Heterologous polypeptide of the TNF family
View Patent ↗A newly identified heteromeric ligand of the Tumor Necrosis Factor (TNF)-family, referred to hereinafter as “APBF” has been identified.
1. An isolated polypeptide comprising:
(a) an APRIL subunit selected from:
(i) human APRIL (SEQ ID NO:2) or partial human APRIL having an amino acid terminus starting at any one of amino acids 105 to 135 of human APRIL (SEQ ID NO:2);
(ii) an amino acid substitution variant of human APRIL (SEQ ID NO:2) or partial human APRIL having an amino acid terminus starting at any one of amino acids 105 to 135 of human APRIL (SEQ ID NO:2), wherein said variant and said APRIL or partial APRIL share at least 75% sequence similarity; or
(iii) a splice variant of human APRIL (SEQ ID NO:2), said variant missing amino acids 113 to 128 of human APRIL (SEQ ID NO:2);
(b) linked via a non-covalent interaction to a BAFF subunit selected from:
(i) human BAFF (SEQ ID NO:6) or partial human BAFF having an amino acid terminus starting at any one of amino acids 134 to 164 of human BAFF (SEQ ID NO:6);
(ii) an amino acid substitution variant of human BAFF (SEQ ID NO:6) or partial human BAFF having an amino acid terminus starting at any one of amino acids 134 to 164 of human BAFF (SEQ ID NO:6), wherein said variant and said BAFF or partial BAFF share at least 75% sequence similarity; or
(iii) a splice variant of human BAFF (SEQ ID NO:6), said variant missing amino acids 142 to 160 of human BAFF (SEQ ID NO:6), wherein said polypeptide induces proliferation of B cells.
2. An isolated polypeptide comprising:
(a) an APRIL subunit encoded by a DNA that hybridizes under high stringency conditions to the complement of a human APRIL cDNA comprising SEQ ID NO:1
(b) linked via a non-covalent interaction to a BAFF subunit encoded by a DNA that hybridizes under high stringency conditions to the complement of a human BAFF cDNA comprising (SEQ ID NO:5
wherein said stringent hybridization conditions comprise hybridization at 65° C. in the presence of 0.5 M NaHPO 4 , 7% SDS and 1 mM EDTA, followed by washing at 68° C. in the presence of 0.1×SSC and 0.1% SDS, and wherein said polypeptide induces proliferation of B cells.
3. An isolated polypeptide comprising:
(a) an APRIL subunit selected from:
(i) murine APRIL (SEQ ID NO:4) or partial murine APRIL having an amino acid terminus starting at amino acid 87 of murine APRIL having (SEQ ID NO:4); or
(ii) an amino acid substitution variant of murine APRIL having (SEQ ID NO:4) or partial murine APRIL having an amino acid terminus starting at amino acid 87 of murine APRIL (SEQ ID NO:4), wherein said variant and said APRIL or partial APRIL share at least 75% sequence similarity;
(b) linked via a non-covalent interaction to a BAFF subunit selected from:
(i) murine BAFF having (SEQ ID NO:8);
(ii) an amino acid substitution variant of the murine BAFF having (SEQ ID NO:8) wherein said variant and said BAFF share at least 75% sequence similarity; or
(iii) a splice variant of murine BAFF having (SEQ ID NO:8), said variant missing amino acids 166 to 184 of murine BAFF (SEQ ID NO:8), and wherein said polypeptide induces proliferation of B cells.
4. An isolated polypeptide comprising:
(a) an APRIL subunit encoded by a DNA that hybridizes under high stringency conditions to the complement of a murine APRIL cDNA having SEQ ID NO:3
(b) linked via a non-covalent interaction to a BAFF subunit encoded by murine BAFF cDNA having SEQ ID NO:7
wherein said stringent hybridization conditions comprise hybridization at 65° C. in the presence of 0.5 M NaHPO 4 , 7% SDS and 1 mM EDTA, followed by washing at 68° C. in the presence of 0.1×SSC and 0.1% SDS, and wherein said polypeptide induces proliferation of B cells.
5. A pharmaceutical composition comprising a therapeutically effective amount of the isolated polypeptide according to claim 1 or 2 and a pharmaceutically acceptable carrier.
6. The isolated polypeptide according to claims 1 or 3 , wherein said sequence similarity is at least 80%.
7. The isolated polypeptide according to claims 1 or 3 , wherein said sequence similarity is at least 85%.
8. The isolated polypeptide according to claims 1 or 3 , wherein sequence similarity is at least 90%.
9. The isolated polypeptide according to claims 1 or 3 , wherein said sequence similarity is at least 95%.
10. The isolated polypeptide according to claim 1 or 2 , wherein said APRIL subunit is a partial human APRIL having an amino acid terminus starting at an amino acid selected from the group consisting of: ammo acid 105 of human APRIL (SEQ ID NO:2), amino acid 110 of human APRIL (SEQ ID NO:2) and amino acid 115 of human APRIL (SEQ ID NO:2).
11. The isolated polypeptide according to claim 1 or 2 , wherein said BAFF subunit is a partial human BAFF having an amino acid terminus starting at amino acid 134 of human BAFF (SEQ ID NO:6) or amino acid 136 of human BAFF (SEQ ID NO:6).
12. The isolated polypeptide according to any one of claims 1 to 4 , further comprising more than one APRIL subunit linked non-covalently to said BAFF subunit.
13. The isolated polypeptide according to claim 12 , wherein two APRIL subunits are linked non-covalently to said BAFF subunit.
14. The isolated polypeptide according to any one of claims 1 to 4 , further comprising more than one BAFF subunit linked non-covalently to said APRIL subunit.
15. The isolated polypeptide according to claim 13 , wherein two BAFF subunits are linked non-covalently to said APRIL subunit.
16. The isolated polypeptide according to any one of claims 1 to 4 , wherein said polypeptide is soluble.