Tracking T-Cell Antigen Receptors | Imaris

Tracking T-Cell Antigen Receptors During Formation of Immunological Synapses

Immunology

Peter Beemiller, Jordan Jacobelli, Matthew F. Krummel, University of California - San Francisco

T-cell activation is an important part of the immune response. It occurs when T-cell antigen receptors (TCRs) bind with antigen-presenting cells, forming what is known as an immunological synapse. Researchers at the University of California in San Francisco are using total internal fluorescence microscopy and Imaris image analysis software to learn more about the dynamic processes involved when immunological synapses form during T-cell motility.

Peter Beemiller, Jordan Jacobelli and Matthew F. Krummel

During activation T-cell antigen receptors collect into microclusters, which are then reorganized into macromolecular domains as part of their regulation. The researchers illustrated how these domains, called supramolecular activation clusters (SMACs), are generated in motile T-cells. “This brings our understanding of the formation of these domains in line with in vivo imaging that shows that T cells are quite dynamic during their interactions with antigen presenting cells,” said Dr. Peter Beemiller, who led the research team.

The researchers used total internal reflection microscopy to image motile synapses formed during T-cell motility. The Imaris XT interface allowed them to work with data from other sources. They identified TCR microclusters outside of Imaris, and then transferred data on the location of the microclusters to Imaris for tracking. They could then switch between MATLAB and Imaris to analyze various track characteristics, such as time of formation.

“Imaris XT let us work with the data in Imaris and extended what we did beyond the rote routines that exist in all image analysis software,” Dr. Beemiller said. “This isn’t limited just to the primary image data, and isn’t limited to using Imaris as a one-way data server for other programs. Analyses from MATLAB can return data to Imaris, allowing us to quickly visualize the output of those routines in the context of our original data.”

The analysis revealed that TCR microclusters in motile synapses did not flow directly into SMACs. Instead, they moved toward an F-actin-poor ‘sink’ region. Inward microcluster flow often followed collapse of the leading edge, which suggested that actin depolymerization regulates microcluster flow and the formation of SMACs. The researchers continue to look for and study actin cytoskeletal proteins that control TCR signaling domain formation.

“By letting people get their data out of Imaris, the XT interface is a great way to make Imaris more valuable,” Dr. Beemiller said.

Research Paper: Integration of the movement of signaling microclusters with cellular motility in immunological synapses. Beemiller P, Jacobelli J, Krummel MF. Nat Immunol. 2012 Jul 1;13(8):787-95. doi: 10.1038/ni.2364.

Multimedia Library
Application Images (3)
Publications Database
Caveolin-1 deficiency induces a MEK-ERK1/2-Snail-1-dependent epithelial–mesenchymal transition and fibrosis during peritoneal dialysis
The chemokine receptors ACKR2 and CCR2 reciprocally regulate lymphatic vessel density
Host DNA released in response to aluminum adjuvant enhances MHC class II-mediated antigen presentation and prolongs CD4 T-cell interactions with dendritic cells
Three-Dimensional Visualization of the Mouse Thymus Organization in Health and Immunodeficiency
Molecular and functional evidence of HCN4 and caveolin-3 interaction during cardiomyocyte differentiation from human embryonic stem cells
Shigella impairs T lymphocyte dynamics in vivo
Constitutive Lymphocyte Transmigration across the Basal Lamina of High Endothelial Venules Is Regulated by the Autotaxin/Lysophosphatidic Acid Axis
Sphingosine-1-phosphate-mediated osteoclast precursor monocyte migration is a critical point of control in antibone-resorptive action of active vitamin D
Opposing chemokine gradients control human thymocyte migration in situ
Motile invaded neutrophils in the small intestine of Toxoplasma gondii-infected mice reveal a potential mechanism for parasite spread
In vivo imaging of CD8+ T cell-mediated elimination of malaria liver stages
Chemokine Receptor CXCR6-Dependent Hepatic NK T Cell Accumulation Promotes Inflammation and Liver Fibrosis
Tetraspanin CD37 contributes to the initiation of cellular immunity by promoting dendritic cell migration
Perivascular Mast Cells Dynamically Probe Cutaneous Blood Vessels to Capture Immunoglobulin
Rgs13 Constrains Early B Cell Responses and Limits Germinal Center Sizes
Elimination of Self-Reactive T Cells in the Thymus: A Timeline for Negative Selection
NLRP10 is a NOD-like receptor essential to initiate adaptive immunity by dendritic cells
Expansion of Cortical and Medullary Sinuses Restrains Lymph Node Hypertrophy during Prolonged Inflammation
MT1-MMP regulates the PI3K (delta) Mi-2/NuRD-dependent control of macrophage immune function
Intravital imaging of CTLs killing islet cells in diabetic mice

Sign up for the Bitplane Newsletter!

Receive articles like this one, release notes, product launches, press releases and more with our regular newsletter. It's free to subscribe, will be sent every 6 weeks, and you can opt out at any time.

NAME
EMAIL


Try Imaris FREE for 10 days?