Flow cytometry analysis of the circulating haemocytes from Biomphalaria glabrata and Biomphalaria tenagophila following Schistosoma mansoni infection

Parasitology ◽  
2009 ◽  
Vol 136 (1) ◽  
pp. 67-76 ◽  
Author(s):  
R. L. MARTINS-SOUZA ◽  
C. A. J. PEREIRA ◽  
P. M. Z. COELHO ◽  
O. A. MARTINS-FILHO ◽  
D. NEGRÃO-CORRÊA
Keyword(s):  
Schistosoma Mansoni ◽  
Flow Cytometric Analysis ◽  
Cell Subset ◽  
Flow Cytometry Analysis ◽  
Forward Scatter ◽  
Flow Cytometric ◽  
Orange Fluorescence ◽  
Labelling Procedure ◽  
Dual Colour ◽  
Cellular Components

SUMMARYAiming to further characterize the haemocyte subsets in Biomphalaria snails, we have performed a detailed flow cytometric analysis of whole haemolymph cellular components using a multiparametric dual colour labelling procedure. Ethidium bromide/acridine orange fluorescence features were used to first select viable haemocytes followed by flow cytometric morphometric analysis based on the laser scatter properties (forward scatter-FSC and side scatter-SSC). Our findings demonstrated that B. glabrata (BG-BH, highly susceptible to S. mansoni) and 2 strains of B. tenagophila (BT-CF, moderately susceptible and BT-Taim, resistant to S. mansoni) have 3 major circulating haemocyte subsets, referred to as small, medium and large haemocytes. The frequency of small haemocytes was higher in BG-BH, while medium haemocytes were the most abundant cell-type in both B. tenagophila strains. Schistosoma mansoni infection resulted in early reduction of large and medium circulating haemocytes followed by an increase of small haemocytes. Although parasite infection induced haemocyte alterations in all Biomphalaria strains, the response was particularly intense in BT-Taim, the parasite-resistant snail. Interestingly, the trematode infection induces changes in haemocytes with less granular rather than in those with more granular profile. The results indicated that, in B. tenagophila of Taim strain, circulating haemocytes, especially the medium and high subset with less granular profile, are very reactive cells upon S. mansoni infection, suggesting that this cell subset would participate in the early parasite destruction observed in this snail strain.

scholarly journals Analysis of Circulating Haemocytes fromBiomphalaria glabratafollowingAngiostrongylus vasorumInfection Using Flow Cytometry

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Thales A. Barçante ◽  
Joziana M. P. Barçante ◽  
Ricardo T. Fujiwara ◽  
Walter S. Lima
Keyword(s):  
Control Strategies ◽  
Flow Cytometric Analysis ◽  
Intermediate Hosts ◽  
Flow Cytometric ◽  
Wide Range ◽  
Activation Mechanisms ◽  
Labelling Procedure ◽  
Dual Colour ◽  
Cellular Defence ◽  
Cellular Components

Angiostrongylus vasorumis an emerging parasite of dogs and related to carnivores that have an indirect life cycle, with a wide range of terrestrial and aquatic gastropods as the obligatory intermediate host. Unfortunately, the relationship betweenA. vasorumand their snail hosts remains poorly understood. Circulating haemocytes are the main line of cellular defence involved in the destruction of helminths in snails. Aiming to further characterize the haemocyte subsets inBiomphalariasnails, we have performed a flow cytometric analysis of whole haemolymph cellular components using a multiparametric dual colour labelling procedure. Our findings demonstrated thatB. glabratainfected withA. vasorumhave two major circulating haemocyte subsets, referred to as small and large haemocytes. Differences in the cell proportion occurred over time. The development of better invertebrate infection control strategies would certainly result in the better control of human diseases caused by other species of the genusAngiostrongylus. Such knowledge will assist in the establishment of novel control strategies aimed at parasites that use molluscs as intermediate hosts and clarify new aspects of the parasite-host relationship regarding cell recognition and activation mechanisms, which are also found in the innate response of vertebrates.

Ibudilast, a nonselective phosphodiesterase inhibitor, regulates Th1/Th2 balance and NKT cell subset in multiple sclerosis

2004 ◽  
Vol 10 (5) ◽  
pp. 494-498 ◽  
Author(s):  
Juan Feng ◽  
Tatsuro Misu ◽  
Kazuo Fujihara ◽  
Saburo Sakoda ◽  
Yuji Nakatsuji ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Flow Cytometric Analysis ◽  
Cell Subset ◽  
Phosphodiesterase Inhibitor ◽  
Interleukin 4 ◽  
Clinical Effects ◽  
Autoimmune Encephalomyelitis ◽  
Flow Cytometric ◽  
Mrna Ratio ◽  
Killer T Cells

We investigated the immunoregulatory effects of ibudilast, a nonselective phosphodiesterase inhibitor, at a clinically applicable dose (60 mg/day p.o. for four weeks) in multiple sclerosis (MS) patients. Sensitive real-time PCR for quantifying cytokine mRNA in the blood CD4- cells revealed that the ibudilast monotherapy significantly reduced tumour necrosis factor-a and interferon (IFN)-g mRNA and the IFN-g/interleukin-4 mRNA ratio, suggesting a shift in the cytokine profile from Th1 toward Th2 dominancy. In a flow cytometric analysis, natural killer T cells, which have been reported to relate to Th2 responses in MS and its animal model (experimental autoimmune encephalomyelitis), increased significantly after the therapy. None of the significant immunological changes were seen in healthy subjects or untreated MS patients. Ibudilast may be a promising therapy for MS and its clinical effects warrant further study.

scholarly journals GPR18 is required for a normal CD8αα intestinal intraepithelial lymphocyte compartment

2014 ◽  
Vol 211 (12) ◽  
pp. 2351-2359 ◽  
Author(s):  
Xiaoming Wang ◽  
Hayakazu Sumida ◽  
Jason G. Cyster
Keyword(s):  
T Cells ◽  
Lamina Propria ◽  
Flow Cytometric Analysis ◽  
Cell Subset ◽  
Intraepithelial Lymphocytes ◽  
Orphan Receptor ◽  
Flow Cytometric ◽  
Intestinal Intraepithelial Lymphocyte ◽  
Mixed Chimeras ◽  
G Protein Coupled

Intraepithelial lymphocytes (IELs) play an important role in maintaining the physiology of the small intestine. The majority of mouse IELs express CD8αα and are either γδ or αβ T cells. Although the development and homing of CD8αα IELs have been studied in some detail, the factors controlling their homeostasis and positioning are incompletely understood. Here we demonstrate that G protein–coupled receptor 18 (GPR18) is abundantly expressed in CD8αα IELs and that mice lacking this orphan receptor have reduced numbers of γδT IELs. Mixed bone marrow chimera experiments reveal a markedly reduced contribution of GPR18-deficient cells to the CD8αα IEL compartment and a reduction in the CD8αβ T cell subset. These defects could be rescued by transduction with a GPR18-expressing retrovirus. The GPR18-deficient γδT IELs that remained in mixed chimeras had elevated Thy1, and there were less granzyme B+ and Vγ7+ cells, indicating a greater reduction in effector-type cells. Flow cytometric analysis indicated GPR18 deficiency more strongly affected the CD8αα cells in the intraepithelial compared with the adjacent lamina propria compartment. These findings establish a requirement for GPR18 in CD8αα and CD8αβ IELs, and we suggest the receptor has a role in augmenting the accumulation of CD8 T cells in the intraepithelial versus lamina propria compartment.

scholarly journals Flow Cytometry Analysis of Recurrent or Persistent Lymphadenopathy in Patients with Nodular Lymphocyte-Predominant Hodgkin Lymphoma

2021 ◽  
pp. 1-6
Author(s):  
James Z. Huang ◽  
Savanah D. Gisriel ◽  
Kristle Haberichter ◽  
Sara Huang ◽  
James Z. Huang
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Hodgkin Lymphoma ◽  
Expression Patterns ◽  
Flow Cytometric Analysis ◽  
Flow Cytometry Analysis ◽  
Flow Cytometric ◽  
Multiple Biopsies ◽  
Highly Sensitive ◽  
Reactive Hyperplasia

Objectives: We recently examined the utility of flow cytometric analysis in the diagnosis of nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) by examining reactive T-cell features. This study aims to compare these features in sequential biopsies of persistent or recurrent lymphadenopathy in patients with NLPHL. Methods: We reanalysed the histopathology and flow cytometry findings of 9 patients with multiple biopsies for persistent or recurrent lymphadenopathy and either initial or recurrent NLPHL. A flow cytometry signature was considered suggestive of NLPHL if ≥12% of T-cells expressed CD57 or ≥3% of T-cells co-expressed CD4 and CD8. Results: A flow cytometry signature considered suggestive of NLPHL was seen in 18 of 20 specimens. Based on histopathology, 11 were diagnosed as NLPHL, 3 were initially underdiagnosed as atypical lymphoid proliferation, and 4 were initially incorrectly diagnosed as negative or progressive transformation of germinal centers. Flow cytometry showed similar expression patterns of CD57 and CD4/CD8 in T-cells between initial and subsequent biopsies. The remaining 2 specimens lacked the flow cytometry signature suggestive of NLPHL and were histopathologically diagnosed as reactive hyperplasia. Conclusion: Flow cytometry analysis based on our criteria is highly sensitive in detecting NLPHL. Correlation with the cytospin cytology may increase the diagnostic specificity. A negative flow essentially ruled out the possibility of NHLPHL.

scholarly journals COVID-19 infection induces readily detectable morphological and inflammation-related phenotypic changes in peripheral blood monocytes, the severity of which correlate with patient outcome

2020 ◽  
Author(s):  
Dan Zhang ◽  
Rui Guo ◽  
Lei Lei ◽  
Hongjuan Liu ◽  
Yawen Wang ◽  
...  
Keyword(s):  
Patient Outcome ◽  
Flow Cytometry ◽  
Peripheral Blood ◽  
Flow Cytometric Analysis ◽  
Blood Monocytes ◽  
Forward Scatter ◽  
Inflammatory Monocytes ◽  
Flow Cytometric ◽  
Functional Differences ◽  
Inflammatory Phenotype

AbstractBackgroundExcessive monocyte/macrophage activation with the development of a cytokine storm and subsequent acute lung injury, leading to acute respiratory distress syndrome (ARDS) is a feared consequence of infection with COVID-19. The ability to recognize and potentially intervene early in those patients at greatest risk of developing this complication could be of great clinical utility.MethodsWe performed detailed flow cytometric analysis of peripheral blood samples from 28 COVID-19 patients treated at Xian No.8 Hospital and the First Affiliated Hospital of Xian Jiaotong University in early 2020 in an attempt to identify factors that could help predict severity of disease and patient outcome.FindingsWhile we did not detect significant differences in the number of monocytes between patients with COVID-19 and normal healthy individuals,we did identify significant morphological and functional differences, which are more pronounced in patients requiring prolonged hospitalization and ICU admission. Patients with COVID-19 have larger than normal monocytes, easily identified on forward scatter, side scatter analysis by routine flow cytometry,with the presence of a distinct population of monocytes with high forward scatter (FSC-high). On more detailed analysis, these FSC-high monocytes are CD11b+, CD14+, CD16+, CD68+, CD80+, CD163+, CD206+ and secrete IL-6, IL-10 and TNF-alpha, consistent with an inflammatory phenotype.ConclusionsThe detection and serial monitoring of this subset of inflammatory monocytes using flow cytometry could be of great help in guiding the prognostication and treatment of patients with COVID-19 and merits further evaluation.

scholarly journals Flow cytometric analysis and purification of airway epithelial cell subsets

2020 ◽  
Author(s):  
Luke R. Bonser ◽  
Kyung Duk Koh ◽  
Kristina Johansson ◽  
Semil P. Choksi ◽  
Dan Cheng ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Flow Cytometric Analysis ◽  
Cell Subset ◽  
Airway Epithelial Cells ◽  
Secretory Cells ◽  
Airway Epithelial ◽  
Specific Expression ◽  
Flow Cytometric ◽  
Micro Rnas ◽  
Epithelial Biology

AbstractThe human airway epithelium is essential in homeostasis, and epithelial dysfunction contributes to chronic airway disease. Development of flow cytometric methods to characterize subsets of airway epithelial cells will enable further dissection of airway epithelial biology. Leveraging single cell RNA-sequencing (scRNA-seq) data in combination with known cell type-specific markers, we developed panels of antibodies to characterize and isolate the major airway epithelial subsets (basal, ciliated, and secretory cells) from human bronchial epithelial cell cultures. We also identified molecularly distinct subpopulations of secretory cells and demonstrated cell subset-specific expression of low abundance transcripts and micro-RNAs that are challenging to analyze with current scRNA-seq methods. These new tools will be valuable for analyzing and separating airway epithelial subsets and interrogating airway epithelial biology.

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