Detection of dimers of dimers of human leukocyte antigen (HLA)-DR on the surface of living cells by single-particle fluorescence imaging

The Journal of Cell Biology
R J CherryN Fernandez

Abstract

The technique of single-particle fluorescence imaging was used to investigate the oligomeric state of MHC class II molecules on the surface of living cells. Cells transfected with human leukocyte antigen (HLA)-DR A and B genes were labeled at saturation with a univalent probe consisting of Fab coupled to R-phycoerythrin. Analysis of the intensities of fluorescent spots on the cell surface revealed the presence of single and double particles consistent with the simultaneous presence of HLA-DR heterodimers and dimers of dimers. The proportion of double particles was lower at 37 degrees C than at 22 degrees C, suggesting that the heterodimers and dimers of dimers exist in a temperature-dependent equilibrium. These results are discussed in the context of a possible role for HLA-DR dimers of dimers in T cell receptor-MHC interactions. The technique is validated by demonstrating that fluorescence imaging can distinguish between dimers and tetramers of human erythrocyte spectrin deposited from solution onto a solid substrate. The methodology will have broad applicability to investigation of the oligomeric state of immunological and other membrane-bound receptors in living cells.

References

Aug 1, 1978·European Journal of Biochemistry·E Ungewickell, W Gratzer
Dec 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·R J PoljakF Saul
Jan 1, 1983·Annual Review of Biochemistry·A N Glazer
Oct 15, 1984·European Journal of Biochemistry·V Ohanian, W Gratzer
Jul 3, 1995·Proceedings of the National Academy of Sciences of the United States of America·T SakihamaE L Reinherz
Sep 1, 1994·Biophysical Journal·I E MorrisonR J Cherry
Feb 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·S DamjanovichT M Jovin
Jan 1, 1994·Annual Review of Immunology·A SetteH M Grey
Jan 28, 1994·Cell·A Weiss, D R Littman
Jul 1, 1993·Nature·H Ploegh, P Benaroch
Apr 2, 1996·Proceedings of the National Academy of Sciences of the United States of America·T SchmidtH Schindler
Jul 1, 1996·Immunology Today·B A Fields, R A Mariuzza
Jun 1, 1996·Immunological Reviews·R LechlerL Lightstone
Jan 1, 1997·Methods in Enzymology·J Matko, M Edidin
Jul 8, 1997·Proceedings of the National Academy of Sciences of the United States of America·A JeneiS Damjanovich
Aug 1, 1997·Biochemical Society Transactions·R J CherryN Fernández

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Citations

Oct 24, 1998·Immunology Today·V HorejsíP Angelisová
Feb 1, 2003·Experimental Gerontology·G J Schütz, P Hinterdorfer
Jan 16, 2002·Current Opinion in Immunology·Rolf König
Jul 21, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Lindsay C C ElliottPaul W Bohn
Mar 21, 2012·Proceedings of the National Academy of Sciences of the United States of America·Yiyuan YinRoy A Mariuzza
May 24, 2000·Proceedings of the National Academy of Sciences of the United States of America·G VerebS Damjanovich
Dec 28, 2006·Proceedings of the National Academy of Sciences of the United States of America·Julia J UnternaehrerIra Mellman
May 29, 2000·Molecular Membrane Biology·G J SchützH Schindler
Nov 13, 2007·Brain Research Reviews·Luigi F AgnatiAlexander O Tarakanov
Jun 15, 2007·Seminars in Immunology·Kaushik Choudhuri, P Anton van der Merwe
Nov 28, 2007·Biopolymers·Christopher W Cairo, David E Golan
May 8, 2007·Microscopy Research and Technique·Rasim GuldikenAhmed A Busnaina
Sep 29, 2005·Journal of Biotechnology·Li SunZhiyong Qiao
Jan 25, 2011·Physical Chemistry Chemical Physics : PCCP·Lindsay C C ElliottPaul W Bohn
Oct 12, 2000·Biophysical Journal·M Goulian, S M Simon
Jun 15, 1999·Biochimica Et Biophysica Acta·P J O'TooleR J Cherry
Jun 26, 2001·Biophysical Journal·Q Tang, M Edidin
Aug 26, 2004·Biochimica Et Biophysica Acta·P H M LommerseT Schmidt
May 14, 1999·International Immunology·K DrbalV Horejsí
Aug 15, 2012·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Ioannis KarakikesNelson Fernandez
Jan 6, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·R LindstedtR Lechler
Mar 7, 2008·The Journal of Immunology : Official Journal of the American Association of Immunologists·Valerie R ShaiklyNelson Fernández
Jun 15, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Adam G SchrumEd Palmer
Jul 15, 2005·Molecular and Cellular Biochemistry·John D Hayball, Richard A Lake
Jun 1, 2001·The Journal of Biological Chemistry·J R CochranL J Stern

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Methods Mentioned

BETA
x-ray crystallography
immunoprecipitation
fluorescence microscopy
fluorescence imaging
size exclusion chromatography
flow cytometry
CLIP
imaging technique
confocal microscopy

Software Mentioned

ValueChrom ®
AT1

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