Control of Antigen Presentation with a Photoreleasable Agonist Peptide

Demond, Andrew L.; Starr, Toby; Dustin, Michael L.; Groves, Jay T.

doi:10.1021/ja065304l

Cited by 35 publications

(38 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Indeed, photoactivatable systems have been used to investigate the kinetics of second-messenger activity through caged calcium (22) and caged cAMP (23). Additionally, the T-cell receptor was studied using major histocompatibility complex (MHC) presenting photoactivatable peptide (24,25).In this report, we dissected the dynamic responses during the initiation of B-cell activation by using a photoactivatable antigen based experimental system in combination with high-resolution high Significance B-cell receptor (BCR) and antigen engagement induces several responses resulting in B-cell activation. However, it has been difficult to study these responses due to their dynamic nature.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Utilization of a photoactivatable antigen system to examine B-cell probing termination and the B-cell receptor sorting mechanisms during B-cell activation

Wang

Tang

Wan

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Antigen binding to the B-cell receptor (BCR) induces several responses, resulting in B-cell activation, proliferation, and differentiation. However, it has been difficult to study these responses due to their dynamic, fast, and transient nature. Here, we attempted to solve this problem by developing a controllable trigger point for BCR and antigen recognition through the construction of a photoactivatable antigen, caged 4-hydroxy-3-nitrophenyl acetyl (caged-NP). This photoactivatable antigen system in combination with live cell and single molecule imaging techniques enabled us to illuminate the previously unidentified B-cell probing termination behaviors and the precise BCR sorting mechanisms during B-cell activation. B cells in contact with caged-NP exhibited probing behaviors as defined by the unceasing extension of membrane pseudopods in random directions. Further analyses showed that such probing behaviors are cell intrinsic with strict dependence on F-actin remodeling but not on tonic BCR signaling. B-cell probing behaviors were terminated within 4 s after photoactivation, suggesting that this response was sensitive and specific to BCR engagement. The termination of B-cell probing was concomitant with the accumulation response of the BCRs into the BCR microclusters. We also determined the Brownian diffusion coefficient of BCRs from the same B cells before and after BCR engagement. The analysis of temporally segregated single molecule images of both BCR and major histocompatibility complex class I (MHC-I) demonstrated that antigen binding induced trapping of BCRs into the BCR microclusters is a fundamental mechanism for B cells to acquire antigens.he immune system uses immune receptors to sense and acquire antigens. Antigen binding induces a series of dynamic changes in the biophysical behaviors and biochemical features of the immune receptors, and these changes determine the fate of a cell (1-3). However, it has been difficult to accurately capture and thus comprehensively investigate these changes because they usually occur very quickly after immune recognition (1, 4). For example, recent live cell imaging studies showed that the B lymphocytes swiftly accumulate the surface-expressed B-cell receptors (BCRs) into the contact interface between the B cells and the antigen-presenting substrates to form a specialized membrane structure, the B-cell immunological synapse (IS) (1, 4). Moreover, both our studies and those of others showed that these accumulation events are sensitive to the biochemical and biophysical features of the antigens that B cells likely encounter in vivo (4, 5). These features include but are not limited to antigen density (6, 7), antigen affinity (6, 7), antigen valency (8-13), the mobility of the antigen (14-17), the stiffness of the substrates presenting the antigen (18, 19) and the mechanical forces delivered to the BCRs by the antigens (20, 21). These facts highlight a long-standing question in immunology: how can the initiation of B-cell activation process the information of antigen s...

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

Utilization of a photoactivatable antigen system to examine B-cell probing termination and the B-cell receptor sorting mechanisms during B-cell activation

Wang

Tang

Wan

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Light-sensitive [97][98][99] photo-switchable molecules 98,100,101 or local uncaging [102][103][104] combined with diffraction-limited lasers enable high spatial and temporal control over protein activity. Optically controlled uncaging of T cell-activating peptide can stimulate the cells with high temporal and possibly spatial resolution 102,103 . Both of these types of approach can cause spatially inhomogeneous activation of cells and, when combined with readouts of signalling 105,106 , can elucidate mechanisms of the spatial control of signalling.…”

Section: Direct Spatial Pattern On Solid Surfacesmentioning

confidence: 99%

Spatial organization and signal transduction at intercellular junctions

Manz

Groves

2010

Nat Rev Mol Cell Biol

Self Cite

122

114

View full text Add to dashboard Cite

The coordinated organization of cell membrane receptors into diverse micrometre-scale spatial patterns is emerging as an important theme of intercellular signalling, as exemplified by immunological synapses. Key characteristics of these patterns are that they transcend direct protein-protein interactions, emerge transiently and modulate signal transduction. Such cooperativity over multiple length scales presents new and intriguing challenges for the study and ultimate understanding of cellular signalling. As a result, new experimental strategies have emerged to manipulate the spatial organization of molecules inside living cells. The resulting spatial mutations yield insights into the interweaving of the spatial, mechanical and chemical aspects of intercellular signalling.Cell-to-cell communication is mediated by various methods. Endocrine signals are secreted and reach distant target cells, paracrine signals are secreted and reach targets in the vicinity, and autocrine signals are secreted and received by the same cell. By contrast, in juxtacrine signalling, surfaces of interacting cells come into direct contact and receptor-ligand recognition at this interface triggers intracellular signalling. Cell-cell interactions involve multiple adhesion and signalling molecules, the collective behaviour of which regulates signal transduction 1 . Also intrinsic to juxtacrine signalling configurations are large physical constraints on molecular movement and assembly. Genetic and biochemical approaches have been invaluable in identifying the molecular components of signal transduction pathways in juxtacrine signalling and in characterizing the biochemical interactions among them. Despite this wealth of information, in many cases it remains impossible to describe the behaviour of a signalling system in terms of the individual molecular properties of its components. Protein-protein inter actions and the formation of molecular clusters are widely implicated in signal transduction and contribute to a first level of cooperativity [2][3][4] . Recently, the coordinated organization of cell membrane receptors into micrometre-scale patterns has emerged as a broadly important theme of intercellular signalling 1,[5][6][7][8][9] .A paradigm for the interplay of spatial patterns and signal transduction is the junction between T cells and their target cells, termed the immunological synapse [8][9][10][11][12][13] . Spatial HHMI Author Manuscript HHMI Author Manuscript HHMI Author Manuscriptpatterns of proteins at the cell-cell interface develop as hundreds of receptors recognize their cognate ligands on the apposed cell membrane. Multiple signalling and adhesion molecules also become organized into distinctive spatial patterns at the interface between the two cells 8,9,13,14 (FIG. 1). The patterns create long-range interactions and seem to have specific purposes in signal transduction 8,9 . They host the local enrichment or depletion of key signalling components, which can bias biochemical cascades towards different functional o...

show abstract

“…2). 10,11 T cells expressing the 5C.C7 TCR bind specifically to a peptide derived from moth cytochrome C (MCC, a.a. 88-103) in the context of the mouse class II MHC protein I-E k . We attached a photocleavable ortho-nitrophenylethyl damaging the surrounding tissue.…”

Section: T Cell Receptor Photoactivation Provides Spatiotemporal Controlmentioning

confidence: 99%

Lymphocyte polarity, the immunological synapse and the scope of biological analogy

Huse¹

2011

BioArchitecture

View full text Add to dashboard Cite

We have developed a photoactivation and imaging strategy that enables highresolution analysis of cytoskeletal dynamics in individual T cells. Using this approach, we have demonstrated that the lipid second messenger diacylglycerol plays a crucial role in promoting MTOC reorientation by recruiting three members of the protein kinase C family to the synapse. Here, I will discuss these results along with studies from other labs, which have explored the role of polarityinducing protein complexes after synapse formation. I will also propose a two-step model for MTOC reorientation in lymphocytes that reflects what we now know about the subject. Finally, I will consider the extent to which lymphocyte polarity resembles analogous cell polarity systems in other cell types.Cell polarity is a precondition of multicellular lifestyle. Polarized cells interact with their surroundings in a fundamentally anisotropic manner, which is crucial for establishing systems, such as neuronal circuits, in which there is directional

show abstract

Control of Antigen Presentation with a Photoreleasable Agonist Peptide

Cited by 35 publications

References 14 publications

Utilization of a photoactivatable antigen system to examine B-cell probing termination and the B-cell receptor sorting mechanisms during B-cell activation

Utilization of a photoactivatable antigen system to examine B-cell probing termination and the B-cell receptor sorting mechanisms during B-cell activation

Spatial organization and signal transduction at intercellular junctions

Lymphocyte polarity, the immunological synapse and the scope of biological analogy

Contact Info

Product

Resources

About