Interaction of HLA-DR and CD74 at the cell surface of antigen-presenting cells by single particle image analysis

Ioannis Karakikes; Ian E G Morrison; Peter O'Toole; Gergana Metodieva; Cristina V Navarrete; Jesus Gomez; Jose M Miranda-Sayago; Richard J Cherry; Metodi Metodiev; Nelson Fernandez

doi:10.1096/fj.12-211466

Interaction of HLA-DR and CD74 at the cell surface of antigen-presenting cells by single particle image analysis

Ioannis Karakikes, Ian E G Morrison, Peter O'Toole, Gergana Metodieva, Cristina V Navarrete, Jesus Gomez, Jose M Miranda-Sayago, Richard J Cherry, Metodi Metodiev, Nelson Fernandez

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Major histocompatibility complex (MHC) class II-associated antigen presentation involves an array of interacting molecules. CD74, the cell surface isoform of the MHC class II-associated invariant chain, is one such molecule; its role remains poorly defined. To address this, we have employed a high-resolution single-particle imaging method for quantifying the colocalization of CD74 with human leukocyte antigen (HLA)-DR molecules on human fibroblast cells known for their capacity to function as antigen-presenting cells. We have also examined whether the colocalization induces internalization of HLA-DR using HA(307-319), a "universal" peptide that binds specifically to the peptide-binding groove of all HLA-DR molecules, irrespective of their alleles. We have determined that 25 ± 1.3% of CD74 and 17 ± 0.3% of HLA-DR are colocalized, and the association of CD74 with HLA-DR and the internalization of HLA-DR are both inhibited by HA(307-319). A similar inhibition of HLA-DR internalization was observed in freshly isolated monocyte-derived dendritic cells. A key role of CD74 is to translocate HLA-DR molecules to early endosomes for reloading with peptides prior to recycling to the cell surface. We conclude that CD74 regulates the balance of peptide-occupied and peptide-free forms of MHC class II at the cell surface.

Original language	English
Pages (from-to)	4886-4896
Journal	The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume	26
Issue number	12
DOIs	https://doi.org/10.1096/fj.12-211466
Publication status	Published - Dec 2012

Keywords

Adult
Algorithms
Antigen-Presenting Cells
Antigens, Differentiation, B-Lymphocyte
Cell Line
Cell Membrane
Cells, Cultured
Dendritic Cells
Endocytosis
Flow Cytometry
Fluorescence Resonance Energy Transfer
HLA-DR Antigens
Hemagglutinin Glycoproteins, Influenza Virus
Histocompatibility Antigens Class II
Humans
Imaging, Three-Dimensional
Microscopy, Confocal
Microscopy, Fluorescence
Peptide Fragments
Protein Binding

Access to Document

10.1096/fj.12-211466

http://www.fasebj.org/content/26/12/4886

Cite this

Karakikes, I., Morrison, I. E. G., O'Toole, P., Metodieva, G., Navarrete, C. V., Gomez, J., Miranda-Sayago, J. M., Cherry, R. J., Metodiev, M., & Fernandez, N. (2012). Interaction of HLA-DR and CD74 at the cell surface of antigen-presenting cells by single particle image analysis. The FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 26(12), 4886-4896. https://doi.org/10.1096/fj.12-211466

@article{3e12fcef9a534234b41574c9fc23c9df,

title = "Interaction of HLA-DR and CD74 at the cell surface of antigen-presenting cells by single particle image analysis",

abstract = "Major histocompatibility complex (MHC) class II-associated antigen presentation involves an array of interacting molecules. CD74, the cell surface isoform of the MHC class II-associated invariant chain, is one such molecule; its role remains poorly defined. To address this, we have employed a high-resolution single-particle imaging method for quantifying the colocalization of CD74 with human leukocyte antigen (HLA)-DR molecules on human fibroblast cells known for their capacity to function as antigen-presenting cells. We have also examined whether the colocalization induces internalization of HLA-DR using HA(307-319), a {"}universal{"} peptide that binds specifically to the peptide-binding groove of all HLA-DR molecules, irrespective of their alleles. We have determined that 25 ± 1.3% of CD74 and 17 ± 0.3% of HLA-DR are colocalized, and the association of CD74 with HLA-DR and the internalization of HLA-DR are both inhibited by HA(307-319). A similar inhibition of HLA-DR internalization was observed in freshly isolated monocyte-derived dendritic cells. A key role of CD74 is to translocate HLA-DR molecules to early endosomes for reloading with peptides prior to recycling to the cell surface. We conclude that CD74 regulates the balance of peptide-occupied and peptide-free forms of MHC class II at the cell surface.",

keywords = "Adult, Algorithms, Antigen-Presenting Cells, Antigens, Differentiation, B-Lymphocyte, Cell Line, Cell Membrane, Cells, Cultured, Dendritic Cells, Endocytosis, Flow Cytometry, Fluorescence Resonance Energy Transfer, HLA-DR Antigens, Hemagglutinin Glycoproteins, Influenza Virus, Histocompatibility Antigens Class II, Humans, Imaging, Three-Dimensional, Microscopy, Confocal, Microscopy, Fluorescence, Peptide Fragments, Protein Binding",

author = "Ioannis Karakikes and Morrison, {Ian E G} and Peter O'Toole and Gergana Metodieva and Navarrete, {Cristina V} and Jesus Gomez and Miranda-Sayago, {Jose M} and Cherry, {Richard J} and Metodi Metodiev and Nelson Fernandez",

year = "2012",

month = dec,

doi = "10.1096/fj.12-211466",

language = "English",

volume = "26",

pages = "4886--4896",

journal = "The FASEB journal : official publication of the Federation of American Societies for Experimental Biology",

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Karakikes, I, Morrison, IEG , O'Toole, P, Metodieva, G, Navarrete, CV, Gomez, J, Miranda-Sayago, JM, Cherry, RJ, Metodiev, M & Fernandez, N 2012, 'Interaction of HLA-DR and CD74 at the cell surface of antigen-presenting cells by single particle image analysis', The FASEB journal : official publication of the Federation of American Societies for Experimental Biology, vol. 26, no. 12, pp. 4886-4896. https://doi.org/10.1096/fj.12-211466

Interaction of HLA-DR and CD74 at the cell surface of antigen-presenting cells by single particle image analysis. / Karakikes, Ioannis; Morrison, Ian E G ; O'Toole, Peter et al.
In: The FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 26, No. 12, 12.2012, p. 4886-4896.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Interaction of HLA-DR and CD74 at the cell surface of antigen-presenting cells by single particle image analysis

AU - Karakikes, Ioannis

AU - Morrison, Ian E G

AU - O'Toole, Peter

AU - Metodieva, Gergana

AU - Navarrete, Cristina V

AU - Gomez, Jesus

AU - Miranda-Sayago, Jose M

AU - Cherry, Richard J

AU - Metodiev, Metodi

AU - Fernandez, Nelson

PY - 2012/12

Y1 - 2012/12

N2 - Major histocompatibility complex (MHC) class II-associated antigen presentation involves an array of interacting molecules. CD74, the cell surface isoform of the MHC class II-associated invariant chain, is one such molecule; its role remains poorly defined. To address this, we have employed a high-resolution single-particle imaging method for quantifying the colocalization of CD74 with human leukocyte antigen (HLA)-DR molecules on human fibroblast cells known for their capacity to function as antigen-presenting cells. We have also examined whether the colocalization induces internalization of HLA-DR using HA(307-319), a "universal" peptide that binds specifically to the peptide-binding groove of all HLA-DR molecules, irrespective of their alleles. We have determined that 25 ± 1.3% of CD74 and 17 ± 0.3% of HLA-DR are colocalized, and the association of CD74 with HLA-DR and the internalization of HLA-DR are both inhibited by HA(307-319). A similar inhibition of HLA-DR internalization was observed in freshly isolated monocyte-derived dendritic cells. A key role of CD74 is to translocate HLA-DR molecules to early endosomes for reloading with peptides prior to recycling to the cell surface. We conclude that CD74 regulates the balance of peptide-occupied and peptide-free forms of MHC class II at the cell surface.

AB - Major histocompatibility complex (MHC) class II-associated antigen presentation involves an array of interacting molecules. CD74, the cell surface isoform of the MHC class II-associated invariant chain, is one such molecule; its role remains poorly defined. To address this, we have employed a high-resolution single-particle imaging method for quantifying the colocalization of CD74 with human leukocyte antigen (HLA)-DR molecules on human fibroblast cells known for their capacity to function as antigen-presenting cells. We have also examined whether the colocalization induces internalization of HLA-DR using HA(307-319), a "universal" peptide that binds specifically to the peptide-binding groove of all HLA-DR molecules, irrespective of their alleles. We have determined that 25 ± 1.3% of CD74 and 17 ± 0.3% of HLA-DR are colocalized, and the association of CD74 with HLA-DR and the internalization of HLA-DR are both inhibited by HA(307-319). A similar inhibition of HLA-DR internalization was observed in freshly isolated monocyte-derived dendritic cells. A key role of CD74 is to translocate HLA-DR molecules to early endosomes for reloading with peptides prior to recycling to the cell surface. We conclude that CD74 regulates the balance of peptide-occupied and peptide-free forms of MHC class II at the cell surface.

KW - Adult

KW - Algorithms

KW - Antigen-Presenting Cells

KW - Antigens, Differentiation, B-Lymphocyte

KW - Cell Line

KW - Cell Membrane

KW - Cells, Cultured

KW - Dendritic Cells

KW - Endocytosis

KW - Flow Cytometry

KW - Fluorescence Resonance Energy Transfer

KW - HLA-DR Antigens

KW - Hemagglutinin Glycoproteins, Influenza Virus

KW - Histocompatibility Antigens Class II

KW - Humans

KW - Imaging, Three-Dimensional

KW - Microscopy, Confocal

KW - Microscopy, Fluorescence

KW - Peptide Fragments

KW - Protein Binding

U2 - 10.1096/fj.12-211466

DO - 10.1096/fj.12-211466

M3 - Article

C2 - 22889831

SN - 0892-6638

VL - 26

SP - 4886

EP - 4896

JO - The FASEB journal : official publication of the Federation of American Societies for Experimental Biology

JF - The FASEB journal : official publication of the Federation of American Societies for Experimental Biology

IS - 12

ER -