IL-5 Receptor

10 Pages·0.101 MB·English

by  Bagley Ch.J., Tavernier J., Woodcock J.M.

IL-5 Receptor
Christopher J. Bagley1,2,*, Jan Tavernier3, Joanna M. Woodcock4,2
and Angel F. Lopez4,2
1Protein Laboratory, Hanson Centre for Cancer Research, Frome Road, Adelaide,
SA, 5000, Australia
2Human Immunology, Institute of Medical and Veterinary Science, Frome Road,
Adelaide, SA, 5000, Australia
3Department of Medical Protein Research, Flanders’ Interuniversity Institute for Biotechnology,
University of Ghent, Ghent, Belgium
4Cytokine Receptor Laboratory, Hanson Centre for Cancer Research, Frome Road,
Adelaide, SA, 5000, Australia
*corresponding author tel: (cid:135)61-8-82223714, fax: (cid:135)61-8-82324092, e-mail: [email protected]
DOI: 10.1006/rwcy.2000.20002.
SUMMARY Tavernieretal.(1991)whoalsoidentifiedthe(cid:12) chain
of the GM-CSF receptor ((cid:12)c, cloned by Hayashida
et al., 1990) as a component of the IL-5R.
ThereceptorforIL-5iscomprisedoftwochains:an(cid:11)
chainthatbindsIL-5withmoderateaffinitybutalone
is unable to mediate signaling, and a (cid:12) chain that
Alternative names
represents the major signaling component of the
receptor. The (cid:12) chain is unable to bind IL-5 alone,
but when expressed together with the IL-5 receptor (cid:11) IL-5R (cid:11) chain: IL-5R(cid:11), Cdw125.
chain (IL-5R(cid:11)) a high-affinity receptor is formed IL-5R(cid:12) chain:IL-5R(cid:12),Cdw131,common(cid:12) chainof
(K (cid:136)0.15nM).The(cid:12)chainissharedwiththerelated the GM-CSF, IL-3 and IL-5 receptors, (cid:12)c.
d
receptors for granulocyte–macrophage colony-stimu-
lating factor (GM-CSF) and IL-3 and many of the
events in IL-5-induced signal transduction are Structure
paralleled in the GM-CSF and IL-3 receptors. The
IL-5R(cid:11)isexpressedonarestrictedrangeofcelltypes, The IL-5 receptor consists of two chains, denoted
principally eosinophils, basophils, and their immedi- (cid:11) and (cid:12). The (cid:11) subunit has specific ligand-binding
ate precursors confining the actions of IL-5 to these activity whereas the (cid:12) subunit does not bind IL-5 by
cells. Thus, the IL-5 receptor is considered to be a itself but enhances the binding of IL-5 and provides
target in the development of treatments for allergic the major determinants of signaling capacity of the
inflammatory diseases such as asthma. receptor. The (cid:12) subunit is also a component of the
high-affinity receptors for GM-CSF and IL-3.
BACKGROUND
Main activities and
Discovery pathophysiological roles
ThetwocomponentsoftheIL-5Rwerefirstidentified Stimulation of the IL-5 receptor in vivo results in the
in crosslinking studies (Mita et al., 1989). The cDNA stimulation of production of eosinophils and baso-
forIL-5R(cid:11)wasclonedbyTakakietal.(1990)andby phils which exacerbates allergic inflammation.
1904 Christopher J. Bagley, Jan Tavernier, Joanna M. Woodcock and Angel F. Lopez
GENE IL-5R(cid:11) produced in Escherichia coli has been
reported to bind IL-5 with near normal affinity
Accession numbers (Monahan et al., 1997).
Theextracellularpartoftheproteinfoldsintothree
fibronectintypeIIIstructuraldomains,eachhavinga
GenBank:
seven (cid:12) sheet scaffold. The juxtamembrane domain
HumanIL-5R(cid:11):M75914,M96651,M96652,X61176,
contains a canonical Trp-Ser-Xaa-Trp-Ser motif
X61177, X61178, X62156
(WSXWS box) and forms together with the central
Human IL-5R(cid:12): M59941, M38275
domain, which itself is characterized by four con-
servedcysteines,aso-calledcytokinereceptormodule
Sequence (CRM) (Goodall et al., 1993) also described as a
cytokine-binding domain (CBD). The N-terminal
Both the murine IL-5R(cid:11) gene (Imamura et al., 1994) domain is more related to the WSXWS box domain
and IL-5R(cid:12) gene (Gorman et al., 1992) have been (Tuypensetal.,1992),andcontainstwoCysresidues,
mapped. which may form an intradomain disulfide bond
(Cornelisetal.,1995a).WiththeexceptionofaBox-1
motif(Pro-Pro-Xaa-Pro),noconsensussequencesare
PROTEIN foundin the cytoplasmicdomain. Ligandrecognition
involvesresiduesinthehingebetweenthecentraland
Accession numbers membrane-proximal domains as well as a region in
the N-terminal domain (Cornelis et al., 1995b).
The nucleotide sequence of the human IL-5R(cid:12)
See Table 1.
cDNApredictsapolypeptideof897aminoacids.The
protein is characterized by a 16 residue N-terminal
Description of protein
signal peptide, followed by a 422 amino acid extra-
cellular domain, a membrane anchor spanning 22
ThenucleotidesequenceofthehumanIL-5R(cid:11)cDNA residues, and a 437 amino acid cytoplasmic tail. The
predicts a polypeptide of 420 amino acids. It is char- predicted molecular mass for the (cid:12) chain is 97.3kDa,
acterized by a 20 residue N-terminal signal peptide, indicating that N-linked glycosylation and probably
followed by a 322 amino acid extracellular domain, O-glycosylation contribute to the apparent molecular
a membrane anchor spanning 20 residues, and a 58 mass of approximately 130kDa. The extracellular
amino acid cytoplasmic tail. The predicted molecular partoftheproteinfoldsintofourfibronectintypeIII
mass for the (cid:11) chain is 45.5kDa, indicating that structural domains grouped into two CRMs (Bagley
N-linked glycosylation of one or more of the six et al., 1997). The membrane-proximal CRM contains
potential N-glycosylation sites (and perhaps O-glyco- the main determinants for interaction with IL-5,
sylation)contributestotheapparentmolecularmassof principally in the loops between the B and C and F
60kDa. The biological role of glycosylation remains and G strands of domain four (Woodcock et al.,
unresolved since deglycosylation appears to lead to 1994, 1996). The cytoplasmic domain contains a
loss of IL-5 binding (Johanson et al., 1995) although Box-1 motif (Pro-Pro-Xaa-Pro) responsible for
Table 1 Protein accession numbers
ID (Swiss) AC (Swiss) AC (GenPept)
Human IL-5R(cid:11) IL5R_HUMAN Q01344
Mouse IL-5R(cid:11) IL5R_MOUSE P21183
Guinea pig IL-5R(cid:11) U55215
Human (cid:12)c CYRB_HUMAN P32927
Mouse (cid:12)c CYRB_MOUSE P26955
Guinea pig (cid:12)c U94688
Rat IL-3-specific Q64146
(or common) (cid:12) chain
IL-5 Receptor 1905
interaction with JAK kinases, eight tyrosine residues expressionleveliscontrolledatdifferentlevels.Inthe
that are susceptible to phosphorylation, and a mouse, in addition to these cell types, the IL-5R(cid:11) is
recently reported phosphoserine-containing site that alsofoundprominentlyonBcellsbelongingtotheB1
binds 14-3-3 proteins (Stomski et al., 1999). lineage(Hitoshietal.,1990).Althoughtherehasbeen
no report of direct measurement of IL-5R on human
B cells, IL-5 appears to augment terminal differentia-
Relevant homologies and
tion of mitogen-stimulated B cells in some cases.
species differences
Regulation of receptor expression
BothchainsoftheIL-5Raremembersofthecytokine
receptorfamily.TheIL-5R(cid:11)ismostcloselyrelatedto
the GM-CSFR(cid:11) and IL-3R(cid:11) and more distantly Two promoters, P1 and P2, have been identified in
relatedtotheIL-13R(cid:11)2.ThetwoCRMsof(cid:12)carenot the hIL-5R(cid:11) gene (Sun et al., 1995; Zhang et al.,
closelyrelatedtoeachother,nortoothermembersof 1997). P1 and P2 precede the first and the second
the cytokine receptor family. exon respectively, and show no significant sequence
Apart from the immediate membrane-proximal similarities. P1 is myeloid and eosinophil lineage-
region, the cytoplasmic domain of (cid:12)c is not signi- specific, whilst the use of P2 is restricted to eosino-
ficantly related to those of other cytokine receptors. philicHL60-C15cells.Itisatpresentunclearwhether
Human and murine IL-5R (cid:11) chains are 68% iden- differential use and regulation of both promoters
tical (extracellular 71%; intracellular 58%). Human occur during eosinophilic differentiation.
and murine common (GM-CSF and IL-5 receptor) (cid:12) The P1 promoter contains multiple consensus
chains are 56% identical (extracellular 57%; intracel- binding sites for AP-1, C/EBP, GATA, and PU.1.
lular 55%). In addition, the region between (cid:255)432 and (cid:255)398 was
showntocontainauniquecis-element(EOS1),neces-
sary and sufficient for the expression in eosinophilic
Affinity for ligand(s)
cell lines (Sun et al., 1995). The putative myeloid- or
eosinophil-specific binding factor(s) has not been
The IL-5R(cid:11) subunit is ligand-specific and binds with identified so far. Involvement of the adjacent AP-1
intermediate affinity in humans (K (cid:136)(cid:24)0.5–1nM) elementatposition(cid:255)440to(cid:255)432intheexpressionin
d
andlowaffinityinthemouse(K (cid:136)(cid:24)5–10nM).Upon eosinophilic HL-60 cells has been demonstrated
d
association with the (cid:12) subunit, similar high-affinity (Baltus et al., 1998), suggesting cooperation between
binding (K (cid:136)150pM) is observed for both species. the cognate binding transcription factors. Supershift
d
The (cid:12) subunit does not have any detectable affinity analysis experiments showed the presence of cJun,
for IL-5 by itself (Takaki et al., 1990, 1991; Devos CREB, and CREM in the AP-1-binding complexes.
et al., 1991; Tavernier et al., 1991; Murata et al., The P2 promoter sequence shows the presence of
1992). The dissociation rate of mIL-5 from the (cid:11)/(cid:12) AP-1, C/EBP, GATA, CLEO (IL-5), and its con-
complex is considerably slower (t >1 hour) than sensus binding sites. A unique functional motif was
1=2
from the low-affinity (cid:11)-binding site (t <2 min) identified between positions (cid:255)19 and (cid:255)14. It is
1=2
(Devos et al., 1991). IL-5 binds to its receptor with involved in the binding of a hitherto unidentified
unidirectional species-specificity: mIL-5 binds with eosinophilic HL-60-C15-specific transcription fac-
comparableaffinitiestobothmurineandhuman(cid:11)sub- tor(s). Alternatively, it may also serve as a non-
units, but hIL-5 displays 100-fold lower binding affi- canonical TATA box, since such a motif is lacking in
nityforthemurine(cid:11)chain,comparedwithitshuman the P2 promoter.
counterpart, a cross-species pattern that is mirrored Rapiddownregulation(maximuminhibitionwithin
in their biological activities (Lopez et al., 1986). 2hours) of the hIL-5R(cid:11) mRNA is induced in peri-
pheral blood eosinophils upon treatment with IL-3,
Cell types and tissues expressing IL-5,orGM-CSF.Incontrast,similartreatmentleads
to upregulation of the IL-3R(cid:11), GM-CSFR(cid:11), and (cid:12)c
the receptor
mRNA. The mechanisms involved were shown to be
promoteractivationandreducedmRNAdegradation,
ExpressionoftheIL-5Rinhumansismostprominent respectively, indicating differential regulation (Wang
on eosinophils and basophils (Lopez et al., 1991). Its et al., 1998). It is at present unclear whether the
appearance on multipotential myeloid progenitors is downmodulation occurs for P1, P2, or both.
criticalfortheirdevelopmenttowardstheeosinophilic The promoter of the mIL-5R(cid:11) contains consensus
lineage. It remains expressed on mature cells, but the binding sites for AP-1, GATA, NF-IL-6, NF(cid:20)B, and
1906 Christopher J. Bagley, Jan Tavernier, Joanna M. Woodcock and Angel F. Lopez
SP-1. Little is known about transcription factors SIGNAL TRANSDUCTION
controlling cell type-specific expression of the mIL-
5R(cid:11) chain.
Signal transduction via the IL-5 receptor involves
ligand binding and receptor dimerization, require-
ments shared by other cytokine receptors (Bagley
Release of soluble receptors
et al., 1997). The structural elements utilized by the
IL-5 receptor (cid:11) and (cid:12) chains to bind IL-5 have been
Human eosinophils express through alternative discussed above. Following binding of IL-5, dimer-
splicing different transcripts from the same IL-5R(cid:11) ization of the IL-5 receptor ensues, a process which
locus(Tavernieretal.,1992;Tuypensetal.,1992).As shares certain events with the cytokine receptor
a result, in addition to the membrane-anchored superfamily at large but which has some features
receptor, two soluble isoforms can be produced. more limited to the IL-5, GM-CSF, and IL-3
One of these soluble variants is the predominant subfamily of receptors.
(>90%) transcript detected in eosinophilic HL-60- Dimerization of the IL-5 receptor involves the
C15cellsandineosinophilsobtainedfromcordblood associationoftheIL-5receptor(cid:11)chainwith(cid:12)c.This
cultures. Variable isoform mRNA expression has takesplacebynoncovalentaswellascovalentmeans.
beenobservedineosinophilspurifiedfromperipheral The covalent linkage of IL-5R(cid:11) and (cid:12)c is probably
blood. The forced expiratory volume in 1 second in the most functionally relevant one as it is associated
patients with mild asthma has been reported to be with tyrosine phosphorylation of the receptor
inversely correlated with the expression of the (Stomski et al., 1998). The cysteines involved are
membrane-anchored isoform and directly correlated Cys86 and Cys91 in the most N-terminal domain of
with the soluble isoform in eosinophils in endobron- (cid:12)c (Stomski et al., 1998). These cysteines interact
chial biopsies (Yasruel et al., 1997). with a Cys in the IL-5 receptor (cid:11) chain which has
In mouse B cells, transcripts encoding secreted not yet been determined, however, since this (cid:11) chain
variants are also generated through alternative has an odd number of cysteines and all cysteines
splicing. In contrast to humans, there is no evidence except Cys86 appear to form intramolecular bonds
for a similar soluble variant-specific exon. Rather, (based on alignment with other members of the
these soluble variant-specific transcripts are formed cytokine receptor superfamily); Cys86 is the prime
by skipping of the membrane anchor exon (Takaki candidate. The related GM-CSF and IL-3 receptor (cid:11)
et al., 1990; Tavernier et al., 1992; Imamura et al., chains also exhibit an odd number of cysteines
1994). consistent with them also forming high-order com-
The soluble hIL-5R(cid:11) isoform has antagonistic plexes with (cid:12)c.
propertiesinvitro.ItbindsoneIL-5dimerinsolution. Both the IL-5R(cid:11) and (cid:12)c subunits are required for
It inhibits various IL-5 activities, including induced signaling. Deletion of the cytoplasmic domains of
tyrosine phosphorylation of JAK2 and (cid:12)c, prolifera- either chain leads to complete loss of signaling,
tion of IL-5-dependent cell lines, and eosinophilic without altered ligand binding (Sakamaki et al.,
differentiation and survival (Tavernier et al., 1991; 1992; Takaki et al., 1994; Cornelis et al., 1995b).
Monahan et al., 1997), suggesting a role in the IL-5 bindingleads tothe rapid tyrosine phosphoryla-
regulation of eosinophilia in vivo. No IL-5-potentiat- tion of a multitude of cytoplasmic proteins.
ing effects have been observed in in vitro assays, Mutations at position 13 of IL-5 cause diminished
underscoring its anti-inflammatory potential. So far, or abrogated activation of the (cid:12)c chain and may
however, neither translation of the message encoding yield variants with antagonistic activity (Tavernier
this soluble variant in vitro in eosinophils, nor et al., 1995; Bagley et al., 1999). An E to K
circulatingsolublehIL-5R(cid:11)invivohasbeenreported. substitution at residue 13 leads to loss of detectable
One possible explanation might be the thermolability phosphorylation of (cid:12)c in eosinophils. Yet, whilst
of this soluble receptor. Alternatively, this splice being deficient in inducing TF-1 proliferation and
regulation could merely serve a regulatory function eosinophil adhesion, this IL-5 mutein still retains the
driving transcription into a nonproductive pathway, capacity to support eosinophil survival, indicating
reducing the expression level of the membrane- that the different signaling pathways and functional
associated receptor. Soluble murine IL-5R(cid:11) also has responses can be segregated (McKinnon et al., 1997).
antagonistic properties in vitro, albeit to a lesser The expression level of the IL-5R(cid:11) subunit may
degreethanitshumancounterpart,consistentwithits control this agonist/antagonist balance (van Ostade
lower affinity for IL-5. et al., 1999).
IL-5 Receptor 1907
Associated or intrinsic kinases is also phosphorylated on serine residues such as
Ser585. This allows (cid:12) to bind to the 14-3-3(cid:16) adapter
protein and presumably associate with other mole-
Neither subunit of the IL-5 receptor possesses intrin-
cules (Stomski et al., 1999). The biological signifi-
sic tyrosine kinase activity.
cance of this is being determined.
Studies in cell lines showed association of JAK2
and JAK1 (or JAK2) with the IL-5R(cid:11) and (cid:12)c sub-
units, respectively, and rapid tyrosine phosphoryla-
DOWNSTREAM GENE
tion upon IL-5-binding (Ogata et al., 1998). This
activationcriticallydependsonthepresenceofintact, ACTIVATION
membrane-proximalproline-richmotifs(Box-1motif)
in both chains (Quelle et al., 1994; Takaki et al., Transcription factors activated
1994).
STAT1 (Pazdrak et al., 1995; van der Bruggen et al.,
1995) and STAT5, c-fos, c-myc (Mui et al., 1995),
Cytoplasmic signaling cascades
NF-AT (Jinquan et al., 1999).
In addition tothe JAK kinases that associate directly
Genes induced
with IL-5R, other kinases such as the Src family
kinasesLynandFyn,andtheBrutontyrosinekinase,
Btk, are activated by IL-5, suggesting a cascade of Activation of STAT5 leads to rapid induction of
tyrosine phosphorylation events. Btk has been impli- immediate early response genes, including Cis, OSM,
cated in IL-5 signaling in B cells only. Mutations in Id, pim-1, c-fos (Mui et al., 1995).
the btk gene lead to B cell deficiencies in humans Other genes induced include c-myc, VEGF
(X-linked agammaglobulinemia) and mice (X-linked (Horiuchi and Weller, 1997), (cid:12) integrin (Palframan
2
immunodeficiency) (Hitoshi et al., 1993; Koike et al., et al., 1998), PAF (Kishimoto et al., 1996), Bcl-xL
1995). Btk functions in concert with the Src family (Dibbert et al., 1998) and Bcl2 (Ochiai et al., 1997;
kinases Lyn and Fyn (Cheng et al., 1994; Appleby Dewson et al., 1999). The expression of the IL-3R(cid:11),
et al., 1995). GM-CSFR(cid:11), and (cid:12)c chains is induced, whereas that
Multiple tyrosine residues become phosphorylated of the IL-5R(cid:11) is repressed (Wang et al., 1998).
on the (cid:12)c subunit and provide docking sites for
signaling molecules. Both STAT1 (Pazdrak et al.,
1995; van der Bruggen et al., 1995) and STAT5 (Mui BIOLOGICAL CONSEQUENCES
et al., 1995) can become phosphorylated and activ-
OF ACTIVATING OR INHIBITING
ated by IL-5 treatment. STAT1 appears to be the
RECEPTOR AND
major STAT activated by IL-5 in eosinophils. In the
case of STAT5, a high degree of redundancy in PATHOPHYSIOLOGY
recruitment sites has been reported (van Dijk et al.,
1997). Unique biological effects of
Via rapid recruitment and phosphorylation of the
activating the receptors
adapters Shc or SHP2, IL-5 signaling can be coupled
totheRaspathway(Pazdraketal.,1997).Downstream
effector molecules of Ras include PI-3 kinase and Although the IL-5 receptor exhibits the same bio-
MAP kinase. PI-3 kinase plays a critical role in the logical activities as the GM-CSF and IL-3 receptors
inductionbyIL-5ofachemokineticresponseinbone in a given cell type by virtue of their sharing the (cid:12)
marrow eosinophils (Palframan et al., 1998). The subunit, its unique biological effect stems from the
downstream targets of PI-3 kinase remain unclear. spectrum of cells types that express the receptor.
ActivationofMAPkinaseisrequiredforinductionof Thus, in contrast to the GM-CSF receptor and to a
members of the AP-1 family, including the c-fos and lesserdegreetheIL-3receptor,whicharemorewidely
c-jun proto-oncogenes. Members of this family are distributed, the expression of the IL-5 receptor is
involved in myeloid differentiation (Foletta et al., largely restricted to eosinophils and basophils in
1998). humans (and to B cells also in the mouse).
Ras activation has also been implicated in Stimulation of the IL-5 receptor in vivo thus results
suppression of apoptosis of eosinophils by IL-5. The in selective stimulation of eosinophil and basophil
(cid:12)c not only undergoes tyrosine phosphorylation but production which exacerbates allergic inflammation.
1908 Christopher J. Bagley, Jan Tavernier, Joanna M. Woodcock and Angel F. Lopez
Phenotypes of receptor knockouts GM-CSF and IL-3, of eosinophils and basophils, the
major inflammatory cell types in allergy. The
and receptor overexpression mice
antibody BION-1 has recently been developed and
thisbindstotheB-Cloopofthefourthdomainof(cid:12)c
Disruption of the IL-5R(cid:11) gene in mice leads to andblockstheproduction,survival,andactivationof
decreased levels of IgA in mucosal secretions (Hiroi eosinophils in response to IL-5, GM-CSF, and IL-3
et al., 1999) and an inability toinduce eosinophilia in (Sun et al., 1999).
response to parasitic infection (Sugaya et al., 1997).
Overexpression of the IL-5R(cid:11) does not lead to
increased levels of eosinophils or B cells, although References
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IL-5 Receptor 1911
LICENSED PRODUCTS
See Table 2.
Table 2 Suppliers of anti-IL-5R(cid:11) and anti-IL-5R(cid:12)
antibodies
Type Class Clone/ID Supplier
Anti-IL-5R(cid:11) antibodies
Polyclonal (goat) IgG AF-253-NA R&D Systems
Monoclonal IgG1 A14 Pharmingen
Anti-IL-5R(cid:12) antibodies
Monoclonal IgG1 MAB1008 Chemicon
International
Monoclonal IgG1 202325 Stratagene
Monoclonal IgG2b S16 Santa Cruz
Monoclonal IgG1k AR-1635 Maine
Biotechnology
Services
Monoclonal IgG1 3D7 Pharmingen
Monoclonal IgG1 4F3 Amrad