scholarly journals A Chimera of Interleukin 2 and a Binding Variant of Aerolysin Is Selectively Toxic to Cells Displaying the Interleukin 2 Receptor

2007 ◽  
Vol 283 (3) ◽  
pp. 1572-1579 ◽  
Author(s):  
Milan Osusky ◽  
Lisa Teschke ◽  
Xiaoying Wang ◽  
Kevin Wong ◽  
J. Thomas Buckley
Keyword(s):  
Cell Death ◽  
Mammalian Cells ◽  
Therapeutic Agent ◽  
Interleukin 2 ◽  
Bacterial Toxin ◽  
Hybrid Protein ◽  
High Affinity ◽  
Interleukin 2 Receptor ◽  
Hybrid Molecules ◽  
N Terminus

Aerolysin is a bacterial toxin that binds to glycosylphosphatidylinositol-anchored proteins (GPI-AP) on mammalian cells and oligomerizes, inserting into the target membranes and forming channels that cause cell death. We have made a variant of aerolysin, R336A, that has greatly reduced the ability to bind to GPI-AP, and as a result it is only very weakly active. Fusion of interleukin 2 (IL2) to the N terminus of R336A-aerolysin results in a hybrid that has little or no activity against cells that do not have an IL2 receptor because it cannot bind to the GPI-AP on the cells. Strikingly, the presence of the IL2 moiety allows this hybrid to bind to cells displaying high affinity IL2 receptors. Once bound, the hybrid molecules form insertion-competent oligomers. Cell death occurs at picomolar concentrations of the hybrid, whereas the same cells are insensitive to much higher concentrations of R336A-aerolysin lacking the IL2 domain. The targeted channel-forming hybrid protein may have important advantages as a therapeutic agent.

scholarly journals Interleukin 2 receptor-targeted cytotoxicity. Interleukin 2 receptor-mediated action of a diphtheria toxin-related interleukin 2 fusion protein.

1988 ◽  
Vol 167 (2) ◽  
pp. 612-622 ◽  
Author(s):  
P Bacha ◽  
D P Williams ◽  
C Waters ◽  
J M Williams ◽  
J R Murphy ◽  
...  
Keyword(s):  
T Cells ◽  
Protein Synthesis ◽  
T Cell ◽  
Diphtheria Toxin ◽  
Interleukin 2 ◽  
Elongation Factor ◽  
High Affinity ◽  
Interleukin 2 Receptor ◽  
Human T Cell ◽  
Inhibition Of Protein Synthesis

The IL-2 toxin-mediated inhibition of protein synthesis in high affinity IL-2-R-positive murine and human T cell lines has been examined. Both excess free IL-2 and mAb to the Tac epitope of the p55 subunit of IL-2-R are shown to block the action of IL-2 toxin; whereas, agents that interact with other receptors or antigens on the T cell surface have no effect. We show that IL-2 toxin, like diphtheria toxin, must pass through an acidic vesicle in order to intoxicate target T cells. Finally, we demonstrate that the IL-2 toxin-mediated inhibition of protein synthesis in both human and murine T cells that bear the high affinity IL-2-R is due to the classic diphtheria toxin fragment A-catalyzed ADP ribosylation of elongation factor 2.

scholarly journals Solution assembly of the pseudo-high affinity and intermediate affinity interleukin-2 receptor complexes

2008 ◽  
Vol 8 (3) ◽  
pp. 482-489 ◽  
Author(s):  
Zining Wu ◽  
Byron Goldstein ◽  
Thomas M. Laue ◽  
Stefano F. Liparoto ◽  
Michael J. Nemeth ◽  
...  
Keyword(s):  
Interleukin 2 ◽  
Receptor Complexes ◽  
High Affinity ◽  
Interleukin 2 Receptor

Stepwise formation of the high-affinity complex of the interleukin 2 receptor

1990 ◽  
Vol 2 (12) ◽  
pp. 1167-1177 ◽  
Author(s):  
Yuji Saito ◽  
Toshihiko Ogura ◽  
Masanori Kamio ◽  
Hisataka Sabe ◽  
Takashi Uchiyama ◽  
...  
Keyword(s):  
Interleukin 2 ◽  
High Affinity ◽  
Interleukin 2 Receptor ◽  
Stepwise Formation

Novel interleukin-2 receptor subunit detected by cross-linking under high-affinity conditions

Science ◽  
1986 ◽  
Vol 234 (4778) ◽  
pp. 859-863 ◽  
Author(s):  
M Sharon ◽  
R. Klausner ◽  
B. Cullen ◽  
R Chizzonite ◽  
W. Leonard
Keyword(s):  
Interleukin 2 ◽  
Cross Linking ◽  
Receptor Subunit ◽  
High Affinity ◽  
Interleukin 2 Receptor

scholarly journals Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kyra See ◽  
Tetsuya Kadonosono ◽  
Kotaro Miyamoto ◽  
Takuya Tsubaki ◽  
Yumi Ota ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Dissociation Constants ◽  
Surface Display ◽  
Interleukin 2 ◽  
Peptide Identification ◽  
Cell Surface Display ◽  
Affinity Maturation ◽  
High Affinity ◽  
Binding Peptides ◽  
Target Binding

AbstractSmall antibody mimetics that contain high-affinity target-binding peptides can be lower cost alternatives to monoclonal antibodies (mAbs). We have recently developed a method to create small antibody mimetics called FLuctuation-regulated Affinity Proteins (FLAPs), which consist of a small protein scaffold with a structurally immobilized target-binding peptide. In this study, to further develop this method, we established a novel screening system for FLAPs called monoclonal antibody-guided peptide identification and engineering (MAGPIE), in which a mAb guides selection in two manners. First, antibody-guided design allows construction of a peptide library that is relatively small in size, but sufficient to identify high-affinity binders in a single selection round. Second, in antibody-guided screening, the fluorescently labeled mAb is used to select mammalian cells that display FLAP candidates with high affinity for the target using fluorescence-activated cell sorting. We demonstrate the reliability and efficacy of MAGPIE using daclizumab, a mAb against human interleukin-2 receptor alpha chain (CD25). Three FLAPs identified by MAGPIE bound CD25 with dissociation constants of approximately 30 nM as measured by biolayer interferometry without undergoing affinity maturation. MAGPIE can be broadly adapted to any mAb to develop small antibody mimetics.

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