CD30 Ligand

Abstract

There is disclosed a polypeptide (CD30-L) and DNA sequences, vectors and transformed host cells useful in providing CD30-L polypeptides. The CD30-L polypeptide binds to the receptor known as CD30, which is expressed on a number of cell types, among which are Hodgkin's Disease tumor cells, large cell anaplastic lymphoma cells, adult T-cell leukemia (T-ALL) cells, and a number of other malignant cell types. CD30-L polypeptides find use as carriers for delivering diagnostic and cytotoxic agents to cells expressing the CD30 receptor.

1 . An isolated DNA sequence encoding a CD30-L polypeptide, wherein said CD30-L comprises an amino acid sequence selected from the group consisting of amino acids 1-220 of SEQ ID NO:19, amino acids 1-215 of SEQ ID NO:23, amino acids 1-239 of SEQ ID NO:6, and amino acids 1-234 of SEQ ID NO:8. 
     
     
         2 . An isolated DNA sequence encoding a soluble CD30-L polypeptide capable of binding CD30, wherein said CD30-L comprises an amino acid sequence selected from the group consisting of amino acids 49-220 of SEQ ID NO:19 and amino acids z-215 of SEQ ID NO:23, wherein z is selected from the group consisting of 44, 45, 46, and 47. 
     
     
         3 . An isolated DNA comprising a DNA sequence that encodes an Fc polypeptide derived from an antibody, fused directly or through a peptide linker-encoding sequence to the 5′ end of a soluble CD30-L-encoding DNA according to  claim 2 . 
     
     
         4 . A DNA sequence according to  claim 2 , wherein said DNA sequence comprises a nucleotide sequence selected from the group consisting of nucleotides 130-645, 133-645, 136-645, and 139-645 of SEQ ID NO:22. 
     
     
         5 . An isolated DNA capable of hybridizing to a DNA sequence of  claim 1  under highly stringent conditions, wherein said isolated DNA encodes a CD30-L polypeptide capable of binding CD30. 
     
     
         6 . An isolated DNA sequence encoding a CD30-L polypeptide capable of binding CD30, wherein said CD30-L comprises an amino acid sequence selected from the group consisting of amino acids x to 239 of SEQ ID NO:6, wherein x is 1-19, and amino acids y to 234 of SEQ ID NO:8, wherein y is 1-19. 
     
     
         7 . An expression vector comprising a DNA sequence according to  claim 1 . 
     
     
         8 . An expression vector comprising a DNA sequence according to  claim 2 . 
     
     
         9 . An expression vector comprising a DNA sequence according to  claim 3 . 
     
     
         10 . An expression vector comprising a DNA sequence according to  claim 5 . 
     
     
         11 . A process for preparing a CD30-L polypeptide, comprising culturing a host cell transformed with a vector according to  claim 7  under conditions promoting expression of CD30-L, and recovering the CD30-L polypeptide. 
     
     
         12 . A process for preparing a CD30-L polypeptide, comprising culturing a host cell transformed with a vector according to  claim 8  under conditions promoting expression of CD30-L and recovering the CD30-L polypeptide. 
     
     
         13 . A process for preparing a soluble CD30-L/Fc fusion protein, comprising culturing a host cell transformed with a vector according to  claim 9  under conditions promoting expression of CD30-L/Fc, and recovering the CD30-L/Fc polypeptide. 
     
     
         14 . A process for preparing a CD30-L polypeptide, comprising culturing a host cell transformed with a vector according to  claim 10  under conditions promoting expression of CD30-L, and recovering the CD30-L polypeptide. 
     
     
         15 . A substantially homogeneous purified biologically active CD30-L protein, wherein said CD30-L is selected from the group consisting of murine CD30-L comprising the N-terminal amino acid sequence Met-Gln-Val-Gln-Pro-Gly-Ser-Val-Ala-Ser-Pro-Trp or Met-Glu-Pro-Gly-Leu-Gln-Gln-Ala-Gly-Ser-Cys-Gly, and human CD30-L comprising the N-terminal amino acid sequence Met-His-Val-Pro-Ala-Gly-Ser-Val-Ala-Ser-His-Leu or Met-Asp Pro-Gly-Leu-Gln-Gln-Ala-Leu-Asn-Gly-Met. 
     
     
         16 . A purified CD30-L according to  claim 15 , wherein said CD30-L comprises an amino acid sequence selected from the group consisting of amino acids 1-220 of SEQ ID NO:19, amino acids 1-215 of SEQ ID NO:23, amino acids 1-239 of SEQ ID NO:6, and amino acids 1-234 of SEQ ID NO:8. 
     
     
         17 . A substantially homogeneous soluble CD30-L polypeptide, wherein said soluble CD30-L comprises an amino acid sequence selected from the group consisting of amino acids 49-220 of SEQ ID NO:19 and amino acids z-215 of SEQ ID NO:23, wherein z is selected from the group consisting of 44, 45, 46, and 47. 
     
     
         18 . A substantially homogeneous purified CD30-L protein capable of binding CD30, wherein said CD30-L is encoded by a DNA sequence that will hybridize to the nucleotide sequence presented in SEQ ID NO:19 or SEQ ID NO:23 under severely stringent conditions. 
     
     
         19 . A CD30-L protein according to  claim 18 , wherein said protein is a soluble human CD30-L protein. 
     
     
         20 . Purified CD30-L according to  claim 18 , wherein said CD30-L comprises an amino acid sequence selected from the group consisting of amino acids x to 239 of SEQ ID NO:6, wherein x is 1-19, and amino acids y to 234 of SEQ ID NO:8, wherein y is 1-19. 
     
     
         21 . A fusion protein comprising a soluble human CD30-L according to  claim 19  and an Fc polypeptide derived from an antibody. 
     
     
         22 . An antibody immunoreactive with a CD30-L polypeptide according to  claim 18 . 
     
     
         23 . An antibody according to  claim 22  wherein said antibody is a monoclonal antibody. 
     
     
         24 . An isolated nucleic acid molecule comprising a sequence of at least about 14 nucleotides of the DNA sequence of SEQ ID NO:5 or SEQ ID NO:7, or the DNA or RNA complement thereof. 
     
     
         25 . A conjugate comprising a diagnostic or therapeutic agent attached to a CD30-L polypeptide according to  claim 18 . 
     
     
         26 . A conjugate comprising a diagnostic or therapeutic agent attached to a soluble CD30-L polypeptide according to  claim 19 . 
     
     
         27 . A method of delivering a diagnostic or therapeutic agent to CD30 +  cells, comprising contacting said cells with a conjugate according to  claim 25 . 
     
     
         28 . A method of delivering a diagnostic or therapeutic agent to CD30 +  cells, comprising contacting said cells with a conjugate according to  claim 26 . 
     
     
         29 . A method according to  claim 28 , wherein said cells are malignant and said conjugate is administered in an effective amount to a human afflicted with said malignant cells. 
     
     
         30 . A method according to  claim 29 , wherein said cells are CD30 +  lymphoma cells. 
     
     
         31 . A method of treating large cell anaplastic lymphoma (LCAL), comprising administering an effective amount of a soluble CD30-L according to  claim 19  to a human afflicted with LCAL.