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Intercellular transfer and supramolecular organization of human leukocyte antigen C at inhibitory natural killer cell immune synapses.

Carlin LM, Eleme K, McCann FE, Davis DM - J. Exp. Med. (2001)

Bottom Line: This unexpected intercellular transfer of HLA-C is dependent on NK receptor recognition, since HLA-Cw6 or -Cw4 but not -Cw3 transfer to an NK transfectant expressing killer Ig-like receptor (KIR)2DL1.However, the intercellular transfer of HLA-C is not dependent on active polymerization of the actin cytoskeleton.In addition, different arrangements of HLA-C are seen at inhibitory NK immune synapses, and these alter as NK synapses mature, but in a fashion distinct from that seen upon T cell activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Sir Alexander Fleming Building, Imperial College of Science, Technology, and Medicine, South Kensington, London SW7 2AZ, United Kingdom.

ABSTRACT
After accumulation of target cell human leukocyte antigen (HLA)-C at inhibitory natural killer (NK) cell immune synapses, some HLA-C transfers from target cells to NK cell plasma membranes and cytoplasm. This unexpected intercellular transfer of HLA-C is dependent on NK receptor recognition, since HLA-Cw6 or -Cw4 but not -Cw3 transfer to an NK transfectant expressing killer Ig-like receptor (KIR)2DL1. Strikingly, live-cell time-lapse laser scanning confocal microscopy shows vesicles containing target cell green fluorescent protein-tagged HLA-C migrating away from immune synapses into NK cells. Unlike clustering of HLA-C at the immune synapse, intercellular transfer of HLA-C is dependent on NK cell ATP, but not target cell ATP. However, the intercellular transfer of HLA-C is not dependent on active polymerization of the actin cytoskeleton. In addition, different arrangements of HLA-C are seen at inhibitory NK immune synapses, and these alter as NK synapses mature, but in a fashion distinct from that seen upon T cell activation.

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Related in: MedlinePlus

Intercellular transfer of HLA-C can be inhibited by depleting NK cell ATP but not by depleting target cell ATP or by impairing both cells' actin cytoskeleton. Plot shows percentage of cell conjugates exhibiting intercellular transfer of HLA-Cw6-GFP as a percentage of those where clustering of HLA-Cw6-GFP was observed at the immune synapse. Cells were incubated with or without the following drugs as labeled (a) 50 mM sodium azide, (b) 13 μM antimycin-A, (c) 10 μM cytochalasin D, or 20 mM BDM. Each plot is representative of >100 conjugates over three independent experiments.
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fig5: Intercellular transfer of HLA-C can be inhibited by depleting NK cell ATP but not by depleting target cell ATP or by impairing both cells' actin cytoskeleton. Plot shows percentage of cell conjugates exhibiting intercellular transfer of HLA-Cw6-GFP as a percentage of those where clustering of HLA-Cw6-GFP was observed at the immune synapse. Cells were incubated with or without the following drugs as labeled (a) 50 mM sodium azide, (b) 13 μM antimycin-A, (c) 10 μM cytochalasin D, or 20 mM BDM. Each plot is representative of >100 conjugates over three independent experiments.

Mentions: Sodium azide and antimycin-A have been shown to deplete ATP in the cell lines used by 94 and 97%, respectively (9). Here, the number of conjugates that exhibited clustering of HLA-Cw6-GFP at the immune synapse and the transfer of HLA-Cw6-GFP from 221-Cw6-GFP to YTS/KIR2DL1 cells were quantified with or without the addition of these drugs. Previously, clustering of MHC at the NK immune synapse was observed to be independent of ATP depletion (9). Here however, live-cell confocal microscopy showed that ATP is required by the NK cell for intercellular transfer of HLA-C to take place (Fig. 5 a and b). This infers that intercellular transfer of HLA-C is an active process by the NK cell, independent of the target cell ATP.


Intercellular transfer and supramolecular organization of human leukocyte antigen C at inhibitory natural killer cell immune synapses.

Carlin LM, Eleme K, McCann FE, Davis DM - J. Exp. Med. (2001)

Intercellular transfer of HLA-C can be inhibited by depleting NK cell ATP but not by depleting target cell ATP or by impairing both cells' actin cytoskeleton. Plot shows percentage of cell conjugates exhibiting intercellular transfer of HLA-Cw6-GFP as a percentage of those where clustering of HLA-Cw6-GFP was observed at the immune synapse. Cells were incubated with or without the following drugs as labeled (a) 50 mM sodium azide, (b) 13 μM antimycin-A, (c) 10 μM cytochalasin D, or 20 mM BDM. Each plot is representative of >100 conjugates over three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193674&req=5

fig5: Intercellular transfer of HLA-C can be inhibited by depleting NK cell ATP but not by depleting target cell ATP or by impairing both cells' actin cytoskeleton. Plot shows percentage of cell conjugates exhibiting intercellular transfer of HLA-Cw6-GFP as a percentage of those where clustering of HLA-Cw6-GFP was observed at the immune synapse. Cells were incubated with or without the following drugs as labeled (a) 50 mM sodium azide, (b) 13 μM antimycin-A, (c) 10 μM cytochalasin D, or 20 mM BDM. Each plot is representative of >100 conjugates over three independent experiments.
Mentions: Sodium azide and antimycin-A have been shown to deplete ATP in the cell lines used by 94 and 97%, respectively (9). Here, the number of conjugates that exhibited clustering of HLA-Cw6-GFP at the immune synapse and the transfer of HLA-Cw6-GFP from 221-Cw6-GFP to YTS/KIR2DL1 cells were quantified with or without the addition of these drugs. Previously, clustering of MHC at the NK immune synapse was observed to be independent of ATP depletion (9). Here however, live-cell confocal microscopy showed that ATP is required by the NK cell for intercellular transfer of HLA-C to take place (Fig. 5 a and b). This infers that intercellular transfer of HLA-C is an active process by the NK cell, independent of the target cell ATP.

Bottom Line: This unexpected intercellular transfer of HLA-C is dependent on NK receptor recognition, since HLA-Cw6 or -Cw4 but not -Cw3 transfer to an NK transfectant expressing killer Ig-like receptor (KIR)2DL1.However, the intercellular transfer of HLA-C is not dependent on active polymerization of the actin cytoskeleton.In addition, different arrangements of HLA-C are seen at inhibitory NK immune synapses, and these alter as NK synapses mature, but in a fashion distinct from that seen upon T cell activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Sir Alexander Fleming Building, Imperial College of Science, Technology, and Medicine, South Kensington, London SW7 2AZ, United Kingdom.

ABSTRACT
After accumulation of target cell human leukocyte antigen (HLA)-C at inhibitory natural killer (NK) cell immune synapses, some HLA-C transfers from target cells to NK cell plasma membranes and cytoplasm. This unexpected intercellular transfer of HLA-C is dependent on NK receptor recognition, since HLA-Cw6 or -Cw4 but not -Cw3 transfer to an NK transfectant expressing killer Ig-like receptor (KIR)2DL1. Strikingly, live-cell time-lapse laser scanning confocal microscopy shows vesicles containing target cell green fluorescent protein-tagged HLA-C migrating away from immune synapses into NK cells. Unlike clustering of HLA-C at the immune synapse, intercellular transfer of HLA-C is dependent on NK cell ATP, but not target cell ATP. However, the intercellular transfer of HLA-C is not dependent on active polymerization of the actin cytoskeleton. In addition, different arrangements of HLA-C are seen at inhibitory NK immune synapses, and these alter as NK synapses mature, but in a fashion distinct from that seen upon T cell activation.

Show MeSH
Related in: MedlinePlus