Detection of Dimers of Dimers of Human Leukocyte Antigen (HLA)–DR on the Surface of Living Cells by Single-Particle Fluorescence Imaging

Cherry, Richard J.; Wilson, Keith; Triantafilou, Kathy; O’Toole, Peter; Morrison, Ian; Smith, Patricia R.; Fernández, Nelson

doi:10.1083/jcb.140.1.71

Cited by 61 publications

(38 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, it is possible that the peptide maps reflected the composition of the MHC class II molecules and not that of MHC class II-associated peptides. Spontaneous MHC class II association has also been suggested by single-particle fluorescence imaging and scanning force microscopy using living cells (16,31). However, the data reported here (Fig.…”

Section: Discussioncontrasting

confidence: 56%

“…High-molecularmass bands containing MHC class II molecules in Western blots of murine and human cell lysates have been interpreted as preformed MHC class II double dimers (12)(13)(14)(15). Furthermore, using biophysical techniques, preformed double dimers have been described in MHC class II-transfected mouse fibroblasts (16). However, the conclusion from experiments involving soluble MHC class II and TCR molecules was that dimerization/oligomerization was driven by cognate interactions between the TCR and MHC class II:peptide complexes (4).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Amino Acid Substitutions in the Putative MHC Class II “Dimer of Dimers” Interface Inhibit CD4+ T Cell Activation

Lindstedt

Monk

Lombardi

et al. 2001

The Journal of Immunology

View full text Add to dashboard Cite

Activation of T lymphocytes is dependent on multiple ligand-receptor interactions. The possibility that TCR dimerization contributes to T cell triggering was raised by the crystallographic analysis of MHC class II molecules. The MHC class II molecules associated as double dimers, and in such a way that two TCR (and two CD4 molecules) could bind simultaneously. Several subsequent studies have lent support to this concept, although the role of TCR cross-linking in T cell activation remains unclear. Using DRA cDNAs modified to encode two different C-terminal tags, no evidence of constitutive double dimer formation was obtained following immunoprecipitation and Western blotting from cells transiently transfected with wild-type DRB and tagged DRA constructs, together with invariant chain and HLA-DM. To determine whether MHC class II molecules contribute actively to TCR-dependent dimerization and consequent T cell activation, panels of HLA-DR1β and H2-Ek cDNAs were generated with mutations in the sequences encoding the interface regions of the MHC class II double dimer. Stable DAP.3 transfectants expressing these cDNAs were generated and characterized biochemically and functionally. Substitutions in either interface region I or III did not affect T cell activation, whereas combinations of amino acid substitutions in both regions led to substantial inhibition of proliferation or IL-2 secretion by human and murine T cells. Because the amino acid-substituted molecules were serologically indistinguishable from wild type, bound antigenic peptide with equal efficiency, and induced Ag-dependent CD25 expression indicating TCR recognition, the reduced ability of the mutants to induce full T cell activation is most likely the result of impaired double dimer formation. These data suggest that MHC class II molecules, due to their structural properties, actively contribute to TCR cross-linking.

show abstract

Section: Discussioncontrasting

confidence: 56%

mentioning

confidence: 99%

Amino Acid Substitutions in the Putative MHC Class II “Dimer of Dimers” Interface Inhibit CD4+ T Cell Activation

Lindstedt

Monk

Lombardi

et al. 2001

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…SPFI allows visualization of individual molecules at the cell surface within a spatial resolution of ϳ25 nm (31). This technique permits the determination of positional information of molecules and the single diffraction limited spot intensity determines the number of molecules.…”

Section: Discussionmentioning

confidence: 99%

“…The SPFI methodology has previously been used to investigate molecules on the cell surface, in adherent cell lines grown in culture, where it is possible to locate the focal plane of single molecules at the cell surface (30,31). When adapting SPFI methodology for this study, it was found that focusing on single particles on embryo blastomeres was difficult due to the large three-dimensional shape of the cells.…”

Section: Spfi To Investigate the Spatial Distribution Of Hla-g In Prementioning

confidence: 99%

Analysis of HLA-G in Maternal Plasma, Follicular Fluid, and Preimplantation Embryos Reveal an Asymmetric Pattern of Expression

Shaikly

Morrison

Taranissi³

et al. 2008

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

Soluble HLA-G (sHLA-G) secretion by human preimplantation embryos in culture has been associated with successful embryo development, and therefore has potential to serve as a noninvasive marker of embryo viability. We have examined the spatial and temporal expression of HLA-G in embryos of varying developmental competence and the role of maternal factors in human embryonic HLA-G expression. Embryos that reached blastocyst stage on day 5 showed a higher frequency of sHLA-G secretion than those at morula or arrested stages (p < 0.05). There was no significant difference in sHLA-G secretion between normal embryos and those diagnosed as chromosomally abnormal by preimplantation genetic diagnosis. HLA-G detected in maternal plasma and follicular fluid did not appear to correlate with HLA-G expressed in the embryo or embryo supernatants. Confocal microscopy analysis indicated that HLA-G protein expression in embryos was not homogeneous; mostly, it was confined to blastocysts localized on trophectoderm and trophectoderm projections. Single-particle fluorescent imaging analysis of HLA-G on the cell surface of JEG-3 cells showed that HLA-G particles were mostly monomeric, but dimeric and higher order oligomers were also observed. These results suggest that HLA-G play an important role in preimplantation embryo development. However, the observed expression of HLA-G in arrested and chromosomally abnormal embryos indicates that HLA-G testing should be used with caution and in conjunction with conventional methods of embryo screening and selection.

show abstract

“…Both FPR 26,28,53 and SPT 30,54,55 studies have shown that the MHC protein is a highly mobile cell surface receptor, and its mobility is in part determined by its cytoplasmic tail. FRET [56][57][58] and single-molecule fluorescence imaging 59 studies have shown that MHC proteins also reorganize into oligomeric clusters. The lateral mobility of the lymphocyte receptor CD2 has been examined by FPR and SPT.…”

mentioning

confidence: 99%

T cell adhesion mechanisms revealed by receptor lateral mobility

Cairo¹,

Golan²

2007

Biopolymers

View full text Add to dashboard Cite

Cell surface receptors mediate the exchange of information between cells and their environment. In the case of adhesion receptors, the spatial distribution and molecular associations of the receptors are critical to their function. Therefore, understanding the mechanisms regulating the distribution and binding associations of these molecules is necessary to understand their functional regulation. Experiments characterizing the lateral mobility of adhesion receptors have revealed a set of common mechanisms that control receptor function and thus cellular behavior. The T cell provides one of the most dynamic examples of cellular adhesion. An individual T cell makes innumerable intercellular contacts with antigen presenting cells, the vascular endothelium, and many other cell types. We review here the mechanisms that regulate T cell adhesion receptor lateral mobility as a window into the molecular regulation of these systems, and we present a general framework for understanding the principles and mechanisms that are likely to be common among these and other cellular adhesion systems. We suggest that receptor lateral mobility is regulated via four major mechanisms—reorganization, recruitment, dispersion, and anchoring—and we review specific examples of T cell adhesion receptor systems that utilize one or more of these mechanisms. © 2007 Wiley Periodicals, Inc. Biopolymers 89: 409–419, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

show abstract

Detection of Dimers of Dimers of Human Leukocyte Antigen (HLA)–DR on the Surface of Living Cells by Single-Particle Fluorescence Imaging

Cited by 61 publications

References 46 publications

Amino Acid Substitutions in the Putative MHC Class II “Dimer of Dimers” Interface Inhibit CD4+ T Cell Activation

Amino Acid Substitutions in the Putative MHC Class II “Dimer of Dimers” Interface Inhibit CD4+ T Cell Activation

Analysis of HLA-G in Maternal Plasma, Follicular Fluid, and Preimplantation Embryos Reveal an Asymmetric Pattern of Expression

T cell adhesion mechanisms revealed by receptor lateral mobility

Contact Info

Product

Resources

About