Interleukin-5: A Proeosinophil Cytokine Mediator of Inflammation in Asthma and a Target for Antisense Therapy

Weltman, Joel K.; Karim, Aftab

doi:10.2500/108854198778557782

Cited by 16 publications

(11 citation statements)

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“…[1][2][3] The effects of IL-5 are mediated through binding to its receptor. The IL-5 receptor is composed of ␣-and ␤-chains, each containing a glycosylated extracellular domain with repeating fibronectin-like sequences, a transmembrane domain, and a cytoplasmic domain.…”

Section: Introductionmentioning

confidence: 99%

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

et al. 2004

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The cyclic peptide AF17121 (Ac-VDECWRIIASHTWFCAEE) that inhibits interleukin 5 (IL-5) function and IL-5 receptor alpha-chain (IL-5Ralpha) binding has been derived from recombinant random peptide library screening and follow-up synthetic variation. To better understand the structural basis of its antagonist activity, AF17121 and a series of analogs of the parent peptide were prepared by solid phase peptide synthesis. Sequence variation was focused on the charged residues Asp(2), Glu(3), Arg(6), Glu(17), and Glu(18). Two of those residues, Glu(3) and Arg(6), form an EXXR motif that was found to be common among library-derived IL-5 antagonists. The E and R in the EXXR motif have a proximity similar to charged residues in a previously identified receptor alpha binding region, the beta-strand between the C- and D-helices of human IL-5. Optical biosensor interaction kinetics and cell proliferation assays were used to evaluate the antagonist activities of the purified synthetic peptides, by measuring competition with the highly active single chain IL-5. Analogs in which acidic residues (Asp(2), Glu(3), Glu(17), and Glu(18)) were replaced individually by Ala retained substantial competition activity, with multiple replacements in these residues leading to fractional loss of potency at most. In contrast, R6A analogs had strongly reduced competition activity. The results reveal that the arginine residue is crucial for the IL-5Ralpha binding of AF17121, while the acidic residues are not essential though likely complex-stabilizing particularly in the Asp(2)-Glu(3) region. By CD, AF17121 exhibited mostly disordered structure with evidence for a small beta-sheet content, and replacement of the arginine had no influence on the observed secondary structure of the peptides. The dominance of Arg(6) in AF17121 activity corresponds to previous findings of dominance of the positive charge balance in the antiparallel beta-sheet of IL-5 composed of (88)EERRR(92) in one strand of the CD turn region of IL-5 and with Arg(32) in the neighboring beta-strand. These results argue that AF17121 and related library-derived peptides function by mimicking the CD turn receptor alpha recognition epitope in IL-5 and open the way to small molecule antagonist design.

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Section: Introductionmentioning

confidence: 99%

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

et al. 2004

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“…(4)(5)(6)(7)(8) Helminth-induced as well as aeroallergen-induced eosinophilia and airway hyperresponiveness are abrogated in IL-5 knockout mice. (9,10) Because of its critical role in eosinophil activation and function, IL-5 has been the target for experimental treatment of diseases associated with excessive eosinophils.…”

Section: Introductionmentioning

confidence: 97%

“…Reagents that can potentially block IL-5 activity include IL-5-specific neutralizing antibodies, modified antisense oligonucleotides, and soluble receptors. (4)(5)(6)(7)(8) Increased numbers of eosinophils have been observed in a number of allergic diseases in dogs. For example, Olivry et al (11) reported that the skin infiltrate of canine atopic dermatitis consists of mast cells, dendritic cells, memory helper T lymphocytes, neutrophils, and eosinophils.…”

Section: Introductionmentioning

confidence: 99%

Canine Interleukin-5: Molecular Characterization of the Gene and Expression of Biologically Active Recombinant Protein

Yang

Sellins

Weber

et al. 2001

Journal of Interferon & Cytokine Research

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Interleukin-5 (IL-5), which is produced primarily by type 2 T helper lymphocytes (Th2), is an eosinophil differentiation and activation factor. Increased numbers of eosinophils in peripherial blood or tissues (eosinophilia) are observed in asthmatic human patients, in animals with helminth infections, and in dogs with allergic diseases. Antagonism of IL-5 activity is being explored as a potential treatment of a number of disease conditions associated with eosinophils in animal models. In order to study the expression and function of this cytokine in the dog, we have isolated and characterized the canine IL-5 gene. The canine IL-5 polypeptide deduced from the cDNA is composed of 134 amino acids that share varying degrees of homology with IL-5 isolated from several mammals. The genomic structure of the canine IL-5 gene consists of four exons and three introns in the coding region, similar to that of the previously characterized human and mouse IL-5 genes. Recombinant canine IL-5 protein, expressed in Pichia pastoris, is biologically active in a cell proliferation assay. Canine IL-5 gene sequences and the biologically active protein described in this study will be useful reagents for future studies of this cytokine in physiologic processes and in pathologic conditions of the dog.

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“…IL5 induces eosinophil production, stimulates the growth and differentiation of eosinophils, regulates the proinflammatory function of eosinophils by promoting the release of the eosinophil granular proteins and prolongs the survival time of eosinophils (Karlen et al, 1998;Weltman & Karim, 1998). IL5 plays a major role in the development of antigen-induced eosinophilic diseases such as asthma and other allergic disorders (Gibson et al, 1998;Gleich & Kita, 1997;Mori et al, 1997;Sehmi et al, 1997;Till et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Coiled coil miniprotein randomization on phage leads to charge pattern mimicry of the receptor recognition determinant of interleukin 5

Plugariu

Bajgier

et al. 2002

J of Molecular Recognition

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Phage display was used to identify sequences that mimic structural determinants in interleukin5 (IL5) for IL5 receptor recognition. A coiled coil stem loop (CCSL) miniprotein scaffold library was constructed with its turn region randomized and panned for binding variants against human IL5 receptor alpha chain (IL5Ralpha). Competition enzyme-linked immunosorbent assays identified CCSL-phage selectants for which binding to IL5Ralpha was competed by IL5. The most frequently selected and IL5-competed CCSL-phage contain charged residues Arg and Glu in their turn sequences, in this regard resembling a beta strand sequence in the 'CD turn' region, of IL5, that has been proposed to present a key determinant for IL5 receptor alpha chain recognition. The most dominant CCSL-phage selectant sequence, PVEGRV, contains a negative/positive charge pattern similar to that seen in the original CD turn. To test the relatedness of CCSL-phage selectant sequences to the IL5 receptor recognition epitope, PVEGRV was grafted into the sequence 87--92 of a monomeric IL5. The resulting IL5 variant, [(87)PVEGRV(92)]GM1, was able to bind to IL5Ralpha in biosensor assays, to elicit TF-1 cell proliferation and to induce STAT5 phosphorylation in TF-1 cells. The results help discern sequence patterns in the IL5 CD turn region which are key in driving receptor recognition and demonstrate the utility of CCSL miniprotein scaffold phage display to identify local IL5 mimetic sequence arrangements that may ultimately lead to IL5 antagonists.

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Interleukin-5: A Proeosinophil Cytokine Mediator of Inflammation in Asthma and a Target for Antisense Therapy

Cited by 16 publications

References 0 publications

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

Canine Interleukin-5: Molecular Characterization of the Gene and Expression of Biologically Active Recombinant Protein

Coiled coil miniprotein randomization on phage leads to charge pattern mimicry of the receptor recognition determinant of interleukin 5

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