Design of human interleukin‐4 antagonists inhibiting interleukin‐4‐dependent and interleukin‐13‐dependent responses in T‐cells and B‐cells with high efficiency

Tony, Hans Peter; Shen, Bo; Reusch, Petra; Sebald, Walter

doi:10.1111/j.1432-1033.1994.00659.x

Cited by 110 publications

(99 citation statements)

References 24 publications

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“…Although hIL13 and hIL4 are dissimilar at the primary amino acid level, they share similar functions by binding to and signaling through a shared receptor complex (11,12). The shared IL13/4 receptor complex is a heterodimer comprised of an approximate 140-kDa subunit, p140 (13), and an approximate 52-kDa subunit, ␣Ј also termed IL13R␣1 (14 -16).…”

Section: Human Interleukin 13 (Hil13)mentioning

confidence: 99%

Mutants of Interleukin 13 with Altered Reactivity toward Interleukin 13 Receptors

Thompson

Debinski

1999

Journal of Biological Chemistry

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Interleukin 13 (IL13) belongs to a family of cytokines whose members exhibit structural homology, despite amino acid sequence dissimilarity. For example, while of limited sequence homology, IL13 and IL4 share a signaling receptor, IL13/4 receptor, on a variety of human normal cells. However, a subclass of IL4-independent IL13 receptors is overexpressed on certain transformed cells, including human malignant gliomas. We introduced mutations into human (h) IL13 to determine the site(s) involved in interaction with the shared receptor and/or the glioma-associated receptor. This analysis identified at least three protein regions that are needed for signaling through the shared receptor. These regions were localized to ␣-helices A, C, and D and were mainly separate from the region(s) needed to interact with the glioma-associated receptor. Glutamic acids at positions 13 and 16 in hIL13 ␣-helix A, arginine and serine at positions 66 and 69 in helix C, and arginine at position 109 in helix D were found to be important in inducing biological signaling since their specific mutation resulted in loss and/or gain of function phenomena. We demonstrate that the molecular requirements of hIL13 to interact with its respective receptors are generally distinct and can be controlled by mutagenesis of the cytokine.

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Section: Human Interleukin 13 (Hil13)mentioning

confidence: 99%

Mutants of Interleukin 13 with Altered Reactivity toward Interleukin 13 Receptors

Thompson

Debinski

1999

Journal of Biological Chemistry

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“…The IL-4 receptor comprises two chains, the IL-4Ra and the yc [7][8][9][10]. Neither of these two chains binds IL-13 [5], but recent reports have shown that IL-4Ra contributes to the IL-13 receptor [11][12][13].…”

Section: Introductionmentioning

confidence: 99%

Cloning of the human IL‐13Rα1 chain and reconstitution with the IL‐4Rα of a functional IL‐4/IL‐13 receptor complex

Miloux¹,

Laurent²,

Bonnin³

et al. 1997

FEBS Letters

200

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The human homologue of the recently cloned murine IL-13 binding protein (IL-13Ral) was cloned from a cDNA library derived from the carcinoma cell line CAKI-1. The cloned cDNA encodes a 427 amino acid protein with two consensus patterns characteristic of the hematopoietic cytokine receptor family and a short cytoplasmic tail. The human protein is 74% identical to the murine IL-13Ral, and 27% identical to the human IL-13Ra2. CHO cells expressing recombinant hlL13Ral specifically bind IL-13 (Äd~4 nM) but not IL-4. Coexpression of the cloned cDNA with that of IL-4Ra resulted in a receptor complex that displayed high affinity for IL-13 (Κ^ ~ 30 pM), and that allowed cross-competition of IL-13 and IL-4. Electrophoretic mobility shift assay showed that IL-13 and IL-4 were able to activate Stat6 in cells expressing both IL-4Roc and IL-13Ral, while no activation was observed in cells expressing either one or the other alone.

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“…Recently the structure of this complex has been resolved by x-ray crystallography revealing specific amino acid residues of IL-4 that are involved in the binding of each of the receptor chains (7). Interestingly, preceding this structural knowledge of the IL-4 cytokine/cytokine receptor complex, IL-4 antagonists had been identified through systematic mutagenesis, seeking to abolish binding to the signaling receptor chain ␥C while preserving high affinity binding to the IL-4R␣ chain (8).…”

Section: Interleukin (Il)mentioning

confidence: 99%

Rational Design of Interleukin-21 Antagonist through Selective Elimination of the γC Binding Epitope

Kang

Bondensgaard

et al. 2010

Journal of Biological Chemistry

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The cytokine interleukin (IL)-21 exerts pleiotropic effects acting through innate as well as adaptive immune responses. The activities of IL-21 are mediated through binding to its cognate receptor complex composed of the IL-21 receptor private chain (IL-21R␣) and the common ␥-chain (␥C), the latter being shared by IL-2, IL-4, IL-7, IL-9, and IL-15. The binding energy of the IL-21 ternary complex is predominantly provided by the high affinity interaction between IL-21 and IL-21R␣, whereas the interaction between IL-21 and ␥C, albeit essential for signaling, is rather weak. The design of IL-21 analogues, which have lost most or all affinity toward the signaling ␥C chain, while simultaneously maintaining a tight interaction with the private chain, would in theory represent candidates for IL-21 antagonists. We predicted the IL-21 residues, which compose the ␥C binding epitope using homology modeling and alignment with the related cytokines, IL-2 and IL-4. Next we systematically analyzed the predicted binding epitope by a mutagenesis study. Indeed two mutants, which have significantly impaired ␥C affinity with undiminished IL-21R␣ affinity, were successfully identified. Functional studies confirmed that these two novel hIL-21 double mutants do act as hIL-21 antagonists. Interleukin (IL),2 -21 which is produced by activated CD4ϩ T cells, T-follicular helper cells, and natural killer T (NKT) cells, (1) has been demonstrated to exert pleiotropic effects on the proliferation, differentiation, and cytotoxicity of various classes of lymphoid cells. More recently, IL-21 has furthermore been shown to play a crucial role in the differentiation of CD4 ϩ T cells into T-helper 17 (TH 17 ) cells, a subset of T cells associated with development of inflammatory conditions and autoimmune diseases (2, 3).IL-21 signals through its receptor complex composed of the private chain IL-21R␣ and the common chain, ␥C, the latter of which is shared by five other cytokines: IL-2, IL-4, IL-7, IL-9, and IL-15 (1). Together these cytokines constitute the so-called ␥C family of cytokines. Despite the rather limited sequence homology (on average 15% sequence identity), these ␥C cytokines share a highly conserved overall four-helix bundle topology.Previously, we have reported the three-dimensional structure of human IL-21 (hIL-21) resolved by heteronuclear NMR spectroscopy (4). The structure of hIL-21 was shown to comprise a typical up-up-down-down four-␣-helical-bundle topology similar to that observed for several other type I cytokines, including the ␥C cytokines IL-2, IL-4, and IL-15. Noteworthy, a segment of hIL-21, including the helix C that by modeling is presumed to be important for IL-21R␣ binding, was demonstrated to represent a structurally unstable segment, not observed in the structures of other ␥C cytokines. A chimeric variant in which this flexible segment of hIL-21 had been exchanged with the corresponding IL-4 sequence was constructed and demonstrated to have a 10-fold increase in potency in a cell-based assay (4).Functional receptor ...

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Design of human interleukin‐4 antagonists inhibiting interleukin‐4‐dependent and interleukin‐13‐dependent responses in T‐cells and B‐cells with high efficiency

Cited by 110 publications

References 24 publications

Mutants of Interleukin 13 with Altered Reactivity toward Interleukin 13 Receptors

Mutants of Interleukin 13 with Altered Reactivity toward Interleukin 13 Receptors

Cloning of the human IL‐13Rα1 chain and reconstitution with the IL‐4Rα of a functional IL‐4/IL‐13 receptor complex

Rational Design of Interleukin-21 Antagonist through Selective Elimination of the γC Binding Epitope

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