MONOCYTE-DERIVED NUCLEIC ACIDS AND RELATED COMPOSITIONS AND METHODS

Abstract: Nucleic acids encoding various monocyte-derived proteins and related compositions, including purified proteins and specific antibodies are described. Methods of using such composition are also provided.

Description: MONOCYTE-DERIVED NUCLEIC ACIDS AND RELATED COMPOSITIONS AND METHODS Field Of The Invention The present invention is directed to compositions related to genes found in monocytes, cells which function in the immune system. These genes function in controlling development, differentiation, and/or physiology of the mammalian immune system. In particular, the invention provides nucleic acids, proteins, antibodies, and methods of using them. Background Of The Invention Monocytes are phagocytic cells that belong to the mononuclear phagocyte system and reside in the circulation. These cells originate in the bone marrow and remain only a short time in the marrow compartment once they differentiate. They then enter the circulation and can remain there for a relatively long period of time, e.g., a few days. Monocytes can enter the tissues and body cavities by a process known as diapedesis, where they differentiate into macrophages and possibly into dendritic cells. In an inflammatory response, the number of monocytes in the circulation may double, and many of the increased number of monocytes diapedese to the site of inflammation. For a review of monocytes and their functions, see, e.g., Gallin, et al. (eds), 1988, Inflammation: Basic Principles and Clinical Correlates, Raven Press, NY; van Furth (ed), 1985, Mononuclear Phagocytes: Characteristics, Physiology and Function, Martinus Nijhoff, Dordrecht, Netherlands. Antigen presentation refers to the cellular events in which a proteinaceous antigen is taken up, processed by antigen presenting cells (APC), and then recognized to initiate an immune response. The most active antigen presenting cells have been characterized as the macrophages, which are direct developmental products from monocytes; dendritic cells; and certain B cells. Macrophages are found in most tissues and are highly active in internalization of a wide variety of protein antigens and microorganisms. They have a highly developed endocytic activity, and secrete many products important in the initiation of an immune response. For this reason, it is believed that many genes expressed by monocytes or induced by monocyte activation are important in antigen uptake, processing, presentation, or regulation of the resulting immune response. Despite the importance of monocytes to immune system function, these cells remain poorly characteπzed, both in terms of the proteins they express and in terms of many of their functions, in particular, the processes and mechanisms related to the mitiation of an immune response, including antigen processing and presentation There is thus a need in the art for agents useful m the diagnosis and treatment of medical conditions caused by, e g , the inappropriate regulation, development, or physiology of antigen presenting cells Summary Of The Invention The present invention fulfills this need by providing compositions and methods for determining the presence, amount, distribution and normalcy of certain gene products and for facilitating the discovery of agents for treating certain disease states The invention is based upon the discovery of novel genes and gene products isolated from activated monocytes The invention provides isolated nucleic acid sequences comprising at least about 12, preferably at least about 18, most preferably at least about 20-35, and most preferably 35-55 or more consecutive nucleotides shown in SEQ ID NO 1, 3, 5, 7, or 9, or which encode an ammo acid sequence shown in SEQ ID NO 2, 4, 6, 8 or 10, including complete protein coding sequences, and complements thereof The invention encompasses sequence-conserv ativ e vaπants and function-conservative variants of these sequences The nucleic acids may be DNA, RNA, DNNRNA duplexes, protein-nucleic acid (PNA), or derivatives thereof The invention also encompasses recombmant DNA v ectors (including DNA expression v ectors) composing these sequences, cells comprising such vectors, including bacterial, fungal, plant, insect, and mammalian cells, and methods for producing expression products composing RNA and polypeptides encoded by the sequences Polypeptide sequences of the inv ention comprise at least eight, preferably at least about 10, and more preferably at least about 12 or more consecutive ammo acid residues deriv ed from SEQ ID NO 2 4 6, 8 or 10 Function-conservative vaπants and homologs are included in the scope of the inv ention The invention further prov ides binding compositions, m particular antibodies, most particularly monoclonal antibodies, which specifically bind to polypeptides having an ammo acid sequence show n in SEQ ID NO 2, 4, 6, 8 or 10 or function conserved v aπants or homologs thereof Methods are also provided for producing antibodies having the desired binding specificitv in a host animal Detailed Description Of The Invention All patent applications, patents, and literature references cited in this specification are hereby incorporated herein by reference in their entirety In practicing the present invention, many conventional techniques m molecular biology, microbiology, and recombmant DNA. are used Such techniques are well known and are explained fully in, for example, Sambrook et al , 1989, Molecular Cloning A Laboratory Manual, Second Edition, Cold Spπng Harbor Laboratory Press, Cold Spπng Harbor, New York, DNA Cloning A Practical Approach, Volumes I and II, 1985 (D N Glover ed ), Ohgonucleotide Synthesis, 1984, (M L Gait ed ), Nucleic Acid Hybridization, 1985, (Hames and Higgms), Transcription and Translation, 1984 (Hames and Higgins eds ), Animal Cell Culture, 1986 (R I Freshney ed ), fmmobilized Cells and Enzymes, 1986 (IRL Press), Perbal, 1984, A Practical Guide to Molecular Cloning, the seπes, Methods in Enz\molog\ (Academic Press, Inc ), Gene Transfer Vectors for Mammalian Cells, 1987 (J H Miller and M P Calos eds , Cold Spπng Harbor Laboratory), and Methods in Enz\ molog\ Vol 154 and Vol 155 (Wu and Grossman, and Wu, eds , respectively) Definitions 1 A "monocyte-deπv ed" nucleic acid or polypeptide refers to the source from which the sequence was originally isolated 2 "Nucleic acid" or "polynucleotide" refers to puπne- and pyπmidine- contaimng polymers of anv length, either polyπbonucleotides or polydeoxynbonucleotides or mixed polvπbo-polydeoxyπbo nucleotides This includes single- and double-stranded molecules, l e , DNA-DNA, DNA-RNA and RNA-RNA hvbπds, as well as "protein nucleic acids" (PNA) formed by conjugating bases to an amino acid backbone This also includes nucleic acids containing modified bases 3 A "coding sequence" or a "protem-coding sequence" is a polynucleotide sequence capable of being transcribed into mRNA and/or capable of being translated into a polypeptide The boundaries of the coding sequence are typically determined by a translation start codon at the 5'-termιnus and a translation stop codon at the 3'-termιnus 4 A "complement" of a nucleic acid sequence refers to the "antisense" sequence that participates in W atson-Cπck base-paiπng with the oπginal sequence 5 An "isolated" nucleic acid or polypeptide refers to component that is removed from its oπginal environment (for example, its natural environment if it is naturally occurring) An isolated nucleic acid or polypeptide preferably contains less than about 50%, more preferably less than about 75%, and most preferably less than about 90%, of the cellular components with which it was oπgmally associated 6 A nucleic acid or polypeptide sequence that is "deπved from" a designated sequence refers to a sequence that coπesponds to a region of the designated sequence For nucleic acid sequences, this encompasses sequences that are homologous or complementary to the sequence, as well as "sequence-conservative vaπants" and "function-conservative vaπants " For polypeptide sequences, this encompasses "function- conservative vaπants " Sequence-conservative vaπants are those in which a change of one or more nucleotides in a given codon position results m no alteration in the ammo acid encoded at that position Function-conservative vaπants are those in which a given ammo acid residue m a polypeptide has been changed without substantially alteπng the overall conformation and function of the native polypeptide, including, but not limited to, replacement of an amino acid with one having similar physico-chemical properties (such as, for example, acidic, basic, hydrophobic, and the like) "Function-conservative" v aπants also include any polypeptides that have the ability to elicit antibodies specific to a designated polypeptide 7 A "probe" refers to a nucleic acid or ohgonucleotide that forms a hybπd structure with a sequence in a target region due to complementaπty of at least one sequence in the probe with a sequence in the target 8 Nucleic acids are "hvbπdizable" to each other when at least one strand of nucleic acid can anneal to another nucleic acid strand under defined stπngency conditions Stπngency of hybridization is determined, e g , by a) the temperature at w hich hybπdization and/or w ashing is performed, and b) the ionic strength and polarity (e g , formamide) of the hybridization and w ashing solutions, as well as other parameters Hybridization requires that the tw o nucleic acids contain substantially complementary sequences, depending on the stπngencv of hybπdization, however, mismatches may be tolerated The appropπate stπngencv for hybridizing nucleic acids depends on the length of the nucleic acids and the degree of complementaπty, vaπables well known in the art 9 An "lmmunogenic component" is a moiety that is capable of eliciting a humoral and/or cellular immune response in a host animal 10 An "antigemc component" is a moiety that binds to its specific antibody w ith sufficiently high affinity to form a detectable antigen-antibody complex 11 A "sample" refers to a biological sample, such as, for example, tissue or fluid isolated from an individual or from an in vitro cell culture constituents, as well as samples obtained from laboratory procedures The invention provides nucleic acid sequences encoding mammalian proteins expressed on monocytes While specific human monocyte-deπved genes and gene products are descπbed herein, the invention encompasses structurally (e g , sequence) related embodiments from other sources or mammalian species, including polymorphic or individual vaπants These will include, e g , proteins which exhibit relatively few changes in sequence, e g , less than about 5%, and number, e g , less than 20 residue substitutions, typically less than 15, preferably less than 10, and more preferably less than 5 substitutions These will also include versions which are truncated from full length and fusion proteins containing substantial segments of these sequences A gene/gene product, isolated from human monocyte cell library and designated FDF03, has been previously descπbed in published International application WO 98 24906, the disclosure of which is incorporated herein in its entirety by reference The FDF03 gene encodes a type I transmembrane protein compπsmg an extracellular portion characteπzed by Ig-hke domains, indicating that this gene encodes a receptor member of the Ig superfamily SEQ NO 1 shows the nucleic acid sequence encoding human FDF03 protein The amino acid sequence of the FDF03 protein is shown in SEQ ID NO 2 The putative coding region runs from about nucleotide 154 to nucleotide 1062 An N-terminal hydrophobic sequence coπesponding to a putaive signal sequence runs from about amino acid residue -19 (Met) to amino acid residue -1 (Leu) An internal hydrophobic sequence corresponding to a putativ e transmembrane segment runs from about residue 177 (Ala) to residue 199 (Leu) The extracellular region is about 170 amino acids, with a potential Ig- like domain structure The intracellular region is about 80 residues Sequence analysis indicates similaπty to GenBank clones H26010 and R50327 from humans Four human FDF03 homologs hav e now been discovered FDF03-ΔTM The second human clone, designated FDF03-deltaTM (FDF03-ΔTM), appears to be a soluble form of human FDF03 generated by alternative splicing The nucleic acid sequence encoding FDF03-ΔTM is shown in SEQ NO 3 The amino acid sequence of the FDF03-ΔTM protein is shown in SEQ ID NO 4 cDNA of the FDF03-ΔTM molecule was amplified along with that of FDF03 duπng the analysis of human FDF03 expression by RT-PCR Using pπmers designed in the 5'-UTR and 3'-UTR of FDF03 gene (FDF03-U25 5'-ACAGCCCTCTTC- GGAGCCTCA (SEQ ID NO 11) and FDF03-L1166 5'-AAGCTGGCCCTGAACT CCTGG (SEQ ID NO 12)), an approximately 200 base pair shorter band as amplified by RT-PCR from PMA/ionomycin activated PBL cDNA, then gel puπfied, cloned and sequenced Different clones contained an identical insert of 943 base pairs with an open reading frame encoding a type I protein of 230 ammo acids The deduced ammo acid sequence of FDF03-ΔTM matched perfectly with that of FDF03, but contained a gap of 73 amino acids that deleted the extracellular threomne-πch region and the transmembrane domain of FDF03 This resulted m a protein with a potential hydrophobic signal peptide followed by the extracellular Ig like-domam linked to the mtracytoplasmic domain of FDF03 cDNA alignments with FDF03 sequence identified a deletion of 219 nucleotides in the FDF03-ΔTM sequence (FDF03 nucleotide 608 to 827) that did not introduce premature stop codons, suggesting that this molecule is the product of an alternative splicing This molecule is believed to be a secreted soluble form of FDF03 and believed to bind to the same hgand(s) as FDF03 The protein alignment of the FDF03 (SEQ ID NO 2) and FDF03-ΔTM (SEQ ID NO 4) is shown below MGRPLLLPLLPLLLPPAFLQPSGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03 MGRPLLLPLLPLLLPPAFLQPSGSTGSGPSY YGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03- -ΔTM ATAPDVRISWRRGHFHGQSFYSTRPPSIHKDYVNRLF NWTEGQKSGFLRISNLQKQDQ FDF03 LATAPDVRISWRRGHFHGQSFYSTRPPSIHKDYVNRLFLN TEGQKSGFLRISNLQKQDQ FDF03- -ΔTM SVYFCRVE DTRSSGRQQWQSIEGT SITQAVTTTTQRPSSMTTTWRLSSTTTTTGLRV FDF03 SVYFCRVE DTRSSGRQQWQSIEGTKLSITQ FDF03 ΔTM TQGKRRSDSWHISLETAVGVAVAVTVLGIMILGLICLLRWRRRKGQQRTKATTPAREPFQ FDF03 -GQQRTKATTPAREPFQ FDF03 ΔTM NTEEPYENIRNEGQNTDPKLNPKDDGIVYASLA SSSTSPRAPPSHRPLKSPQNETLYSV FDF03 NTEEPYENIRNEGQNTDPK NPKDDGIVYAS A SSSTSPRAPPSHRPLKSPQNETLYSV FDF03- -ΔTM LKA FDF03 LKA FDF03- -ΔTM ( deletion) FDF03-S1 The third clone, designated FDF03-Shortl (FDF03-S1), is an Ig- ke molecule homologous to FDF03 but with a short mtracytoplasmic domain and a charged residue in transmembrane domain. Comparative DNA and protein analysis suggests the presence of different genes for FDF03 and FDF03S 1, rather than alternatively spliced products. The nucleic acid sequence encoding FDF03-S1 is shown in SEQ NO: 5. The amino acid sequence of the FDF03-S1 protein is shown in SEQ ID NO: 6. FDF03-S1 is a type I transmembrane protein belonging to the Ig superfamily. FDF03-S1 contains a hydrophobic leader sequence followed by an extracellular region (-170 residues) with a V-type Ig domain structure homologous to that of FDF03 (88% homology at the amino acid level). Unlike FDF03, FDF03-S1 possesses a transmembrane domain with a charged amino acid (K), and a small intracellular tail (15 residues) without ITIM or internalization motif. FDF03-S1 is believed to represent an activation isoform of FDF03 and may associate with ITIM-bearing molecules such as DAP 12. The amino acid sequence is shown below, wherein the signal peptide and transmembrane domain are underlined. The charged amino acid, lysine (K), residue (arrow) in the transmembrane domain may permit association with another chain, for example DAP12. MGRPLLLPLLLLLOPPAFLOPGGSTGSOPSYLYGVTOPKHLSASMGGSVEIPFSFYYPWEL AIVPNVRISWRRGHFHGQSFYSTRPPSIHKDYVNRLFLN TEGQESGFLRISNLRKEDQSV YFCRVELDTRRSGRQQLQSIKGTILTITQAVTTTTTWRPΞSTTTIAGLRVTESKGHSESWH LSLDTAIRVALAVAVLKTVILGLLCLLLLWWRRRKGSRAPSSDF ( SEQ ID NO : 6) T The protein alignment of FDF03 (SEQ ID NO: 2) and FDF03-S1 (SEQ ID NO: 6) is shown below. MGRPLLLPLLPLLLPPAFLQPSGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03 MGRPLLLPLLLLLQPPAFLQPGGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03-S1 LATAPDVRISWRRGHFHGQSFYSTRPPSIHKDYVNRLFLNWTEGQKSGFLRISNLQKQDQ FDF03 LAIVP RISWRRGHFHGQΞFYSTRPPSIH DYVΗRLFLNWTEGQEΞGFLRISNLRKEDQ FDF03-S1 SVYFCRVELDTRSSGRQQWQSIEGTKLSITQATTTTQRPSSMTTTWRLSSTTTTTGLRV FDF03 SVYFCRVELDTRRSGRQQLQSIKGTKLTITQAVTTTT TWRPSSTTTIAGLRV FDF03-S1 + + + + ++++++++ + ++ TQGKRRSDSWHISLETAVGVAVAVTVLGIMILGLICLL.. RWRRRKGQQRTKATTPAREP FDF03 TESKGHΞESWHLSLDTAIRVALAVAVLKTVILGLLCLLLLWWRRRKGSRAPSSDF FDF03-S1 ++ ++ + + + ++ + + +++ + +++ ++++++++ FQNTEEPYENIRNEGQNTDPKLNPKDDGIVYAΞLALSSSTSPRAPPSHRPLKSPQNETLY FDF03 SVLKA FDF03 + : residue different or gap Distribution studies (RT-PCR) shows strong expression in B cells (pool resting + activated), T cells and PBL Lower expression was observed m monocytes, dendπtic cells and granulocytes FDF03-M14 The fourth clone, desginated FDF03-M14, is a potential soluble form of human FDF03 generated by alternative splicing The nucleic acid sequence encoding FDF03- Ml 4 is shown in SEQ ID NO 7 The amino acid sequence of the FDF03-M- 14 protein is shown m SEQ ID NO 8 cDNA of this molecule was amplified along with that of FDF03 duπng the analysis of human FDF03 expression by RT-PCR Using pπmers designed in the 5'-UTR and 3'-UTR of FDF03 gene (FDF03-U25 5'-ACAGCCC- TCTTCGGAGCCTCA (SEQ ID NO 1 1) and FDF03-L 1 166 5'-AGCTGGCCCTGA- ACTCCTGG (SEQ ID NO 12)), an approximately 200 base pair shorter band was amplified by RT-PCR from activated PBL cDNA, then gel puπfied, cloned and sequenced One clone (Ml 4) contained an insert of 908 base pairs with an ORF encoding a type I protein of 175 amino acids cDNA alignments with FDF03 sequence identified a deletion of 253 nucleotides in FDF03-M 14 sequence (FDF03 nucleotide 608 to 861) that deleted the sequences encoding the extracellular threomne-πch region, the transmembrane domain and the start of the mtracytoplasmic domain of FDF03, and that introduced a prematured stop codon at position 655 of FDF03-M14 The deduced amino acid sequence of FDF03-M 14 resulted in a protein with a potential hydrophobic signal peptide followed by an extracellular Ig like-domain that matched perfectly with that of FDF03, but that was linked to a COOH-terminal 24 amino acid sequence different from FDF03 This molecule may be the product of an alternative splicing of FDF03 mRNA Like FDF03-ΔTM, this molecule mav represent a secreted soluble form of FDF03 and may bind to the same hgand(s) as FDF03 The amino acid sequence is shown below, wherein the signal sequence is underlined MGRPLLLPLLPLLLPPAFLOPSGSTGSGPSYLYGVTOPKHLSASMGGSVEI PFSFYYPWEL ATAPDVRISWRRGHFHGQS FYΞTP PPS I HKΩYWRLFLNWTEGQKSGFLRISNLQKQDQSV YFCRVELDTRSSGRQQWQS I EGTKLS I TQGNPΞKTQRSHMRISGMRDKIQIPS ( SEQ ID NO 8 ) The protein alignment of FDF03 (SEQ ID NO 2) and FDF03-M14 (SEQ ID NO 8) is shown below MGRPJ-.LLPLLPLLLPPAFLQPSGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03 1 MGRPLLLPLLPLLLPPAFLQPSGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03 - 14 6. ATAPDVRISWRRGHFHGQS^YSTRPPSIHKDYVNRLFLN TEGQKSGFLRISNLQKQDQ FDF03 6^ ATAPDVRISWRRGHFHGQSFYSTRPPSIHKDYV RLF N TEGQKSGFLRISNLQKQDQ FDF03-M14 121 SVYFCRVELDTRSSGR QQ WQSIEGTKISITQAVTTTTQRPSSMTTTWRLSSTTTTTGLRV FDF03 121 SVYFCRVELDTRSSGRQQ QSIEGTKLSITQGNPSKTQRSHMRISGMRDKIQIPS FDF03 -M14 **** * ******** ******* 181 T Q GKRRSDSWHISLETAVGVAVAVTVLGIMILGLICLLR RRRKGQQRTKATTPAREPFQ FDF03 241 NTEEPYENIRNEGQNTDPK1NPKDDGIVYASLALSSSTSPRAPPSHRPLKSPQNETLYSV KA FDF03 * residue dif ferent FDF03-S2 The fifth clone, designated FDF03-S2 is an Ig- ke molecule homologous to FDF03 but with a short mtracytoplasmic domain and a charged residue in transmembrane domain This molecule is highly homologous to FDF03-S1 and is a potential DAP 12- associated protein The nucleic acid sequence encoding FDF03-S2 is shown is SEQ ID NO 9 The amino acid sequence of the FDF03-S2 protein is shown in SEQ ID NO 10 cDNA of this molecule was amplified using pπmers specific for FDF03-S2 Specificity is obtained with forward pπmer designed in 5'UTR of FDF03-S2 FDF03-S2- forward 5'-CAAGG- GATAAAAAGGCAC (SEQ ID NO 13) (does not amplify FDF03, FDF03ΔTM or FDF03-S 1 ) FDF03-S2-reverse 5'-AACTCTCCTCCAGTCGGT (SEQ ID NO 14) (can amplify FDF03-S 1 , but not FDF03 or FDF03deltaTM) FDF03-S2 is a type I transmembrane protein belonging to the Ig superfamily FDF03-S2 contains a hydrophobic leader sequence followed by an extracellular region (-170 residues) with one V-type Ig domain structure homologous to that of FDF03 (-85% homology at the ammo acid level) nhke FDF03, FDF03-S2 possesses a transmembrane domain with a charged amino acid (K), and a small intracellular tail (15 residues) without ITIM or lnternahzation motif FDF03-S2 is highly homologous to FDF03-S1 (3 ammo acid difference in the extracellular domain and one amino acid missing in the transmembrane domain) Like FDF03-S 1 , FDF03-S2 may represent an activation isoform of FDF03 and may associate with ITAM-beaπng molecules such as DAP 12 There are two putative start codons in frame (position 1 17 and 309) The first one is not contained within a typical Kozak sequence The sequence shown below is deduced from the second start codon (nucleotide 309), as starting at the first start codon in frame (position 1 17) does not encode for a hydrophobic sequence followed by another Ig-hke domain In the sequence sho n below, the signal peptide and transmembrane domain are underlined The charged amino acid, lysine (K) residue (arrow) in transmembrane domain may permit association with another chain, for example DAP 12 MGRPLLLPLLLLLOPPAFLOPGGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE LATAPDVRISWRRGHFHGQSFYSTRPPSIHrDYVNRLFLNWTEGQESGFLRISNLRKEDQ SVYFCRVELDTRRSGRQQLQSIKGTKLTITQAVTTTTTWRPSSTTTIAGLRVTESKGHSE SWHLSLDTAIRVALAVAVLKTVILGLLCLLL WRRRKGSRAPSSDF T The protein alignments of FDF03, FDF03-S1 and FDF03-S2 is shown below. 1 MGRPLLLPLLPLLLPPAFLQPSGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03 1 MGRPLLLPLLLLLQPPAFLQPGGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03-S1 1 MGRPLLLPLLLLLQPPAFLQPGGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWE FDF03-S2 61 LATAPDVRISWRRGHFHGQSFYSTRPPSIHKDYVNRLFLNWTEGQKSGFLRISNLQKQDQ FDF03 61 LAIVPNVRISWRRGHFHGQSFYSTRPPSIHKDYWRLFLNWTEGQESGFLRISNLRKEDQ FDF03-S1 61 LATAPDVRISWRRGHFHGQSFYSTRPPSIHKDYVNRLFLNWTEGQESGFLRISNLRKEDQ FDF03-S2 121 SVYFCRVELDTRSSGRQQWQSIEGTKLSITQAVTTTTQRPSSMTTTWRLSSTTTTTGLRV FDF03 121 SVYFCRVELDTRRSGRQQ QSIKGTKLTITQAVTTTT TWRPSSTTTIAGLRV FDF03-S1 121 SVYFCRVELDTRRSGRQQLQSIKGTKLTITQAVTTTT TWRPSSTTTIAGLRV FDF03-S2 + + + + ++++++++ + ++ 181 TQGKRRSDSWHISLETAVGVAVAVTVLGIMILGLICLL.. RWRRRKGQQRTKATTPAREP FDF03 1"3 TESKGHSESWHLSLDTAIRVALAVAVLKTVILGLLCLLLLWWRRRKGSRAPSSDF FDF03-S1 173 TESKGHSESWHLSLDTAIRVALAVAVLKTVILGLLCLLL.WWRRRKGSRAPSSDF FDF03-S2 ++ ++ + + + ++ + + +++ + +*+ ++++++++ 239 FQNTEEPYENIRNEGQNTDPKLNPKDDGIVYASLALSSSTSPRAPPSHRPLKSPQNETLY FDF03 299 SVLKA FDF03 + : residue different or gap between FDF03 -S2/FDF03-S1 and FDF03 * • residue different or gap between FDF03 -S2/FDF03 and FDF03-S1 Distribution studies (RT-PCR) shows expression in activated dendritic cells (CD34-derived), PBMC, monocytes and tonsil B cells Alignment with human IgV domains and TCR V domain is given below. This alignment shows the conserved VDJ structure of FDF03. Ig V region QVQ . LQESGPG . LVKPSET SLTCTVSGGSVSSGSYYWSW . IRQAPGKGLEWIG TCR human QVQ . LQESGPG . LVKPSΞTLSLTCTVSGYSISSG . YYWG . IRQPPGKGLEWIG FDF03 QPSGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWELATAPDVRISWRR FDF03-S1 QPGGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWELAIVPNVRISWRR FDF03-S2 QPGGSTGSGPSYLYGVTQPKHLSASMGGSVEIPFSFYYPWELATAPDVRISWRR Ig V region YIYYSGSTNY. NRSHKSPVNIS VDTAKNQFSLKLSSVSTADTAVYYCARIT TCP human SIYHSGSTYY. NPSLKSRVTIS . VDTSKNQFSLKLSSVTAADTAVYYCARVR FDF03 GHFH.GQΞFYSTRPPSIHKDYVNRLFLNWTEGQKSGF.LRISNLQKQDQSVYFC RVE FDF03 -SI GHFH . GQSFYSTRPPSIHKDYVNRLFLNWTEGQESGF . LRISNLRKEDQSVYFC RVE FDF03 -S2 GHFH . GQSFYSTRPPSIHKDYVNRLFLNWTEGQESGF . LRISNLRKEDQSVYFC . RVE Ig V region TTVPSSWYYYYMDVWDKGTTVTVSS TCR human RRYSSSAS ... KI I FGSGTRLSIR FDF03 LDTRSSGRQQWQS IEGTKLSITQ FDF03-S1 LDTRRSGRQQLQS . IKGTKLTITQ FDF03-S2 LDTRRSGRQQLQS IKGTKLTITQ Studies of human genomic DNA clones show that chromosome 7 contains both FDF03-S1 and FDF03 specific sequences, confirming that the two molecules are encoded by two different genes. These studies also suggest that FDF03-S1 and -S2 genes are two different alleles of the same gene. In addition, PCR from intronic sequence surrounding the areas of difference between SI and S2 on genomic DNA from different donors shows the existence of homozygotes and S1/S2 heterozygotes at this locus. It is thus likely that these two cDNAs are from different alleles. The genomic organization of the FDF03 gene confirms that FDF03-ΔTM is produced by alternative splicing (deletion of exon 3 coding for the hinge region and TM domain). This is also the case for FDF03-M14 (deletion of exons 3 and 4). The two forms of FDF03-S1/2 may be advantageously used as population markers. The two forms of this protein will either not bind the same ligand (e.g., as in the case of the NK receptor family) or will bind at different affinities, thus potentially giving individuals a different response to receptor/ligand interaction. The localization of the genes encoding FDF03 (including the ΔTM and M14 forms) and FDF03-S1 on human chromosome 7q22 is interesting because this region is frequently deleted in myelodystrophic syndromes such as Acute Myeloid Leukemia (AML). The implication of the possible deletion of a myeloid inhibitory receptor in a proliferative disease leads to a possibile use in gene therapy. Nucleic Acids, Vectors, and Host Cells The invention provides nucleic acid sequences, in particular the nucleic acid sequences shown in SEQ ID NO: 1 , 5, 7 or 9 or nucleic acid sequences which encode an amino acid sequences shown in SEQ ID NO: 2, 4, 6, 8 or 10. The invention encompasses isolated nucleic acid fragments comprising all or part of the individual nucleic acid sequences disclosed herein. The nucleic acid sequences of the invention comprise at least about 12, preferably at least about 18, more preferably at least about 20-35 and most preferably at least about 35-55 or more consecutive nucleotides, including complete protein-coding sequences, or complements thereof. The invention encompasses sequence-conservative variants and function-conservative variants of these sequences. Nucleic acids comprising any of the sequences disclosed herein or subsequences thereof can be prepared by standard methods using the nucleic acid sequence information provided in SEQ ID NO: 1 , 3, 5, 7 and 9. For example, nucleic acids can be chemically synthesized using, e.g., the phosphoramidite solid support method of Matteucci et al, 1981, J Am Chem Soc 103 3185, the method of Yoo et al , 1989, J Bwl Chem 764 17078, or other well known methods This can be done by sequentially linking a seπes of ohgonucleotide cassettes compπsmg pairs of synthetic ohgonucleotides The nucleic acids may be isolated directly from cells Alternatively, the polymerase chain reaction (PCR) method can be used to produce the nucleic acids of the invention, using either chemically synthesized strands or genomic mateπal as templates Pπmers used for PCR can be synthesized using the sequence information provided herein and can further be designed to introduce appropπate new restπction sites, if desirable, to facilitate incorporation into a given vector for recombinant expression Of course, due to the degeneracy of the genetic code, many different nucleotide sequences can encode polypeptides having the ammo acid sequences defined by SEQ ID NO 2, 4, 6, 8 or 10 subsequences thereof The codons can be selected for optimal expression m prokaryotic or eukaryotic systems Such degenerate vaπants are also encompassed by this invention The encoded polypeptides may be expressed by using many known vectors such as pUC plasmids, pET plasmids (Novagen, Inc , Madison, WI), or pRSET or pREP (Inv itrogen, San Diego, CA), and many appropπate host cells such as Esche ichia coh, Saccharomyces cerevtsiae, and insect and mammalian cell lines using methods known to those skilled in the art The particular choice of vector/host is not cπtical to the practice of the invention The nucleic acids of the present invention find use, e g , as templates for the recombinant production of peptides or polypeptides, as probes and pπmers for the detection of the human genes descπbed herein, for chromosome mapping, and as probes or to design PCR pπmers to identify homologous genes in other mammalian species Homology may be determined expeπmentally Alternatively, homology analysis may be performed computationally In practicing the present invention, a gene that shares at least about 70% DNA sequence homology at the nucleotide level with the genome of another mammalian species is considered to be present in that species The determination that a gene is present in another mammal may be achieved using any technique known m the art Appropπate techniques include without limitation hybπdization to genomic DNA, colonv, hybπdization to a genomic or cDNA library, polymerase chain reaction (PCR) using degenerate primers or gene-specific pπmers and genomic DNA as a template, genetic complementation, antibody cross-reactivity, or biochemical complementation in vitro In applying these techniques, conditions are established that discπminate different levels of homology between probe and template For example, for hybπdization of a probe to immobilized DNA (whether in a Southern blot, dot blot, or colony hybridization format), varying the SSC concentration in the buffer allows the detection of hybrids having different levels of homology (IX SSC is 0.15 M NaCl - 0.015 M Na citrate). In a wash buffer containing 6M urea and 0.4% sodium dodecyl sulfate, the presence of 2X SSC, 0.5X SSC, 0.1X SSC, and 0.05X SSC allows the formation of hybrids having threshold homologies of at least 55% + 5%, 65% + 5%, 75% + 5%, and >85%, respectively. Preferably, once a gene has been identified m another organism by hybridization or PCR, the DNA sequence of the gene is determined directly. It will be understood that some methods that detect homologous sequences may result in the identification or isolation of only a portion of the entire protein-coding sequence of a particular gene. The entire protein-coding sequence can be isolated and identified, for example, by using an isolated nucleic acid encoding the known portion of the sequence, or fragments thereof, to pπme a sequencing reaction with cDNA as template; this is followed by sequencing the amplified product. The isolated nucleic acid encoding the disclosed sequence, or fragments thereof, can also be hybridized to appropπate cDNA libraπes to identify clones containing additional complete segments of the protein-coding sequence of which the shorter sequence forms a part. Then, the entire protein-coding sequence, or fragments thereof, or nucleic acids encoding all or part of the sequence, or sequence-conservative or function-conservative variants thereof, may be employed in practicing the present invention In a similar manner, additional sequences derived from the 5' and 3' flanking regions of sequence encoding the protein, including regulatory sequences, may be isolated, and the nucleotide sequence determined. Polypeptides Both the naturally occurring and recombinant forms of the polypeptides described herein, including both glycosylated and non-glycosylated forms are encompassed by the invention. The polypeptides of the present invention, including function-conservative variants, may be isolated from human monocytes, or from heterologous organisms or cells (e.g., bacteπa, fungi, insect, plant, and mammalian cells) into which a protein- coding sequence has been introduced and expressed. The proteins described herein, or portions thereof, also may be expressed as fusions with other proteins. The polypeptides may be chemically synthesized by commercially available automated procedures, including, without limitation, exclusive solid phase synthesis, partial solid phase methods, fragment condensation or classical solution synthesis The polypeptides can also, advantageously, be made by in vitro translation Methods for polypeptide puπfication are well-known in the art, including, without limitation, preparative disc-gel electrophoresis, isoelectπc focusing, sucrose density gradient centπfugation, HPLC, reversed-phase HPLC, gel filtration, ion exchange and partition chromatography, and countercurrent distπbution For some purposes, it is preferable to produce the polypeptide in a recombinant system m which the protein contains an additional sequence tag that facilitates puπfication, such as, but not limited to, a polyhistidine sequence The polypeptide can then be puπfied from a crude lysate of the host cell by chromatography on an appropπate solid-phase matπx Alternatively, antibodies produced against a protein or against peptides deπved therefrom can be used as puπfication reagents Other puπfication methods are possible The present invention also encompasses deπvatives and homologues of the polypeptides specifically disclosed herein For some purposes, nucleic acid sequences encoding the peptides may be altered by substitutions, additions, or deletions that provide for functionally equivalent molecules, 1 e , function-conservative vaπants For example, one or more amino acid residues within the sequence can be substituted by another amino acid of similar properties, such as, for example, positively charged amino acids (arginine, lysine, and histidine), negatively charged amino acids (aspartate and glutamate), polar neutral amino acids, and non-polar amino acids The isolated polypeptides may be modified by, for example, phosphorylation, sulfation, acylation, or other protein modifications They may also be modified with a label capable of providing a detectable signal, either directly or indirectly, including, but not limited to, radioisotopes and fluorescent compounds The polypeptides of the inv ention find use, e g , for binding studies, for construction and expression of modified molecules, for structure/function studies and for the preparation of polyclonal and monoclonal antibodies Polypeptides useful as lmmunogenic components for prepaπng antibodies or as targets for binding agent studies are at least five or more residues in length Preferably, the polypeptides comprise at least about 12, more preferably at least about 20, and most preferably at least about 30 or more residues Methods for obtaining these polypeptides are well known and are explained in Immunochemical Methods in Cell and Molecular Biology, 1987 (Mayer and Waler, eds, Academic Press, London), Scopes. 1987, Protein Purification Principles and Practice, Second Edition (Spπnger-Verlag, N Y ) and Handbook of Experimental Immunology , 1986, Volumes I-IV (Weir and Blackwell, eds ) Having isolated one member of a binding partner of a specific interaction, methods exist for isolating the counter-partner. See, e.g., Gearing et al, 1989, EMBO J 8:3667-3676. Many methods of screening for binding activity are known by those skilled in the art and may be used to practice the invention. For example, an expression library can be screened for specific binding to the protein, e.g., by cell sorting, or other screening to detect subpopulations which express such a binding component. See, e.g., Ho et al., 1993, Proc. Natl. Acad. Sci USA 90:1 1267-11271. Alternatively, a panning method may be used. See, e.g., Seed and Aruffo, 1987, Proc. Natl. Acad. Sci. USA 84:3365-3369. A two-hybrid selection system may also be applied making appropriate constructs with the available protein sequences. See, e.g., Fields and Song, 1989, Nature 340:245-246 Several methods of automated assays have been developed in recent years so as to permit screening of tens of thousands of compounds in a short period of time. Physical Variants This invention also encompasses proteins or peptides having substantial amino acid sequence similarity with an amino acid sequence of a SEQ ID NO. 2, 4, 6, 8 or 10. Vaπants exhibiting substitutions, e.g., 20 or fewer, preferably 10 or fewer, and more preferably 5 or fewer substitutions, are encompassed. Where the substitutions are conservative substitutions, the vaπants will share lmmunogenic or antigemc similarity or cross-reactivity with a corresponding natural sequence protein Natural vaπants include individual, allehc, polymorphic, strain, or species vaπants. Amino acid sequence similaπty, or sequence identity, is determined by optimizing residue matches, if necessary', by introducing gaps as required This changes when consideπng conservative substitutions as matches. Conservative substitutions typically include substitutions within the following groups: glycme, alamne; vahne, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine, seπne, threonine; lysine, arginine; and phenylalamne, tyrosine Homologous ammo acid sequences include natural allehc and interspecies vaπations in each respective protein sequence. Typical homologous proteins or peptides w ill have from 50-100% similaπty (if gaps can be introduced), to 75-100% similarity (if conservative substitutions are included) with the amino acid sequence of the relevant protein Identity measures will be at least about 50%, generally at least 60%, more generally at least 65%, usually at least 70%, more usually at least 75%, preferably at least 80%, and more preferably at least 80%, and in particularly preferred embodiments, at least 85% or more. See also Needleham et al., 1970, J. Mol. Biol 48.443-453; Sankoff et al, 1983, Time Warps, String Edits, and Macromolecules The Theory and Practice of Sequence Comparison Chapter One, Addison- Wesley, Reading, MN, and software packages from IntelliGenetics, Mountain View, CN; and the University of Wisconsin Genetics Computer Group (GCG), Madison, WI Nucleic acids encoding the corresponding proteins will typically hybπdize to SEQ ID NO 1, 3, 5, 7 or 9 under stπngent conditions For example, nucleic acids encoding the respective proteins will typically hybπdize to the nucleic acid of SEQ ID NO 1 , 3, 5, 7 or 9 under stπngent hybπdization conditions, while providing few false positive hybπdization signals Generally, stnngent conditions are selected to be about 10° C lower than the thermal melting point (Tm) for the sequence being hybπdized to at a defined ionic strength and pH The Tm is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybπdizes to a perfectly matched probe Typically, stπngent conditions will be those in which the salt concentration in wash is about 0 02 molar at pH 7 and the temperature is at least about 50° C Other factors may significantly affect the stπngency of hybπdization, including, among others, base composition and size of the complementary strands, the presence of organic solvents such as formamide, and the extent of base mismatching A preferred embodiment will include nucleic acids that will bind to disclosed sequences in 50% formamide and 20-50 mM NaCl at 42° C An isolated nucleic acid can be readily modified by nucleotide substitutions, nucleotide deletions, nucleotide insertions, and inv ersions of nucleotide stretches These modifications result in nov el DNA sequences which encode these antigens, their deπvatives, or proteins hav ing highly similar physiological, lmmunogenic, or antige c activity Modified sequences can be used to produce mutant antigens or to enhance expression Enhanced expression ma involve gene amplification, increased transcπption, increased translation, and other mechanisms Such mutant protein derivatives include predetermined or site-specific mutations of the respective protein or its fragments "Mutant protein" encompasses a polypeptide otherwise falling within the homology definition of the proteins as set forth above, but having an amino acid sequence which differs from that of the protein as found in nature, whether by way of deletion, substitution, or insertion In particular, "site specific mutant protein" generally includes proteins having significant similarity with a protein having a sequence of SEQ ID NO 2, 4, 6, 8 or 10 Generally, the variant will share many physicochemical and biological activities, e.g , antigemc or lmmunogenic, with those sequences, and m preferred embodiments contain most or all of the disclosed sequence Glycosylation alterations are included, e g , made by modifying the glycosylation patterns of a polypeptide duπng its synthesis and processing, or in further processing steps Particularly preferred means for accomplishing this are by exposing the polypeptide to glycosylatmg enzymes deπved from cells that normally provide such processing, e g , mammalian glycosylation enzymes Deglycosylation enzymes are also contemplated Also embraced are versions of the same pπmary amino acid sequence which have other minor modifications, including phosphorylated ammo acid residues, e g , phosphotyrosme, phosphoseπne, or phosphothreonme, or other moieties, including πbosyl groups or cross-linking reagents Also, proteins compπsmg substitutions are encompassed, which should retain substantial immunogenicity, to produce antibodies that recognize a protein of SEQ ID NO 2, 4, 6, 8 or 10 Typically, these proteins will contain less than 20 residue substitutions from the disclosed sequence, more typically less than 10 substitutions, preferably less than 5, and more preferably less than three Alternatively, proteins that begin and end at structural domains will usually retain antigemcity and cross immunogenicity A major group of deπv atives are covalent conjugates of the proteins descπbed herein or fragments thereof with other proteins or polypeptides These deπvatives can be synthesized in recombinant culture such as N- or C-termmal fusions or by the use of agents known in the art for their usefulness in cross-linking proteins through reactive side groups Prefeπed protein deπvatization sites with cross-linking agents are at free amino groups, carbohydrate moieties, and cysteine residues Fusion polypeptides between these proteins and other homologous or heterologous proteins are also prov ided Heterologous polypeptides may be fusions between different surface markers, resulting in, e g , a hybπd protein Likewise, heterologous fusions may be constructed which would exhibit a combination of properties or activities of the deπvative proteins Typical examples are fusions of a reporter polypeptide, e g , luciferase, with a segment or domain of a protein, e g , a receptor-binding segment, so that the presence or location of the fused protein may be easily determined See, e g , U S Patent No 4,859,609 Other gene fusion partners include bacteπal β-galactosidase, trpE, Protein A, β-lactamase, alpha amylase, alcohol dehydrogenase, and yeast alpha mating factor See, e g , Godowski et al , 1988, Science 241 812-816 Such polypeptides may also have amino acid residues that have been chemically modified by phosphorylation, sulfonation, biotinylation, or the addition or removal of other moieties, particularly those that have molecular shapes similar to phosphate groups. In some embodiments, the modifications will be useful labeling reagents, or serve as purification targets, e.g., affinity ligands. This invention also contemplates the use of derivatives of these proteins other than variations in amino acid sequence or glycosylation. Such derivatives may involve covalent or aggregative association with chemical moieties. These derivatives generally fall into the three classes: (1) salts, (2) side chain and terminal residue covalent modifications, and (3) adsorption complexes, for example with cell membranes. Such covalent or aggregative derivatives are useful as immunogens, as reagents in immunoassays, or in purification methods such as for affinity purification of ligands or other binding ligands. For example, a protein antigen can be immobilized by covalent bonding to a solid support such as cyanogen bromide-activated Sepharose, by methods which are well known in the art, or adsorbed onto polyolefin surfaces, with or without glutaraldehyde cross-linking, for use in the assay or purification of antibodies. The proteins can also be labeled with a detectable group, e.g., radioiodinated by the chloramine T procedure, covalently bound to rare earth chelates, or conjugated to another fluorescent moiety for use in diagnostic assays. Purification of these proteins may be accomplished by immobilized antibodies. Antibodies The immunogenic components of this invention, as discribed above, are useful as antigens for preparing antibodies by standard methods. Such immunogenic components can be produced by proteolytic cleavage of larger polypeptides or by chemical synthesis or recombinant technology and are thus not limited by proteolytic cleavage sites. Preferably, smaller immunogenic components will first be rendered more immunogenic by cross-linking or by coupling to an immunogenic carrier molecule (i.e., a macromolecule having the property of independently eliciting an immunological response in a host animal, to which the immunogenic components of the invention can be covalently linked). Cross-linking or conjugation to a carrier molecule may be required because small polypeptide fragments sometimes act as haptens (molecules which are capable of specifically binding to an antibody but incapable of eliciting antibody production, i.e., they are not immunogenic). Conjugation of such fragments to an immunogenic carrier molecule renders them immunogenic through what is commonly known as the "carrier effect". Antibodies according to the present invention include polyclonal and monoclonal antibodies. The antibodies may be elicited in an animal host by immunization with immunogenic components of the invention or may be formed by in vitro immunization (sensitization) of immune cells The immunogenic components used to elicit the production of antibodies may be isolated from human cells (e.g., human monocytes) or chemically synthesized. The antibodies may also be produced in recombinant systems programmed with appropπate antibody-encoding DNA. Alternatively, the antibodies may be constructed by biochemical reconstitution of purified heavy and light chains. The antibodies of this invention can be puπfied by standard methods, including but not limited to preparative disc-gel electrophoresis, lsoelectric focusing, HPLC, reversed-phase HPLC, gel filtration, ion exchange and partition chromatography, and countercurrent distribution Puπfication methods for antibodies are disclosed, e.g., in The Art of Antibody Purification, 1989, Amicon Division, W.R. Grace & Co. General protein puπfication methods are descπbed in Protein Purification Principles and Practice, R.K. Scopes, Ed., 1987, Spπnger-Verlag, New York, NY. Suitable adjuvants for the vaccination of animals include but are not limited to Adjuvant 65 (containing peanut oil. mannide monooleate and aluminum monostearate); Freund's complete or incomplete adjuvant, mineral gels such as aluminum hydroxide, aluminum phosphate and alum, surfactants such as hexadecylamine, octadecylamine, lysolecithin, dime hyldioctadecyl-ammonium bromide, N,N-dιoctadecyl-N',N'-bιs(2- hydroxymethyl) propane-diamine, methoxyhexadecylglycerol and pluronic polyols, polyanions such as pyran, dextran sulfate, poly IC, polyacryhc acid and carbopol, peptides such as muramyl dipeptide, dimethylglycine and tuftsin; and oil emulsions. The immunogenic components could also be administered following incorporation into hposomes or other microcarπers Information concerning adjuvants and vaπous aspects of immunoassays are disclosed, e g . in the seπes by P Tijssen, 1987, Practice and Theon of Enzyme Immunoassaxs, 3rd Edition, Elsevier, New York. Serum produced from animals thus immunized can be used directly Alternatively, the IgG fraction can be separated from the serum using standard methods such as plasmaphoresis or adsorption chromatography using IgG specific adsorbents such as immobilized Protein A Hybπdomas of the invention used to make monoclonal antibodies against the immunogenic components of the invention are produced by well-known techniques Usually, the process involves the fusion of an immortalizing cell line with a B- lymphocyte that produces the desired antibody Alternatively, non-fusion techniques for generating immortal antibody-producmg cell lines are possible, and come within the purview of the present invention, e.g., virally-induced transformation, Casali et al , 1986, Science 234 476 Immortalizing cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine, and human oπgm Most frequently, rat or mouse myeloma cell lines are employed as a matter of convenience and availability Techniques for obtaining the appropπate lymphocytes from mammals injected with the immunogenic components are well known Generally, peπpheral blood lymphocytes (PBLs) are used if cells of human oπgm are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired A host animal is injected with repeated dosages of a preferably puπfied immunogenic component, and the animal is permitted to generate the desired antibody-producmg cells before these are harvested for fusion with the immortalizing cell line Techniques for fusion are also well known in the art, and in general involve mixing the cells with a fusing agent, such as polyethylene glycol Hybπdomas are selected by standard procedures, such as HAT (hypoxanthine- aminopteπn-thymidine) selection From among these hybπdomas, those secreting the desired antibody are selected by assaying their culture medium by standard immunoassays, such as Western blotting, ELISA (enzyme-linked immunosorbent assay), RIA (radioimmunoassay), or the like Antibodies are recovered from the medium using standard protein purification techniques, Tijssen, 1985, Practice and Theory of Enzyme Immunoassays, Elsevier, Amsterdam Many references are av ailable for guidance in applying any of the above techniques Kohler et al , 1980, Hx rtdoma Techniques, Cold Spnng Harbor Laboratory, New York, Tijssen, 1985, Piacticc and Theory of Enzyme Immunoassays, Elsevier, Amsterdam, Campbell, 1984, Monoclonal Antibody Technolog , Elsevier, Amsterdam, Hurrell, 1982, Monoclonal Hy bndoma Antibodies Techniques and Applications, CRC Press, Boca Raton, FL Monoclonal antibodies can also be produced using well known phage library systems The use and generation of antibody fragments is also well known, e g , Fab fragments Tijssen, 1985, Practice and Theory of Enzyme Immunoassays, Elsevier, Amsterdam, Fv fragments Hochman et al , 1973, Biochemistry 12 1 130, Sharon et al , 1976, Biochemistry 15 1591 , Ehrhch et al , U S Patent No 4,355,023, and antibody half molecules: Auditore-Hargreaves, U.S. Patent No. 4,470,925. These also may be useful in immunoassays. These antibodies, whether polyclonal or monoclonal, can be used, e.g., in an immobilized form bound to a solid support by well known methods, to isolate and purify the immunogenic components by immunoaffmity chromatography. The antibodies are useful as probes to distinguish tissue and cell type distribution. The antibodies may be used to screen expression libraπes for particular expression products. Usually the antibodies used in such a procedure will be labeled with a moiety allowing easy detection of presence of antigen by antibody binding. Antibodies to proteins may be used for the analysis or, or identification of specific cell population components which express the respective protein. By assaying the expression products of cells expressing the proteins described herein it is possible to diagnose disease, e.g., immune-compromised conditions, monocyte depleted conditions, or overproduction of monocytes. Antibodies raised against the proteins will also be useful to raise anti-idiotypic antibodies. These will be useful in detecting or diagnosing vaπous immunological conditions related to expression of the respective antigens The present invention encompasses antibodies that specifically recognize monocyte-deπved immunogenic components. Such antibodies can be used conventionally, e.g., as reagents for purification of monocyte cell components, or in diagnostic applications. Diagnostic Applications The invention encompasses compositions, methods, and kits useful in clinical settings for the qualitative or quantitative diagnosis, i.e., detection of specific components in a biological sample These applications utilize nucleic acids, peptides/polypeptides, or antibodies specific for the components descπbed herein. Both antibody-based and nucleic acid-based diagnostic methods, including PCR-based diagnostic methods are contemplated. Detection of the level of monocyte cells present in a sample is important for diagnosis of certain aberrant disease conditions. For example, an increase in the number of monocytes in a tissue or the lymph system can be indicative of the presence of a monocyte hyperplasia, tissue or graft rejection, or inflammation A low monocyte population can indicate an abnormal reaction to, e.g., a bacteπal or viral infection, which may require an appropriate treatment to normalize the monocyte response. Both the naturally occumng and the recombinant form of the proteins of this invention are particularly useful in kits and assay methods which are capable of screening compounds for binding activity to the proteins In nucleic-acid-type diagnostic methods, the sample to be analyzed may be contacted directed with the nucleic acid probes. Probes include oligonucleotides at least 12 nucleotides, preferably at least 18, and most preferably 20-35 or more nucleotides in length. Alternatively, the sample may be treated to extract the nucleic acids contained therein. It will be understood that the particular method used to extract DNA will depend on the nature of the biological sample. The resulting nucleic acid from the sample may be subjected to gel electrophoresis or other size separation techniques, or, the nucleic acid sample may be immobilized on an appropriate solid matrix without size separation or used for PCR. Kits suitable for antibody-based diagnostic applications typically include one or more of the following components: (i) Antibodies: The antibodies may be pre-labeled; alternatively, the antibody may be unlabelled and the ingredients for labeling may be included in the kit in separate containers, or a secondary, labeled antibody is provided; and (ii) Reaction components: The kit may also contain other suitably packaged reagents and materials needed for the particular immunoassay protocol, including solid- phase matrices, if applicable, and standards. Kits suitable for nucleic acid-based diagnostic applications typically include the following components: (i) Probe DNA: The probe DNA may be pre-labeled; alternatively, the probe DNA may be unlabelled and the ingredients for labeling may be included in the kit in separate containers; and (ii) Hybridization reagents' The kit may also contain other suitably packaged reagents and materials needed for the particular hybridization protocol, including solid- phase matrices, if applicable, and standards. PCR based diagnostic kits are also contemplated and are encompassed by the invention. The kits refeπed to above may include instructions for conducting the test. Furthermore, in preferred embodiments, the diagnostic kits are adaptable to high- throughput and/or automated operation. Therapeutic Applications The invention also provides reagents that may exhibit significant therapeutic value. The proteins (naturally occurring or recombinant), fragments thereof, and antibodies thereto, along with compounds identified as having binding affinity to the proteins, may be useful m the treatment of conditions associated with abnormal physiology or development For example, a disease or disorder associated with abnormal expression or abnormal signaling by a monocyte, e g , as an antigen presenting cell, is a target for an agonist or antagonist of the protein The proteins likely play a role in regulation or development of hematopoietic cells, e g , lymphoid cells, which affect lmmunological responses, e g , antigen presentation and the resulting effector functions Other abnormal developmental conditions are known in cell types shown to possess monocyte protein mRNA by northern blot analysis See Berkow (ed ) The Merck Manual of Diagnosis and Therapy, Merck & Co , Rahway, NJ, and Thorn, et al Hainson's Pπnciples of Internal Medicine, McGraw-Hill, NY Developmental or functional abnormalities, e g , of the immune system, cause significant medical abnormalities and conditions that may be susceptible to prevention or treatment using compositions provided herein Recombinant monocyte-deπved proteins or antibodies of the invention may be puπfied and then administered to a patient These reagents can be combined for therapeutic use with additional active or inert ingredients, e g , in conventional pharmaceutically acceptable earners or diluents, e g , immunogenic adjuvants, along with physiologically innocuous stabilizers and excipients In particular, these may be useful in a vaccine context, where the antigen is combined with one of these therapeutic versions of agonists or antagonists These combinations can be steπle filtered and placed into dosage forms as by lyophihzation in dosage vials or storage in stabilized aqueous preparations This inv ention also contemplates use of antibodies or binding fragments thereof, including forms which are not complement binding Drug screening using antibodies or receptor or fragments thereof can identify compounds having binding affinity to these monocyte-deπved proteins, including isolation of associated components Subsequent biological assays can then be utilized to determine if the compound blocks or antagonizes the activity of the protein Likewise, a compound having intπnsic stimulating activity might activate the cell through the protein and is thus an agonist This inv ention further contemplates the therapeutic use of antibodies to the proteins as antagonists The quantities of reagents necessary for effective therapy will depend upon many different factors, including means of administration, target site, physiological state of the patient, and other medicants administered Thus, treatment dosages should be titrated to optimize safety and efficacy Typically, dosages used in vitro may provide useful guidance in the amounts useful for in situ administration of these reagents Animal testing of effective doses for treatment of particular disorders will provide further predictive indication of human dosage. Various considerations are described, e.g., in Gilman, et al. (eds.) (1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics (8th ed.) Pergamon Press; and (1990) Remington's Pharmaceutical Sciences (17th ed.) Mack Publishing Co., Easton, PA. Methods for administration are discussed therein and below, e.g., for oral, intravenous, intraperitoneal, or intramuscular administration, transdermal diffusion, and others. Pharmaceutically acceptable carriers will include water, saline, buffers, and other compounds described, e.g., in the Merck Index, Merck & Co., Rahway, NJ. Dosage ranges would ordinarily be expected to be in amounts lower than 1 mM concentrations, typically less than about 10 μM concentrations, usually less than about 100 nM, preferably less than about 10 pM (picomolar), and most preferably less than about 1 fM (femtomolar), with an appropriate carrier. Slow release formulations, or a slow release apparatus will often be utilized for continuous administration. The proteins, antagonists, and agonists could be administered directly to the host to be treated or, depending on the size of the compounds, it may be desirable to conjugate them to carrier proteins such as ovalbumin or serum albumin prior to their administration. Therapeutic formulations may be administered in many conventional dosage formulations. While it is possible for the active ingredient to be administered alone, it is preferable to present it as a pharmaceutical formulation. Formulations typically comprise at least one active ingredient, as defined above, together with one or more acceptable carriers thereof. Each carrier should be both pharmaceutically and physiologically acceptable in the sense of being compatible with the other ingredients and not injurious to the patient. Formulations include those suitable for oral, rectal, nasal, or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. See, e.g., Gilman, et al. (eds.) ( 1990) Goodman and Gilman's: The Pharmacological Bases of Therapeutics (8th ed.) Pergamon Press; and (1990) Remington's Pharmaceutical Sciences (17th ed.) Mack Publishing Co., Easton, PA; Avis, et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosgae Forms: Tablets Dekker, NY; and Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems Dekker, NY. The therapy of this invention may be combined with or used in association with other chemotherapeutic or chemopreventive agents. Many modifications and variations of this invention can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. The specific embodiments described herein are offered by way of example only, and the invention is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled.

Claims: WHAT IS CLAIMED IS: 1. An isolated polypeptide comprising an amino acid sequence derived from SEQ ID NO: 2, 4, 6, 8 or 10. 2. The polypeptide of claim 1 comprising the amino acid sequence of the mature protein. 3. An isolated nucleic acid comprising a nucleotide sequence encoding an amino acid sequence derived from SEQ ID NO: 2. 4, 6, 8 or 10. 4. The nucleic acid of claim 3 wherein the nucleotide sequence encodes the mature protein. 5. The nucleic acid of claim 4 comprising the nucleotide sequence shown in SEQ ID: NO 1. 3. 5. 7 or 9. 6. A fusion protein comprising the polypeptide of claim 1. 7. A binding compound which specifically binds to the polypeptide of claim 1. 8. The binding compound of claim 7 which is an antibody or antibody fragment. 9. The binding compound of claim 8 wherein the anitbody is a monoclonal antibody. 10. An expression vector comprising the nucleic acid of claim 3. 1 1. A host cell comprising the vector of claim 10. 1 . A process for recombinatlv producing a polypeptide comprising culturing the host cell of claim 1 1 under conditions in which the polypeptide is expressed. 13. A method for detecting a specific nucleic acid sequence in a sample, said method comprising the steps of: (i) contacting a sample suspected to contain a specific nucleic acid sequence with a probe comprising a nucleic acid sequence comorising at least 8 consecutive nucleotides selected from SEQ ID NO: 1, 3, 5, 7, or 9 under conditions in which a hybrid can form between said probe and the specific nucleic acid in said sample; and (ii) detecting any hybrid formed in step (i), wherein detection of said hybrid indicates the presence of the specific nucleic acid sequence in said sample. 14. The method of claim 13 further comprising amplifying said specific sequence in said sample prior to said detecting step. 15. A method for detecting a specific antigenic component in a sample, said method comprising the steps of: (i) contacting a sample suspected to contain a specific antigenic component encoded by an amino acid sequence derived from SEQ ID NO: 2, 4. 6, 8, or 10 with an antibody specific for said component, under conditions in which a stable antigen-antibody complex can form between said antibody and said antigenic component in said sample; and (ii) detecting any antigen-antibody complex formed in step (i), wherein detection of an antigen-antibody complex indicates the presence of said antigenic component in said sample. 16. A method of screening for candidate therapeutic agents comprising: selecting as a target sequence a polypeptide having an amino acid sequence derived from SEQ ID NO: 2. 4. 6, 8 or 10; contacting a test compound with said target sequence; and selecting as candidate therapeutic agent those test compounds which bind to the target sequence.

Inventors: BATES ELIZABETH, CHAULUS LIONEL, FOURNIER NATHALIE, GARRONE PIERRE, BATES, ELIZABETH, CHAULUS, LIONEL, FOURNIER, NATHALIE, GARRONE, PIERRE
Applicants: BATES ELIZABETH, CHAULUS LIONEL, FOURNIER NATHALIE, GARRONE PIERRE, SCHERING CORP, BATES, ELIZABETH, CHAULUS, LIONEL, FOURNIER, NATHALIE, GARRONE, PIERRE, SCHERING CORPORATION
Application year: 1999
Main classification: C12N15
IPC codes: C07K14, C07K16, C07K19, C12N1, C12N5, C12N15, C12P21, C12Q1, G01N33