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(12) Patent Application Publication (10) Pub. No.: US 2013/0136768 A1 Picker Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2013/0136768 A1 Picker Et Al US 2013 O136768A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2013/0136768 A1 Picker et al. (43) Pub. Date: May 30, 2013 (54) RECOMBINANT HCMV AND RHCMV Publication Classification VECTORS AND USES THEREOF (51) Int. Cl. Applicant: Oregon Health & Science University, CI2N 5/869 (2006.01) (71) A6139/08 (2006.01) Portland, OR (US) A6139/00 (2006.01) A639/2 (2006.01) (72) Inventors: Louis Picker, Portland, OR (US); Jay A. A 6LX39/275 (2006.01) Nelson, Lake Oswego, OR (US); Klaus A639/2I (2006.01) Frueh, Portland, OR (US); Michael A. A639/13 (2006.01) Jarvis, Portland, OR (US); Scott G. A639/45 (2006.01) Hansen, Portland, OR (US) (52) U.S. Cl. CPC .............. CI2N 15/869 (2013.01); A61K 39/13 (2013.01); A61 K39/08 (2013.01); A61 K Assignee: Oregon Health & Science University, 39/145 (2013.01); A61 K39/12 (2013.01); (73) A61K 39/275 (2013.01); A6IK 39/21 Portland, OR (US) (2013.01); A61 K39/00II (2013.01) USPC ..................................... 424/199.1: 435/320.1 (21) Appl. No.: 13/694,280 (57) ABSTRACT Described herein are recombinant rhesus cytomegalovirus (RhCMV) and human cytomegalovirus (HCMV) vectors encoding heterologous antigens, such as pathogen-specific (22) Filed: Nov. 14, 2012 antigens or tumor antigens. The recombinant vectors elicit and maintain high level cellular and humoral immune responses specific for the heterologous antigen. The recom Related U.S. Application Data binant RhCMV and HCMV vectors may be used, for example, for the treatment or prevention of infectious disease (63) Continuation-in-part of application No. PCT/US 11/ or cancer. In some examples, the recombinant RhCMV or 36657, filed on May 16, 2011, which is a continuation HCMV vectors may include deletions in genes encoding in-part of application No. PCT/US 11/29930, filed on immunomodulatory proteins. In some examples, the recom Mar. 25, 2011. binant RhCMV or HCMV vectors may be deficient or impaired in their ability to replicate within a cell, disseminate (60) Provisional application No. 61/376,911, filed on Aug. within the host or spread among hosts by including a deletion 25, 2010, provisional application No. 61/334,976, in one or more genes essential or augmenting for CMV rep filed on May 14, 2010. lication, dissemination or spread. Patent Application Publication May 30, 2013 Sheet 1 of 40 US 2013/O136768A1 % responding of total PBT cells: CD4+ A CD8+ 4 CMV Status: - us % responding of memory PBT cells: CD4 CD8t CMV Status: - s FIG. 1 Patent Application Publication May 30, 2013 Sheet 2 of 40 US 2013/O136768A1 90Taen 68"in Awooqoqwa CD4+ T cells CD8-- Cels §3 94"In 84"Tn CD8+ T cells FIG. 2 Patent Application Publication May 30, 2013 Sheet 3 of 40 US 2013/O136768A1 s Patent Application Publication May 30, 2013 Sheet 4 of 40 US 2013/O136768A1 A d it. t be ce e pit al e W . ON - 0(...) e re Sapuods. , Patent Application Publication May 30, 2013 Sheet 5 of 40 US 2013/O136768A1 SêN:S??001+8°CIO Patent Application Publication May 30, 2013 Sheet 6 of 40 US 2013/013.6768A1 a put 23 Sodoo Patent Application Publication May 30, 2013 Sheet 7 of 40 US 2013/O136768A1 RhCMV WLOxp SVimac239gag cassette F.G. 7 Patent Application Publication May 30, 2013 Sheet 8 of 40 US 2013/O136768A1 D co 21. Sa O t N s s O e tf e ve E Y SN &N 3ve n c 75 a it v. ix. see is: Yw 50 - FIG. 8 Patent Application Publication May 30, 2013 Sheet 9 of 40 US 2013/O136768A1 Pre-inoculation a-a-a-e- - noSA- Ad -whole CMV 99.5aq 5mers ... : 0.02% --TNF--- Day 7 post-first RhCMV(gag) re-infection O Ag' whole CMW gag 15mers F.G. 9 Patent Application Publication May 30, 2013 Sheet 10 of 40 US 2013/O136768A1 CD4 T cellgated FIG 10 Patent Application Publication May 30, 2013 Sheet 11 of 40 US 2013/O136768A1 00€0s,00zOst0010s0-00s00?ost09 9960;&# 0 0 w rt a rs it - re. (s ce as es e a e (a.k.st Autist) up bills y, Patent Application Publication May 30, 2013 Sheet 12 of 40 US 2013/013.6768A1 5 O O. 6. 0. O St 100 50 200 250 300 time (days) post initial re-infection with RCAYOgag) FIG. 12 Patent Application Publication May 30, 2013 Sheet 13 of 40 US 2013/013.6768 A1 Frequencies of gag-specific T cells 3 2: SIV(Anef) vaccinees 2. 5 RhCMV(gag) vaccinees ahs 0. is: CD4+ CD8+ CD4+ CD8+ Lung Lavage Blood FIG. 13 Patent Application Publication May 30, 2013 Sheet 14 of 40 US 2013/O136768A1 CD3t Small Lymphocyte Gated ; 3 FSC TNFox (PECY-7) TNFa. PE CY-7) CD4 (AmCyan) CD4 T-Cell Gated FIG. 14 Patent Application Publication May 30, 2013 Sheet 15 of 40 US 2013/O136768A1 RCMV BAC FIG. 15 Patent Application Publication May 30, 2013 Sheet 16 of 40 US 2013/O136768A1 RhCMW RS2 R1S6 X cro H 3 S. 9 P h Dox - Dox gll NA2 FIG. 16 Patent Application Publication May 30, 2013 Sheet 17 of 40 US 2013/O136768A1 RCMV X Rhio H + Dox - DOX Ns A2 s PS or Y a 2. : is FIG. 17 Patent Application Publication May 30, 2013 Sheet 18 of 40 US 2013/O136768A1 RhCMV FIG. 18 Patent Application Publication May 30, 2013 Sheet 19 of 40 US 2013/O136768A1 RhCMV BAC a N EIT Recorn.& -- l s V Flp Recom. t t FIG. 19 Patent Application Publication May 30, 2013 Sheet 20 of 40 US 2013/O136768A1 & sh r eSuodsej Auoueu% Patent Application Publication May 30, 2013 Sheet 21 of 40 US 2013/O136768A1 miR-142-3p MEF C-21 BMDC PB DC FIG. 21 Patent Application Publication May 30, 2013 Sheet 22 of 40 US 2013/O136768A1 S.$ 5 days123456, post infection 8 FIG. 22 Patent Application Publication May 30, 2013 Sheet 23 of 40 US 2013/O136768A1 106 C-21 E3-142 s 0 1 2 3 4 5 6 7 8 S days post infection FIG. 23 Patent Application Publication May 30, 2013 Sheet 24 of 40 US 2013/O136768A1 15 3T3 is 20 (rg25 Day 1 9:30 Day 5 s 35 (9 4. O IE3-015 E3-142 E3-015 FIG. 24 Patent Application Publication May 30, 2013 Sheet 25 of 40 US 2013/O136768A1 s 35 Ego 5 E3-142 Esos TIE-42 FIG. 25 Patent Application Publication US 2013/O136768A1 Patent Application Publication May 30, 2013 Sheet 27 of 40 US 2013/O136768A1 Patent Application Publication May 30, 2013 Sheet 28 of 40 US 2013/O136768A1 88.88:38.88&. Patent Application Publication May 30, 2013 Sheet 29 of 40 US 2013/O136768A1 MCMVIZEBOWNP NCNWPSA MCMV WT C (5A1, mouse 1) 901 4.08 3 C 4 s TNF"re Q.54 “TNFr"saw-----ry-rri---- 0. l FG, 28C Patent Application Publication May 30, 2013 Sheet 30 of 40 US 2013/O136768A1 3. 5 3. O 2. 5 2. O 1 5 1 O 0. 5 4. Time (weeks) FIG. 29 Patent Application Publication May 30, 2013 Sheet 31 of 40 US 2013/O136768A1 y inne Weeks) O O 4. 8 Civ2EBOVA MichiwitzebowAt t -- 6 acAW26Bow (D1) CMVizesgypt Ne -- s McAv wt. McMy WT s J. -- gigka s 4 Harvast inoculation boost pro-OOO5 vs. MCMV WT) EBO PSA M38 M45 FIG. 30 Patent Application Publication May 30, 2013 Sheet 32 of 40 US 2013/O136768A1 -go-Mock -- MCMV WT -0-MCMVIZEBOV (A1) -0-MCMVIZEBOv(D1) -(- WSWAGIZEBOWGP 4. 6 8 10 28 Time after Challenge (Days) A. 6 8 10 12 14 Time after Challenge (Days) (1.7x106) (3.3x106) Oe WSWAG MCMV MCAVE MCMV WT Mock ZEBOVGP ZEBOVA1) ZEBOW(D1) FIG. 31 Patent Application Publication May 30, 2013 Sheet 33 of 40 US 2013/O136768A1 Patent Application Publication May 30, 2013 Sheet 34 of 40 US 2013/O136768A1 go O 2 4. 6 8 10 12 Time (days) FIG. 33 Patent Application Publication May 30, 2013 Sheet 35 of 40 US 2013/O136768A1 ? §§. Patent Application Publication May 30, 2013 Sheet 36 of 40 US 2013/O136768A1 Patent Application Publication May 30, 2013 Sheet 37 of 40 US 2013/O136768A1 Multi-step Growth Curve of RhCMWW and RhCMVARh10 on telomerized Rhesus Fibroblasts -0-RhCMV WT on trfs was RhCMVARh 10 retanefon tRFs Days post infection Multi-step Growth Curve of RhCMV WT and RhCMVARh110 on telomerized Rhesus Fibroblasts+pp71HA (tet) s --RhCMVWT on tPs + pp.71 (tet) --RhCMV ARh110 on trfs+ pp.71 (tet) re-RhCMV ARh 110 retanefon tRFs+pp71 (tet) 9 Days post infection FIG. 36 Patent Application Publication May 30, 2013 Sheet 38 of 40 US 2013/O136768A1 3-833X.3888 : : :8.: pig. 3 Patent Application Publication May 30, 2013 Sheet 39 of 40 US 2013/O136768A1 six. 38 Patent Application Publication May 30, 2013 Sheet 40 of 40 US 2013/O136768A1 US 2013/013.6768 A1 May 30, 2013 RECOMBINANT HCMV AND RHCMV with non-replicating viral vector expression systems (prime VECTORS AND USES THEREOF boost) (Barouch, D. H. 2008. Challenges in the development of an HIV-1 vaccine. Nature 455:613-619). One of the goals RELATED APPLICATIONS AND of this project is to generate an effective and safe HIV vaccine INCORPORATION BY REFERENCE using HCMV as a vaccine vector. 0001. This application is a continuation-in-part applica 0006 CMV is an ubiquitous virus and a member of the tion of international patent application PCT/US2011/036657 beta subclass of the herpesvirus family.
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