scholarly journals Label-Free, High-Throughput Assay of Human Dendritic Cells from Whole-Blood Samples with Microfluidic Inertial Separation Suitable for Resource-Limited Manufacturing

Micromachines ◽  
2020 ◽  
Vol 11 (5) ◽  
pp. 514
Author(s):  
Mohamed Yousuff Caffiyar ◽  
Kue Peng Lim ◽  
Ismail Hussain Kamal Basha ◽  
Nor Hisham Hamid ◽  
Sok Ching Cheong ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Point Of Care ◽  
Scale Up ◽  
Academic Research ◽  
Cell Types ◽  
Label Free ◽  
Resource Limited ◽  
Human Blood Cells ◽  
Human Dendritic Cells ◽  
High Throughput Assay

Microfluidics technology has not impacted the delivery and accessibility of point-of-care health services, like diagnosing infectious disease, monitoring health or delivering interventions. Most microfluidics prototypes in academic research are not easy to scale-up with industrial-scale fabrication techniques and cannot be operated without complex manipulations of supporting equipment and additives, such as labels or reagents. We propose a label- and reagent-free inertial spiral microfluidic device to separate red blood, white blood and dendritic cells from blood fluid, for applications in health monitoring and immunotherapy. We demonstrate that using larger channel widths, in the range of 200 to 600 µm, allows separation of cells into multiple focused streams, according to different size ranges, and we utilize a novel technique to collect the closely separated focused cell streams, without constricting the channel. Our contribution is a method to adapt spiral inertial microfluidic designs to separate more than two cell types in the same device, which is robust against clogging, simple to operate and suitable for fabrication and deployment in resource-limited populations. When tested on actual human blood cells, 77% of dendritic cells were separated and 80% of cells remained viable after our assay.

scholarly journals Label-free, High-throughput Assay of Human Dendritic Cells from Whole-blood Samples with Microfluidic Inertial Separation Suitable for Resource-limited Manufacturing

2020 ◽  
Author(s):  
Caffiyar Mohd Yousuff ◽  
Kue Peng Lim ◽  
Ismail Hussain Kamal Basha ◽  
Nor Hisham Hamid ◽  
Sok Ching Cheong ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Point Of Care ◽  
Academic Research ◽  
Cell Types ◽  
Disease Diagnosis ◽  
Label Free ◽  
Resource Limited ◽  
Human Blood Cells ◽  
Human Dendritic Cells ◽  
High Throughput Assay

Microfluidics technology has not impacted the delivery and accessibility of point of care health services like diagnosis of infectious disease diagnosis, monitoring health or delivering interventions. Most microfluidics prototypes from academic research are not easy to manufacture with industrial scale fabrication techniques and cannot be operated without complex manipulations of supporting equipment and additives such as labels or reagents. We propose a label- and reagent-free inertial spiral microfluidic device to separate red blood, white blood and dendritic cells from blood fluid for applications in health monitoring and immunotherapy. We demonstrate that using larger channel widths in the range of 200 to 600 µm allows separation of cells into multiple streams according to different size ranges and we utilize a novel technique to collect the closely separated focused cell streams without constricting the channel. When tested on actual human blood cells, 77% of dendritic cells were separated and 80% of cells remained viable after our assay. Our contribution is a method to adapt spiral inertial microfluidic designs to separate more than two cell types in the same device which is robust against clogging, simple to operate and suitable for fabrication and deployment in resource-limited populations.

scholarly journals Human Dendritic Cells Mediate Cellular Apoptosis via Tumor Necrosis Factor–Related Apoptosis-Inducing Ligand (Trail)

1999 ◽  
Vol 190 (8) ◽  
pp. 1155-1164 ◽  
Author(s):  
Neil A. Fanger ◽  
Charles R. Maliszewski ◽  
Ken Schooley ◽  
Thomas S. Griffith
Keyword(s):  
Dendritic Cells ◽  
Tumor Cells ◽  
Cell Types ◽  
Target Cells ◽  
Functional Difference ◽  
Cellular Apoptosis ◽  
Soluble Trail ◽  
Innate Effector ◽  
Human Dendritic Cells ◽  
Necrosis Factor

TRAIL (TNF-related apoptosis-inducing ligand) is a member of the TNF family that induces apoptosis in a variety of cancer cells. In this study, we demonstrate that human CD11c+ blood dendritic cells (DCs) express TRAIL after stimulation with either interferon (IFN)-γ or -α and acquire the ability to kill TRAIL-sensitive tumor cell targets but not TRAIL-resistant tumor cells or normal cell types. The DC-mediated apoptosis was TRAIL specific, as soluble TRAIL receptor blocked target cell death. Moreover, IFN-stimulated interleukin (IL)-3 receptor (R)α+ blood precursor (pre-)DCs displayed minimal cytotoxicity toward the same target cells, demonstrating a clear functional difference between the CD11c+ DC and IL-3Rα+ pre-DC subsets. These results indicate that TRAIL may serve as an innate effector molecule on CD11c+ DCs for the elimination of spontaneously arising tumor cells and suggest a means by which TRAIL-expressing DCs may regulate or eliminate T cells responding to antigen presented by the DCs.

scholarly journals Attenuated Interferon and Proinflammatory Response in SARS-CoV-2–Infected Human Dendritic Cells Is Associated With Viral Antagonism of STAT1 Phosphorylation

2020 ◽  
Vol 222 (5) ◽  
pp. 734-745 ◽  
Author(s):  
Dong Yang ◽  
Hin Chu ◽  
Yuxin Hou ◽  
Yue Chai ◽  
Huiping Shuai ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Clinical Manifestations ◽  
Cell Types ◽  
Host Responses ◽  
Interferon Response ◽  
Proinflammatory Response ◽  
Innate And Adaptive Immunity ◽  
Virus Shedding ◽  
Human Dendritic Cells ◽  
Productive Virus

Abstract Clinical manifestations of coronavirus disease 2019 (COVID-19) vary from asymptomatic virus shedding, nonspecific pharyngitis, to pneumonia with silent hypoxia and respiratory failure. Dendritic cells and macrophages are sentinel cells for innate and adaptive immunity that affect the pathogenesis of severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). The interplay between SARS-CoV-2 and these cell types remains unknown. We investigated infection and host responses of monocyte-derived dendritic cells (moDCs) and macrophages (MDMs) infected by SARS-CoV-2. MoDCs and MDMs were permissive to SARS-CoV-2 infection and protein expression but did not support productive virus replication. Importantly, SARS-CoV-2 launched an attenuated interferon response in both cell types and triggered significant proinflammatory cytokine/chemokine expression in MDMs but not moDCs. Investigations suggested that this attenuated immune response to SARS-CoV-2 in moDCs was associated with viral antagonism of STAT1 phosphorylation. These findings may explain the mild and insidious course of COVID-19 until late deterioration.

scholarly journals The potential of SERS as an AST methodology in clinical settings

Nanophotonics ◽  
2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Ota Samek ◽  
Silvie Bernatová ◽  
Fadi Dohnal
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Physiological State ◽  
Analytical Techniques ◽  
Cell Types ◽  
Rapid Identification ◽  
Clinical Diagnostics ◽  
Current Status ◽  
Label Free ◽  
Purine Bases

Abstract The ability to identify and characterize microorganisms from tiny sample volumes in a rapid and reliable way is the first and crucial step in the diagnostics of microbial infections. Ideal analytical techniques would require minimal and low-cost sample preparation, permit automatic analysis of many serial samples, and allow rapid classification of present microorganisms against a stable database. Current practice, however, is far from this ideal; a typical analytical procedure might require a few days. Delayed laboratory results might lead, for example, to progress/spread of the infection, more serious condition of the patient, even death, prescription of inappropriate antibiotics that could be ineffective against causative agents and may as well contribute to the emerging problem of drug resistance in microorganisms. Several studies confirmed that surface enhanced Raman scattering (SERS) is capable of a rapid identification and discrimination of biological samples including medically relevant bacteria. A typical spectrum contains a wealth of information indicative of the cellular content of nucleic acids, purine bases, proteins, carbohydrates, and lipids. Such a spectrum functions as a cellular ‘fingerprint’ and serves as a sensitive indicator of the physiological state of the cell which in turn enables to differentiate cell types, actual physiological states, nutrient conditions, and phenotype changes. Consequently, the focus of this review is on the SERS spectra of bacteria which result from secreted metabolic substances – the purine bases – which are a common feature in the label-free SERS research related to clinical diagnostics of pathogens. Here is the review of the current status of SERS applications on bacteria. A special attention is given to the efforts of profiling antimicrobial susceptibility at clinically relevant species, which in turn has a great potential for use in routine point-of-care (POC) tests. Thus, early and accurate infection disease management can be provided at the bedside or at remote care centres.

scholarly journals Human Cytomegalovirus Gene Products US2 and US11 Differ in Their Ability To Attack Major Histocompatibility Class I Heavy Chains in Dendritic Cells

2002 ◽  
Vol 76 (10) ◽  
pp. 5043-5050 ◽  
Author(s):  
Armin Rehm ◽  
Arne Engelsberg ◽  
Domenico Tortorella ◽  
Ida J. Körner ◽  
Insa Lehmann ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Mhc Class I ◽  
Mhc Class Ii ◽  
Human Cytomegalovirus ◽  
Cell Types ◽  
Class I ◽  
Major Histocompatibility ◽  
Heavy Chains ◽  
Histocompatibility Complex ◽  
Human Dendritic Cells

ABSTRACT Human cytomegalovirus (HCMV) encodes several proteins that inhibit major histocompatibility complex (MHC) class I-dependent antigen presentation. The HCMV products US2 and US11 are each sufficient for causing the dislocation of human and murine MHC class I heavy chains from the lumen of the endoplasmic reticulum to the cytosol, where the heavy chains are readily degraded. The apparent redundancy of US2 and US11 has been probed predominantly in cultured cell lines, where differences in their specificities were shown for murine and human MHC class I locus products. Here, we expressed US11 and US2 via adenovirus vectors and show that US11 exhibits a superior ability to degrade MHC class I molecules in primary human dendritic cells. MHC class II complexes are unaffected by US2- and US11-mediated attack. We suggest that multiple HCMV-encoded immunoevasions have evolved complementary functions in response to diverse host cell types and tissues.

scholarly journals Human Dendritic Cells with Th2-Polarizing Capacity: Analysis Using Label-Free Quantitative Proteomics

2017 ◽  
Vol 174 (3-4) ◽  
pp. 170-182 ◽  
Author(s):  
Leonie Hussaarts ◽  
Maria Mardalena Martini Kaisar ◽  
Arzu Tugce Guler ◽  
Hans Dalebout ◽  
Bart Everts ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Quantitative Proteomics ◽  
Capacity Analysis ◽  
Label Free ◽  
Human Dendritic Cells

scholarly journals Characterisation of secretome-based immune responses of human leukocytes infected with various Mycobacterium tuberculosis lineages

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11565
Author(s):  
Benjawan Kaewseekhao ◽  
Sittiruk Roytrakul ◽  
Yodying Yingchutrakul ◽  
Marut Laohaviroj ◽  
Kanin Salao ◽  
...  
Keyword(s):  
Immune Response ◽  
Dendritic Cells ◽  
Mycobacterium Tuberculosis ◽  
B Cells ◽  
Immune Responses ◽  
Antigen Processing ◽  
Immune Cell ◽  
Cell Types ◽  
Label Free ◽  
Related Proteins

Background Differences in immune responses against different lineages of Mycobacterium tuberculosis (Mtb), and by different types of immune cell, are still poorly understood. We aimed to compare the secretome-based immune responses among three Mtb lineages and among immune-cell types. The immune responses were also investigated during infection and when the bacilli had been eliminated from the immune cells. Methods Human primary leukocytes were infected with strains representing three lineages of Mtb (East-Asian, Indo-Oceanic and Euro-American). Label-free GeLC MS/MS proteomic analysis of secretomes was performed. The response of each immune-cell type was compared with the appropriate interactome database for each. Results The expression pattern of proteins secreted by Mtb-infected leukocytes differed among Mtb lineages. The ancestral lineage (IO lineage) had a greater ability to activate MMP14 (associated with leukocyte migration) than did the more recent lineages (EA and EuA). During infection, proteins secreted by macrophages, dendritic cells, neutrophils and B-cells were associated with cell proliferation. Following clearance of Mtb, proteins associated with interferon signaling were found in macrophages, dendritic cells and neutrophils: proteins associated with antigen processing were found in B-cells and regulatory T-cells. Expression of immune response-related proteins from many immune-cell types might be suppressed by Mtb infection. Our study has provided a better insight into the host-pathogen interaction and immune response against different Mtb lineages.

Smartphone-Based Mobile Learning with Physician Trainees in Botswana

2012 ◽  
Vol 4 (2) ◽  
pp. 1-14 ◽  
Author(s):  
Aileen Y. Chang ◽  
Ryan Littman-Quinn ◽  
Dineo Ketshogileng ◽  
Amit Chandra ◽  
Taatske Rijken ◽  
...  
Keyword(s):  
Mobile Learning ◽  
Point Of Care ◽  
Scale Up ◽  
Healthcare Providers ◽  
Technology Support ◽  
University Of Botswana ◽  
Resource Limited ◽  
Access To Health ◽  
Information And Communication ◽  
The University

In recent years, mobile learning in medicine has been utilized to increase healthcare providers’ access to health information. This has improved healthcare providers’ ability to make appropriate clinical decisions at point-of-care, particularly in resource-limited settings. Mobile phones facilitate information and communication technology support for patient care and collaboration amongst providers. In this paper, the authors describe a smartphone-based mobile learning initiative with physician trainees at the University of Botswana School of Medicine, focusing on the authors’ experiences with recent scale-up efforts to remote areas of Botswana. The authors also explore the potential impact of mobile learning in developing health capacity.

scholarly journals Aminopeptidase N (CD13) Is Involved in Phagocytic Processes in Human Dendritic Cells and Macrophages

2013 ◽  
Vol 2013 ◽  
pp. 1-12 ◽  
Author(s):  
Mónica I. Villaseñor-Cardoso ◽  
Dulce A. Frausto-Del-Río ◽  
Enrique Ortega
Keyword(s):  
Dendritic Cells ◽  
Tumor Invasion ◽  
Cell Types ◽  
Innate Immune ◽  
Aminopeptidase N ◽  
Igg Antibodies ◽  
E Coli ◽  
Monocytes And Macrophages ◽  
Human Dendritic Cells ◽  
Human Monocytes And Macrophages

Aminopeptidase N (APN or CD13) is a membrane ectopeptidase expressed by many cell types, including myelomonocytic lineage cells: monocytes, macrophages, and dendritic cells. CD13 is known to regulate the biological activity of various peptides by proteolysis, and it has been proposed that CD13 also participates in several functions such as angiogenesis, cell adhesion, metastasis, and tumor invasion. We had previously reported that, in human monocytes and macrophages, CD13 modulates the phagocytosis mediated by receptors for the Fc portion of IgG antibodies (FcγRs). In this work, we analyzed the possible interaction of CD13 with other phagocytic receptors. We found out that the cross-linking of CD13 positively modulates the phagocytosis mediated by receptors of the innate immune system, since a significant increase in the phagocytosis of zymosan particles or heat-killedE. coliwas observed when CD13 was cross-linked using anti-CD13 antibodies, in both macrophages and dendritic cells. Also, we observed that, during the phagocytosis of zymosan, CD13 redistributes and is internalized into the phagosome. These findings suggest that, besides its known functions, CD13 participates in phagocytic processes in dendritic cells and macrophages.

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