) cells are a class of large granular lymphocytes that perform killing and immunomodulatory functions. Our previous study demonstrated that NK cells played the “protective” role in SAA, which is weakened. However, the mechanism remains unclear.
MethodsPeripheral blood NK cells from SAA patients and normal controls were sorted and total proteins were extracted. Then, mass spectrometry was performed to screen differentially expressed proteins (DEPs).
ResultsSignificant differences in the expression levels of 93 proteins were observed in NK cells of SAA patients compared with normal controls. Among them, 48 were upregulated proteins, including histone H1.2, histone H1.3, heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1), and interferon regulatory factor 1 (IRF‐1), and 45 were downregulated proteins, including actin‐related complex (ARP2/3), histone H3, histone H4, phosphoglycerate kinase 1 (PGK1), talin‐1. Gene Ontology (GO) function indicated that the DEPs most involved were vesicle‐mediated transport, innate immune response, and DNA binding. KEGG analysis showed 3 upregulated and 12 downregulated pathways, in which cell endocytosis and FC‐γ receptor‐mediated phagocytosis were most closely related to NK cell functions.
ConclusionOur study is the first analysis of proteomic profile in NK cells in SAA and found many DEPs involving in dysfunction of NK cells, which provides potential targets for deeper research of inadequate immunomodulation.
Routine blood biomarkers for the detection of multiple myeloma using machine learning Tập 44 Số 3 - Trang 558-566 - 2022
Gaowei Fan, Ruifang Cui, Qian Zhang, Shunli Zhang, Ruipeng Guo, Yixuan Zhai, Yuhong Yue, Qingtao Wang
AbstractIntroductionPrimary laboratory tests performed in the diagnosis of multiple myeloma (MM) include bone marrow examination and free light chain assay; however, these may only be ordered after clinical suspicion of disease. In contrast, routine blood test results are readily available.
MethodsMachine learning algorithms (ML) combined with routine blood tests were used to detect MM. Feature selection was performed to achieve improved classification performance. The robustness of the classification models was assessed in an internal and external validation data set. To minimize the divergence, the training and validation data sets were combined and used to assess the performance of the ML algorithms.
ResultsThe AdaBoost‐DecisionTable produced the best performance (accuracy =94.75%, sensitivity =87.70%, positive predictive value (PPV) =92.50%, F‐measure =90.00%, and areas under the receiver operating characteristic curves (AUC) =97.50%) in the training data set using a 10‐fold cross‐validation. Performance in the validation data sets was affected by the divergence of the data sets, with accuracy greater than 85% and AUC greater than 90% in the validation data sets. The ML algorithm achieved a high accuracy of 92.61%, high AUC (96.80%), a sensitivity value of 85.20%, a PPV value of 88.50%, and an F‐measure of 86.80% in a test set that was randomly selected from the combined data set.
ConclusionsCombining ML and routine serum biomarkers hold a potential benefit in MM diagnosis.
