Humanized antibodies

Abstract

Humanized antibodies that bind ICAM-1 are provided. Antibodies include those selected from: SEQ ID NO:1 and 3 (HumA); SEQ ID NO:5 and 7 (HumB); SEQ ID NO:9 and 11 (HumC); SEQ ID NO:13 and 15 (HumD); SEQ ID NO:17 and 19 (HumE); SEQ ID NO:21 and 23 (HumF); SEQ ID NO:25 and 27 (HumG); SEQ ID NO:29 and 31 (HumH); and SEQ ID NO:33 and 35 (HumI). Subsequences of the humanized antibodies capable of binding an ICAM-1 epitope are also provided. Methods of inhibiting pathogen infection (e.g., HRV) of a cell employing humanized antibodies capable of binding an ICAM-1 epitope are further provided.

What is claimed is:  
     
         1 . A humanized antibody that binds ICAM-1, said antibody selected from: SEQ ID NO:1 and 3 (HumA); SEQ ID NO:5 and 7 (HumB); SEQ ID NO:9 and 11 (HumC); SEQ ID NO:13 and 15 (HumD); SEQ ID NO:17 and 19 (HumE); SEQ ID NO:21 and 23 (HumF); SEQ ID NO:25 and 27 (HumG); SEQ ID NO:29 and 31 (HumH); and SEQ ID NO:33 and 35 (HumI).  
     
     
         2 . A subsequence of the antibody of  claim 1 , said antibody subsequence capable of binding an ICAM-1 epitope.  
     
     
         3 . The humanized antibody of  claim 2 , wherein the antibody subsequence comprises a single chain, Fab, Fab′ or (Fab) 2  fragment.  
     
     
         4 . The humanized antibody of  claim 1 , said antibody having one or more amino acid substitutions, provided that said antibody is capable of binding an ICAM-1 epitope.  
     
     
         5 . A humanized antibody that binds ICAM-1 and inhibits pathogen infection of cells expressing ICAM-1.  
     
     
         6 . The humanized antibody of  claim 5 , said antibody having a protective efficacy at least 2 times greater than the non-humanized antibody.  
     
     
         7 . The humanized antibody of  claim 5 , said antibody having a protective efficacy at least 5 times greater than the non-humanized antibody.  
     
     
         8 . The humanized antibody of  claim 5 , said antibody having a protective efficacy at least 10 times greater than the non-humanized antibody.  
     
     
         9 . The humanized antibody of  claim 5 , said antibody having a protective efficacy at least 20 times greater than the non-humanized antibody.  
     
     
         10 . The humanized antibody of  claim 5 , said antibody having a protective efficacy at least 30 times greater than the non-humanized antibody.  
     
     
         11 . The humanized antibody of  claim 5 , wherein the pathogen is human rhinovirus (HRV).  
     
     
         12 . The humanized antibody of  claim 5 , wherein the pathogen is coxackie A virus, respiratory syncytial virus, or malaria.  
     
     
         13 . The humanized antibody of  claim 5 , wherein the antibody is an intact immunoglobulin molecule comprising 2 full-length heavy chains and 2 full-length light chains.  
     
     
         14 . The humanized antibody of  claim 5 , wherein the antibody is an antibody subsequence that binds to ICAM-1.  
     
     
         15 . The humanized antibody of  claim 14 , wherein the antibody subsequence comprises a single chain, Fab, Fab′ or (Fab) 2  fragment.  
     
     
         16 . The humanized antibody of  claim 5 , wherein the antibody is multispecific or multifunctional.;  
     
     
         17 . The humanized antibody of  claim 5 , wherein the antibody is linked to one or more identical or different antibodies to form a multimer.  
     
     
         18 . The humanized antibody of  claim 17 , wherein the multimer comprises a homo- or hetero-dimer, trimer, or tetramer.  
     
     
         19 . The humanized antibody of  claim 17 , wherein the multimer is formed via a multimerization domain.  
     
     
         20 . The humanized antibody of  claim 19 , wherein the multimerization domain comprises a human amino acid sequence.  
     
     
         21 . The humanized antibody of  claim 19 , further comprising a linker located between the multimerization domain and the antibody.  
     
     
         22 . A humanized antibody that inhibits human rhinovirus (HRV) infection of cells comprising the amino acid sequence set forth in any of SEQ ID NO:1 and 3 (HumA); SEQ ID NO:5 and 7 (HumB); SEQ ID NO:9 and 11 (HumC); SEQ ID NO:13 and 15 (HumD); SEQ ID NO:17 and 19 (HumE); SEQ ID NO:21 and 23 (HumF); SEQ ID NO:25 and 27 (HumG); SEQ ID NO:29 and 31 (HumH); and SEQ ID NO:33 and 35 (HumI); or a subsequence thereof.  
     
     
         23 . The humanized antibody of  claim 22 , wherein the antibody is an immunoglobulin molecule comprising 2 full-length heavy chain polypeptides and 2 full-length light chain polypeptides.  
     
     
         24 . The humanized antibody of  claim 22 , wherein the subsequence comprises a single chain, Fab, Fab′ or (Fab) 2  fragment.  
     
     
         25 . The humanized antibody of  claim 22 , wherein the antibody is linked with other identical or different antibodies to form a multimer.  
     
     
         26 . The humanized antibody of  claim 25 , wherein the multimer comprises a homo- or hetero-dimer, trimer, or tetramer.  
     
     
         27 . The humanized antibody of  claim 25 , wherein the different antibodies are human, humanized or non-human.  
     
     
         28 . A nucleic acid sequence encoding a humanized antibody of  claim 1  or  22  or a subsequence thereof.  
     
     
         29 . An expression cassette comprising the nucleic acid sequence of  claim 28  operably linked to an expression control element.  
     
     
         30 . A vector comprising the nucleic acid sequence of  claim 29 .  
     
     
         31 . The vector of  claim 29 , wherein the nucleic acid sequence is operably linked to an expression control element.  
     
     
         32 . A cell comprising the nucleic acid sequence of  claim 28 .  
     
     
         33 . The cell of  claim 31 , wherein the cell is prokaryotic or eukaryotic.  
     
     
         34 . A pharmaceutical composition comprising a humanized antibody of  claim 1  or  5 , and a pharmaceutically acceptable carrier.  
     
     
         35 . The pharmaceutical composition of  claim 34 , wherein the carrier is compatible with inhalation or nasal delivery to a subject.  
     
     
         36 . A method of inhibiting pathogen infection of a cell comprising contacting a pathogen or a cell with an amount of a humanized antibody of claims  1  or  5 , sufficient to inhibit pathogen infection of the cell.  
     
     
         37 . The method of  claim 36 , wherein the cell is present in a subject.  
     
     
         38 . The method of  claim 37 , wherein the cell is an epithelial cell.  
     
     
         39 . The method of  claim 37 , wherein the cell expresses ICAM-1.  
     
     
         40 . A method of inhibiting HRV infection of a cell comprising contacting HRV or a cell susceptible to HRV infection with an amount of a humanized antibody of  claim 21  effective to inhibit HRV infection of the cell.  
     
     
         41 . The method of  claim 40 , wherein the cell is present in a subject.  
     
     
         42 . The method of  claim 41 , wherein the subject has or is at risk of having asthma.  
     
     
         43 . The method of  claim 40 , wherein the antibody binds to an antigen present on the surface of the cell.  
     
     
         44 . The method of  claim 40 , wherein the cell expresses ICAM-1.  
     
     
         45 . The method of  claim 40 , wherein the cell is an epithelial cell.  
     
     
         46 . The method of  claim 40 , wherein the humanized antibody is administered locally.  
     
     
         47 . The method of  claim 40 , wherein the humanized antibody is administered via inhalation or intranasaly.  
     
     
         48 . A method of inhibiting HRV infection, inhibiting HRV progression or treating HRV infection of a subject comprising administering to a subject having or at risk of having HRV infection an amount of a humanized antibody of  claim 21  effective to inhibit, inhibit progression or treat HRV infection of the subject.  
     
     
         49 . The method of  claim 48 , wherein the humanized antibody is administered locally.  
     
     
         50 . The method of  claim 48 , wherein the humanized antibody is administered via inhalation or intranasaly.  
     
     
         51 . The method of  claim 48 , wherein the subject has or is at risk of having asthma.  
     
     
         52 . The method of  claim 48 , wherein the subject is a newborn or between the ages of 1 to 5, 5 to 10 or 10 to 18.  
     
     
         53 . A method of decreasing or inhibiting one or more symptoms of the common cold in a subject comprising administering to a subject having a common cold an amount of a humanized antibody of  claim 21  effective to decrease or inhibit one or more symptoms of the common cold in the subject.  
     
     
         54 . The method of  claim 53 , wherein the humanized antibody is administered locally.  
     
     
         55 . The method of  claim 53 , wherein the humanized antibody is administered via inhalation or intranasaly.  
     
     
         56 . The method of  claim 53 , wherein the subject has or is at risk of having asthma.  
     
     
         57 . The method of  claim 53 , wherein the subject is a newborn or between the ages of 1 to 5, 5 to 10 or 10 to 18.