Compositions, kits, and methods for the diagnosis, prognosis, and monitoring of immune disorders using galectin-1

Abstract

The present invention is based, in part, on the discovery that galectin-1 (Gal1) plays a role in immune disorders, including Hodgkin lymphoma, anaplastic large cell lymphoma, or MLL+ pre B-cell ALL. Accordingly, the invention relates to compositions, kits, and methods for diagnosing, prognosing, and monitoring immune disorders, e.g., Hodgkin lymphoma, anaplastic large cell lymphoma, or MLL+ pre B-cell ALL.

1 . A method for detecting the presence of a Gal1 polypeptide or fragment thereof; or a Gal1 polynucleotide or fragment thereof in a sample comprising:
 a) contacting the sample with a compound which selectively binds to a Gal1 polypeptide or fragment thereof, optionally wherein the compound is an antibody, an antibody derivative, or an antibody fragment; or a nucleic acid probe or primer which selectively hybridizes to a Gal1 polynucleotide or fragment thereof, and is contacted with a nucleic acid probe, optionally wherein the sample comprises a transcribed polynucleotide, or a portion thereof; and   b) determining whether the compound binds to a Gal1 polypeptide or fragment thereof in the sample to thereby detect the presence of a Gal1 polypeptide or fragment thereof; or determining whether the nucleic acid probe or primer binds to a nucleic acid molecule in the sample to thereby detect the presence of a Gal1 polynucleotide or fragment thereof;   optionally wherein the transcribed polynucleotide is mRNA or cDNA; optionally wherein the step of detecting further comprises amplifying the transcribed polynucleotide; optionally wherein the level of expression of Gal1 is assessed by detecting the presence in the sample of a transcribed polynucleotide which anneals with a Gal1 polynucleotide or anneals with a portion of a Gal1 polynucleotide, under stringent hybridization conditions.   
     
     
         2 - 4 . (canceled) 
     
     
         5 . A method for identifying a compound which binds to a Gal1 polypeptide or fragment thereof comprising:
 a) contacting a Gal1 polypeptide or fragment thereof, or a cell expressing said polypeptide with a test compound; and   b) determining whether said polypeptide binds to the test compound.   
     
     
         6 . The method of  claim 5 , wherein the binding of the test compound to a Gal1 polypeptide or fragment thereof is detected by a method selected from the group consisting of:
 a) detection of binding by direct detection of test compound/polypeptide binding;   b) detection of binding using a competition binding assay; and   c) detection of binding using an assay for Gal1 activity.   
     
     
         7 . The method of  claim 1 , further comprising assessing whether a subject has a condition that would benefit from upregulation of an immune response, the method comprising comparing:
 a) the level of expression of Gal1 in a subject sample; and   b) the normal level of expression of Gal1 in a control sample,   
       wherein a significant increase in the level of expression of Gal1 in the subject sample relative to the normal level is an indication that the subject is afflicted with said condition;
 optionally wherein the sample comprises cells obtained from the subject, optionally wherein the cells are in a fluid selected from the group consisting of whole blood fluid, serum fluid, plasma fluid, interstitial fluid, cerebrospinal fluid, lymph fluid, saliva, stool, and urine. 
 
     
     
         8 . The method of  claim 7 , wherein the condition is an immune disorder or cancer. 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 8 , wherein the condition is Hodgkin lymphoma, anaplastic large cell lymphoma, or MLL +  pre B-cell ALL. 
     
     
         11 - 19 . (canceled) 
     
     
         20 . The method of  claim 7 , wherein said significant increase comprises an at least two fold increase between the level of expression of Gal1 in the subject sample relative to the normal level of expression of Gal1 in the sample from the control subject. 
     
     
         21 . The method of  claim 20 , wherein said significant increase comprises an at least five fold increase between the level of expression of Gal1 in the subject sample relative to the normal level of expression of Gal1 in the sample from the control subject. 
     
     
         22 . The method of  claim 1 , further comprising monitoring the progression of an immune disorder in a subject, the method comprising:
 a) detecting in a subject sample at a first point in time the expression of Gal1;   b) repeating step a) at a subsequent point in time; and   c) comparing the level of expression of said Gal1 detected in steps a) and b) to monitor the progression of the immune disorder;   optionally wherein between the first point in time and the subsequent point in time, the subject has undergone treatment to ameliorate the immune disorder; optionally wherein said treatment comprises chemotherapy.   
     
     
         23 . (canceled) 
     
     
         24 . The method of  claim 1 , further comprising assessing the efficacy of a test compound for inhibiting an immune disorder in a subject, the method comprising comparing:
 a) the level of expression of Gal1 in a first sample obtained from the subject and exposed to the test compound; and   b) the level of expression of Gal1 in a second sample obtained from the subject, wherein the second sample is not exposed to the test compound, and a significantly lower level of expression of Gal1, relative to the second sample, is an indication that the test compound is efficacious for inhibiting an immune disorder in the subject;   optionally wherein the first and second samples are portions of a single sample obtained from the subject or portions of pooled samples obtained from the subject.   
     
     
         25 . (canceled) 
     
     
         26 . The method of  claim 1 , further comprising predicting the clinical outcome of a patient, the method comprising:
 a) determining the level of expression of Gal1 in a patient sample   b) determining the level of expression of Gal1 in a sample from a control subject having a good clinical outcome; and   c) comparing the level of expression of Gal1 in the patient sample and in the sample from the control subject;   
       wherein a significantly higher level of expression in the patient sample as compared to the expression level in the sample from the control subject is an indication that the patient has a poor clinical outcome. 
     
     
         27 . The method of  claim 1 , further comprising assessing the efficacy of a therapy for inhibiting an immune disorder in a subject, the method comprising comparing:
 a) the level of expression of Gal1 in the first sample obtained from the subject prior to providing at least a portion of the therapy to the subject, and   b) the level of expression of Gal1 in a second sample obtained from the subject following provision of the portion of the therapy,   
       wherein a significantly lower level of expression of Gal1 in the second sample, relative to the first sample, is an indication that the therapy is efficacious for inhibiting the immune disorder in the subject. 
     
     
         28 . The method of  claim 5 , further comprising making an isolated hybridoma which produces an antibody that specifically binds to a Gal1 polypeptide or a fragment thereof, the method comprising:
 a) immunizing a mammal using a composition comprising a Gal1 polypeptide or a fragment thereof;   b) isolating splenocytes from the immunized mammal;   c) fusing the isolated splenocytes with an immortalized cell line to form hybridomas; and   d) screening individual hybridomas for production of an antibody which specifically binds with said polypeptide thereof to isolate the hybridoma.   
     
     
         29 . An antibody produced by a hybridoma according to the method of  claim 28 . 
     
     
         30 . An isolated antibody or antigen binding portion thereof that specifically binds to a polypeptide selected from the group consisting of:
 a) a fragment of a polypeptide comprising the amino acid sequence of human Gal1;   b) a polypeptide which is encoded by a nucleic acid comprising a nucleotide sequence which is at least 80% homologous to a nucleic acid comprising the nucleotide sequence of human Gal1; and   c) a polypeptide comprising an amino acid sequence which is at least 80% homologous to the amino acid sequence of human Gal1;   
       optionally wherein the antibody or antigen binding portion thereof is a monoclonal antibody, a polyclonal antibody, a chimeric antibody, or a humanized antibody; optionally wherein the antibody or antigen binding portion thereof comprises an effector domain; optionally wherein the antibody or antigen binding portion thereof comprises an Fc domain; optionally wherein the antibody or antigen binding portion thereof is a single-chain antibody; and/or optionally wherein the antibody or antigen binding portion thereof is a Fab fragment. 
     
     
         31 - 33 . (canceled) 
     
     
         34 . The antibody or antigen binding portion thereof of  claim 30  which is detectably labeled. 
     
     
         35 . The antibody or antigen binding portion thereof of  claim 30 , wherein the antibody or antigen binding portion thereof inhibits Hodgkin lymphoma, anaplastic large cell lymphoma, or MLL +  pre B-cell ALL in a subject. 
     
     
         36 . The antibody or antigen binding portion thereof of  claim 30 , wherein the antibody or antigen binding portion thereof specifically binds a Gal1 epitope comprising the carbohydrate binding domain, optionally wherein the antibody or antigen binding portion thereof specifically binds a Gal1 epitope comprising amino acids 62-86. 
     
     
         37 - 42 . (canceled) 
     
     
         43 . A kit comprising an agent which selectively binds to a Gal1 polypeptide or fragment thereof produced by the method according to  claim 5 , and instructions for use. 
     
     
         44 . A kit for use in  claim 1 , comprising an agent which selectively hybridizes to a Gal1 polynucleotide or fragment thereof and instructions for use.