Do microscopy imaging frequency and experiment duration impact the analysis of T cell movement?

2021 ◽  
Author(s):  
Viktor Zenkov ◽  
James O'Connor ◽  
Hayley McNamara ◽  
Ian Cockburn ◽  
Vitaly Ganusov
Keyword(s):  
T Cell ◽  
Cell Movement ◽  
Microscopy Imaging ◽  
Experiment Duration

Electrical Properties and Anion Permeability of Doubly Rectifying Junctions in the Leech Central Nervous System

1988 ◽  
Vol 137 (1) ◽  
pp. 1-11
Author(s):  
Susan E. Acklin
Keyword(s):  
Central Nervous System ◽  
T Cells ◽  
Nervous System ◽  
T Cell ◽  
Action Potentials ◽  
Sodium Pump ◽  
Current Flow ◽  
Cell Movement ◽  
Voltage Dependent ◽  
Electrical Connections

A study has been made of the electrical connections between touch sensory (T) neurones in the leech central nervous system (CNS) which display remarkable double rectification: depolarization spreads in both directions although hyperpolarization spreads poorly. Tests were made to determine whether this double rectification was a property of the junctions themselves or whether it resulted from changes in the length constants of processes intervening between the cell body and the junctions. Following trains of action potentials, T cells and their fine processes within the neuropile became hyperpolarized through the activity of an electrogenie sodium pump. When any T cell was hyperpolarized by 25 mV by repetitive stimulation, hyperpolarization failed to spread to the T cells to which it was electrically coupled. Further evidence for double rectification of junctions linking T cells was provided by experiments in which Cl− was injected electrophoretically. Cl− injection into one T cell caused inhibitory potentials recorded in it to become reversed. After a delay, Cl− spread to reverse IPSPs in the coupled T cell. Movement of Cl−, like current flow, was dependent on membrane potential. When the T cell into which Cl− was injected was kept hyperpolarized, Cl− failed to move into the adjacent T cell. Upon release of the hyperpolarization in the injected T cell, Cl− moved and reversed IPSPs in the coupled T cell. Together these results indicate that the gating properties of channels linking T cells are voltage-dependent, such that depolarization of either cell allows channels to open whereas hyperpolarization causes them to close.

Classification of T cell movement tracks allows for prediction of cell function

2014 ◽  
Vol 7 (2/3) ◽  
pp. 113 ◽  
Author(s):  
Reka K. Kelemen ◽  
Gengen F. He ◽  
Hannah L. Woo ◽  
Thomas Lane ◽  
Caroline Rempe ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Function ◽  
Cell Movement

scholarly journals Longitudinal confocal microscopy imaging of solid tumor destruction following adoptive T cell transfer

2013 ◽  
Vol 2 (11) ◽  
pp. e26677 ◽  
Author(s):  
Andrea Schietinger ◽  
Ainhoa Arina ◽  
Rebecca B Liu ◽  
Sam Wells ◽  
Jianhua Huang ◽  
...  
Keyword(s):  
T Cell ◽  
Confocal Microscopy ◽  
Solid Tumor ◽  
Cell Transfer ◽  
Microscopy Imaging ◽  
Adoptive T Cell Transfer ◽  
Tumor Destruction ◽  
T Cell Transfer ◽  
Confocal Microscopy Imaging

scholarly journals A minority of liver-localized CD8 T cells demonstrate difficult-to-detect attraction to Plasmodium-infected hepatocytes

2020 ◽  
Author(s):  
Viktor S Zenkov ◽  
James O'Connor ◽  
Ian A Cockburn ◽  
Vitaly V Ganusov
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cd8 T Cells ◽  
Cell Movement ◽  
Stochastic Simulations ◽  
Cell Vaccine ◽  
Infection Site ◽  
T Cell Vaccine ◽  
Von Mises ◽  
Sterilizing Immunity

Malaria is a disease caused by parasites from genus Plasmodium resulting in over 200 million infections and 400,000 deaths every year. A critical step of malaria infection is when mosquito-injected sporozoites travel to the liver and form liver stages. Several malaria vaccine candidates tested in mice induce high levels of malaria-specific CD8 T cells which are able to eliminate all liver stages, thus providing sterilizing immunity against the disease. However, how CD8 T cells locate the site of infection is not well understood. We generated and analyzed data from intravital microscopy experiments in mice in which the movement of T cells relative to the liver stage was recorded in several different settings. To detect attraction of T cells towards the infection site, we developed a novel metric based on the Von Mises-Fisher (VMF) distribution, which is more powerful than previously used metrics. Our results suggested that the majority (∼ 70 – 95%) of malaria-specific CD8 T cells and T cells of irrelevant specificity did not display any attraction towards the parasite when the parasite was not found by T cells, but some T cells displayed strong attraction when there was a large cluster of Plasmodium-specific CD8 T cells near the parasite. We found that the speed of T cell movement (and small turning angles) correlated with the bias of T cell movement towards the infection site (but several other parameters did not) suggesting that a deeper understanding of what determines the speed of T cell movement in tissues may help with improving T cell vaccine efficacy. Stochastic simulations suggested that a small movement bias towards the parasite dramatically reduces the number of CD8 T cells needed for a complete elimination of the malaria liver stages from the liver, and yet, to detect such attraction exhibited by individual cells requires extremely long imaging experiments. We thus have established a framework for how attraction of moving cells towards a particular location can be rigorously evaluated.

scholarly journals Metabolic programs define dysfunctional immune responses in severe COVID-19 patients

2020 ◽  
Author(s):  
Elizabeth A. Thompson ◽  
Katherine Cascino ◽  
Alvaro A. Ordonez ◽  
Weiqiang Zhou ◽  
Ajay Vaghasia ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Severe Disease ◽  
Suppressor Cells ◽  
Anion Channel ◽  
Metabolic Phenotype ◽  
Hexokinase Ii ◽  
Voltage Dependent Anion Channel ◽  
Microscopy Imaging

ABSTRACTIt remains unclear why some patients infected with SARS-CoV-2 readily resolve infection while others develop severe disease. To address this question, we employed a novel assay to interrogate immune-metabolic programs of T cells and myeloid cells in severe and recovered COVID-19 patients. Using this approach, we identified a unique population of T cells expressing high H3K27me3 and the mitochondrial membrane protein voltage-dependent anion channel (VDAC), which were expanded in acutely ill COVID-19 patients and distinct from T cells found in patients infected with hepatitis c or influenza and in recovered COVID-19. Increased VDAC was associated with gene programs linked to mitochondrial dysfunction and apoptosis. High-resolution fluorescence and electron microscopy imaging of the cells revealed dysmorphic mitochondria and release of cytochrome c into the cytoplasm, indicative of apoptosis activation. The percentage of these cells was markedly increased in elderly patients and correlated with lymphopenia. Importantly, T cell apoptosis could be inhibited in vitro by targeting the oligomerization of VDAC or blocking caspase activity. In addition to these T cell findings, we also observed a robust population of Hexokinase II+ polymorphonuclear-myeloid derived suppressor cells (PMN-MDSC), exclusively found in the acutely ill COVID-19 patients and not the other viral diseases. Finally, we revealed a unique population of monocytic MDSC (M-MDSC) expressing high levels of carnitine palmitoyltransferase 1a (CPT1a) and VDAC. The metabolic phenotype of these cells was not only highly specific to COVID-19 patients but the presence of these cells was able to distinguish severe from mild disease. Overall, the identification of these novel metabolic phenotypes not only provides insight into the dysfunctional immune response in acutely ill COVID-19 patients but also provide a means to predict and track disease severity as well as an opportunity to design and evaluate novel metabolic therapeutic regimens.GRAPHICAL ABSTRACT

From Microbiology to Microcontrollers: Robot Search Patterns Inspired by T Cell Movement

2013 ◽  
Author(s):  
G. Matthew Fricke ◽  
François Asperti-Boursin ◽  
Joshua Hecker ◽  
Judy Cannon ◽  
Melanie Moses
Keyword(s):  
T Cell ◽  
Cell Movement ◽  
Search Patterns

scholarly journals Characterizing T Cell Movement within Lymph Nodes in the Absence of Antigen

2007 ◽  
Vol 178 (9) ◽  
pp. 5505-5512 ◽  
Author(s):  
Catherine Beauchemin ◽  
Narendra M. Dixit ◽  
Alan S. Perelson
Keyword(s):  
T Cell ◽  
Lymph Nodes ◽  
Cell Movement

scholarly journals T-Cell-Independent Immune Responses Do Not Require Cxc Ligand 13-Mediated B1 Cell Migration

2010 ◽  
Vol 78 (9) ◽  
pp. 3950-3956 ◽  
Author(s):  
Matthew J. Colombo ◽  
Guizhi Sun ◽  
Kishore R. Alugupalli
Keyword(s):  
Cell Migration ◽  
T Cell ◽  
Specific Antigen ◽  
Cell Movement ◽  
Antibody Responses ◽  
Wild Type ◽  
Coelomic Cavity ◽  
Borrelia Hermsii ◽  
Anatomical Compartment ◽  
B1 Cells

ABSTRACT The dynamic movement of B cells increases the probability of encountering specific antigen and facilitates cell-cell interactions required for mounting a rapid antibody response. B1a and B1b cells are enriched in the coelomic cavity, contribute to T-cell-independent (TI) antibody responses, and increase in number upon antigen exposure. B1 cell movement is largely governed by Cxc ligand 13 (Cxcl13), and mice deficient in this chemokine have a severe reduction in peritoneal B1 cells. In this study, we examined the role of Cxcl13-dependent B cell migration using Borrelia hermsii infection or intraperitoneal immunization with pneumococcal polysaccharide or 4-hydroxy-3-nitrophenyl-acetyl (NP)-Ficoll, all of which induce robust antibody responses from B1b cells. Surprisingly, we found that antibody responses to B. hermsii or to FhbA, an antigenic target of B1b cells, and the resolution of bacteremia were indistinguishable between wild-type and Cxcl13 −/− mice. Importantly, we did not observe an expansion of peritoneal B1b cell numbers in Cxcl13 −/− mice. Nonetheless, mice that had resolved infection were resistant to reinfection, indicating that the peritoneal B1b cell reservoir is not required for controlling B. hermsii. Furthermore, despite a reduced peritoneal B1b compartment, immunization with pneumococcal polysaccharide vaccine yielded comparable antigen-specific antibody responses in wild-type and Cxcl13 −/− mice and conferred protection against Streptococcus pneumoniae. Likewise, immunization with NP-Ficoll elicited similar antibody responses in wild-type and Cxcl13 −/− mice. These data demonstrate that homing of B1 cells into the coelomic cavity is not a requirement for generating protective TI antibody responses, even when antigen is initially localized to this anatomical compartment.

scholarly journals Cell responses to simulated microgravity and hydrodynamic stress can be distinguished by using comparative transcriptomic analysis

2021 ◽  
Author(s):  
Nikolai V Kouznetsov
Keyword(s):  
T Cells ◽  
T Cell ◽  
Immune Cell ◽  
Cell Movement ◽  
T Cell Response ◽  
Human Immune System ◽  
Hydrodynamic Stress ◽  
Cytoskeletal Network ◽  
Cultural Medium ◽  
Cell Transcriptome

The human immune system is compromised in microgravity (MG) conditions during an orbital flight and upon return to Earth. T cells are critical for the immune response and execute their functions via actin mediated immune cell-cell interactions that could be disturbed by MG conditions. Here, we have applied two rotational platforms to simulate MG conditions: fast rotating clinostat (CL) and random positioning machine (RPM) followed by global T cell transcriptome analysis using RNA sequencing. We demonstrate that the T cell transcriptome profile in response to simulated MG treatment was clearly distinguishable from the T cell transcriptome response to hydrodynamic stress (HS) induced by shear forces upon cell movement in cultural medium. Gene expression profiling of genes related to or involved in actin cytoskeleton networks using RT-qPCR confirmed two sets of differentially regulated genes in the T cell response to MG or to HS. Several key genes potentially involved in T cell gravisensing (Fam163b, Dnph1, Trim34, Upk-1b) were identified. A number of candidate biomarker genes of the response to MG (VAV1, VAV2, VAV3, and NFATC2) and of the response to HS (ITGAL, ITGB1, ITGB2, RAC1 and RAC2) could be used to distinguish between these processes on the gene transcription level. Together, MG induces changes in the overall transcriptome of T cells leading to specific shifts in expression of cytoskeletal network genes.

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