Characterization of Extracellular Nucleotide-Induced Mac-1 (αMβ2Integrin) Surface Expression on Peripheral Blood Leukocytes

1997 ◽  
Vol 233 (1) ◽  
pp. 71-75 ◽  
Author(s):  
G.K.Mohammed Akbar ◽  
David C.B. Mills ◽  
Satya P. Kunapuli
Keyword(s):  
Peripheral Blood ◽  
Surface Expression ◽  
Peripheral Blood Leukocytes ◽  
Extracellular Nucleotide ◽  
Blood Leukocytes

scholarly journals Hyperexpression of TLR2 and TLR4 in patients with ischemic stroke in acute period of the disease

2020 ◽  
Vol 22 (4) ◽  
pp. 665-674
Author(s):  
L. V. Gankovskaya ◽  
L. V. Stakhovskaya ◽  
V. V. Grechenko ◽  
E. A. Koltsova ◽  
O. S. Uvarova ◽  
...  
Keyword(s):  
Gene Expression ◽  
Innate Immunity ◽  
Ischemic Stroke ◽  
Peripheral Blood ◽  
Surface Expression ◽  
Control Group ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Healthy Donors ◽  
Tlr4 Gene

Pathogenesis of ischemic stroke  is actively  involved  in the  system  of innate immunity. Under conditions of cerebral  ischemia, a number of biologically  active  substances are  released  that  interact with innate immunity receptors, in particular TLR2  and  TLR4, which  exacerbate inflammation in brain  tissue. Identification of predictor markers  at the level of the innate immunity system may foresee the clinical course of ischemic stroke and ensure timely treatment. Our objective was to study expression of TLR2 and TLR4 receptors in peripheral blood leukocytes  in patients with ischemic stroke in the dynamics of the disease. 27 people  were included in the study. The main  group consisted of patients with ischemic stroke of varying severity (n = 19). Patients of the main  group were divided into two subgroups:  with an NIHSS index value of < 10 (n = 10) and > 10 (n = 9). The control group included healthy  donors  with no history  of acute  and chronic inflammatory diseases (n = 8). Peripheral blood  leukocytes  were used as the  test material. To determine expression  of the TLR2  and TLR4  genes, RT-PCR in real time was used. Surface  expression  of TLRs was determined by flow cytometry. A study of the TLR2 and TLR4 gene expression showed that on the 1st, 3rd  and 7th  day post-stroke, the TLR4 gene expression  in patients was significantly  increased, when compared to the control group (p < 0.01), whereas TLR2 gene expression on the 3rd  day of the disease was not statistically different from the control group. A study of surface expression  of receptors showed that the average TLR2 fluorescence intensity on the patients’ peripheral blood monocytes was significantly  increased on the 1st  and 3rd  day of disease when compared to the control group.  The  surface  expression  of TLR4  on monocytes has a statistically significant  increase  only on day 7. Assessment  of surface expression  of TLRs in subgroups  with different  severity values by NIHSS showed that  patients with a NIHSS index > 10 had a significantly  higher  level of surface of TLR2  expression  over the observation period, while the largest difference in TLR4  expression  in the subgroups  was observed  on the 1st day of the disease (p < 0.05). Patients with ischemic stroke showed an increase  in TLR2 and TLR4 expression at the gene and protein level, compared to healthy  donors. These indices can be considered possible predictors for clinical  prognosis  of ischemic stroke.

scholarly journals The Characterization of peripheral blood leukocytes during the development of adjuvant arthritis in rats.

Ensho ◽  
1992 ◽  
Vol 12 (1) ◽  
pp. 55-61
Author(s):  
Yoshihide Segawa ◽  
Masakatsu Nozaki ◽  
Kaito Tsurumi ◽  
Shunro Kohbata
Keyword(s):  
Peripheral Blood ◽  
Adjuvant Arthritis ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes

Leukocyte Integrin CD18 Expression Mediates Transient Neutropenia Following G-CSF Administration.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3587-3587
Author(s):  
Laura Tuschong ◽  
Catherine E. Dejesus ◽  
Meredith Adams ◽  
Aylin C. Bonifacino ◽  
Dennis D. Hickstein ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Conflicts Of Interest ◽  
Leukocyte Adhesion ◽  
Surface Expression ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Cell Counts ◽  
Transient Loss ◽  
The Mean ◽  
White Blood Cell Counts

Abstract Abstract 3587 Poster Board III-524 The mechanism whereby neutrophils traffic from the circulation in response to G-CSF has remained unclear despite the observation of ourselves and others that there is a dramatic, yet transient, loss of circulating neutrophils shortly following the administration of G-CSF in humans, non-human primates, and mice (Gordon BC, et al. Exp Hematol. 35:872-8, 2007). To determine the role of the CD18 leukocyte integrin on neutrophils in the egress of neutrophils from the circulation, we used dogs with canine leukocyte adhesion deficiency (CLAD), a genetic disease in which a mutation in CD18 prevents CD18 surface expression. We selected CLAD dogs who had 5-10% CD18+ neutrophils following either matched littermate allogeneic transplant or autologous gene therapy for CLAD. Three CLAD dogs meeting these criteria were evaluated. Three carrier dogs served as controls. G-CSF was administered at 10μg/kg SQ to all six animals. Peripheral blood samples (EDTA) were taken immediately prior to G-CSF administration, and at 15, 30, 60, 120, 240 minutes, and 24 hours following G-CSF administration. Total white blood cell counts, neutrophil counts, and the number and percentage of CD18+ peripheral blood leukocytes were assessed. As anticipated, the control dogs had a 60% decrease in circulating neutrophils 30 minutes following G-CSF administration: the mean +/− standard of deviation (SD) absolute neutrophil baseline count decreased from 6806+/−1072/μL to 2727+/−767/μL. In five control animals the neutrophil nadir occurred at 30 minutes post-G-CSF, and in one control dogs it occurred 15 minutes following G-CSF administration. Experimental CLAD dogs had only a 35% decline in neutrophil numbers at 30 minutes, from a mean baseline of 6777+/− 672/μL to 4433+/−265/μL. In these dogs the neutrophil count returned to pre-G-CSF levels by 60 minutes post-G-CSF. By 24 hours after G-CSF, the neutrophil level was increased 3-fold from baseline. Immunophenotyping using an anti-CD18 and a canine specific anti-neutrophil PE conjugated antibody indicated that only the CD18+ neutrophils disappeared from the circulation following G-CSF administration. At baseline the transplanted CLAD dogs had a mean of 15.2+/−3.9% CD18+ peripheral blood leukocytes, of these 50.7+/−7.1% were CD18+ neutrophils. Thirty minutes following G-CSF administration the mean+/−SD percentage of CD18+ leukocytes declined to 13.7+/−3.7% with 33.4+/−5.8% being neutrophils. There was also a slight decline in CD14+CD18+ monocytes from 6.2 +/− 1.5% to 4.0 +/− 1.2%, which was not observed in the controls. There was no change in CD18- leukocyte numbers. The percentage of CD18+ neutrophils returned to baseline by 60 minutes and remained there at subsequent time points. These results demonstrate that the CD18 leukocyte integrin on circulating neutrophils mediates the transient neutropenia associated with G-CSF administration. Disclosures: No relevant conflicts of interest to declare.

High-Throughput Sequencing for Diagnosis, Prognosis and Monitoring of Lymphoid Malignancies

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3779-3779
Author(s):  
Scott D Boyd ◽  
Jason D. Merker ◽  
James L. Zehnder ◽  
Andrew Z Fire
Keyword(s):  
High Throughput ◽  
Peripheral Blood ◽  
Read Length ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Normal Peripheral Blood ◽  
Dilution Series ◽  
Lymphoid Malignancies ◽  
Immune Receptors

Abstract New high-throughput DNA sequencing methods have many advantages for the analysis of rearranged immunoglobulin (Ig) and T cell receptor (TCR) loci in suspected lymphoid malignancies. Instead of primarily analyzing the size distribution of rearranged immune receptors using gel or capillary electrophoresis, sequencing complex nucleic acid pools provides much more complete characterization of the receptor sequences present. Currently, separate assays are used to evaluate Ig and TCR clonality, Ig hypermutation status, and to test for residual malignant cells after treatment. We used multiplexed PCR amplification of immunoglobulin heavy chain loci for high-throughput pyrosequencing with the Roche 454 platform to evaluate the detection of clonal IgH sequences, determination of hypermutation status, and sensitivity of detection of low levels of a known clonal receptor. Our method also evaluated the feasibility of physical “bar-coding” of DNA molecules derived from different samples to enable cost-effective sequencing of sample pools. 23 pooled specimens were sequenced including: known lymphoma specimens (4); peripheral blood leukocytes from normal individuals (4); specimens with indeterminate or discordant results by conventional clonality assays (7); and a dilution series of a clonal CLL/SLL specimen into normal peripheral blood leukocytes. Our initial experiment characterized 522,099 IgH sequences from these specimens, with a mean read length of 242 base pairs. We have employed IgBlast (NCBI) for initial alignment of rearranged IgH sequences to germline V, D and J gene segments. Our data demonstrate that clonal IgH populations can be readily detected and characterized as to their germline or hypermutated status and unique V-D and D-J junctions with this approach. In our dilution series experiment, the clonal receptor sequence could be reliably observed after 1000-fold dilution. Normal patient specimens show expected extensive sequence diversity, and we are exploring methods for optimally summarizing and representing these data. Some PCR artifacts, as well as non-immunoglobulin sequences commonly amplified by current IgH primer sets were also detected, and suggest avenues for further improvement of the methods used for detection and characterization of clonal immune receptors. More broadly, these methods also offer a novel approach to monitoring normal immune responses such as the response of patients to vaccination, and of characterizing the immune system in patients with autoimmune diseases.

scholarly journals Functional characterization of neotropical snakes peripheral blood leukocytes subsets: Linking flow cytometry cell features, microscopy images and serum corticosterone levels

2017 ◽  
Vol 74 ◽  
pp. 144-153 ◽  
Author(s):  
Marcelo Pires Nogueira de Carvalho ◽  
Nicolle Gilda Teixeira Queiroz-Hazarbassanov ◽  
Cristina de Oliveira Massoco ◽  
Sávio Stefanini Sant’Anna ◽  
Mariana Mathias Lourenço ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Peripheral Blood ◽  
Functional Characterization ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Serum Corticosterone ◽  
Microscopy Images
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