Antibody cytokine engrafted compositions and methods of use for immunoregulation

Abstract

The present disclosure provides antibody cytokine engrafted proteins that bind to and stimulate intracellular signaling through interleukin 10 receptor. Provided antibody cytokine engrafted proteins find use in enhancing anti-inflammatory cell responses, and reducing pro-inflammatory effects in the treatment, amelioration and prophylaxis of immune related disorders. Additionally provided are polynucleotides and vectors that encode antibody cytokine engrafted proteins and host cells capable of producing antibody cytokine engrafted proteins, as well as methods of combining antibody cytokine engrafted proteins with other therapeutics in treating immune related disorder.

1 - 36 . (canceled) 
     
     
         37 . A method of activating monocytes in a patient in need thereof, comprising administering an antibody cytokine engrafted protein comprising:
 (a) a heavy chain variable region (VH), comprising Complementarity Determining Regions (CDR) HCDR1, HCDR2, HCDR3; and   (b) a light chain variable region (VL), comprising LCDR1, LCDR2, LCDR3; and   (c) an Interleukin 10 (IL10) monomeric molecule engrafted into a CDR of the VH or the VL.   
     
     
         38 . The method of  claim 37 , wherein monocytes are activated and T cells or NK cells are not activated. 
     
     
         39 . The method of  claim 37 , wherein administration of the antibody cytokine engrafted protein reduces TNFα production. 
     
     
         40 - 46 . (canceled) 
     
     
         47 . The method of  claim 37 , wherein the IL10 monomeric molecule is engrafted into a heavy chain CDR. 
     
     
         48 . The method of  claim 47 , wherein the heavy chain CDR is HCDR1, HCDR2 or HCDR3. 
     
     
         49 . The method of  claim 48 , wherein the IL10 monomeric molecule is engrafted into the HCDR1. 
     
     
         50 . The method of  claim 37 , wherein the IL10 monomeric molecule is engrafted into a light chain CDR. 
     
     
         51 . The method of  claim 50 , wherein the light chain CDR is LCDR1, LCDR2 or LCDR3. 
     
     
         52 . The method of  claim 51 , wherein the IL10 monomeric molecule is engrafted into the LCDR1. 
     
     
         53 . The method of  claim 37 , wherein the antibody cytokine engrafted protein is capable of less activation of T cells or NK cells when compared to recombinant human IL10. 
     
     
         54 . The method of  claim 37 , wherein the antibody cytokine engrafted protein has a longer half-life than rhIL10. 
     
     
         55 . The method of  claim 37 , wherein the IL10 monomeric molecule consists of SEQ ID NO:209. 
     
     
         56 . The method of  claim 37 , wherein the antibody cytokine engrafted protein further comprises an IgG class antibody heavy chain. 
     
     
         57 . The method of  claim 56 , wherein the IgG class heavy chain is selected from IgG1, IgG2, or IgG4. 
     
     
         58 . The method of  claim 37 , wherein the binding specificity of the CDRs to a target protein is reduced in the presence of the engrafted IL10 monomeric molecule. 
     
     
         59 . The method of  claim 37 , wherein the binding specificity of the CDRs to a target protein is retained in the presence of the engrafted IL10 monomeric molecule. 
     
     
         60 . The method of  claim 59 , wherein the binding specificity of the CDRs is distinct from the cytokine-receptor binding specificity of the IL10 monomeric molecule. 
     
     
         61 . The method of  claim 37 , wherein the antibody cytokine engrafted protein comprises:
 (i) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO: 193, (b) a HCDR2 of SEQ ID NO:194, (c) a HCDR3 of SEQ ID NO:195 and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:196, (e) a LCDR2 of SEQ ID NO:197, and (f) a LCDR3 of SEQ ID NO:198;   (ii) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:97, (b) a HCDR2 of SEQ ID NO:98, (c) a HCDR3 of SEQ ID NO:99; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:100, (e) a LCDR2 of SEQ ID NO:101, and (f) a LCDR3 of SEQ ID NO:102;   (iii) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:1, (b) a HCDR2 of SEQ ID NO:2, (c) a HCDR3 of SEQ ID NO:3; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:4, (e) a LCDR2 of SEQ ID NO:5, and (f) a LCDR3 of SEQ ID NO:6;   (iv) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:17, (b) a HCDR2 of SEQ ID NO:18, (c) a HCDR3 of SEQ ID NO:19; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:20, (e) a LCDR2 of SEQ ID NO:21, and (f) a LCDR3 of SEQ ID NO:22;   (v) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:33, (b) a HCDR2 of SEQ ID NO:34, (c) a HCDR3 of SEQ ID NO:35; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:36, (e) a LCDR2 of SEQ ID NO:37, and (f) a LCDR3 of SEQ ID NO:38;   (vi) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:49, (b) a HCDR2 of SEQ ID NO:50, (c) a HCDR3 of SEQ ID NO:51; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:52, (e) a LCDR2 of SEQ ID NO:53, and (f) a LCDR3 of SEQ ID NO:54;   (vii) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:65, (b) a HCDR2 of SEQ ID NO:66, (c) a HCDR3 of SEQ ID NO:67; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:68, (e) a LCDR2 of SEQ ID NO:69, and (f) a LCDR3 of SEQ ID NO:70;   (viii) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:81, (b) a HCDR2 of SEQ ID NO:82, (c) a HCDR3 of SEQ ID NO:83; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:84, (e) a LCDR2 of SEQ ID NO:85, and (f) a LCDR3 of SEQ ID NO:86;   (ix) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:113, (b) a HCDR2 of SEQ ID NO:114, (c) a HCDR3 of SEQ ID NO:115; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:116, (e) a LCDR2 of SEQ ID NO:117, and (f) a LCDR3 of SEQ ID NO:118;   (x) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:129, (b) a HCDR2 of SEQ ID NO:130, (c) a HCDR3 of SEQ ID NO:131; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:132, (e) a LCDR2 of SEQ ID NO:133, and (f) a LCDR3 of SEQ ID NO:134;   (xi) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:145, (b) a HCDR2 of SEQ ID NO:146, (c) a HCDR3 of SEQ ID NO:147; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:148, (e) a LCDR2 of SEQ ID NO:149, and (f) a LCDR3 of SEQ ID NO:150;   (xii) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:161, (b) a HCDR2 of SEQ ID NO:162, (c) a HCDR3 of SEQ ID NO:163; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:164, (e) a LCDR2 of SEQ ID NO:165, and (f) a LCDR3 of SEQ ID NO:166;   (xiii) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:177, (b) a HCDR2 of SEQ ID NO:178, (c) a HCDR3 of SEQ ID NO:179; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:180, (e) a LCDR2 of SEQ ID NO:181, and (f) a LCDR3 of SEQ ID NO:182;   (xiv) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:210, (b) a HCDR2 of SEQ ID NO:211, (c) a HCDR3 of SEQ ID NO:212; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:213, (e) a LCDR2 of SEQ ID NO:214, and (f) a LCDR3 of SEQ ID NO:215; or   (xv) a heavy chain variable region that comprises (a) a HCDR1 of SEQ ID NO:226, (b) a HCDR2 of SEQ ID NO:227, (c) a HCDR3 of SEQ ID NO:228; and a light chain variable region that comprises: (d) a LCDR1 of SEQ ID NO:229, (e) a LCDR2 of SEQ ID NO:230, and (f) a LCDR3 of SEQ ID NO:231.   
     
     
         62 . The method of  claim 37 , wherein the antibody cytokine engrafted protein comprises:
 (i) a heavy chain variable region (VH) that comprises SEQ ID NO:205, and a light chain variable region (VL) that comprises SEQ ID NO:206;   (ii) a heavy chain variable region (VH) that comprises SEQ ID NO: 109, and a light chain variable region (VL) that comprises SEQ ID NO: 110;   (iii) a heavy chain variable region (VH) that comprises SEQ ID NO:13, and a light chain variable region (VL) that comprises SEQ ID NO:14;   (iv) a heavy chain variable region (VH) that comprises SEQ ID NO:29, and a light chain variable region (VL) that comprises SEQ ID NO:30;   (v) a heavy chain variable region (VH) that comprises SEQ ID NO:45, and a light chain variable region (VL) that comprises SEQ ID NO:46;   (vi) a heavy chain variable region (VH) that comprises SEQ ID NO:61, and a light chain variable region (VL) that comprises SEQ ID NO:62;   (vii) a heavy chain variable region (VH) that comprises SEQ ID NO:77, and a light chain variable region (VL) that comprises SEQ ID NO:78;   (viii) a heavy chain variable region (VH) that comprises SEQ ID NO:93, and a light chain variable region (VL) that comprises SEQ ID NO:94;   (ix) a heavy chain variable region (VH) that comprises SEQ ID NO:125, and a light chain variable region (VL) that comprises SEQ ID NO:126;   (x) a heavy chain variable region (VH) that comprises SEQ ID NO:141, and a light chain variable region (VL) that comprises SEQ ID NO:142;   (xi) a heavy chain variable region (VH) that comprises SEQ ID NO:157, and a light chain variable region (VL) that comprises SEQ ID NO:158;   (xii) a heavy chain variable region (VH) that comprises SEQ ID NO:173, and a light chain variable region (VL) that comprises SEQ ID NO:174;   (xiii) a heavy chain variable region (VH) that comprises SEQ ID NO:189, and a light chain variable region (VL) that comprises SEQ ID NO:190;   (xiv) a heavy chain variable region (VH) that comprises SEQ ID NO:222, and a light chain variable region (VL) that comprises SEQ ID NO:223; or   (xv) a heavy chain variable region (VH) that comprises SEQ ID NO:238, and a light chain variable region (VL) that comprises SEQ ID NO:239.