SDF-1 Charles R. Mackay* The Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst (Sydney), New South Wales, 2010, Australia *corresponding author tel: 61-2-92958405, fax: 61-2-92958404, e-mail: [email protected] DOI: 10.1006/rwcy.2000.10010. SUMMARY proliferation which was distinct from IL-7 and stem cell factor (Nagasawa et al., 1994). A cDNA clone encoding this growth-stimulating factor was isolated SDF-1 is a CXC chemokine that has several by expression cloning. In the third study, a partial distinguishing features. It widely expressed and has cDNA sequence, termed TPAR1 (TPA repressed a broad range of actions, affecting T cell chemotaxis, gene 1), was isolated from a cDNA library con- B cell chemotaxis, B cell development, CD34(cid:135) stem structed from mouse embryo fibroblasts treated with cellmigration,andneuraldevelopment,aswellasthe TPA. Full-length clones were later isolated (Jiang development and vascularization of various other et al., 1994). tissues. SDF-1 appears to be particularly important Two alternatively spliced SDF-1 mRNAs encode for basal trafficking of cells, for instance naı¨ve T cell SDF-1(cid:11) (68 residues) and SDF-1(cid:12) (72 residues). The traffic through lymph nodes, rather than the four additional residues of SDF-1(cid:12) are located at the trafficking of effector leukocytes to inflammatory C-terminus (Shirozu et al., 1995). A form that had lesions. SDF-1 is highly conserved between mouse beenprocessedattheC-terminalendtogeneratea67 and human, and binds to the chemokine receptor residue protein was purified from stromal cells (Bleul CXCR4. SDF-1 is able to inhibit T-tropic HIV-1 etal.,1996a).AnunusualfeatureofSDF-1isthatthe entry into CXCR4-bearing cell lines, and SDF-1 human and mouse molecules are virtually identical; expression may have an important bearing on AIDS there is a single conservative change: Val to Ile at pathogenesis, since a polymorphism in the human position 18 (Shirozu et al., 1995). This conservation SDF-1 gene affects the onset of AIDS. has led to speculation that SDF-1 has fundamental functions, and may be a primordial chemokine. BACKGROUND Discovery Alternative names SDF-1 was discovered by three groups independently SDF-1(cid:11) (and its alternative form SDF-1(cid:12)) were (Tashiro et al., 1993; Jiang et al., 1994; Nagasawa described as such in 1993 (Tashiro et al., 1993), etal.,1994).Inonestudy,amethodwasdevelopedto although this molecule was also reported as ‘pre-B- clone cDNAs that carry N-terminal signal sequences cell growth-stimulating factor’ (PBSF) (Nagasawa (signal sequence Trap) (Tashiro et al., 1993). An etal.,1994),aswellas‘TPAR-1’(TPArepressedgene1) expression cDNA library was constructed from a (Jiang et al., 1994). SDF-1 is now the recognized and bone marrow stromal cell line, and two cDNAs that universally used name. encoded chemokines, stromal cell-derived factor-1(cid:11) (SDF-1a) and SDF-1b were cloned (Tashiro et al., Structure 1993). In another study, SDF-1 was found by virtue of its ability to stimulate early B cell progenitors (Nagasawaetal.,1994).Astromalcellline,PA6,was SDF-1 is a member of the (CXC) chemokines, found to produce a soluble mediator for B cell althoughitdoesnotcontaintheELRmotiftypicalof 1120 Charles R. Mackay the IL-8-like chemokines. The amino acid sequence lymphocytes,forinstancethroughlymphnodes,rather contains four cysteine residues conserved by most of than to inflammatory lesions. SDF-1 is one of four theCXCchemokines.TheN-terminusofSDF-1may chemokines that is able to induce firm adhesion and be particularly important for activity, since peptides arrest of rolling T cells under flow conditions corresponding to the N-terminal 9 residues of SDF-1 (Campbell et al., 1998). SDF-1, as well as 6C-kine were able to induce intracellular calcium and chemo- and MIP-3(cid:12), induced adhesion of most circulating taxis in T lymphocytes, and bind to CXCR4 lymphocytes to ICAM-1 under flow (Campbell et al., (Loetscher et al., 1998). The crystal structure of 1998). SDF-1 is discussed below. CD34 Cell Migration Main activities and Hematopoietic progenitor cells were shown to mig- rate in vitro and in vivo towards a gradient of SDF-1 pathophysiological roles producedbystromalcells.CD34(cid:135)migratingtoSDF- 1 included cells with a more primitive (CD34(cid:135)/ Much of this discussion is also relevant for CXCR4 CD38(cid:255) or CD34(cid:135)/DR(cid:255)) phenotype as well as function and pathological role, since SDF-1 and CD34(cid:135) cells phenotypically committed to the ery- CXCR4 are a monogamous receptor ligand pair. throid, lymphoid, and myeloid lineages (Aiuti et al., The receptor for SDF-1, CXCR4, is widely 1997). expressedonleukocytes,maturedendriticcells,endo- thelial cells, brain cells, and megakaryocytes, hence Brain Cells theactivitiesofSDF-1arenumerousandvaried.This A role for SDF-1, and its receptor CXCR4, in chemokine,morethananyotherchemokineidentified neuronal development is inferred from studies with thus far, has the widest range of biological functions, CXCR4-deficient mice (Zou et al., 1998), as well as especially outside of the immune system. The most immunohistochemical localization of SDF-1 and its significant functional effects of SDF-1 are outlined receptor to various cell types in the brain. One study below. showedthatSDF-1coulddirectlyinduceapoptosisin a neuronal cell line (Hesselgesser et al., 1998). B Cell Development and Migration SDF-1 was originally identified through its ability to HIV Infection support the proliferation of a stromal cell-dependent CXCR4 serves as a coreceptor for T-tropic strains of pre-B cell clone and augment the growth of bone HIV-1, and SDF-1 is able to block T-tropic HIV-1 marrow B cell progenitors in the presence of IL-7 entry into CXCR4-bearing cells (Bleul et al., 1996b; (Nagasawaetal.,1994).Thisresult,togetherwiththe Oberlin et al., 1996). The significance of CXCR4 for analysis of SDF-1- and CXCR4-deficient mice show HIV entry and AIDS pathogenesis is discussed more that SDF-1 is critical for B cell development. fully in the chapter on CXCR4 (Moore et al., 1997). SDF-1isalsoaBcellchemoattractant,andCXCR4 SDF-1 expression may have an important bearing on isexpressedonBcellsatvariousstagesofmaturation AIDS pathogenesis. A polymorphism was identified oreffectorfunction(D’Apuzzoetal.,1997;Bleuletal., in an evolutionarily conserved region of the 30 UTR 1998).Inhumans,SDF-1attractsnaı¨veandmemory, of the human SDF-1 gene. In the homozygous state, butnotgerminalcenterBcells(Bleuletal.,1998).The this polymorphism delayed the onset of AIDS, and receptor for SDF-1, CXCR4, was found on respon- this protective effect was twice as strong as the sive (as well as some unresponsive) B cell subsets. dominant genetic restriction of AIDS conferred by mRNA for SDF-1 was detected by in situ hybridiza- CCR5 and CCR2 chemokine receptor variants tioninalayerofcellssurroundingthegerminalcenter, (Winkler et al., 1998). suggesting that SDF-1 is important for the position- ing of B cells within secondary lymphoid tissue. GENE AND GENE REGULATION T Cell Chemotaxis Accession numbers SDF-1 is one of the most efficacious T cell chemo- attractants (Bleul et al., 1996b).This probably relates to the wide expression of CXCR4 on many T cells, Human: 1352728, 730278, 3914966, U16752 particularly naı¨ve T cells (Bleul et al., 1997). SDF-1 Mouse: D21072, D43804, D43805, 539916, L12029, may be more important for basal trafficking of L12030, S74318 SDF-1 1121 Chromosome location Sequence ThehumanSDF-1genemapstochromosome10qby See Figure 1. fluorescence in situ hybridization (Shirozu et al., 1995), which is unusual given that genes encoding Discussion of crystal structure other members of the chemokine family are localized on chromosome 4q or 17q. The three-dimensional structure of SDF-1 was deter- mined by NMR spectroscopy (Crump et al., 1997). SDF-1wasfoundtobeamonomerwithadisordered Regulatory sites and corresponding N-terminal region (residues 1–8), and differed from transcription factors other chemokines in the packing of the hydrophobic core and surface charge distribution. This study showed that residues 12–17 of the loop region A GC-rich sequence in the 50-flanking region of the (RFFESH motif) formed a receptor-binding site, SDF-1 gene, similar to other ‘housekeeping’ genes, which was proposed as an important initial docking may relate to the ubiquitous expression of SDF-1. site for SDF-1 with its receptor (Crump et al., 1997). Other regulatory elements are unknown. A crystal structure of a variant SDF-1(cid:11) ([N33A]- SDF-1(cid:11), prepared by chemical synthesis) has been refined to 2.2A˚ resolution (Dealwis et al., 1998). Cells and tissues that express SDF-1(cid:11) shows the typical chemokine (cid:12)-(cid:12)-(cid:12)-(cid:11) the gene topology, although the packing of the (cid:11) helix against the (cid:12) sheet was found to be strikingly different. Structural analysis also revealed a positively An unusual feature of SDF-1 is that it is expressed charged surface that may be important for binding constitutively by many cell types (Shirozu et al., to the negatively charged extracellular loops of 1995), in contrast to many other chemokines which CXCR4 (Dealwis et al., 1998). are induced by inflammatory cytokines. This prob- ablyrelatestothebroadfunctionalactivityofSDF-1, Important homologies in particular its role in nonimmune cell migration, positioning or survival. The exact function of SDF-1 is still poorly understood, and so the significance of SDF-1 is a member of the chemokine family, its wide expression is currently speculative. By simple althoughitisdistinctiveinthatthemouseandhuman northernblot,mRNAforSDF-1isexpressedinbrain, proteins are highly conserved (99%). Although SDF- heart, liver, bone marrow, kidney, spleen, stomach, 1 is a CXC chemokine, it is relatively distinct from and gastrointestinal tract. Stromal cells express high otherbranchesofthisfamily,suchastheIL-8family, levelsofSDF-1,particularlythosethatsupportBcell and the IP-10, MIG, I-TAC family. lymphopoiesis. CELLULAR SOURCES AND TISSUE EXPRESSION PROTEIN Cellular sources that produce Accession numbers SDF-1 is produced by a broad range of cell types, in SwissProt: P40224, P48061 particular by bone marrow stromal cells, as well as Figure 1 Amino acid sequence of SDF-1(cid:11) and SDF-1(cid:12); the signal peptide is underlined. SDF-1a MNAKVVVVLV LVLTALCLSD G KPVSLSYRC PCRFFESHVA RANVKHLKIL NTPNCALQIV ARLKNNNRQV CIDPKLKWIQ EYLEKALNK SDF-1b MNAKVVVVLV LVLTALCLSD G KPVSLSYRC PCRFFESHVA RANVKHLKIL NTPNCALQIV ARLKNNNRQV CIDPKLKWIQ EYLEKALNKR FKM 1122 Charles R. Mackay numerous others (Shirozu et al., 1995) (see above), cell progenitors in mutant embryos were severely althoughtheseanalyseshavebeenmostlyattheRNA reduced in fetal liver and bone marrow, and myeloid level. There have been few reports on SDF-1 protein progenitors were reduced in the bone marrow (but localization. not in the fetal liver). In addition, the mutants had a cardiacventricularseptaldefect,whichmaybeoneof the reasons for lethality (Nagasawa et al., 1996). In another study, SDF-1- (and CXCR4)-deficient mice RECEPTOR UTILIZATION werefoundtohaveseverelyreducedBlymphopoiesis, reduced or absent myelopoiesis in fetal liver or bone SDF-1 binds the chemokine receptor CXCR4 marrow, but normal T lymphopoiesis. Moreover, the (Bleul et al., 1996b; Oberlin et al., 1996). SDF-1 and cerebellum was found to develop abnormally in this CXCR4 are monogamous partners. study (Ma et al., 1998). In another study by the Japanese group, a role for SDF-1 (and its receptor CXCR4) was implicated in the vascularization of IN VITRO ACTIVITIES various tissues, particularly the gastrointestinal tract (Tachibana et al., 1998). In another study using CXCR4-deficient mice, hematopoietic and cardiac In vitro findings defectsidenticaltothoseofSDF-1-deficientmicewere noted, as well as altered fetal cerebellar development, SDF-1 supports the proliferation of a stromal cell- indicating a role for CXCR4 (and its ligand) in dependent pre-B cell clone and augments the growth neuronal cell migration and patterning in the central of bone marrow B cell progenitors in the presence of nervous system (Zou et al., 1998). IL-7 (Nagasawa et al., 1994). SDF-1 is also a B cell chemoattractant, attracting naı¨ve and memory, but notgerminalcenterBcells (Bleulet al.,1998).SDF-1 PATHOPHYSIOLOGICAL ROLES is one of the most efficacious T cell chemoattractants (Bleul et al., 1996a). SDF-1 is able to induce firm IN NORMAL HUMANS AND adhesion and arrest of rolling T cells under flow DISEASE STATES AND conditions (Campbell et al., 1998). CD34(cid:135) hemato- DIAGNOSTIC UTILITY poieticprogenitorcellswereshowntomigrateinvitro andin vivotowardsagradientofSDF-1 producedby stromal cells (Aiuti et al., 1997). SDF-1 blocks the Role in experiments of nature and infection in vitro of T cells with T-tropic strains of disease states HIV-1 (Bleul et al., 1996b; Oberlin et al., 1996). Disease association has not been defined, although high levels of SDF-1 may have a bearing on the Bioassays used spread of T-tropic HIV-1. The most common bioassay used for SDF-1 is a chemotaxis assay (Qin et al., 1996), although SDF-1 IN THERAPY also elicits other responses such as calcium flux. Effects of therapy: Cytokine, IN VIVO BIOLOGICAL antibody to cytokine inhibitors, etc. ACTIVITIES OF LIGANDS IN The therapeutic utility of SDF-1 revolves more ANIMAL MODELS around its antagonism, rather than its stimulation of various cell types. For instance, small molecule Knockout mouse phenotypes antagonists have been developed that block SDF-1 binding to its receptor, CXCR4 (Donzella et al., In the two studies that have been reported, SDF-1 is 1998). The utility of such drugs would be for inhi- essential for embryonic development, since mice bition of HIV infection. Such drugs may find wider lacking SDF-1 died perinatally (Nagasawa et al., applications, although at present such applications 1996;Maetal.,1998).Inthefirststudy,numbersofB are not obvious. 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