Substances and their uses

Details Substances and their uses Substances and their uses

2000-04-06

2004-06-01

Eyler, Yvonne (Department: 1646)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

C435S069100, C435S007100, C435S173300, C435S235100, C435S325000, C435S320100, C435S069700, C530S300000, C530S351000, C536S023100, C536S023500, C536S023400, C536S024300

Reexamination Certificate

active

06743604

ABSTRACT:

IL4 and IL13 are related cytokines that show a significant sequence identity [1,2] and share numerous biological activities. Both have been shown to be important in the induction of IgE and IgG4 synthesis in human B cells [3-6] and the differentiation of Th cells into a Th2 phenotype [8,11]. Among the events leading to IgE synthesis by B cells, induction of germline &egr; RNA transcription, which precedes the class switching to the corresponding H chain C region, has been shown to be triggered by IL4 and IL13 [6,9,10]. Th cells can be subdivided into two major subtypes according to their cytokine production capacities [11]. The major distinction between the two phenotypes are the capacity of Th1 cells to secrete IFN&ggr; and Th2 cells to produce IL4 and IL5 [11]. Th2 cells are thought to be implicated in the development of atopy, allergy and some forms of asthma [12,13]. The differentiation of Th cells into the Th2 phenotype can be induced by IL4 [11]. IL13 was first considered as to be inactive on T cells [2]. However it has been shown recently to induce the differentiation of murine Th cells into the Th2 phenotype [8].
In addition to their effects on lymphocytes, IL4 and IL13 share the ability to inhibit the production of inflammatory cytokines by macrophages [1,2] and to up-regulate the expression of the vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells [16,17,18] leading to adhesion and trans-endothelial migration of very late antigen 4 (VLA4) expressing leukocytes [19]. This provides a basis for selective extravasation of eosinophils, the hallmark of the inflammatory pathology seen in allergy and asthma [20,58,59].
These two cytokines activate common cytokine receptor signaling pathways involving 4PS/IRS-1 [21-25] and the signal transducer and activator of transcription 6 (STAT-6; ref. 26-28). In the murine system, inactivation of STAT-6 has been demonstrated to affect both IL4 and IL13 signaling and to block IL4 and IL13-induced IgE synthesis or Th2 differentiation [29-31]
Studies have been conducted to examine if these two cytokines share a receptor or receptor subunits [2,18,32-34]. The IL4R is composed of two chains, the IL4R&agr; chain and the common &ggr; chain (&ggr;
c
) The &ggr;
c
is shared with the receptors of many of the other 4-helix bundle cytokines such as L2, IL4, IL7, IL9 and IL15 [35,36]. The IL4R&agr; chain alone forms a tight complex with its ligand, whereas the &ggr;
c
was thought to be mainly responsible for signal transduction. However, IL4- and IL-3-induced responses could be observed in cells which naturally do not express &ggr;
c
or in lymphocytes obtained from severe combined immunodeficiency (SCID) patients who are deficient for &ggr;
c
[34,37-39]. It has therefore been proposed that a second form of an IL4R exists which would be activated by both IL4 and IL13 (IL4R type II/IL13R: ref. 40).
A cDNA encoding for a human IL-13 receptor &agr; chain (IL-13 R&agr;) has been cloned (Caput et al [42]). IL-13 R &agr; has been shown to participate in the type II IL-4 receptor which also contains an IL-4 R &agr; chain.
The present inventors have now cloned from a human tissue source a cDNA encoding a polypeptide capable of binding human IL-13 which has not previously been identified. This polypeptide has only 27% sequence identity with the polypeptide identified by Caput et al. Furthermore the expression pattern of mRNAs encoding the molecule identified by the present inventors is very different from that of the molecule identified by Caput er al.
According to one aspect of the present invention there is provided a polypeptide which is capable of binding human IL-13 and/or of binding human IL-4 in the presence of IL4 R &agr;; which:
a) comprises the amino acid sequence shown in
FIG. 1
;
b) has one or more amino acid substitutions, deletions or insertions relative to a polypeptide as defined in a) above; or
c) is a fragment of a polypeptide as defined in a) or b) above which is at least ten amino acids long and which is preferably at least fifty amino acids long.
The term “polypeptide” is used herein in a broad sense to indicate that there are a plurality of peptide bonds present. It therefore includes within its scope substances which may sometimes be referred to in the literature as peptides, polypeptides or proteins.
It can be determined by using techniques known in the art whether or not a particular polypeptide is capable of binding human IL-13. For example, by binding the polypeptide to radiolabelled or tagged forms of human IL-13 or by competitive inhibition of the binding of radiolabelled or tagged forms of human IL-13 to its natural receptor. The binding affinity of the polypeptide for human IL-13 is preferably less than 10 &mgr;M, more preferably less than 1 &mgr;M (when determined at 37° C.).
Polypeptides within the scope of the present invention may be capable of binding human IL-4. This can also be determined by using techniques known to those skilled in the art. For example, by binding the polypeptide to radiolabelled or tagged forms of human IL-4 or by competitive inhibition of the binding of radiolabelled or tagged forms of human IL-4 to its natural receptor. The affinity of the polypeptide would be in the &mgr;M. ideally nM range.
Preferred polypeptides of the present invention form a moiety when combined with the IL-4 R &agr; chain which is capable of binding IL-4 and/or IL-13. This moiety is within the scope of the present invention and is preferably membrane bound. It may represent a new form of IL-4 receptor (referred to herein as IL-4 type II receptor) and be useful in studying the structure and function of said receptor.
The polypeptides of the present invention which are capable of binding human IL-13 and/or human IL-4 are useful for a number of other purposes.
For example, they can be used to bind human IL-4 or human IL-13 and thereby to act as inhibitors by interfering with the interaction between human IL-13 or human IL-4 and their natural receptors. This is useful in medicine since it can be used in the treatment of diseases in which human IL-4 or human IL-13 are responsible (at least partially) for adverse effects in a patient. For example, polypeptides of the present invention could be used to inhibit IL-13 or IL-4 induced IgE synthesis in B cells. This is useful in the treatment of diseases where IgE or Th2 differentiation plays a role—e.g. in the treatment of atopy, atopic dermatitis, allergies, rhinitis, eczema, asthma or AIDS.
Polypeptides of the present invention may therefore be used in the treatment of a human or non-human animal. The treatment may be prophylactic or may be in respect of an existing condition. Examples of particular disorders which can be treated are discussed supra.
Thus a polypeptide of the present invention may be used in the manufacture of a medicament for the treatment of one or more disorders.
The medicament will usually be supplied as part of a pharmaceutical composition. which may include a pharmaceutically acceptable carrier. This pharmaceutical composition will usually be sterile and can be in any suitable form, (depending upon the desired method of administering it to a patient).
It may be provided in unit dosage form, will generally be provided in a sealed container, and can be provided as part of a kit. Such a kit is within the scope of the present invention. It would normally (although not necessarily) include instructions for use. It may include a plurality of said unit dosage forms.
Pharmaceutical compositions within the scope of the present invention may be adapted for administration by any appropriate route, for example by the oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or tansdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) route. Such a composition may be prepared by any

Affiliated with

Also associated with

Rating

Substances and their uses does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Substances and their uses, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Substances and their uses will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3364392