AL-2 neurotrophic factor

Abstract

The present invention provides nucleic acids encoding AL-2 protein, as well as AL-2 protein produced by recombinant DNA methods. Such AL-2 protein and nucleic acid are useful in preparing antibodies and antagonists and in diagnosing and treating various neuronal disorders and disorders or conditions associated with angiogenesis.

What is claimed is:  
     
         1 . An isolated nucleic acid, which encodes a polypeptide having an amino acid sequence that is at least 75% identical to the amino acid sequence for mature human AL-2 shown in FIGS.  1 A- 1 B or FIGS.  2 A- 2 B.  
     
     
         2 . The isolated nucleic acid according to  claim 1 , which encodes a polypeptide having an amino acid sequence that is at least 85% identical to the amino acid sequence for mature human AL-2 shown in FIGS.  1 A- 1 B or FIGS.  2 A- 2 B.  
     
     
         3 . The isolated nucleic acid of  claim 2 , comprising a nucleotide sequence encoding the amino acid sequence shown in FIGS.  1 A- 1 B for mature AL-21.  
     
     
         4 . The isolated nucleic acid of  claim 2 , comprising a nucleotide sequence encoding the amino acid sequence shown in FIGS.  2 A- 2 B for mature AL-2s.  
     
     
         5 . The isolated nucleic acid of  claim 2 , which encodes a polypeptide having an amino acid sequence that is at least 75% homologous to the amino acid sequence of the extracellular domain of AL-2 shown in FIGS.  1 A- 1 B.  
     
     
         6 . The isolated nucleic acid of  claim 5 , which encodes a polypeptide having the amino acid sequence of the extracellular domain shown in FIGS.  1 A- 1 B for AL-2.  
     
     
         7 . The isolated nucleic acid of  claim 1 , wherein AL-2 is joined to an immunoglobulin.  
     
     
         8 . The isolated nucleic acid of  claim 7 , which encodes AL-2-IgG.  
     
     
         9 . The isolated nucleic acid of  claim 1 , wherein AL-2 is fused to a tag polypeptide.  
     
     
         10 . The isolated nucleic acid of  claim 1 , which hybridizes to DNA encoding mature human AL-21 of FIGS.  1 A- 1 B or mature human AL-2s of FIGS.  2 A- 2 B under stringent conditions, and which encodes a polypeptide that is antigenically cross-reactive to mature human AL-2s or AL-21.  
     
     
         11 . An expression vector comprising the nucleotide sequence of  claim 1  operably linked to a promoter.  
     
     
         12 . The expression vector of  claim 11 , wherein the nucleotide sequence encodes the amino acid sequence for mature AL-2 shown in FIGS.  1 A- 1 B or FIGS.  2 A- 2 B.  
     
     
         13 . The expression vector of  claim 12 , wherein the nucleotide sequence encoding the amino acid sequence for mature AL-2 is that shown in FIGS.  1 A- 1 B or FIG. 2A- 2 B.  
     
     
         14 . A host cell transformed with the expression vector of  claim 11 .  
     
     
         15 . The host cell of  claim 14 , wherein the nucleotide sequence encodes the amino acid sequence for mature AL-2 shown in FIGS.  1 A- 1 B or FIG. 2A- 2 B.  
     
     
         16 . A method of using the host cell of  claim 14 , which method comprises culturing the host cell under conditions that allow replication of the expression vector.  
     
     
         17 . A process which comprises transforming a host cell with an expression vector capable, in the host cell transformed with the vector, of expressing a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence shown in FIGS.  1 A- 1 B or FIGS.  2 A- 2 B for mature AL-2, and culturing the transformed host cell under conditions such that the AL-2 polypeptide is synthesized.  
     
     
         18 . An isolated polypeptide having an amino acid sequence that is at least 75% homologous to the mature human AL-2 amino acid sequence shown in FIGS.  1 A- 1 B or FIGS.  2 A- 2 B.  
     
     
         19 . The isolated polypeptide of  claim 18  having an amino acid sequence that is at least 85% homologous to the mature human AL-2 amino acid sequence shown in FIGS.  1 A- 1 B or FIGS.  2 A- 2 B.  
     
     
         20 . The isolated polypeptide of  claim 19  having the mature human AL-2 amino acid sequence shown in FIGS.  1 A- 1 B or FIGS.  2 A- 2 B.  
     
     
         21 . The isolated polypeptide of  claim 18  having an amino acid sequence that is at least 75% homologous to the amino acid sequence of the extracellular domain shown in FIGS.  1 A- 1 B for mature human AL-2.  
     
     
         22 . The isolated polypeptide of  claim 21  having the amino acid sequence of the extracellular domain shown in FIGS.  1 A- 1 B for AL-2.  
     
     
         23 . The isolated polypeptide of  claim 18 , wherein AL-2 is joined to an immunoglobulin.  
     
     
         24 . The polypeptide of  claim 23 , wherein the AL-2 extracellular domain is joined to an immunoglobulin constant domain.  
     
     
         25 . The polypeptide of  claim 24 , wherein the constant domain is that of an immunoglobulin heavy chain.  
     
     
         26 . The polypeptide of  claim 23  that is AL-2-IgG.  
     
     
         27 . The polypeptide of  claim 18 , wherein AL-2 is fused to a tag polypeptide.  
     
     
         28 . A pharmaceutical composition comprising the polypeptide of  claim 18  and a physiologically acceptable carrier.  
     
     
         29 . An antibody that specifically binds to a polypeptide having the amino acid sequence shown in FIGS.  1 A- 1 B for mature AL-2.  
     
     
         30 . The antibody of  claim 29  that is a monoclonal antibody.  
     
     
         31 . A method for activating a tyrosine kinase domain of an AL-2-binding Eph-family receptor, comprising contacting an extracellular domain of the receptor with the AL-2 of  claim 1 .  
     
     
         32 . A method of treating a neurologic disease or disorder in a mammal, comprising administering to the mammal a therapeutically effective amount of the composition of  claim 28 .  
     
     
         33 . The method of  claim 32  wherein the neurologic disease or disorder is trauma-induced, surgery-induced, stroke-induced, ischemia-induced, infection-induced, metabolic disease-related, nutritional deficiency-induced, malignancy-induced, neurotoxicity, Alzheimer's disease, amyotrophic lateral sclerosis, Bell's palsy, spinal muscular atrophy or paralysis, Parkinson's disease, epilepsy, multiple sclerosis, Huntington's chorea, Down's Syndrome, nerve deafness, Meniere's disease, post-polio syndrome, Charcot-Marie-Tooth disease, Refsum's disease, Abetalipoproteinemia, Tangier disease, Krabbe's disease, Metachromatic leukodystrophy, Fabry's disease, and Dejerine-Sottas syndrome.  
     
     
         34 . The method of  claim 32  that further comprises administering a therapeutically effective amount of a second neurotrophic factor.  
     
     
         35 . A method for accelerating the neovascularization of a wound, comprising applying to the wound an angiogenically effective amount of the composition of  claim 28 .  
     
     
         36 . A method of modulating angiogenesis associated with a disease condition in a mammal, comprising administering to the mammal an angiogenically-modulating amount of an AL-2 antagonist.  
     
     
         37 . The method of  claim 36 , wherein the angiogenesis-associated disease condition is rheumatoid arthritis or tumor formation.  
     
     
         38 . A method of diagnosing a neurologic disease or disorder, comprising contacting nucleic acid of a sample with a second nucleic acid comprising at least 10 nucleotides of the nucleotide sequence shown in FIGS.  1 A- 1 B or  2 A- 2 B under conditions that allow hybridization of complementary nucleotide sequences, and detecting any hybridization that occurs.  
     
     
         39 . The method of  claim 38 , further comprising amplifying the sample nucleic acid to which the second nucleic acid hybridizes.