Diagnosis, prevention and treatment of calicivirus infection in humans

Abstract

Caliciviruses are herein recognized as potentially important human pathogens, infecting a large percentage (18%) of a sampled human population. A new calicivirus isolate called “San Miguel sea lion virus 5 serotype Homosapien-1” (SMSV-5 Hom-1) is also disclosed. Compositions and methods are disclosed that allow diagnosis, prevention and treatment of calicivirus (including SMSV-5 Hom-1) infection in humans.

1 . A purified protein isolated from calicivirus, and comprising an amino acid sequence selected from the group consisting of: 
 (a) an amino acid sequence selected from the group consisting of SEQ ID NOS: 5, 11, 12, and 13;    (b) amino acid sequence that differ from those specified in (a) by one or more conservative amino acid substitutions; and    (c) amino acid sequences having at least 80% sequence identity to the sequences specified in (a) or (b).    
     
     
         2 . An isolated nucleic acid molecule encoding a protein according to  claim 1 .  
     
     
         3 . An isolated nucleic acid molecule according to  claim 2 , wherein the molecule comprises a sequence selected from the group consisting of: 
 (a) SEQ ID NO: 3;    (b) SEQ ID NO: 4; and    (c) at least 15 consecutive nucleotides of the sequences provided in (a) or (b).    
     
     
         4 . An isolated nucleic acid molecule that hybridizes under high stringency conditions with the nucleic acid molecule of  claim 3 .  
     
     
         5 . A recombinant nucleic acid molecule comprising a promoter sequence operably linked to a nucleic acid sequence according to  claim 2 .  
     
     
         6 . A cell transformed with a recombinant nucleic acid molecule according to  claim 5 .  
     
     
         7 . An immunostimulatory polypeptide comprising at least one polypeptide selected from the group consisting of SEQ ID NO: 5, 11, 12, 13, and fragments thereof.  
     
     
         8 . A method of detecting calicivirus in a tissue sample from a human, the method comprising: 
 contacting the tissue sample with a calicivirus specific binding agent wherein the calicivirus specific binding agent binds at least two different calicivirus serotypes; and    detecting calicivirus or portions of calicivirus that are bound to the specific binding agent.    
     
     
         9 . The method of  claim 8  wherein the human is at risk of having a spontaneous abortion.  
     
     
         10 . The method of  claim 8  wherein the human is at risk of having calicivirus hepatitis.  
     
     
         11 . The method of  claim 8  wherein the human is at risk of having chronic fatigue syndrome.  
     
     
         12 . The method of  claim 8  wherein the human is at risk of having a disorder selected from the group consisting of diarrhea, myocarditis, myositis, encephalitis, hemorrhage, vesicles, pneumonia, blistering and erosion of mucosal membranes, and thymic involution.  
     
     
         13 . A method of generating monoclonal antibodies specific for multiple serotypes of calicivirus, the method comprising; 
 infecting a cell culture specimen with at least one strain of calicivirus;    allowing the calicivirus to initiate replication;    lysing the cell cutlure;    using the lysate to immunize a subject;    harvesting at least one cell from the subject;    fusing the cell from the subject to a myeloma partner; and    identifying hybridomas producing antibodies which bind to at least two different strains of calicivirus.    
     
     
         14 . A hybridoma produced by the method of  claim 13 .  
     
     
         15 . A method of detecting calicivirus antibodies in a human, the method comprising: 
 contacting a tissue sample from the human with at least one calicivirus epitope; and    detecting antibody bound to the epitope.    
     
     
         16 . The method of  claim 15 , wherein the human has had a spontaneous abortion.  
     
     
         17 . The method of  claim 15 , wherein the human has hepititis symptoms.  
     
     
         18 . The method of  claim 15 , wherein the human has chronic fatigue syndrome.  
     
     
         19 . The method of  claim 15 , wherein the human has a disorder selected from the group consisting of diarrhea, myocarditis, myositis, encephalitis, hemorrhage, vesicles, pneumonia, blistering and erosion of mucosal membranes and thymic involution.  
     
     
         20 . The method of  claim 15 , wherein the epitope is found in SMSV-5 Hom-1 (ATCC No. PTA-4762).  
     
     
         21 . The method of  claim 15 , wherein the calicivirus epitope is selected from the group consisting of the polypeptide shown in SEQ ID NO: 5, and fragments thereof.  
     
     
         22 . An isolated polypeptide comprising the amino acid sequence shown in SEQ ID NO: 5.  
     
     
         23 . An isolated calicivirus comprising, the SMSV-5 Hom-1 isolate (ATCC No. PTA-4762).  
     
     
         24 . An isolated nucleic acid sequence comprising the nucleic acid sequence shown in SEQ ID NO: 14.  
     
     
         25 . A method of detecting calicivirus nucleic acid sequences in a human tissue sample, the method comprising: 
 contacting the human tissue sample with the nucleic acid sequence shown is SEQ ID NO:1 and fragments thereof; and    detecting calicivirus nucleic acid sequences bound to the nucleic acid sequence.    
     
     
         26 . A method of inhibiting caliciviral replication, comprising: 
 contacting a cell with a therapeutically effective amount of an antisense or antigene molecule, wherein the antisense or antigene molecule comprises a sequences selected from the group consisting of SEQ ID NOS: 1, 3, 4, and fragments thereof.