Compositions and methods for antibody delivery

Abstract

To provide, as strategies to administer therapeutic antibodies, recombinant nucleic acids encoding antibodies for use in viruses, compositions, formulations, and medicaments, and / or methods suitable for delivering antibodies to one or more sites and / or tissues of a subject.SOLUTION: Certain aspects of the present disclosure relate to a replication-deficient herpes simplex virus that comprises recombinant herpes simplex virus genome comprising one or more polynucleotides encoding an antibody, wherein the antibody is an antibody fragment, the antibody fragment is a Fab, Fab' Fab'-SH, F(ab')2, Fv, scFv, or scFv-Fc fragment, and the antibody is a murine antibody, a chimeric antibody, a humanized antibody, a human antibody, a monoclonal antibody or a multispecific antibody.SELECTED DRAWING: Figure 1-8

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application Serial No. 62 / 692,514, filed June 29, 2018, and U.S. Provisional Application Serial No. 62 / 713,066, filed August 1, 2018, each of which is incorporated herein by reference in its entirety.

[0002] Submitting a sequence listing as an ASCII text file The following submission in an ASCII text file is incorporated herein by reference in its entirety: Sequence Listing in Computer Readable Form (CRF) (Filename: 761342000840SEQLIST.txt, Date Recorded: June 27, 2019, Size: 764KB).

[0003] FIELD OF THE INVENTION The present disclosure relates in part to recombinant nucleic acids (e.g., recombinant herpesvirus genomes) comprising one or more polynucleotides encoding antibodies (or portions thereof), to viruses (e.g., herpesviruses) comprising the recombinant nucleic acids, to compositions comprising the recombinant nucleic acids and / or viruses, to methods of their use (e.g., methods for viral-mediated localized delivery and expression of encoded antibodies), and to articles of manufacture or kits thereof. [Background technology]

[0004] background In recent years, therapeutic antibodies have become one of the most commercially successful classes of biopharmaceuticals. While antibodies have successfully treated several major diseases, including autoimmune diseases, cardiovascular and infectious diseases, cancer, and inflammation, systemic administration of therapeutic antibodies has many functional limitations, including poor pharmacokinetics and tissue accessibility. Furthermore, systemic exposure to certain antibodies has been shown to suppress the immune system, placing patients at significant risk for infections and other complications. Therefore, alternative strategies for administering therapeutic antibodies to patients in need are needed.

[0005] All references cited herein, including patent applications, patent publications, non-patent literature, and UniProtKB / Swiss-Prot accession numbers, are incorporated by reference in their entirety as if each individual reference was specifically and individually indicated to be incorporated by reference. Summary of the Invention

[0006] overview In some embodiments, provided herein are recombinant nucleic acids (e.g., recombinant herpesvirus genomes) encoding antibodies (e.g., full-length antibodies, antibody fragments, etc.) for use in viruses (e.g., herpesviruses), compositions, formulations, drugs, and / or useful methods for delivering antibodies to one or more sites and / or tissues of a subject (e.g., locally, intra-articularly, intravitreally, etc.). The inventors have shown that the recombinant attenuated viruses described herein are capable of 1) expression of both full-length antibodies and antibody fragments (scFv-Fc), 2) encoding and delivering murine, chimeric, and fully human antibodies (of various IgG isotypes), and 3) directing dose-dependent secretion of the encoded antibodies from human cells, with the antibodies being functional (see, e.g., Example 2). Furthermore, the inventors have shown that the viruses described herein can be used to successfully express their encoded antibodies in vivo following local administration (see, e.g., Example 3). Furthermore, the inventors have demonstrated that the viruses described herein can be used to successfully express therapeutic antibodies for treating one or more symptoms of inflammatory skin conditions (atopic dermatitis) following topical administration (see, e.g., Example 3). Without wishing to be bound by theory, it is believed that the recombinant nucleic acids (e.g., recombinant viral genomes), viruses, pharmaceutical compositions, drugs, and / or methods described herein provide a novel system for delivering therapeutic antibodies to patients. Specifically, without wishing to be bound by theory, it is believed that the recombinant herpesviruses described herein provide a unique system for locally administering therapeutic antibodies to a subject with the objectives of: 1) improving antibody pharmacokinetics at the site of interest; 2) improving tissue accessibility and / or penetration of the antibody; 3) reducing the total dose of antibody administered to a subject; 4) providing a minimally invasive or non-invasive method of administering the antibody to a subject; and / or 5) reducing or eliminating a subject's systemic exposure to the antibody (e.g., avoiding one or more side effects (e.g., global immunosuppression) observed following systemic administration of certain antibodies).

[0007] Accordingly, certain aspects of the present disclosure relate to a recombinant herpesvirus genome comprising one or more polynucleotides encoding an antibody. In some embodiments, the antibody is an antibody fragment. In some embodiments, the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, scFv, or scFv-Fc fragment. In some embodiments, the antibody fragment is an scFv-Fc fragment. In some embodiments, the scFv-Fc comprises the Fc region of an IgG antibody (e.g., the Fc region of an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody). In some embodiments, the scFv-Fc comprises the Fc region of an IgG1 antibody. In some embodiments, the scFv-Fc comprises the Fc region of an IgG4 antibody. In some embodiments, the antibody is a full-length antibody. In some embodiments that may be combined with any of the preceding embodiments, the antibody is a murine antibody, a chimeric antibody, a humanized antibody, a human antibody, a monoclonal antibody, or a multispecific antibody. In some embodiments that can be combined with any of the preceding embodiments, the antibody is an IgA antibody, an IgD antibody, an IgE antibody, an IgG antibody, or an IgM antibody. In some embodiments that can be combined with any of the preceding embodiments, the antibody is an IgG antibody. In some embodiments, the IgG antibody is an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. In some embodiments, the IgG antibody is an IgG1 antibody. In some embodiments, the IgG antibody is an IgG4 antibody. In some embodiments that can be combined with any of the preceding embodiments, the antibody is an agonist antibody or an antagonist antibody. In some embodiments, the antibody is an agonist antibody. In some embodiments, the antibody is an antagonist antibody.

[0008] In some embodiments that may be combined with any of the preceding embodiments, the antibody comprises a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3, wherein HVR-H1 comprises a sequence selected from SEQ ID NOs: 1-59, HVR-H2 comprises a sequence selected from SEQ ID NOs: 60-122, and / or HVR-H3 comprises a sequence selected from SEQ ID NOs: 123-185. In some embodiments that may be combined with any of the preceding embodiments, the antibody comprises a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3, wherein HVR-L1 comprises a sequence selected from SEQ ID NOs: 186-242, HVR-L2 comprises a sequence selected from SEQ ID NOs: 243-294, and / or HVR-L3 comprises a sequence selected from SEQ ID NOs: 295-354.

[0009] In some embodiments that can be combined with any of the preceding embodiments, the antibody comprises a heavy chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 355-419 or SEQ ID NOs: 614-865. In some embodiments that can be combined with any of the preceding embodiments, the antibody comprises a light chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 420-482 or SEQ ID NOs: 866-1116. In some embodiments that may be combined with any of the preceding embodiments, the antibody comprises: (a) a heavy chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs:355-419 or SEQ ID NOs:614-865; and (b) a light chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs:420-482 or SEQ ID NOs:866-1116.

[0010] In some embodiments that may be combined with any of the preceding embodiments, the antibody is abagovomab, abciximab, abituzumab, abrezekimab, abrilumab, actoxumab, adalimumab, adecatumumab, aducanumab, afasevikumab, afelimomab, afutuzumab, alacizumab, alemtuzumab, alirocumab, altumomab, amatuximab, anatumomab, andecaliximab, alametuzumab ... Netumab, anifrolumab, anrukinzumab, apolizumab, aprutumab, arcitumomab, asclinical steroids, aselizumab, atezolizumab, atinumab, atolimumab, avelumab, azintuxizumab, bapineuzumab, basiliximab, bavituximab, bectumomab, begelomab, belantamab, belimumab, bemarituzumab, berimumab, bemarituzumab ab), benralizumab, berlimatoxumab, bersanlimab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab, biciromab, bimagrumab, bimekizumab, viltamimab, bivatuzumab, bleselumab, blinatumomab, brontuvetomab, brosozumab, bococizumab, brazikumab, brentuximab, briakinumab, brodalumab, brolucizumab, brontiximab burosumab, cabilalizumab, camidanlumab, camrelizumab, canakinumab, cantuzumab, caplacizumab, capromab, carlumab, carotuximab, catumaxomab, cedelizumab, cemiplimab, sergituzumab, certolizumab, cetrelimab, cetuximab, civisatamab, sitatuzumab, cizutumumab, clazakizumab, clenoliximab, clivatuzumab, codrituzumab, cofetuzumab, coltuximab, conatumumab, concizumab,Cosfrobiximab, crenezumab, crizanlizumab, clotedumab, cusatuzumab, dacetuzumab, daclizumab, dalotuzumab, dapirolizumab, daratumumab, dectrekumab, demcizumab, denintuzumab, denosumab, depatuxizumab, derlotuximab, detumomab, desamizumab, dinutuximab, diridavumab, domagrozumab, dorlimomab dorlimomab), drozitumab, durigotuzumab, dupilumab, durvalumab, dusigizumab, duvortuxizumab, ecromeximab, eculizumab, edovacomab, edrecolomab, efalizumab, efangumab, eldelumab, elezanumab, elgemtumab, elotuzumab, elsilimomab, emactuzumab, emapalumab, emibetuzumab, emicizumab, enapotamab, enabatuzumab, Enfortumab, enlimomab, enoblitzumab, enokizumab, enoticumab, encituximab, epitumomab, epratuzumab, eptinezumab, erenumab, erlizumab, ertumaxomab, etaracizumab, etigilimab, etrolizumab, evinacumab, evolocumab, exbivirumab, fanolesomab, faralimomab, faricimab, farletuzumab, fasinumab, felvizumab, fezakinumab, filipin fibatuzumab, ficlatuzumab, figitumumab, firivumab, framvotumab, fretikumab, flotetuzumab, fontolizumab, foralumab, foravirumab, fremanezumab, fresolimumab, furunvetomab, furanumab, futuximab, galcanezumab, galiximab, gancotamab, ganitumab, gantenerumab, gatipotuzumab, gavilimomab, gezivumab, gemtuzumab, gevokizumab,Gilvetmab, gimsilumab, girentuximab, glembatumumab, golimumab, gomiliximab, goslanemab, guselkumab, ianalumab, ibalizumab, ibritumomab, icrucumab, idarucizumab, ifavotuzumab, igovomab, iradatuzumab, imalumab, imaprelimab, imuciromab, imgatuzumab, inlacumab, indatuximab, indusatumab, Inebilizumab, Inflectra, infliximab, intetumumab, inolimomab, inotuzumab, ipilimumab, iratumumab, isatuximab, iscalimab, istiratumab, itolizumab, ixekizumab, keliximab, labetuzumab, lacnotuzumab, ladiratuzumab, lampalizumab, lanadelumab, landgrozumab, laprituximab, ralcabiximab, lebrikizumab, lemalesomab, and lendalizumab (lendalizumab), lenvervimab, lenzilumab, lerdelimumab, leronlimab, lesofabumab, letolizumab, lexatumumab, ribivirumab, rifastuzumab, ligelizumab, loncastuximab, rosatuxizumab, rilotomab, lintuzumab, lirilumab, rodelcizumab, lokivetmab, lorvotuzumab, lucatumumab, lulizumab umab), lumiliximab, lumuletuzumab, rupartumab, lutikizumab, mapatumumab, marjetuximab, marstacimab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minretumomab, mirikizumab, mirvetuximab, mitumomab, modotuximab, mogamulizumab, monalizumab, morolimumab, mosunetuzumab, motavizumab, moxetumomab,Nacolomab, namilumab, naptumomab, naratuximab, narutuximab, natalizumab, navicixizumab, navivumab, naxitamab, nebacumab, necitumumab, nemolizumab, nerelimomab, nesbacumab, netakimab, nimotuzumab, nirsevimab, nivolumab, nofetumomab, obilutoxaximab, occaratuzumab, ocrelizumab, ozlimomab, ofatumumab, olaratumab, orexin lumab, orendalizumab, olokizumab, omalizumab, onartuzumab, ontuxizumab, onvatilimab, opicinumab, oportuzumab, oregovomab, orticumab, otelixizumab, otilimab, otlertuzumab, oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, pamrevlumab, panitumumab, pancomab, panobacumab, palsatuzumab, pascolizumab, pasotuximab xizumab, pateclizumab, patritumab, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab, pintumomab, placuramb, prosalizumab, pogalizumab, polatuzumab, ponezumab, polgabiximab, prasinezumab, prezalizumab, priliximab, pritoxaximab ab), pritumumab, kirusumab, racotumomab, radretumab, rafivirumab, ralpancizumab, ramucirumab, ranevetomab, ranibizumab, raxibacumab, ravagalimab, ravuturizumab, refanezumab, regavirumab, lemtolumab, reslizumab, rilotumumab, rinucumab, risankizumab, rituximab,Rivazumab, lobatumumab, rolezumab, romilukimab, romozozumab, lontalizumab, rosmantuzumab, rovalpituzumab, rovelizumab, rozanolixizumab, ruplizumab, sacituzumab, samalizumab, samrotamab, sapelizumab, sarilumab, satralizumab, satumomab, secukinumab, selicrelumab, seribantumab, setoxaximab, setrusumab, sevilumab sibrotuzumab, sifalimumab, siltuximab, simtuzumab, siplizumab, sirtratumab, sirkumab, sofituzumab, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab, sulesomab, sputumumab, stimulimab, suvizumab, subratoxumab, tabalumab, tacatuzumab, tadocizumab, talacotaxumab tuzumab, talizumab, tamtubetmab, tanezumab, taplitumomab, talexuzumab, tabolimab, tefibazumab, telimomab, telisotuzumab, tenatumomab, teneliximab, teplizumab, tepoditamab, teprotumumab, tesidolumab, tetulomab, tezepelumab, tiburizumab, tildrakizumab, tigatuzumab, timigituzumab imigutuzumab, timolumab, tiragotumab, tislelizumab, tisotumab, tocilizumab, tomuzotuximab, toralizumab, tosatoxumab, tositumomab, tobetumab, tralokinumab, trastuzumab, tregalizumab, tremelimumab, treboglumab, tucotuzumab, tuvilumab, ublituximab, urocuplumab, urelumab, urtoxazumab, ustekinumab, utomilumab,Vadastuximab, vanalimab, bundletuzumab, vanticizumab, vanucizumab, vapaliximab, varisacumab, varlilumab, vatelizumab, vedolizumab, veltuzumab, bepalimomab (vepa, limomab, besencumab, visilizumab, bovalilizumab, volociximab, bonlerolizumab, bopratelimab, borsetuzumab, votumumab, bunakizumab, xentuzumab, zalutumumab, zanolimumab, zatuximab, zenocutuzumab, ziralimumab, zolbetuximab, and zolimomab.

[0011] In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpesvirus genome is replication-competent. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpesvirus genome is replication-deficient. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpesvirus genome is selected from a recombinant herpes simplex virus genome, a recombinant varicella-zoster virus genome, a recombinant human cytomegalovirus genome, a recombinant herpesvirus 6A genome, a recombinant herpesvirus 6B genome, a recombinant herpesvirus 7 genome, a recombinant Kaposi's sarcoma-associated herpesvirus genome, and any derivative thereof.

[0012] In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes virus genome is a recombinant herpes simplex virus genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome, a recombinant herpes simplex virus type 2 (HSV-2) genome, or any derivative thereof. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation. In some embodiments, the inactivating mutation is in a herpes simplex virus gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the herpes simplex virus gene. In some embodiments, the herpes simplex virus gene is selected from infected cell protein (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL) 41, and / or UL55. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in one or both copies of the ICP4 gene. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL41 gene. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in one or both copies of the ICP0 gene. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene.In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the joint region. In some embodiments, the recombinant herpes simplex virus genome comprises a deletion in the joint region. In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides within one or both ICP4 viral loci.

[0013] In some embodiments that may be combined with any of the preceding embodiments, the recombinant herpesvirus genome has reduced cytotoxicity when introduced into a target cell compared to a corresponding wild-type herpesvirus genome, hi some embodiments, the target cell is a human cell.

[0014] Another aspect of the present disclosure relates to a herpesvirus comprising any of the recombinant herpesvirus genomes described herein. In some embodiments, the herpesvirus is replication-competent. In some embodiments, the herpesvirus is replication-deficient. In some embodiments, the herpesvirus is attenuated. In some embodiments that may be combined with any of the preceding embodiments, the herpesvirus has reduced cytotoxicity compared to a corresponding wild-type herpesvirus. In some embodiments that may be combined with any of the preceding embodiments, the herpesvirus is selected from herpes simplex virus, varicella-zoster virus, human cytomegalovirus, herpesvirus 6A, herpesvirus 6B, herpesvirus 7, and Kaposi's sarcoma-associated herpesvirus. In some embodiments that may be combined with any of the preceding embodiments, the herpesvirus is a herpes simplex virus. In some embodiments, the herpes simplex virus is herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), or any derivative thereof. In some embodiments, the herpes simplex virus is herpes simplex virus type 1 (HSV-1).

[0015] Another aspect of the present disclosure relates to a pharmaceutical composition comprising (a) any of the recombinant herpesvirus genomes described herein and / or any of the herpesviruses described herein, and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is suitable for topical, transdermal, subcutaneous, intradermal, transmucosal, oral, intranasal, intratracheal, sublingual, intranasal, buccal, rectal, intravaginal, intravenous, intraarterial, intramuscular, intracardiac, intraosseous, intraperitoneal, intraorbital, intravitreal, subconjunctival, suprachoroidal, subretinal, intraarticular, periarticular, local, epicutaneous, and / or inhalation administration. In some embodiments, the pharmaceutical composition is suitable for topical administration. In some embodiments, the pharmaceutical composition is suitable for inhalation administration. In some embodiments, the pharmaceutical composition is suitable for injection.

[0016] Another aspect of the present disclosure relates to the use of any of the recombinant herpesvirus genomes, herpesviruses and / or pharmaceutical compositions described herein as a medicament.

[0017] Other aspects of the present disclosure relate to the use of any of the recombinant herpesvirus genomes, herpesviruses and / or pharmaceutical compositions described herein in therapy.

[0018] Another aspect of the present disclosure relates to the use of any of the recombinant herpesvirus genomes, herpesviruses, and / or pharmaceutical compositions described herein in the manufacture of a medicament for treating a disease. In some embodiments, the disease is an inflammatory skin disease (e.g., atopic dermatitis). In some embodiments, the disease is selected from psoriasis, atopic dermatitis, pyoderma gangrenosum, bullous diseases, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behçet's disease, cancer, hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, autoimmune diseases, asthma, thyroid eye disease, infectious diseases, and neurological diseases.

[0019]

[0010] Another aspect of the present disclosure relates to a method of administering an antibody to a subject, the method comprising administering to the subject an effective amount of any of the herpesviruses described herein and / or any of the pharmaceutical compositions described herein. In some embodiments, the herpesvirus or pharmaceutical composition is administered topically, transdermally, subcutaneously, intradermally, transmucosally, orally, intranasally, intratracheally, sublingually, intranasally, bucally, rectally, intravaginally, intravenously, intraarterially, intramuscularly, intracardially, intraosseously, intraperitoneally, intraorbitally, intravitreally, subconjunctivally, epichoroidally, subretinally, intra-articularly, periarticularly, locally, epidermally, or by inhalation.

[0020]

[0010] Another aspect of the present disclosure relates to methods for providing prophylactic, palliative, and / or therapeutic relief of one or more signs or symptoms of a disease in a subject, the methods comprising administering to the subject an effective amount of any of the herpesviruses described herein and / or any of the pharmaceutical compositions described herein. In some embodiments, the herpesvirus or pharmaceutical composition is administered topically, transdermally, subcutaneously, intradermally, transmucosally, orally, intranasally, intratracheally, sublingually, intranasally, bucally, rectally, intravaginally, intravenously, intraarterially, intramuscularly, intracardially, intraosseously, intraperitoneally, intraorbitally, intravitreally, subconjunctivally, epichoroidally, subretinally, intraarticularly, periarticularly, locally, epidermally, or by inhalation. In some embodiments that may be combined with any of the preceding embodiments, the disease is selected from psoriasis, atopic dermatitis, pyoderma gangrenosum, bullous disease, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behcet's disease, cancer, hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, asthma, autoimmune diseases, thyroid eye disease, infectious diseases, and neurological diseases.

[0021] Another aspect of the present disclosure relates to a method of directing antibodies to the epidermis and / or dermis of a subject, the method comprising topically, transdermally, subcutaneously, or intradermally administering to the subject an effective amount of any of the herpesviruses described herein and / or any of the pharmaceutical compositions described herein. In some embodiments, the subject's skin is abraded or otherwise permeabilized prior to administration.

[0022] Another aspect of the present disclosure relates to a method of administering an antibody to the mucosa of a subject, the method comprising administering to the subject topically, mucosally, orally, sublingually, nasally, intranasally, by inhalation, or bucally an effective amount of any of the herpesviruses described herein and / or any of the pharmaceutical compositions described herein.

[0023] Another aspect of the present disclosure relates to a method of administering an antibody to the respiratory tract and / or lungs of a subject, the method comprising administering orally, sublingually, nasally, intranasally, intratracheally, by inhalation, or bucally to the subject an effective amount of any of the herpesviruses described herein and / or any of the pharmaceutical compositions described herein.

[0024] Another aspect of the present disclosure relates to a method of administering an antibody to one or more joints in a subject, the method comprising intra-articular and / or peri-articular administration to the subject of an effective amount of any of the herpesviruses described herein and / or any of the pharmaceutical compositions described herein.

[0025] Another aspect of the present disclosure relates to a method of administering an antibody to one or both eyes of a subject, the method comprising administering to the subject topically, intraorbitally, intravitreally, subconjunctivally, subretinally, or suprachoroidally an effective amount of any of the herpesviruses described herein and / or any of the pharmaceutical compositions described herein.

[0026] In some embodiments that may be combined with any of the preceding embodiments, the subject is a human. In some embodiments that may be combined with any of the preceding embodiments, the subject has not been systemically exposed to the antibody.

[0027] Another aspect of the present disclosure relates to a recombinant herpes simplex virus (HSV) genome comprising one or more polynucleotides encoding an antibody. In some embodiments, the antibody is an antibody fragment. In some embodiments, the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, scFv, or scFv-Fc fragment. In some embodiments, the antibody is a full-length antibody.

[0028] In some embodiments that may be combined with any of the preceding embodiments, the antibody is a murine antibody, a chimeric antibody, a humanized antibody, a human antibody, a monoclonal antibody, or a multispecific antibody. In some embodiments that may be combined with any of the preceding embodiments, the antibody is an IgA antibody, an IgD antibody, an IgE antibody, an IgG antibody, or an IgM antibody. In some embodiments that may be combined with any of the preceding embodiments, the antibody is an IgG antibody. In some embodiments, the IgG antibody is an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. In some embodiments, the antibody is an agonist antibody. In some embodiments, the antibody is an antagonist antibody.

[0029] In some embodiments that may be combined with any of the preceding embodiments, the antibody comprises a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3, wherein HVR-H1 comprises a sequence selected from the group consisting of SEQ ID NOs: 1-59, HVR-H2 comprises a sequence selected from the group consisting of SEQ ID NOs: 60-122, and / or HVR-H3 comprises a sequence selected from the group consisting of SEQ ID NOs: 123-185. In some embodiments, the heavy chain variable region comprises a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 355-419. In some embodiments that may be combined with any of the preceding embodiments, the antibody comprises a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3, wherein HVR-L1 comprises a sequence selected from the group consisting of SEQ ID NOs: 186-242, HVR-L2 comprises a sequence selected from the group consisting of SEQ ID NOs: 243-294, and / or HVR-L3 comprises a sequence selected from the group consisting of SEQ ID NOs: 395-354. In some embodiments, the light chain variable region comprises a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence selected from SEQ ID NOs: 420-482.

[0030] In some embodiments that may be combined with any of the preceding embodiments, the recombinant genome is a recombinant HSV-1 genome, a recombinant HSV-2 genome, or any derivative thereof. In some embodiments that may be combined with any of the preceding embodiments, the recombinant genome comprises an inactivating mutation in a herpes simplex virus gene. In some embodiments, the herpes simplex virus gene is selected from the group consisting of infected cell protein (ICP)0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL)41, and / or UL55. In some embodiments, the recombinant genome comprises an inactivating mutation in one or both copies of the ICP4 gene. In some embodiments, the recombinant genome comprises an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant genome comprises an inactivating mutation in the UL41 gene. In some embodiments, the recombinant genome comprises an inactivating mutation in the ICP0 gene. In some embodiments, the recombinant genome comprises an inactivating mutation in the ICP27 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the gene(s). In some embodiments that may be combined with any of the preceding embodiments, the recombinant genome has reduced cytotoxicity when introduced into a target cell compared to a wild-type herpes simplex virus genome. In some embodiments, the target cell is a human cell. In some embodiments, the target cell is a keratinocyte or a fibroblast.

[0031] In some embodiments that may be combined with any of the preceding embodiments, the recombinant genome comprises one or more polynucleotides in one or more viral loci. In some embodiments, the recombinant genome comprises one or more polynucleotides in one or both ICP4 viral loci. In some embodiments, the recombinant genome comprises one or more polynucleotides in an ICP22 viral locus. In some embodiments, the recombinant genome comprises one or more polynucleotides in a UL41 viral locus.

[0032] Another aspect of the present disclosure relates to a herpes simplex virus (HSV) comprising any of the recombinant genomes described herein. In some embodiments, the HSV is replication-competent. In some embodiments, the HSV is replication-deficient. In some embodiments that may be combined with any of the preceding embodiments, the HSV has reduced cytotoxicity compared to wild-type herpes simplex virus. In some embodiments that may be combined with any of the preceding embodiments, the HSV is herpes simplex virus type 1, herpes simplex virus type 2, or any derivative thereof.

[0033] Another aspect of the present disclosure relates to a pharmaceutical composition comprising any of the recombinant genomes and / or viruses described herein and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is suitable for topical, transdermal, subcutaneous, intradermal, transmucosal, sublingual, nasal, buccal, intraorbital, intravitreal, subconjunctival, suprachoroidal, intraarticular, and / or inhalation administration. In some embodiments, the pharmaceutical composition is suitable for topical administration. In some embodiments that can be combined with any of the preceding embodiments, the pharmaceutical composition comprises hydroxypropyl methylcellulose gel. In some embodiments that can be combined with any of the preceding embodiments, the pharmaceutical composition comprises a phosphate buffer. In some embodiments that can be combined with any of the preceding embodiments, the pharmaceutical composition comprises glycerol. In some embodiments that can be combined with any of the preceding embodiments, the pharmaceutical composition comprises a lipid carrier. In some embodiments that can be combined with any of the preceding embodiments, the pharmaceutical composition comprises a nanoparticle carrier.

[0034] Another aspect of the present disclosure relates to a method of administering an antibody to a subject, the method comprising administering to the subject an effective amount of any of the viruses or pharmaceutical compositions described herein. In some embodiments, the virus or composition is administered topically, transdermally, subcutaneously, intradermally, transmucosally, sublingually, intranasally, bucally, intravitreally, subconjunctivally, suprachoroidally, intraarticularly, or by inhalation. In some embodiments that may be combined with any of the preceding embodiments, the subject is a human. In some embodiments that may be combined with any of the preceding embodiments, the subject has not been systemically exposed to the antibody.

[0035] Other aspects of the present disclosure relate to methods of providing prophylactic, palliative, and / or therapeutic relief of one or more signs or symptoms of a disease in a subject, the method comprising administering to the subject an effective amount of any of the viruses or pharmaceutical compositions described herein. In some embodiments, the virus or composition is administered topically, transdermally, subcutaneously, intradermally, transmucosally, sublingually, intranasally, bucally, intravitreally, subconjunctivally, suprachoroidally, intraarticularly, or by inhalation. In some embodiments that may be combined with any of the preceding embodiments, the disease is selected from the group consisting of psoriasis, atopic dermatitis, pyoderma gangrenosum, bullous disease, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behcet's disease, cancer, hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, autoimmune disease, melanoma, uveal melanoma, and thyroid eye disease. In some embodiments, the disease is not cancer. In some embodiments that may be combined with any of the preceding embodiments, the subject is a human. In some embodiments that may be combined with any of the preceding embodiments, the subject has not been systemically exposed to an antibody.

[0036] Another aspect of the present disclosure relates to a method of administering an antibody to the epidermis and / or dermis of a subject, the method comprising topically, transdermally, or intradermally administering to the subject an effective amount of any of the viruses or pharmaceutical compositions described herein. In some embodiments, the subject's skin is abraded prior to administration. In some embodiments that may be combined with any of the preceding embodiments, the subject is a human. In some embodiments that may be combined with any of the preceding embodiments, the subject has not been systemically exposed to the antibody.

[0037] Another aspect of the present disclosure relates to a method of administering an antibody to the mucosa of a subject, the method comprising administering to the subject topically, mucosally, sublingually, nasally, or bucally an effective amount of any of the viruses or pharmaceutical compositions described herein. In some embodiments, the subject is a human. In some embodiments that may be combined with any of the preceding embodiments, the subject has not been systemically exposed to the antibody.

[0038] Another aspect of the present disclosure relates to a method of administering an antibody to one or more joints of a subject, the method comprising intra-articularly administering to the subject an effective amount of any of the viruses or pharmaceutical compositions described herein. In some embodiments, the subject is a human. In some embodiments that may be combined with any of the preceding embodiments, the subject has not been systemically exposed to the antibody.

[0039] Another aspect of the present disclosure relates to a method of administering an antibody to one or both eyes of a subject, the method comprising topically, intraorbitally, intravitreally, subconjunctivally, or suprachoroidally administering to the subject an effective amount of any of the viruses or pharmaceutical compositions described herein. In some embodiments, the subject is a human. In some embodiments that may be combined with any of the preceding embodiments, the subject has not been systemically exposed to the antibody. [The present invention 1001] A recombinant herpesvirus genome comprising one or more polynucleotides encoding an antibody. [The present invention 1002] 1001. The recombinant herpesvirus genome of the present invention, wherein the antibody is an antibody fragment. [The present invention 1003] 1002. The recombinant herpesvirus genome of the present invention, wherein the antibody fragment is a Fab, Fab', Fab'-SH, F(ab')2, Fv, scFv or scFv-Fc fragment. [The present invention 1004] 1001. The recombinant herpesvirus genome of the present invention, wherein said antibody is a single domain antibody. [The present invention 1005] 1001. The recombinant herpesvirus genome of the present invention, wherein said antibody is a full-length antibody. [The present invention 1006] The recombinant herpesvirus genome of any one of claims 1001 to 1005, wherein the antibody is a mouse antibody, a chimeric antibody, a humanized antibody, a human antibody, a monoclonal antibody, or a multispecific antibody. [The present invention 1007] The recombinant herpesvirus genome of any one of claims 1001 to 1006, wherein the antibody is an IgA antibody, an IgD antibody, an IgE antibody, an IgG antibody, or an IgM antibody. [The present invention 1008] The recombinant herpesvirus genome of any one of claims 1001 to 1007, wherein the antibody is an IgG antibody. [The present invention 1009] The recombinant herpesvirus genome of the present invention 1008, wherein the IgG antibody is an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. [The present invention 1010] The recombinant herpesvirus genome of the present invention 1008 or 1009, wherein the IgG antibody is an IgG1 antibody. [The present invention 1011] The recombinant herpesvirus genome of the present invention 1008 or 1009, wherein the IgG antibody is an IgG4 antibody. [The present invention 1012] The recombinant herpesvirus genome of any one of claims 1001 to 1011, wherein the antibody is an agonist antibody. [The present invention 1013] The recombinant herpesvirus genome of any one of claims 1001 to 1011, wherein the antibody is an antagonist antibody. [The present invention 1014] the antibody comprises a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3; the HVR-H1 comprises a sequence selected from the group consisting of SEQ ID NOs: 1 to 59; the HVR-H2 comprises a sequence selected from the group consisting of SEQ ID NOs: 60 to 122, and / or the HVR-H3 comprises a sequence selected from the group consisting of SEQ ID NOs: 123 to 185; The recombinant herpesvirus genome of any one of 1001 to 1013 of the present invention. [The present invention 1015] the antibody comprises a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3; the HVR-L1 comprises a sequence selected from the group consisting of SEQ ID NOs: 186 to 242; the HVR-L2 comprises a sequence selected from the group consisting of SEQ ID NOs: 243 to 294; and / or the HVR-L3 comprises a sequence selected from the group consisting of SEQ ID NOs: 295 to 354; The recombinant herpesvirus genome of any one of 1001 to 1014 of the present invention. [The present invention 1016] The recombinant herpesvirus genome of any of the present inventions 1001 to 1013, wherein the antibody comprises a heavy chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 355 to 419 or SEQ ID NOs: 614 to 865. [The present invention 1017] The recombinant herpesvirus genome of any of the present inventions 1001 to 1013, wherein the antibody comprises a light chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 420 to 482 or SEQ ID NOs: 866 to 1116. [The present invention 1018] The antibody (a) a heavy chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 355-419 or SEQ ID NOs: 614-865; and (b) a light chain variable region comprising a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 420-482 or SEQ ID NOs: 866-1116. The recombinant herpesvirus genome of any one of claims 1001 to 1013, comprising: [The present invention 1019] The antibody is selected from the group consisting of abagovomab, abciximab, abituzumab, abrezekimab, abrilumab, actoxumab, adalimumab, adecatumumab, aducanumab, afasevikumab, afelimomab, afutuzumab, alacizumab, alemtuzumab, alirocumab, altumomab, amatuximab, anatumomab, andecaliximab, anetuzumab, anifrolumab, anrukinzumab, apolizumab, apultumab, andeccaliximab, anetuzumab, anifrolumab, anrukinzumab, apolizumab, andeccalixim ... aprutumab, arcitumomab, asclinicalvacuumab, acelizumab, atezolizumab, atinumab, atolimumab, avelumab, azintuxizumab, bapineuzumab, basiliximab, bavituximab, bectumomab, begelomab, belantamab, belimumab, bemarituzumab, berimumab, benralizumab, berlimatoxumab ab), bersanlimab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab, biciromab, bimagrumab, bimekizumab, viltamimab, bivatuzumab, bleselumab, blinatumomab, brontuvetmab, brosozumab, bococizumab, brazikumab, brentuximab, briakinumab, brodalumab, brolucizumab, brontixituzumab, burosumab, cabilalizumab, camidanlumab, camrelizumab, kana Kinumab, cantuzumab, caplacizumab, capromab, carlumab, carotuximab, catumaxomab, cedelizumab, cemiplimab, sergituzumab, certolizumab, cetrelimab, cetuximab, civisatamab, sitatuzumab, cizutumumab, clazakizumab, clenoliximab, clivatuzumab, codrituzumab, cofetuzumab, coltuximab, conatumumab, concizumab, cosfrobiximab, crenezumab, crizanlizumab, cloteduumab,Cusatuzumab, dacetuzumab, daclizumab, dalotuzumab, dapirolizumab, daratumumab, dectrekumab, demcizumab, denintuzumab, denosumab, depatuximab, derlotuximab, detumomab, dezamizumab, dinutuximab, diridavumab, domaglotuzumab, dorlimomab, drozitumab, durigotuzumab, dupilumab, dur Valumab, dusigizumab, duvortuxizumab, ecromeximab, eculizumab, edovacomab, edrecolomab, efalizumab, efangumab, eldelumab, elezanumab, elgemtumab, elotuzumab, elsilimomab, emactuzumab, emapalumab, emibetuzumab, emicizumab, enapotamab, enavatuzumab, enfortumab, enlimomab, enoblitzumab, enokizumab, enoticumab, encicumab Ximab, epitumomab, epratuzumab, eptinezumab, erenumab, erlizumab, ertumaxomab, etaracizumab, etigilimab, etrolizumab, evinacumab, evolocumab, exbivirumab, fanolesomab, faralimomab, faricimab, farletuzumab, fasinumab, felvizumab, fezakinumab, fibatuzumab, ficlatuzumab, figitumumab, filibumab ivumab), framvotumab, fretikumab, flotetuzumab, fontolizumab, foralumab, foravirumab, fremanezumab, fresolimumab, furunvetomab, furanumab, futuximab, galcanezumab, galiximab, gancotamab, ganitumab, gantenerumab, gatipotuzumab, gavilimomab, gezivumab, gemtuzumab, gevokizumab, gilvemab, gilvetmab, gimsilumab, girentuximab, glembatumumab, golimumab,Gomiliximab, goslanemab, guselkumab, ianalumab, ibalizumab, ibritumomab, icrucumab, idarucizumab, ifavotuzumab, igovomab, iradatuzumab, imalumab, imaprelimab, imuciromab, imgatuzumab, inlacumab, indatuximab, indusatumab, inebilizumab, inflectra, infliximab, intetumumab, inolimomab, inotuzumab, ipirili tumab, iratumumab, isatuximab, iscalimab, istiratumab, itolizumab, ixekizumab, keliximab, labetuzumab, lacnotuzumab, ladiratuzumab, lampalizumab, lanadelumab, landgrozumab, laprituximab, ralcabiximab, lebrikizumab, lemaresomab, lensalizumab, lenvervimab, lenzilumab, lerdelimumab (lerdelimumab), leronlimab, lesofabumab, letolizumab, lexatumumab, ribivirumab, rifastuzumab, ligelizumab, loncastuximab, rosatuxizumab, rilotomab, lintuzumab, lirilumab, rodelcizumab, lokivetmab, lorvotuzumab, lucatumumab, lulizumab, rumiliximab, lumletuzumab, rupartumab, rutikizumab tikizumab, mapatumumab, marjetuximab, marstacimab, maslimomab, mavrilimumab, matuzumab, mepolizumab, metelimumab, milatuzumab, minretumomab, mirikizumab, mirvetuximab, mitumomab, modotuximab, mogamulizumab, monalizumab, morolimumab, mosunetuzumab, motavizumab, moxetumomab, nacolomab, namilumab, naptumomab, naratuximab,Narutumab, natalizumab, navicixizumab, navivumab, naxitamab, nebacumab, necitumumab, nemolizumab, nerelimomab, nesbacumab, netakimab, nimotuzumab, nirsevimab, nivolumab, nofetumomab, obilutoxaximab, obinutuzumab, occaratuzumab, ocrelizumab, ozlimomab, ofatumumab, olaratumab, oleculab, orendalizumab, olokizumab, omalizumab, onartuzumab, ontuxizumab umab), onvatilimab, opicinumab, oportuzumab, oregovomab, orticumab, otelixizumab, otilimab, otlertuzumab, oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, pamrevlumab, panitumumab, pancomab, panobacumab, palsatuzumab, pascolizumab, pasotuxizumab, pateclizumab, patritumab, pembrolizumab, pemtumomab , perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab, pintumomab, placumab, prozalizumab, pogalizumab, polatuzumab, ponezumab, polgabiximab, prasinezumab, prezalizumab, priliximab, pritoxaximab, pritumumab, kilimab, racotumomab, rad radretumab, rafivirumab, ralpancizumab, ramucirumab, ranevetomab, ranibizumab, raxibacumab, ravagalimab, ravtolumab, refanezumab, regavirumab, remtolumab, reslizumab, rilotumumab, rinucumab, risankizumab, rituximab, rivabazumab, lobatumumab, rolezumab, romilkimab,Lomozozumab, lontalizumab, rosmantuzumab, rovalpituzumab, rovelizumab, rozanolixizumab, ruplizumab, sacituzumab, samalizumab, samrotamab, sapelizumab, sarilumab, satralizumab, satumomab, secukinumab, selicrelumab, seribantumab, setoxaximab, setrusumab, sevilumab, sibrotuzumab, sifalimumab, Siltuximab, simtuzumab, siplizumab, sirtratumab, sirkumab, sofituzumab, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab, sulesomab, sputumumab, stimulimab, suvizumab, subratoxumab, tabalumab, tacatuzumab, tadocizumab, talacotuzumab, talizumab tamtuvetmab, tanezumab, taplitumomab, talexuzumab, tabolimab, tefibazumab, telimomab, telisotuzumab, tenatumomab, teneliximab, teplizumab, tepoditamab, teprotumumab, tesidolumab, tetulomab, tezepelumab, tiburizumab, tildrakizumab, tigatuzumab, thimig Tuzumab (timigutuzumab), timolumab (timolumab), tiragotumab (tiragotumab), tislelizumab, tisotumab, tocilizumab, tomzotuximab (tomuzotuximab), toralizumab, tosatoxumab (tosatoxumab), tositumomab, tobetumab, tralokinumab, trastuzumab, tregalizumab, tremelimumab, treboglumab, tucotuzumab, tuvilumab, ublituximab, urocuplumab, urelumab, Urutoxazumab, ustekinumab, utomilumab, vadastuximab, vanalimab, bundletuzumab, vanticizumab, vanucizumab, vapaliximab, varisacumab, varlilumab, vatelizumab, vedolizumab, veltuzumab, bepalimomab, besencumab, visilizumab, bovalilizumab, volociximab, bonrelozumab Any of the recombinant herpesvirus genomes of 1001 to 1013 of the present invention, selected from the group consisting of tuzumab, bupratelimab, borsetuzumab, votumumab, bunakizumab, xentuzumab, zalutumumab, zanolimumab, zatuximab, zenocutuzumab, ziralimumab, zolbetuximab and zolimomab. [The present invention 1020] The recombinant herpesvirus genome of any one of claims 1001 to 1019, wherein the recombinant herpesvirus genome has replication ability. [The present invention 1021] The recombinant herpesvirus genome of any one of claims 1001 to 1019, wherein the recombinant herpesvirus genome is replication-deficient. [The present invention 1022] The recombinant herpesvirus genome of any of claims 1001 to 1021, wherein the recombinant herpesvirus genome is selected from the group consisting of a recombinant herpes simplex virus genome, a recombinant varicella-zoster virus genome, a recombinant human cytomegalovirus genome, a recombinant herpesvirus 6A genome, a recombinant herpesvirus 6B genome, a recombinant herpesvirus 7 genome, a recombinant Kaposi's sarcoma-associated herpesvirus genome, and any derivative thereof. [The present invention 1023] The recombinant herpesvirus genome of any one of claims 1001 to 1022, wherein the recombinant herpesvirus genome is a recombinant herpes simplex virus genome. [The present invention 1024] The recombinant herpesvirus genome of the present invention 1023, wherein the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome, a recombinant herpes simplex virus type 2 (HSV-2) genome, or any derivative thereof. [The present invention 1025] The recombinant herpesvirus genome of the present invention 1023 or 1024, wherein the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome. [The present invention 1026] The recombinant herpesvirus genome of any one of claims 1022 to 1025, wherein the recombinant herpes simplex virus genome comprises an inactivating mutation. [The present invention 1027] 1026. A recombinant herpesvirus genome of the present invention, wherein said inactivating mutation is present in a herpes simplex virus gene. [The present invention 1028] 1027. The recombinant herpesvirus genome of the present invention, wherein said inactivating mutation is a deletion in the coding sequence of said herpes simplex virus gene. [The present invention 1029] The recombinant herpesvirus genome of the present invention 1027 or 1028, wherein the herpes simplex virus gene is selected from the group consisting of infected cell protein (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL) 41, and UL55. [The present invention 1030] 1029. The recombinant herpes simplex virus genome of the present invention, wherein said recombinant herpes simplex virus genome comprises an inactivating mutation in one or both copies of said ICP4 gene. [The present invention 1031] The recombinant herpesvirus genome of claim 1029 or claim 1030, wherein said recombinant herpes simplex virus genome comprises an inactivating mutation in said ICP22 gene. [The present invention 1032] The recombinant herpesvirus genome of any one of claims 1029 to 1031, wherein the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL41 gene. [The present invention 1033] The recombinant herpesvirus genome of any of claims 1029 to 1032, wherein the recombinant herpes simplex virus genome comprises an inactivating mutation in one or both copies of the ICP0 gene. [The present invention 1034] The recombinant herpesvirus genome of any one of claims 1029 to 1033, wherein the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene. [This invention 1035] The recombinant herpes simplex virus genome of any one of claims 1022 to 1034, wherein the recombinant herpes simplex virus genome comprises the one or more polynucleotides encoding the antibody within one or both ICP4 viral loci. [The present invention 1036] The recombinant herpesvirus genome of any one of claims 1001 to 1035, wherein the recombinant herpesvirus genome has reduced cytotoxicity when introduced into a target cell, compared to the corresponding wild-type herpesvirus genome. [This invention 1037] 1036. The recombinant herpesvirus genome of claim 1036, wherein the target cell is a human cell. [The present invention 1038] A herpesvirus comprising the recombinant herpesvirus genome of any one of the present inventions 1001 to 1037. [This invention 1039] The herpesvirus of claim 1038, wherein the herpesvirus is replication-competent. [The present invention 1040] The herpesvirus of claim 1038, wherein the herpesvirus is replication-deficient. [This invention 1041] The herpesvirus of any one of claims 1038 to 1040, wherein the herpesvirus has reduced cytotoxicity compared to the corresponding wild-type herpesvirus. [The present invention 1042] The herpesvirus of any one of claims 1038 to 1041, wherein the herpesvirus is selected from the group consisting of herpes simplex virus, varicella-zoster virus, human cytomegalovirus, herpesvirus 6A, herpesvirus 6B, herpesvirus 7, and Kaposi's sarcoma-associated herpesvirus. [This invention 1043] The herpesvirus of any one of claims 1038 to 1042, wherein the herpesvirus is a herpes simplex virus. [This invention 1044] The herpesvirus of the present invention 1043, wherein the herpes simplex virus is herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), or any derivative thereof. [This invention 1045] The herpesvirus of the present invention 1043 or 1044, wherein the herpes simplex virus is herpes simplex virus type 1 (HSV-1). [The present invention 1046] a recombinant herpesvirus genome according to any one of claims 1001 to 1037 of the present invention or a herpesvirus according to any one of claims 1038 to 1045 of the present invention; a pharmaceutically acceptable excipient; A pharmaceutical composition comprising: [This invention 1047] 1046. The pharmaceutical composition of the present invention, wherein the pharmaceutical composition is suitable for topical, transdermal, subcutaneous, intradermal, transmucosal, oral, intranasal, intratracheal, sublingual, nasal, buccal, rectal, intravaginal, intravenous, intraarterial, intramuscular, intracardiac, intraosseous, intraperitoneal, intraorbital, intravitreal, subconjunctival, suprachoroidal, subretinal, intraarticular, periarticular, local, epicutaneous, and / or inhalation administration. [This invention 1048] The pharmaceutical composition of claim 1046 or 1047, wherein the pharmaceutical composition is suitable for topical administration. [This invention 1049] A herpesvirus according to any one of claims 1038 to 1045 or a pharmaceutical composition according to any one of claims 1046 to 1048 for use as a drug. [The present invention 1050] A herpesvirus according to any one of claims 1038 to 1045 or a pharmaceutical composition according to any one of claims 1046 to 1048 for use in therapy. [This invention 1051] Use of the herpesvirus of any of claims 1038 to 1045 or the pharmaceutical composition of any of claims 1046 to 1048 in the manufacture of a medicament for treating a disease. [This invention 1052] The use of 1051 of the present invention, wherein the disease is selected from the group consisting of psoriasis, atopic dermatitis, pyoderma gangrenosum, bullous diseases, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behcet's disease, cancer, hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, autoimmune diseases, asthma, thyroid eye disease, infectious diseases and neurological diseases. [This invention 1053] 1. A method of administering an antibody to a subject, comprising: administering to the subject an effective amount of any of the herpesviruses of the present inventions 1038 to 1045 or any of the pharmaceutical compositions of the present inventions 1046 to 1048. The method comprising: [This invention 1054] 1. A method for providing prophylactic, palliative, and / or therapeutic relief of one or more signs or symptoms of a disease in a subject, comprising: administering to the subject an effective amount of any of the herpesviruses of the present inventions 1038 to 1045 or any of the pharmaceutical compositions of the present inventions 1046 to 1048. The method comprising: [This invention 1055] The method of invention 1053 or invention 1054, wherein the herpes virus or the pharmaceutical composition is administered topically, transdermally, subcutaneously, intradermally, transmucosally, orally, intranasally, intratracheally, sublingually, intranasally, bucally, rectally, intravaginally, intravenously, intraarterially, intramuscularly, intracardially, intraosseously, intraperitoneally, intraorbitally, intravitreally, subconjunctivally, epichoroidally, subretinally, intraarticularly, periarticularly, locally, epicutaneously, or by inhalation. [The present invention 1056] The method of invention 1054 or invention 1055, wherein the disease is selected from the group consisting of psoriasis, atopic dermatitis, pyoderma gangrenosum, bullous disease, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behcet's disease, cancer, hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, asthma, autoimmune diseases, thyroid eye disease, infectious diseases and neurological diseases. [This invention 1057] 1. A method of administering an antibody to the epidermis and / or dermis of a subject, comprising: administering an effective amount of any one of the herpesviruses of the present inventions 1038 to 1045 or any one of the pharmaceutical compositions of the present inventions 1046 to 1048 to the subject topically, transdermally, or intradermally; The method comprising: [This invention 1058] The method of claim 1057, wherein the subject's skin is abraded prior to administration. [This invention 1059] 1. A method of administering an antibody to a mucosa of a subject, comprising: administering an effective amount of any of the herpesviruses of the present inventions 1038 to 1045 or any of the pharmaceutical compositions of the present inventions 1046 to 1048 to the subject by topical administration, transmucosal administration, oral administration, sublingual administration, nasal administration, intranasal administration, inhalation, or buccal administration; The method comprising: [The present invention 1060] 1. A method of administering an antibody to the airways or lungs of a subject, comprising: administering an effective amount of any one of the herpesviruses of the present inventions 1038 to 1045 or any one of the pharmaceutical compositions of the present inventions 1046 to 1048 to the subject orally, sublingually, nasally, intranasally, intratracheally, by inhalation, or bucally; The method comprising: [The present invention 1061] 1. A method of administering an antibody to one or more joints in a subject, comprising: an effective amount of the herpesvirus of any one of claims 1038 to 1045 of the present invention or the pharmaceutical composition of any one of claims 1046 to 1048 of the present invention is administered intra-articularly to the subject; The method comprising: [The present invention 1062] 1. A method of administering an antibody to one or both eyes of a subject, comprising: administering an effective amount of any one of the herpesviruses of the present inventions 1038 to 1045 or any one of the pharmaceutical compositions of the present inventions 1046 to 1048 to the subject topically, intraorbitally, intravitreally, subconjunctivally, subretinally, or suprachoroidally; The method comprising: [The present invention 1063] The method of any one of claims 1053 to 1062, wherein the subject is a human. [The present invention 1064] 1064. The method of any of claims 1053 to 1063, wherein said subject has not been systemically exposed to said antibody. [Brief explanation of the drawings]

[0040] This patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0041] [Figure 1-1] 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes: Figure 1A shows the wild-type herpes simplex virus genome. [Figure 1-2]Figures 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes. Figure 1B shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies) and ICP22, where a polynucleotide contains the coding sequence for a single-chain antibody (scFv-Fc) operably linked to a heterologous promoter integrated into each ICP4 locus. Figure 1C shows a modified herpes simplex virus genome containing deletions of the coding sequence for ICP4 (both copies), where a polynucleotide contains the coding sequence for a scFv-Fc operably linked to a heterologous promoter integrated into each ICP4 locus. [Figure 1-3] Figures 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes. Figure 1D shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies) and ICP22, where a polynucleotide is integrated into each ICP4 locus, containing 1) a coding sequence for an antibody heavy chain operably linked to a first heterologous promoter, and 2) a coding sequence for an antibody light chain operably linked to a second heterologous promoter. Both the antibody heavy chain and the antibody light chain are encoded on the same DNA strand. Figure 1E shows a modified herpes simplex virus genome containing deletions of the coding sequence for ICP4 (both copies), where a polynucleotide is integrated into each ICP4 locus, containing 1) a coding sequence for an antibody heavy chain operably linked to a first heterologous promoter, and 2) a coding sequence for an antibody light chain operably linked to a second heterologous promoter. Both the antibody heavy chain and the antibody light chain are encoded on the same DNA strand. [Figure 1-4]Figures 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes. Figure 1F shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies) and ICP22, where a polynucleotide is integrated into each ICP4 locus, containing 1) a coding sequence for an antibody heavy chain operably linked to a first heterologous promoter, and 2) a coding sequence for an antibody light chain operably linked to a second heterologous promoter. The antibody heavy chain and antibody light chain are encoded on opposite DNA strands. Figure 1G shows a modified herpes simplex virus genome containing deletions of the coding sequence for ICP4 (both copies), where a polynucleotide is integrated into each ICP4 locus, containing 1) a coding sequence for an antibody heavy chain operably linked to a first heterologous promoter, and 2) a coding sequence for an antibody light chain operably linked to a second heterologous promoter. The antibody heavy chain and antibody light chain are encoded on opposite DNA strands. [Figure 1-5] Figures 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes. Figure 1H shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies) and ICP22, where a polynucleotide encodes a polycistronic mRNA operably linked to a heterologous promoter integrated into each ICP4 locus. The polycistronic mRNA contains coding sequences for antibody heavy and light chains separated by an internal ribosome entry site (IRES). Figure 1I shows a modified herpes simplex virus genome containing deletions of the coding sequence for ICP4 (both copies), where a polynucleotide encodes a polycistronic mRNA operably linked to a heterologous promoter integrated into each ICP4 locus. The polycistronic mRNA contains coding sequences for antibody heavy and light chains separated by an internal ribosome entry site (IRES). [Figure 1-6]Figures 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes. Figure 1J shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies), ICP22, and UL41, wherein a first polynucleotide contains the coding sequence for an antibody heavy chain operably linked to a heterologous promoter integrated into each ICP4 locus, and a second polynucleotide contains the coding sequence for an antibody light chain operably linked to a heterologous promoter integrated into the UL41 locus and the ICP22 locus. FIG. 1K shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies), ICP22, and UL41, wherein a first polynucleotide contains the coding sequence for an antibody light chain operably linked to a heterologous promoter integrated into each ICP4 locus, and a second polynucleotide contains the coding sequence for an antibody heavy chain operably linked to a heterologous promoter integrated into the UL41 locus and the ICP22 locus. [Figure 1-7]Figures 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes. Figure 1L shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies) and UL41, where a first polynucleotide contains the coding sequence for an antibody heavy chain operably linked to a heterologous promoter integrated into each ICP4 locus, and a second polynucleotide contains the coding sequence for a polycistronic mRNA operably linked to a heterologous promoter integrated into the UL41 locus. The polycistronic mRNA contains two copies of the coding sequence for the antibody light chain separated by an internal ribosome entry site (IRES). Figure 1M shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies) and ICP22, wherein a first polynucleotide contains the coding sequence for an antibody heavy chain operably linked to a heterologous promoter integrated into each ICP4 locus, and a second polynucleotide contains the coding sequence for a polycistronic mRNA operably linked to a heterologous promoter integrated into the ICP22 locus. The polycistronic mRNA contains two copies of the coding sequence for the antibody light chain separated by an internal ribosome entry site (IRES). [Figure 1-8] 1A-N show schematic diagrams of wild-type and modified herpes simplex virus genomes. Figure 1N shows a modified herpes simplex virus genome containing deletions of the coding sequences for ICP4 (both copies), ICP22, and UL41, wherein a first polynucleotide contains a coding sequence for an antibody heavy chain operably linked to a heterologous promoter integrated into the ICP22 locus, and a second polynucleotide contains a coding sequence for an antibody light chain operably linked to a heterologous promoter integrated into the UL41 locus. [Figure 2]Figures 2A-B show antibody concentrations, assessed by ELISA, in cell supernatants harvested from mock-infected (MOI-0) immortalized human keratinocytes (HaCaT) or from HaCaT cells infected at the indicated multiplicity of infection (MOI) with engineered HSV vectors encoding the indicated antibodies. Figure 2A shows antibody concentrations in cell supernatants harvested from HaCaT cells infected with HSV encoding human (Ab1Fc1 or Ab1Fc2) single-chain antibodies or chimeric (Ab2Fc2) single-chain antibodies. Figure 2B shows antibody concentrations in cell supernatants harvested from HaCaT cells infected with HSV encoding murine (Ab66Fc1 or Ab66Fc2) single-chain antibodies. For each condition, data represent two replicates ± SEM. [Figure 3] Figure 1 shows the detectable levels of recombinant human TNFα spiked into cell supernatants harvested from mock-infected (MOI 0) immortalized human keratinocytes (HaCaT) or from HaCaT infected at the indicated multiplicity of infection (MOI) with an engineered HSV vector encoding an anti-TNFα human single-chain antibody (Ab1Fc1), as assessed by ELISA. For each condition, data represent two replicates ± SEM. [Figure 4] Figures 4A-B show the relative fold change in transcript levels of specific markers of atopic dermatitis-like lesions in the ear and dorsal skin of mice treated topically with the vitamin D3 synthetic analog calcipotriol (MC903) for 5 days compared to the ethanol (EtOH) control, as assessed by qRT-PCR analysis. Figure 4A shows the relative fold change in TSLP transcripts compared to the EtOH control in the ear and dorsal skin of mice treated topically with MC903 on days 1-5, with tissues harvested on days 5 or 7. Figure 4B shows the relative fold change in IL-4 transcripts compared to the EtOH control in the ear and dorsal skin of mice treated topically with MC903 on days 1-5, with tissues harvested on days 5, 7, or 9. [Figure 5] Shown is histology of ear skin from representative mice treated with MC903 or EtOH control, as assessed by hematoxylin and eosin (H&E) staining. [Figure 6A]Figures 6A-B show representative immunofluorescence images of human single-chain antibody (Ab1Fc1) expression in ear and dorsal skin biopsies taken from MC903-challenged C57BL / 6J mice treated topically with either HSV-Ab1Fc1 or negative control (vehicle). DAPI staining was used to visualize nuclei. Figure 6A shows Ab1Fc1 expression in the ear and dorsal skin of mice treated with MC903 on days 1-5 and topically with HSV-Ab1Fc1 (or vehicle control) on day 5; tissues were harvested on day 7. [Figure 6B] Figures 6A-B show representative immunofluorescence images of human single-chain antibody (Ab1Fc1) expression in ear and dorsal skin biopsies taken from MC903-challenged C57BL / 6J mice treated topically with either HSV-Ab1Fc1 or negative control (vehicle). Nuclei were visualized using DAPI staining. Figure 6B shows Ab1Fc1 expression in the ear and dorsal skin of mice treated with MC903 on days 1-5 and topically with HSV-Ab1Fc1 (or vehicle control) on day 7; tissues were harvested on day 9. [Figure 7] Figures 7A-C show mouse anti-mouse IL-4Ra antibody (Ab66Fc1) nucleic acid analysis of ear and dorsal skin in an MC903-induced atopic dermatitis model after infection with HSV-Ab66Fc1. Figure 7A shows the levels of Ab66Fc1 DNA present in ear tissue biopsies taken from MC903-treated or ethanol (EtOH)-treated animals after repeated topical applications of HSV-Ab66Fc1 or a solvent control, as measured by qPCR analysis. Figure 7B shows the levels of Ab66Fc1 DNA present in dorsal skin tissue biopsies taken from MC903-treated or ethanol (EtOH)-treated animals after repeated topical applications of HSV-Ab66Fc1 or a solvent control, as measured by qPCR analysis. Figure 7C shows the levels of Ab66Fc1 transcripts present in ear tissue biopsies taken from MC903-treated or ethanol (EtOH)-treated animals after repeated topical applications of HSV-Ab66Fc1 or vehicle control, as measured by qRT-PCR analysis. For each condition in qPCR and qRT-PCR analysis, data represent two replicates ± SEM. [Figure 8] Figures 8A-B show the effects of HSV-Ab66Fc1 or vehicle control on the development of specific ear phenotypes in the MC903-induced atopic dermatitis model. Figure 8A shows the mean ear thickness on days 1-10 in MC903- or ethanol-treated animals after repeated topical application of HSV-Ab66Fc1 or vehicle control. Asterisks indicate statistically significant differences between the MC903 / vehicle and MC903 / Ab66 groups at each time point. Figure 8B shows the mean ear weight on day 10 in MC903- or ethanol-treated animals after repeated topical application of HSV-Ab66Fc1 or vehicle control. For each time point, data represent the mean ± SEM of four ears. Statistics were calculated using an unpaired Student's t-test: *p<0.05; **p<0.01; ***p<0.005. DETAILED DESCRIPTION OF THE INVENTION

[0042] Detailed Description The following description sets forth example methods, parameters, etc. However, it should be recognized that such description is not intended to limit the scope of the present disclosure, but is instead provided as a description of example embodiments.

[0043] I. General techniques The techniques and procedures described or referenced herein are generally well understood by those of skill in the art and may be adapted from conventional methodology, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual 3rd edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, Current Protocols in Molecular Biology (F.M.A.usubel, et al. eds., (2003)), the series Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (M.J.MacPherson, B.D.Hames and G.R.Taylor eds. (1995)), Harlow and Lane, eds. (1988), Oligonucleotide Synthesis (M.J.Gait, ed., 1984), Methods in Molecular Biology, Humana Press, Cell Biology: A Laboratory Notebook (J.E.Cellis, ed., 1998) Academic Press, Animal Cell Culture (RIFreshney), ed., 1987), Introduction to Cell and Tissue Culture (JP Mather and PE Roberts, 1998) Plenum Press, Cell and Tissue Culture: Laboratory Procedures (A. Doyle, JBGriffiths, and DG Newell, eds., 1993-8) J. Wiley and Sons, Gene Transfer Vectors for Mammalian Cells (JMMiller and MP Calos, eds., 1987), PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994) and the widely used methodology described in Short Protocols in Molecular Biology (Wiley and Sons, 1999).

[0044] II. Definition Before describing the present disclosure in detail, it is to be understood that this disclosure is not limited to particular compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0045] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to a "molecule" optionally includes combinations of two or more such molecules, and the like.

[0046] As used herein, the term "and / or" may include any and all combinations of one or more of the associated listed items. For example, the term "a and / or b" can refer to "a only," "b only," "a or b," or "a and b," and the term "a, b and / or c" can refer to "a only," "b only," "c only," "a or b," "a or c," "b or c," "a, b or c," "a and b," "a and c," "b and c," or "a, b and c," etc.

[0047] As used herein, the term "about" refers to a normal range of error for the respective value, which would be readily understood by one of ordinary skill in the art. Reference herein to "about" a value or parameter includes (and describes) embodiments that are directed to the value or parameter itself.

[0048] It is understood that aspects and embodiments of the present disclosure include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0049] As used herein, the terms "polynucleotide," "nucleic acid sequence," "nucleic acid," and variations thereof are intended to refer collectively to polydeoxyribonucleotides (containing 2-deoxy-D-ribose), polyribonucleotides (containing D-ribose), any other type of polynucleotide that is an N-glycoside of a purine or pyrimidine base, and other polymers containing a non-nucleotide backbone, provided that the polymer contains nucleobases in a configuration that allows base pairing and base stacking as found in DNA and RNA. Thus, these terms include known types of nucleic acid sequence modifications, such as substitution of one or more naturally occurring nucleotides with an analog, and internucleotide modifications.

[0050] As used herein, a nucleic acid is "operatively linked" or "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence, or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, "operatively linked" or "operably linked" means that the DNA or RNA sequences being linked are contiguous.

[0051] As used herein, the term "vector" refers to an individual element used to introduce heterologous nucleic acids into cells for either their expression or replication. Expression vectors include vectors capable of expressing nucleic acids operably linked to regulatory sequences, such as promoter regions, which can effect expression of such nucleic acids. Thus, an expression vector can refer to a DNA or RNA construct, such as a plasmid, phage, recombinant virus, or other vector that, when introduced into an appropriate host cell, results in expression of a nucleic acid. Suitable expression vectors are well known to those skilled in the art and include those that are replicable in eukaryotic cells and those that remain episomal or integrate into the host cell genome.

[0052] As used herein, "open reading frame" or "ORF" refers to a contiguous stretch of nucleic acid, either DNA or RNA, that encodes a protein or polypeptide. Typically, the nucleic acid includes a translation initiation signal or start codon, such as ATG or AUG, and a stop codon.

[0053] As used herein, "untranslated region" or "UTR" refers to untranslated nucleic acid at the 5' and / or 3' end of an open reading frame. The inclusion of one or more UTRs in a polynucleotide may affect post-transcriptional regulation, mRNA stability, and / or translation of the polynucleotide.

[0054] As used herein, the term "transgene" refers to a polynucleotide that can be introduced into a cell and then transcribed into RNA, translated, and / or expressed under appropriate conditions. In some aspects, a transgene confers a desired characteristic on the cell into which it is introduced, or otherwise produces a desired therapeutic or diagnostic result.

[0055] As used herein, the terms "polypeptide," "protein," and "peptide" are used interchangeably and may refer to a polymer of two or more amino acids.

[0056] As used herein, the term "antibody" is used in the broadest sense and encompasses a variety of antibody structures, including, for example, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies, trispecific antibodies, etc.), and antibody fragments, so long as they exhibit the desired biological activity. The term "antibody" also encompasses hybrid antibodies, modified antibodies, chimeric antibodies, and humanized antibodies. The term antibody includes hybrid (chimeric) antibody molecules (see, e.g., Winter et al. (1991) Nature 349:293-299, and U.S. Pat. No. 4,816,567); F(ab')2 and F(ab)2 fragments; ... v molecules (non-covalent heterodimers, see e.g., Inbar et al. (1972) Proc Natl Acad Sci USA 69:2659-2662, and Ehrlich et al. (1980) Biochem 19:4091-4096); single-chain Fv molecules (scFv) (see e.g., Huston et al. (1988) Proc Natl Acad Sci USA 85:5879-5883); nanobodies or single-domain antibodies (sdAbs) (see e.g., Wang et al. (2016) Int J Nanomedicine 11:3287-3303, Vincke et al. (2012) Methods Mol Biol 911:15-26); dimeric and trimeric antibody fragment constructs; minibodies (see e.g., Pack et al. (2012) Methods Mol Biol 911:15-26); al. (1992) Biochem 31:1579-1584, Cumber et al. (1992) J Immunology 149B:120-126); humanized antibody molecules (see, e.g., Riechmann et al. (1988) Nature 332:323-327, Verhoeyan et al. (1988) Science 239:1534-1536, and British Patent Publication No. GB ​​2,276,169, published September 21, 1994), as well as any functional fragments derived from such molecules which retain the specific binding properties of the parent antibody molecule.

[0057] The basic four-chain antibody unit is a heterotetrameric glycoprotein consisting of two identical light chains and two identical heavy chains. H ) area and variable light (V L The pairing of these domains together forms an antigen-binding site. The structure and properties of different classes of antibodies are described, for example, in Basic and Clinical Immunology, 8 th See page 71 and Chapter 6 of Ed., Daniel P. Stites, Abba I. Terr, and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994.

[0058] Light chains from any vertebrate species can be assigned to one of two clearly distinct types, called kappa ("κ") (see, e.g., SEQ ID NO:601 for an exemplary human kappa constant domain sequence) and lambda ("λ") (see, e.g., SEQ ID NO:602 for an exemplary human lambda constant domain sequence), based on the amino acid sequence of their constant domain. Depending on the amino acid sequence of the constant domain (CH) of their heavy chain, immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, each with a heavy chain designated alpha ("α"), delta ("δ"), epsilon ("ε"), gamma ("γ"), and mu ("μ"). The gamma and alpha classes are further divided into subclasses (isotypes) based on relatively minor differences in CH sequence and function; for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known and generally described, for example, in Abbas et al., Cellular and Molecular Immunology, 4 th This is described in ed. (WBSaunders Co., 2000).

[0059] As used herein, the term "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of the antibody's heavy or light chain. These domains are generally the most variable parts of an antibody (relative to other antibodies of the same class) and contain the antigen-binding sites.

[0060] As used herein, the term "variable" refers to the fact that certain segments of the variable domains differ extensively in sequence among antibodies. The variable domains mediate antigen binding and define the specificity of a particular antibody for its particular antigen. However, variability is not evenly distributed throughout the variable domains. Rather, it is concentrated in three segments called hypervariable regions (HVRs) in both the light- and heavy-chain variable domains. The more highly conserved portions of the variable domains are called framework regions (FRs).

[0061] As used herein, the term "hypervariable region" or "HVR" refers to a region of an antibody variable domain that is hypervariable in sequence and / or forms structurally defined loops. Antibodies generally contain six HVRs: three in the VH (HVR-H1, HVR-H2, and HVR-H3) and three in the VL (HVR-L1, HVR-L2, and HVR-L3). In natural antibodies, HVR-H3 and HVR-L3 exhibit the highest diversity among these six HVRs, and HVR-H3 in particular is thought to play a unique role in conferring superior specificity to antibodies (see, e.g., Xu et al., Immunity 13:37-45 (2000) and Johnson and Wu, Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa NJ, 2003)). Indeed, natural camelid antibodies consisting only of heavy chains are functional and stable in the absence of light chains (see, e.g., Hamers-Casterman et al. Nature 363:446-448 (1993) and Sheriff et al. Nature Struct. Biol. 3:733-736 (1996)).

[0062] Several HVR descriptions are used and encompassed herein. EU or Kabat complementarity-determining regions (CDRs) HVRs are based on sequence variability and are the most commonly used, while Chothia refers to the location of structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). AbM HVRs represent a compromise between EU or Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. "Contact" HVRs are based on analysis of available complex crystal structures.

[0063] HVRs may include "extended HVRs" as follows: 24-36 or 24-24 (HVR-L1), 46-56 or 50-46 (HVR-L2), and 89-97 or 89-96 (HVR-L3) in VL, and 26-35 (HVR-H1), 50-65 or 49-65 (HVR-H2), and 93-201, 94-102, or 95-102 (HVR-H3) in VH. Variable domain residues are numbered according to EU or Kabat et al. for each of these extended HVR definitions.

[0064] As used herein, the term "framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences of a variable domain generally appear in VH (or VL) in the following sequence: FR1-HVR-H1(L1)-FR2-HVR-H2(L2)-FR3-HVR-H3(L3)-FR4.

[0065] As used herein, the terms "full-length antibody," "intact antibody," or "whole antibody" are used interchangeably to refer to an antibody having a heavy chain having a structure substantially similar to that of a native antibody or containing an Fc region as defined herein. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. In some embodiments, an intact antibody has one or more effector functions.

[0066] As used herein, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, including native-sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is usually defined to stretch from the amino acid residue at position Cys226 or Pro230 to its carboxyl-terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU or Kabat numbering system) can be removed, for example, during antibody production or by recombinantly engineering the nucleic acid encoding the antibody heavy chain. Thus, an intact antibody composition can include an antibody population with all K447 residues removed, an antibody population without the K447 residue removed, and an antibody population having a mixture of antibodies with and without the K447 residue.

[0067] Antibody "effector functions" refer to those biological activities attributable to the Fc region of an antibody (a native sequence Fc region or amino acid sequence variant Fc region), and vary with the antibody isotype.

[0068] As used herein, the term "native antibody" refers to antibodies that are typically heterotetrameric glycoproteins of approximately 150,000 daltons, consisting of two identical light chains and two identical heavy chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, although the number of disulfide bonds varies among heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced interchain disulfide bridges. Each heavy chain has a variable domain (VH) at one end followed by several constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end, with the light chain constant domain aligned with the first constant domain of the heavy chain and the light chain variable domain aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light chain variable domain and the heavy chain variable domain.

[0069] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population. That is, the individual antibodies within the population are identical except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation), which may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against one or more antigenic sites. In some embodiments, the monoclonal antibodies of the present disclosure may be multispecific (e.g., bispecific, trispecific). In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant at one or more antigenic sites. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous antibody population and is not to be construed as requiring production of the antibody by any particular method.

[0070] As used herein, a "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species, as well as fragments of such antibodies, provided they exhibit the desired biological activity. As used herein, "humanized" antibodies are used as a subset of "chimeric" antibodies.

[0071] As used herein, a "humanized" antibody refers to a chimeric antibody comprising amino acid residues derived from non-human HVRs and human FRs. In some embodiments, a humanized antibody comprises substantially all of at least one, and typically two, variable domains, in which all or substantially all HVRs correspond to HVRs of a non-human antibody and all or substantially all FRs correspond to FRs of a human antibody. A humanized antibody may optionally comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody, e.g., a non-human antibody, refers to an antibody that has been humanized. For further details, see, e.g., Jones et al. Nature 321:522-525 (1986), Riechmann et al. Nature 332:323-329 (1988), Presta, Curr. Op. Stuct. Biol. 2:593-596 (1992), Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol 1:105-115 (1998), Harris Biochem. Soc. Transactions 23:1035-1038 (1995), Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994), US 6,982,321 and US 7,087,409.

[0072] As used herein, a "human" antibody refers to an antibody possessing an amino acid sequence that corresponds to that of an antibody produced by a human or human cell, or that corresponds to that of an antibody derived from a human antibody repertoire or other non-human source that utilizes human antibody coding sequences. This definition specifically excludes humanized antibodies that comprise non-human antigen-binding residues.

[0073] As used herein, "human consensus framework" refers to a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the human immunoglobulin VL or VH sequence is selected from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as found in Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda, MD, vols. 1-3.

[0074] As used herein, the term "antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody, preferably the antigen-binding and / or variable region of the intact antibody. Examples of antibody fragments can include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, scFv, SMIP, domain antibodies, di-scFv, scFv-Fc, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), minibodies, diabodies, triabodies, linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0075] Papain digestion of antibodies produces two identical antigen-binding fragments, called Fab fragments, and one residual Fc fragment (a designation reflecting the ability to crystallize readily). The Fab fragment contains the entire light chain and the variable region and first constant domain of one heavy chain (C H1). Each Fab fragment is monovalent with respect to antigen binding, i.e., each Fab fragment has a single antigen-binding site. The Fc fragment contains the carboxy-terminal portions of both heavy chains held together by disulfide bonds. The effector functions of the antibody are determined by the sequence of the Fc region, which is also recognized by Fc receptors (FcRs) found on certain types of cells.

[0076] Pepsin treatment of antibodies yields a single large F(ab')2 fragment, which roughly corresponds to two disulfide-linked Fab fragments with different antigen-binding activities and is still capable of cross-linking antigen. Fab' fragments contain one or more cysteines from the antibody hinge region. H F(ab')2 antibody fragments differ from Fab fragments by having several additional residues at the carboxy terminus of one domain. Fab'-SH is the designation for Fab' in which the cysteine ​​residue(s) in the constant domains bear a free thiol group. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments that have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0077] An "Fv" is the minimum antibody fragment containing a complete antigen-recognition and antigen-binding site. This fragment consists of a dimer of one heavy-chain variable region domain and one light-chain variable region domain in tight, non-covalent association. The folding of these two domains generates six hypervariable loops (three loops each from the heavy and light chains) that provide the amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three antigen-specific HVRs) has the ability to recognize and bind to an antigen, albeit with lower affinity than the entire binding site.

[0078] "Single-chain Fv," also abbreviated as "sFv" or "scFv," is an antibody fragment comprising the VH and VL antibody domains linked in a single polypeptide chain. Preferably, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the scFv to form the desired structure for antigen binding. For a review of scFvs, see, e.g., Plückthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore (eds.), Springer-Verlag, New York, pp. 269-315 (1994); U.S. Pat. No. 6,248,516.

[0079] "Nanobody" refers to a single domain antibody (sdAb) that can selectively bind to an antigen. Nanobodies may contain a heavy chain variable domain and not a light chain variable domain, or vice versa. Nanobodies are also known as single domain antibodies (sdAbs) that are capable of selectively binding to an antigen. H H antibody) or cartilaginous fish (V NAR Alternatively, nanobodies can be obtained by splitting the dimeric variable domain from an antibody, for example an IgG antibody, into monomers.

[0080] Diabodies are antibody fragments with two antigen-binding sites, which may be bivalent or bispecific. The term "diabody" refers to small antibody fragments prepared by constructing scFv fragments using a short linker (approximately 5-10 residues) between the VH and VL domains to achieve inter-chain, rather than intra-chain, V domain pairing, thereby resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. Bispecific diabodies are heterodimers of two "crossover" scFv fragments in which the VH and VL domains of the two antibodies are present on different polypeptide chains. Diabodies are described in further detail, for example, in EP 404097, WO 93 / 11161, Hudson et al. (2003) Nat. Med. 9:129-134, and Hoolinger et al. PNAS USA 90:6444-48 (1993). Triabodies and tetrabodies are also described in Hudson et al. (2003) Nat. Med. 9:129-134.

[0081] As used herein, the terms "specifically recognize" or "specifically bind" refer to a measurable and reproducible interaction, such as attraction or binding, between a target and an antibody that determines the presence of the target in the presence of a heterogeneous population of molecules, including biomolecules. For example, an antibody that specifically or preferentially binds to a target or epitope is one that binds to this target or epitope with higher affinity, avidity, more readily, and / or with longer duration than it binds to other targets or other epitopes of that target. For example, it is understood that an antibody (or moiety) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. Thus, "specific binding" or "preferential binding" does not necessarily require exclusive binding (although it can include exclusive binding).

[0082] An "agonist" or "activating" antibody is an antibody that induces (e.g., increases) one or more activities or functions of an antigen after the antibody binds to the antigen and / or induces (e.g., increases) antigen binding to one or more ligands after the antibody binds to the antigen.

[0083] A "blocking," "antagonist," or "inhibitory" antibody is an antibody that inhibits or reduces (e.g., decreases) antigen binding to one or more ligands and / or inhibits or reduces (e.g., decreases) one or more activities or functions of the antigen after the antibody binds to the antigen. In some embodiments, a blocking, antagonist, or inhibitory antibody substantially or completely inhibits antigen binding to one or more ligands and / or substantially or completely inhibits one or more activities or functions of the antigen.

[0084] As used herein, "subject," "host," or "individual" refers to any animal classified as a mammal, including humans, domestic animals, and livestock, as well as zoo, sport, or pet animals, such as dogs, horses, cats, cows, and animals used in research, such as mice, rats, hamsters, rabbits, and non-human primates. In some embodiments, the mammal is a human.

[0085] As used herein, the term "pharmaceutical formulation" or "pharmaceutical composition" refers to a preparation in a form such that the biological activity of the active ingredient(s) is effective and which does not contain additional components that have unacceptable toxicity to the subject receiving the composition or formulation. A "pharmaceutically acceptable" excipient (e.g., vehicle, additive) is one that can be reasonably administered to a subject and provide an effective dose of the active ingredient(s) for use.

[0086] As used herein, "dermal administration" or "administering dermatologically" refers to the delivery of a composition to a subject by directly or otherwise contacting a formulation containing the composition with all ("systemic") or part ("topical") of the patient's skin. The term encompasses several routes of administration, including, but not limited to, topical and transdermal. Topical administration can be used as a means to deliver a composition to a subject's epidermis or dermis, or specific layers thereof.

[0087] As used herein, an "effective amount" is at least the minimum amount necessary to bring about a measurable improvement or prevention of one or more symptoms of a particular disorder. An "effective amount" may vary depending on factors such as the patient's condition, age, sex, and weight. An effective amount is also one in which the therapeutic beneficial effects outweigh any toxic or adverse effects of the treatment. For prophylactic use, beneficial or desired results include results such as eliminating or reducing the risk of disease, reducing the severity of disease, or delaying the development of the disease, its complications, and intermediate pathological phenotypes manifested during disease development. For therapeutic use, beneficial or desired results include clinical results such as alleviation of one or more symptoms caused by the disease, improving the quality of life of those suffering from the disease, reducing the dose of other medications used to treat the symptoms of the disease, delaying disease progression, and / or extending survival. An effective amount can be administered in one or more administrations. For purposes of this disclosure, an effective amount of a recombinant nucleic acid, virus, and / or pharmaceutical composition is an amount sufficient to achieve prophylactic or therapeutic treatment, either directly or indirectly. As understood in a clinical context, an effective amount of a recombinant nucleic acid, virus, and / or pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administration of one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desired result can be or is achieved.

[0088] As used herein, "treatment" refers to a clinical intervention designed to alter the natural course of the individual or cell being treated during the course of a clinical condition. Desirable effects of treatment include slowing the rate of progression of the disease / disorder / condition, improving or alleviating the disease / disorder / condition, and achieving remission or improving prognosis.

[0089] As used herein, the term "delaying the progression of" a disease / disorder / disorder refers to postponing, preventing, slowing, inhibiting, stabilizing, and / or delaying the onset of the disease / disorder / disorder. This delay may be of different length or duration depending on the history of the disease / disorder / disorder and / or the individual being treated. As will be apparent to one of skill in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease.

[0090] III. Recombinant Nucleic Acids Certain aspects of the present disclosure relate to recombinant nucleic acids (e.g., isolated recombinant nucleic acids) comprising one or more (e.g., one or more, two or more, three or more, four or more, five or more, ten or more, etc.) polynucleotides encoding an antibody. The antibody can be any antibody (in any form) described herein or known in the art. In some embodiments, the antibody is a full-length antibody. In some embodiments, the antibody is an antibody fragment. In some embodiments, the antibody is an agonist antibody. In some embodiments, the antibody is an antagonist antibody.

[0091] In some embodiments, the recombinant nucleic acid is a vector. In some embodiments, the recombinant nucleic acid is a viral vector. In some embodiments, the recombinant nucleic acid is a herpes virus vector. In some embodiments, the recombinant nucleic acid is a herpes simplex virus amplicon. In some embodiments, the recombinant nucleic acid is a recombinant herpes virus genome. In some embodiments, the recombinant nucleic acid is a recombinant herpes simplex virus genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome.

[0092] Polynucleotides encoding antibodies In some embodiments, the present disclosure relates to a recombinant nucleic acid (e.g., a recombinant herpesvirus genome) comprising one or more (e.g., one or more, two or more, three or more, four or more, five or more, ten or more, etc.) polynucleotides encoding an antibody. In some embodiments, at least one of the polynucleotides encodes a single chain antibody (e.g., scFv, scFv-Fc, etc.). In some embodiments, at least one of the polynucleotides comprises multiple expression cassettes encoding an antibody (e.g., a first expression cassette encoding an antibody heavy chain and a second expression cassette encoding an antibody light chain, etc.). In some embodiments, at least one of the polynucleotides encodes a polycistronic mRNA encoding an antibody (e.g., a polycistronic mRNA comprising an ORF encoding an antibody heavy chain and an ORF encoding an antibody light chain separated by an IRES). In some embodiments, at least one of the polynucleotides encodes a chimeric polypeptide (e.g., a polypeptide comprising an antibody heavy chain and an antibody light chain separated by a cleavable linker, etc.). In some embodiments, the recombinant genome comprises one polynucleotide encoding an antibody. In some embodiments, the recombinant genome comprises two or more polynucleotides encoding an antibody (e.g., a first polynucleotide encoding an antibody heavy chain and a second polynucleotide encoding an antibody light chain).

[0093] In some embodiments, a first recombinant nucleic acid of the present disclosure comprises one or more polynucleotides encoding a portion of an antibody (e.g., an antibody heavy chain) and is used in conjunction with a second recombinant nucleic acid comprising one or more polynucleotides encoding a complementary portion of the antibody (e.g., an antibody light chain). In some embodiments, the first and second recombinant nucleic acids are present in a single composition (e.g., comprised in separate herpes simplex viruses formulated as a single pharmaceutical composition). In some embodiments, the first and second recombinant nucleic acids are present in different compositions (e.g., comprised in separate herpes simplex viruses formulated as two separate, single pharmaceutical compositions). In some embodiments, the first recombinant nucleic acid is delivered to a target cell before, along with, or after delivery of the second recombinant nucleic acid to the target cell (e.g., to produce a single full-length antibody in one or more cells of a subject).

[0094] In some embodiments, a recombinant nucleic acid of the disclosure comprises polynucleotides encoding two or more antibodies (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.). In some embodiments, the two or more antibodies are identical. In some embodiments, the two or more antibodies are different.

[0095] antibody Antibodies encoded by one or more of the polynucleotides of the present disclosure may be derived from any suitable species known in the art, including, for example, human antibodies, mouse antibodies, rat antibodies, rabbit antibodies, camel antibodies, chicken antibodies, donkey antibodies, cat antibodies, goat antibodies, sheep antibodies, horse antibodies, hamster antibodies, guinea pig antibodies, shark antibodies, and any chimeric antibodies thereof. In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is a mouse antibody. In some embodiments, the antibody is a chimeric antibody (e.g., a human-mouse chimeric antibody). In some embodiments, the antibody is a humanized antibody.

[0096] Antibodies encoded by one or more of the polynucleotides of the present disclosure can be of any suitable isotype known in the art, including, for example, IgA, IgD, IgE, IgG, IgM, and any combination thereof. In some embodiments, the antibody is an IgG antibody. In some embodiments, the IgG antibody is an IgG1 antibody (see, e.g., SEQ ID NO:596 or SEQ ID NO:597 for exemplary human IgG1 constant region sequences), an IgG2 antibody (see, e.g., SEQ ID NO:598 for exemplary human IgG2 constant region sequences), an IgG3 antibody (see, e.g., SEQ ID NO:599 for exemplary human IgG3 constant region sequences), an IgG4 antibody (see, e.g., SEQ ID NO:600 for exemplary human IgG4 constant region sequences), and any chimeric IgG antibody thereof. In some embodiments, the IgG antibody is an IgG1 antibody.

[0097] In some embodiments, an antibody encoded by one or more polynucleotides of the present disclosure is an antibody fragment. For example, any type or form of antibody fragment known in the art can be encoded by a polynucleotide of the present disclosure, including a Fab fragment, a Fab' fragment, a Fab'-SH fragment, a F(ab')2 fragment, an Fv fragment, an scFv fragment, an scFv-Fc fragment, and any other type or form of antibody fragment described herein or known in the art. In some embodiments, the antibody fragment is a Fab fragment. In some embodiments, the antibody fragment is an scFv. In some embodiments, the antibody fragment is an scFv-Fc. For a review of specific antibody fragments, see, e.g., Hudson et al. (2003) Nat. Med. 9:129-134, Pluckthun The Pharmacology of Monoclonal Antibodies vol. 113, Rosenburg and Moore eds. (Springer-Verlag, New York) pp. 269-315 (1994), WO 93 / 16185, US 5,571,894, US 5,587,458 and US 5,869,046.

[0098] In some embodiments, an antibody encoded by one or more polynucleotides of the present disclosure is a chimeric antibody. Certain chimeric antibodies are described, for example, in US 4,816,567 and Morrison et al. (1984) PNAS USA 81:6851-6855. In some embodiments, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, non-human primate, etc.) and a human constant region. In some embodiments, a chimeric antibody is a "class-switched" antibody, in which the class or subclass is changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof. In some embodiments, a chimeric antibody is a mouse-human chimeric antibody.

[0099] In some embodiments, an antibody encoded by one or more polynucleotides of the present disclosure is a humanized antibody. Typically, non-human antibodies are humanized to reduce immunogenicity in humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody comprises one or more variable domains in which the HVRs (or portions thereof) are derived from a non-human antibody and the FRs (or portions thereof) are derived from human antibody sequences. Optionally, a humanized antibody also comprises at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are substituted with the corresponding residues from the non-human antibody (e.g., the antibody from which the HVR residues are derived) to, for example, restore or improve antibody specificity and / or affinity.

[0100] Humanized antibodies and methods for making them are described, for example, in Almagro and Fransson (2008) Front. Biosci. 13:1619-1633, Riechmann et al. (1988) Nature 332:323-329, Queen et al. (1989) PNAS USA 86:10029-10033, US 5,821,337, US 7,527,791, US 6,982,321, US 7,087,409, Kashmiri et al. (2005) Methods 36:25-34 (describing specificity-determining region (SDR) grafting), Padlam (1991) Mol Immunol 28:489-498 (describing "resurfacing"), Dall'Acqua et al. al. (2005) Methods 36:43-60 (describing "FR shuffling"), Osbourn et al. (2005) Methods 36:61-68, and Klimka et al. (2000) Br J Cancer 83:252-260 (describing a "guided selection" approach to FR shuffling).

[0101] Human framework regions that can be used for humanization include, for example, framework regions selected using the "best-fit" method (see, e.g., Sims et al. (1993) J Immunol 151:2296), framework regions derived from consensus sequences of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. (1992) PNAS USA 89:4285; see also Presta et al. (1993) J Immunol 151:2623), human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson (2008) Front Biosci 13:1619-1633), and framework regions obtained by screening FR libraries (see, e.g., Baca et al. (1997) J Biol Chem 272:10678-10684; see also Rosok et al. (see also, e.g., et al. (1996) J Biol Chem 271:22611-22618).

[0102] In some embodiments, an antibody encoded by one or more polynucleotides of the present disclosure is a human antibody, as generally described in van Dijk and van de Winkel (2001) Curr Opin Pharmacol 5:368-74 and Lonberg (2008) Curr Opin Immunol 20:450-459. Certain details regarding human antibodies can be found, for example, in Hoogenboom and Winter, J. Mol. Biol. 227:381 (1991), Marks et al. J. Mol. Biol. 222:581 (1991), Cole et al. Monoclonal Antibodies and Cancer Therapy, p. 77 (1985), Boerner et al. J. Immunol. 147(1):86-95 (1991), van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5:368-74 (2001), US 6,075,181, US 6,150,584, and Li et al. PNAS USA 103:3557-3562 (2006).

[0103] In some embodiments, an antibody encoded by one or more polynucleotides of the present disclosure is a multispecific antibody (e.g., a bispecific antibody, a trispecific antibody, etc.). Techniques for producing multispecific antibodies include, for example, recombinant coexpression of two immunoglobulin heavy chain-light chain pairs with different specificities (see, e.g., Milstein and Cuello, Nature 305:537 (1983), WO93 / 08829, Traunecker et al. EMBO J. 10:3655 (1991), and "knob-in-hole" engineering (e.g., as described in U.S. Pat. No. 5,731,168)). Multispecific antibodies can also be produced by manipulating electrostatic steering effects to create antibody Fc-heterodimeric molecules (see, e.g., WO2009 / 089004), cross-linking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980; Brennan et al. Science, 229:81 (1985)), using leucine zippers to produce bispecific antibodies (see, e.g., Kostelny et al. J Immunol, 148(5):1547-53 (1992)), using "diabody" technology to create bispecific antibody fragments (see, e.g., Hollinger et al. PNAS USA 90:6444-8 (1993)), using single-chain Fv dimers (see, e.g., Gruber et al. J Immunol, 148(5):1547-53 (1992)), and cross-linking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980; Brennan et al. Science, 229:81 (1985)). Immunol. 152:5368 (1994)), using dual-acting Fabs (see, e.g., US2008 / 0069820), and preparing trispecific or trivalent antibodies (see, e.g., Tutt et al. J Immunol 147:60 (1991), WO2017 / 074878).

[0104] An antibody (or antigen-binding fragment thereof) encoded by one or more polynucleotides of the present disclosure may include (1) HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of any antibody described herein or known in the art, (2) a heavy chain variable region and / or a light chain variable region of any antibody described herein or known in the art, and / or (3) a full-length heavy chain and / or a full-length light chain of any antibody described herein or known in the art. Examples of suitable antibodies that may be encoded by polynucleotides of the present disclosure include, for example, abagovomab, abciximab (V), and avian globulin (AV). H -SEQ ID NO:617, V L -SEQ ID NO:869), abituzumab (V H -SEQ ID NO:618,V L -SEQ ID NO:870), abrezekimab, abrilumab (V H -SEQ ID NO:619,V L -SEQ ID NO:871), actoxumab (V H -SEQ ID NO:620,V L -SEQ ID NO:872), adalimumab (V H -SEQ ID NO:355, V L -SEQ ID NO:420), adecatumumab, aducanumab (V H -SEQ ID NO:621, V L -SEQ ID NO:873), afasevicumab (V H -SEQ ID NO:622, V L -SEQ ID NO:874), afelimomab, afutuzumab, alacizumab (V H -SEQ ID NO:629,V L -SEQ ID NO:881), alemtuzumab (V H -SEQ ID NO:382, V L -SEQ ID NO:447), alirocumab (V H -SEQ ID NO:392, V L -SEQ ID NO:457), altumomab, amatuximab (VH -SEQ ID NO:623, V L -SEQ ID NO:875), anatumomab, andecaliximab (V H -SEQ ID NO:624, V L -SEQ ID NO:876), anetumab (V H -SEQ ID NO:625, V L -SEQ ID NO:877), anifrolumab (V H -SEQ ID NO:387, V L -SEQ ID NO:452), anrukinzumab (V H -SEQ ID NO:626, V L -SEQ ID NO:878), apolizumab, apolizumab (V H -SEQ ID NO:627, V L -SEQ ID NO:879), arcitumomab (V H -SEQ ID NO:628,V L -SEQ ID NO:880), ascribable to bacumab, acelizumab, atezolizumab (V H -SEQ ID NO:374, V L -SEQ ID NO:439), atinumab, atlizumab, atorlimumab, avelumab (V H -SEQ ID NO:630,V L -SEQ ID NO:882), azintuxizumab (V H -SEQ ID NO:631, V L -SEQ ID NO:883), bapineuzumab (V H -SEQ ID NO:632, V L -SEQ ID NO:884), basiliximab (V H -SEQ ID NO:389, V L -SEQ ID NO:454), bavituximab (V H -SEQ ID NO:633, V L -SEQ ID NO:885), bectumomab, begelomab (V H -SEQ ID NO:634,V L -SEQ ID NO:886), belantamab (VH -SEQ ID NO:635, V L -SEQ ID NO:887), belimumab (V H -SEQ ID NO:388, V L -SEQ ID NO:453), bemarituzumab, belimumab, bemarituzumab (V H -SEQ ID NO:636,V L -SEQ ID NO:888), benralizumab (V H -SEQ ID NO:637, V L -SEQ ID NO:889), berlimatoxumab (V H -SEQ ID NO:638,V L -SEQ ID NO:890), versanlimab (V H -SEQ ID NO:639, V L -SEQ ID NO:891), bertilimumab, besilesomab, bevacizumab (V H -SEQ ID NO:357, V L -SEQ ID NO:422), bezlotoxumab (V H -SEQ ID NO:640,V L -SEQ ID NO:892), biciromab, bimagrumab (V H -SEQ ID NO: 415, V L -SEQ ID NO:478), bimekizumab (V H -SEQ ID NO:641, V L -SEQ ID NO:893), vilutamimab (V H -SEQ ID NO:642, V L -SEQ ID NO:894), bivatuzumab, bleselumab (V H -SEQ ID NO:643, V L -SEQ ID NO:895), blinatumomab (V H -SEQ ID NO:644, V L -SEQ ID NO:896), brontuzumab (V H -SEQ ID NO:645, V L -SEQ ID NO:897), brosozumab (V H-SEQ ID NO:646,V L -SEQ ID NO:898), bococizumab (V H -SEQ ID NO:647, V L -SEQ ID NO:899), brazikumab (V H -SEQ ID NO:648,V L -SEQ ID NO:900), brentuximab (V H -SEQ ID NO:649,V L -SEQ ID NO:901), briakinumab (V H -SEQ ID NO:650,V L -SEQ ID NO:902), brodalumab (V H -SEQ ID NO:361, V L -SEQ ID NO:426), brolucizumab (V H -SEQ ID NO:651, V L -SEQ ID NO:903), brontixutuzumab (V H -SEQ ID NO:652, V L -SEQ ID NO:904), burosumab (V H -SEQ ID NO:653, V L -SEQ ID NO:905), cabilalizumab (V H -SEQ ID NO:654, V L -SEQ ID NO:906), camidanlumab (V H -SEQ ID NO:655,V L -SEQ ID NO:907), camrelizumab (V H -SEQ ID NO:656,V L -SEQ ID NO:908), canakinumab (V H -SEQ ID NO:378, V L -SEQ ID NO:443), cantuzumab (V H -SEQ ID NO:657, V L -SEQ ID NO:909), caplacizumab, capromab, carlumab (V H -SEQ ID NO:658,V L -SEQ ID NO:910), carotuximab (VH -SEQ ID NO:659,V L -SEQ ID NO:911), catumaxomab, cedelizumab, cemiplimab (V H -SEQ ID NO:394, V L -SEQ ID NO:459), sergituzumab (V H -SEQ ID NO:660,V L -SEQ ID NO:912), certolizumab (V H -SEQ ID NO:370, V L -SEQ ID NO:435), cetrelimab (V H -SEQ ID NO:661, V L -SEQ ID NO:913), cetuximab (V H -SEQ ID NO:662, V L -SEQ ID NO:914), civisatamab (bispecific:V H1 -SEQ ID NO:660,V L1 -SEQ ID NO:915, V H2 -SEQ ID NO:660,V L2 -SEQ ID NO:912), sitatuzumab (V H -SEQ ID NO:663, V L -SEQ ID NO:916), cizutumumab (V H -SEQ ID NO:664, V L -SEQ ID NO:917), clazakizumab (V H -SEQ ID NO:665, V L -SEQ ID NO:918), clenoliximab, clivatuzumab (V H -SEQ ID NO:666,V L -SEQ ID NO:919), codrituzumab (V H -SEQ ID NO:667, V L -SEQ ID NO:920), cofetuzumab (V H -SEQ ID NO:668,V L -SEQ ID NO:921), coltuximab (V H -SEQ ID NO:669,V L-SEQ ID NO:922), conatumumab (V H -SEQ ID NO:670,V L -SEQ ID NO:923), concizumab (V H -SEQ ID NO:671, V L -SEQ ID NO:924), cosflobiximab (V H -SEQ ID NO:672, V L -SEQ ID NO:925), crenezumab (V H -SEQ ID NO:673, V L -SEQ ID NO:926), crizanlizumab (V H -SEQ ID NO:674, V L -SEQ ID NO:927), cloteduumab (V H -SEQ ID NO:675,V L -SEQ ID NO:928), cusatuzumab (V H -SEQ ID NO:676,V L -SEQ ID NO:929), dacetuzumab (V H -SEQ ID NO:677, V L -SEQ ID NO:930), daclizumab (V H -SEQ ID NO:390,V L -SEQ ID NO:455), dalotuzumab (V H -SEQ ID NO:678,V L -SEQ ID NO:931), dapirolizumab (V H -SEQ ID NO:679,V L -SEQ ID NO:932), daratumumab (V H -SEQ ID NO:680,V L -SEQ ID NO:933), dextrecumab (V H -SEQ ID NO:681, V L -SEQ ID NO:934), demcizumab (V H -SEQ ID NO:682, V L -SEQ ID NO:935), denintuzumab (V H -SEQ ID NO:683, V L-SEQ ID NO:936), denosumab (V H -SEQ ID NO:684,V L -SEQ ID NO:937), depatuxizumab (V H -SEQ ID NO:685,V L -SEQ ID NO:938), dellotuximab (V H -SEQ ID NO:686,V L -SEQ ID NO:939), detumomab, dezamizumab (V H -SEQ ID NO:687,V L -SEQ ID NO:940), dinutuximab (V H -SEQ ID NO:688,V L -SEQ ID NO:941), ziridabu (V H -SEQ ID NO:689,V L -SEQ ID NO:942), domagrozumab (V H -SEQ ID NO:690,V L -SEQ ID NO:943), dorlimomab , drozitumab (V H -SEQ ID NO:691, V L -SEQ ID NO:944), durigotuzumab (V H -SEQ ID NO:692, V L -SEQ ID NO:945), dupilumab (V H -SEQ ID NO:391, V L -SEQ ID NO:456), durvalumab (V H -SEQ ID NO:375, V L -SEQ ID NO:440), dusigitumab (V H -SEQ ID NO:693, V L -SEQ ID NO:946), duvortuxizumab (bispecific:V H1 -SEQ ID NO:694, V L1 -SEQ ID NO:947, V H2 -SEQ ID NO:695,V L2 -SEQ ID NO:948), eclumeximab, eculizumab (VH -SEQ ID NO:385, V L -SEQ ID NO:450), edovacomab, edrecolomab, efalizumab (V H -SEQ ID NO:696,V L -SEQ ID NO:949), efangumab, eldelumab (V H -SEQ ID NO:697, V L -SEQ ID NO:950), elezanumab (V H -SEQ ID NO:698,V L -SEQ ID NO:951), elgemtumab (V H -SEQ ID NO:699,V L -SEQ ID NO:952), elotuzumab (V H -SEQ ID NO:700,V L -SEQ ID NO:953), elcilimomab, emactuzumab (V H -SEQ ID NO:701, V L -SEQ ID NO:954), emapalumab (V H -SEQ ID NO:702, V L -SEQ ID NO:955), emibetuzumab (V H -SEQ ID NO:703, V L -SEQ ID NO:956), emicizumab (bispecific:V H1 -SEQ ID NO:704, V L1 -SEQ ID NO:957, V H2 -SEQ ID NO:705, V L2 -SEQ ID NO:957), enapotamab (V H -SEQ ID NO:706, V L -SEQ ID NO:958), enavatuzumab (V H -SEQ ID NO:707, V L -SEQ ID NO:959), enfortumab (V H -SEQ ID NO:708, V L -SEQ ID NO:960), enlimomab, enoblitzumab (V H -SEQ ID NO:371, VL -SEQ ID NO:436), enokizumab (V H -SEQ ID NO:709, V L -SEQ ID NO:961), enoticum (V H -SEQ ID NO:710,V L -SEQ ID NO:962), ensituximab (V H -SEQ ID NO:711, V L -SEQ ID NO:963), epitumomab, epratuzumab (V H -SEQ ID NO:712, V L -SEQ ID NO:964), eptinezumab (V H -SEQ ID NO:713, V L -SEQ ID NO:965), erenumab (V H -SEQ ID NO: 414, V L -SEQ ID NO:477), erlizumab, ertumaxomab, etaracizumab (V H -SEQ ID NO:714, V L -SEQ ID NO:966), etigilimab (V H -SEQ ID NO:715, V L -SEQ ID NO:967), etrolizumab (V H -SEQ ID NO:716,V L -SEQ ID NO:968), evinacumab (V H -SEQ ID NO: 406, V L -SEQ ID NO:470), evolocumab (V H -SEQ ID NO:717, V L -SEQ ID NO:969), exbivirumab, fanolesomab, faralimomab, faricimab, farletuzumab (V H -SEQ ID NO:718,V L -SEQ ID NO:970), fasinumab (V H -SEQ ID NO: 407, V L -SEQ ID NO:471), felvizumab, fezakinumab (V H -SEQ ID NO:719,VL -SEQ ID NO:971), Fivatuzumab, Ficlatuzumab (V H -SEQ ID NO:720,V L -SEQ ID NO:972), figitumumab (V H -SEQ ID NO:721, V L -SEQ ID NO:973), filibumab (V H -SEQ ID NO:722, V L -SEQ ID NO:974), framvotumab (V H -SEQ ID NO:723, V L -SEQ ID NO:975), fretikumab (V H -SEQ ID NO:724,V L -SEQ ID NO:976), flotetuzumab, fontolizumab, foralumab (V H -SEQ ID NO:725,V L -SEQ ID NO:977), foravirumab (V H -SEQ ID NO:726,V L -SEQ ID NO:978), fremanezumab, fresolimumab (SEQ ID NO:396, V L -SEQ ID NO:461), flunevetomab, fluranumab (V H -SEQ ID NO:727, V L -SEQ ID NO:976), futuximab (V H -SEQ ID NO:728,V L -SEQ ID NO:979), galcanezumab (V H -SEQ ID NO:729,V L -SEQ ID NO:980), galiximab (V H -SEQ ID NO:730,V L -SEQ ID NO:981), gancotamab, ganitumab (V H -SEQ ID NO:731, V L -SEQ ID NO:982), gantenerumab (V H -SEQ ID NO:732, V L-SEQ ID NO:983), gatipotuzumab, gavilimomab, gezivumab, gemtuzumab (V H -SEQ ID NO:733, V L -SEQ ID NO:984), gevokizumab (V H -SEQ ID NO:734, V L -SEQ ID NO:985), gilbetumab, gimsilumab, girentuximab (V H -SEQ ID NO:735, V L -SEQ ID NO:986), glembatumumab (V H -SEQ ID NO:736,V L -SEQ ID NO:987), golimumab (V H -SEQ ID NO:369,V L -SEQ ID NO:434), gomiliximab, goslanemab, guselkumab (V H -SEQ ID NO:362, V L -SEQ ID NO:427), ianalumab, ibalizumab (V H -SEQ ID NO:737, V L -SEQ ID NO:988), ibritumomab, icrucumab (V H -SEQ ID NO:738, V L -SEQ ID NO:989), idarucizumab (V H -SEQ ID NO:739, V L -SEQ ID NO:990), ifavotuzumab, igovomab, iradatuzumab, imalumab (V H -SEQ ID NO:740,V L -SEQ ID NO:991), imaprelimab, imuciromab, imgatuzumab (V H -SEQ ID NO:741, V L -SEQ ID NO:992), inlacumab (V H -SEQ ID NO:742, V L -SEQ ID NO:993), indatuximab (V H -SEQ ID NO:743, V L -SEQ ID NO:994), indusatumab (VH -SEQ ID NO:744, V L -SEQ ID NO:995), Inebilizumab, Inflectra, Infliximab (V H -SEQ ID NO:365,V L -SEQ ID NO:430), intetumumab (V H -SEQ ID NO:745, V L -SEQ ID NO:996), inolimomab, inotuzumab (V H -SEQ ID NO:746, V L -SEQ ID NO:997), ipilimumab (V H -SEQ ID NO:372, V L -SEQ ID NO:437), iratumumab, isatuximab (V H -SEQ ID NO:393, V L -SEQ ID NO:458), iscalimab, istiratumab, itolizumab (V H -SEQ ID NO:747, V L -SEQ ID NO:998), ixekizumab (V H -SEQ ID NO:360,V L -SEQ ID NO:425), keliximab, labetuzumab (V H -SEQ ID NO:748, V L -SEQ ID NO:999), lacnotuzumab, ladiratuzumab, lampalizumab (V H -SEQ ID NO:749, V L -SEQ ID NO:1000), lanadelumab, landgrozumab (V H -SEQ ID NO:750,V L -SEQ ID NO:1001), laprituximab, ralcabiximab, lebrikizumab (V H -SEQ ID NO:751, V L -SEQ ID NO:1002), remaresomab, lendalizumab, lembervimab, lenzilumab (V H -SEQ ID NO:752, V L-SEQ ID NO:1003), lerdelimumab, leronlimab, lesofabumab, letolizumab, lexatumumab, ribivirumab, rifastuzumab (V H -SEQ ID NO:753, V L -SEQ ID NO:1004), ligelizumab (V H -SEQ ID NO: 417, V L -SEQ ID NO:480), loncustuximab, rosatuximab, rilotomab (V H -SEQ ID NO:754, V L -SEQ ID NO:1005), lintuzumab (V H -SEQ ID NO:755,V L -SEQ ID NO:1006), lirilumab (V H -SEQ ID NO:756,V L -SEQ ID NO:1007), roderucizumab (V H -SEQ ID NO:757, V L -SEQ ID NO:1008), lokivetomab (V H -SEQ ID NO:758,V L -SEQ ID NO:1009), lorvotuzumab (V H -SEQ ID NO:759, V L -SEQ ID NO:1010), lucatumumab, lurizumab, lumiliximab (V H -SEQ ID NO:760,V L -SEQ ID NO:1011), lumletuzumab (V H -SEQ ID NO:761, V L -SEQ ID NO:1012), rupartumab, rutikizumab, mapatumumab, marjetuximab (V H -SEQ ID NO:762, V L -SEQ ID NO:1013), marstacimab, maslimomab, mavrilimumab (V H -SEQ ID NO:366,V L -SEQ ID NO:431), matuzumab (V H -SEQ ID NO:763, V L-SEQ ID NO:1014), mepolizumab (V H -SEQ ID N O:764, V L -SEQ ID NO:1015), metelimumab, milatuzumab (V H -SEQ ID NO:765, V L -SEQ ID NO:1016), minletumomab, mirikizumab, mirvetuximab (V H -SEQ ID NO:766,V L -SEQ ID NO:1017), mitumomab, modotuximab (V H -SEQ ID NO:767, V L -SEQ ID NO:1018), mogamulizumab (V H -SEQ ID NO:768,V L -SEQ ID NO:1019), monalizumab (V H -SEQ ID NO:769,V L -SEQ ID NO:1020), morolimumab, mosunetuzumab, motavizumab (V H -SEQ ID NO:770,V L -SEQ ID NO:1021), moxetumomab, muromonab (V H -SEQ ID NO:771, V L -SEQ ID NO:1022), nacolomab, namilumab (V H -SEQ ID NO:772, V L -SEQ ID NO:1023), naptumomab, naratuximab, narutumab (V H -SEQ ID NO:773, V L -SEQ ID NO:1024), natalizumab (V H -SEQ ID NO:384, V L -SEQ ID NO:449), nabicixizumab, nabicixizumab (V H -SEQ ID NO:774, V L -SEQ ID NO:1025), naxitamab, nebacumab, necitumumab (V H -SEQ ID NO:775, V L-SEQ ID NO:1026), nemolizumab (V H -SEQ ID NO:776,V L -SEQ ID NO:1027), nerelimomab, nesbacumab (V H -SEQ ID NO:777, V L -SEQ ID NO:1028), netakimab, nimotuzumab, nirsevimab, nivolumab (V H -SEQ ID NO:376, V L -SEQ ID NO:441), nofetumomab, obiltoxaximab (V H -SEQ ID NO:778,V L -SEQ ID NO:1029), obinutuzumab (V H -SEQ ID NO:779, V L -SEQ ID NO:1030), ocaratu- zumab (V H -SEQ ID NO:780,V L -SEQ ID NO:1031), ocrelizumab (V H -SEQ ID NO:379, V L -SEQ ID NO:444), ozlimomab, ofatumumab (V H -SEQ ID NO:380,V L -SEQ ID NO:445), olaratumab (V H -SEQ ID NO:781, V L -SEQ ID NO:1032), olecurumab, olendalizumab, olokizumab (V H -SEQ ID NO:782, V L -SEQ ID NO:1033), omalizumab (V H -SEQ ID NO: 418, V L -SEQ ID NO:481), onartuzumab (V H -SEQ ID NO:783, V L -SEQ ID NO:1034), ontuxizumab (V H -SEQ ID NO:37,V L -SEQ ID NO:438), onvatilimab, opicinumab (V H -SEQ ID NO:383, VL -SEQ ID NO:448), oportuzumab, oregovomab, olticumab (V H -SEQ ID NO:784, V L -SEQ ID NO:1035), otelixizumab (V H -SEQ ID NO:785,V L -SEQ ID NO:1036), otilimab, otlertuzumab (V H -SEQ ID NO:786,V L -SEQ ID NO:1037), oxelumab (V H -SEQ ID NO:787, V L -SEQ ID NO:1038), ozanezumab (V H -SEQ ID NO:788,V L -SEQ ID NO:1039), ozoralizumab, pagibaximab, palivizumab (V H -SEQ ID NO:789,V L -SEQ ID NO:1040), pamrevlumab (V H -SEQ ID NO:790,V L -SEQ ID NO:1041), panitumumab (V H -SEQ ID NO:791, V L -SEQ ID NO:1042), pancomab, panobacumab (V H -SEQ ID NO:792, V L -SEQ ID NO:1043), palsatuzumab (V H -SEQ ID NO:793, V L -SEQ ID NO:1044), pascolizumab, pasotuxizumab, pateclizumab (V H- SEQ ID NO:794,V L -SEQ ID NO:1045), patritumab (V H -SEQ ID NO:795, V L -SEQ ID NO:1046), pembrolizumab (V H -SEQ ID NO:377, V L -SEQ ID NO:442), pemtumomab, perakizumab (V H-SEQ ID NO:796,V L -SEQ ID NO:1047), pertuzumab (V H -SEQ ID NO:797, V L -SEQ ID NO:1048), pexelizumab, pidilizumab (V H -SEQ ID NO:798, V L -SEQ ID NO:1049), pinatuzumab (V H -SEQ ID NO:799, V L -SEQ ID NO:1050), pintumomab, placurab, prosalizumab (V H -SEQ ID NO:800,V L -SEQ ID NO:1051), pogalizumab, polatuzumab (V H -SEQ ID NO:801, V L -SEQ ID NO:1052), ponezumab (V H -SEQ ID NO:802, V L -SEQ ID NO:1053), polgabiximab, prasinezumab, prezalizumab, priliximab, plitoxaximab (V H -SEQ ID NO:803, V L -SEQ ID NO:1054), Pritumumab, Kirizumab (V H -SEQ ID NO:804, V L -SEQ ID NO:1055), racotumomab (V H -SEQ ID NO:805, V L -SEQ ID NO:1056), radletumab (V H -SEQ ID NO:806,V L -SEQ ID NO:1057), rafivirumab (V H -SEQ ID NO:807, V L -SEQ ID NO:1058), ralpancizumab (V H -SEQ ID NO:808,V L -SEQ ID NO:1059), ramucirumab (V H -SEQ ID NO:809,V L-SEQ ID NO:1060), ranevetomab, ranibizumab (V H -SEQ ID NO: 416, V L -SEQ ID NO:479), raxibacumab, ravagalimab, ravutolizumab, refanezumab (V H -SEQ ID NO:810,V L -SEQ ID NO:1061), regavirumab, lemtolumab, reslizumab (V H -SEQ ID NO:811, V L -SEQ ID NO:1062), rilotumumab (V H -SEQ ID NO:812, V L -SEQ ID NO:1063), linucumab (V H -SEQ ID NO:813, V L -SEQ ID NO:1064), risankizumab (V H -SEQ ID NO:363, V L -SEQ ID NO:428), rituximab (V H -SEQ ID NO:356,V L -SEQ ID NO:421), rivavazumab (V H -SEQ ID NO:814,V L -SEQ ID NO:1065), lobatumumab (V H -SEQ ID NO:815,V L -SEQ ID NO:1066), lorezumab (V H -SEQ ID NO:816,V L -SEQ ID NO:1067), romilukimab, romozozumab (V H -SEQ ID NO:817,V L -SEQ ID NO:1068), lontalizumab (V H -SEQ ID NO:818,V L -SEQ ID NO:1069), rosmantuzumab, rovalpituzumab (V H -SEQ ID NO:819,V L -SEQ ID NO:1070), rovelizumab, rozanolixizumab, ruplizumab, sacituzumab (V H-SEQ ID NO:820,V L -SEQ ID NO:1071), samalizumab (V H -SEQ ID NO:821, V L -SEQ ID NO:1072), samrotamab, sapelizumab, sarilumab (V H -SEQ ID NO:368,V L -SEQ ID NO:433), satralizumab (V H -SEQ ID NO:386,V L -SEQ ID NO:451), satumomab (V H -SEQ ID NO:822, V L -SEQ ID NO:1073), secukinumab (V H -SEQ ID NO:359, V L -SEQ ID NO:424), cericlerumab, seribantumab (V H -SEQ ID NO:823, V L -SEQ ID NO:1074), cetoxaximab (V H -SEQ ID NO:824,V L -SEQ ID NO:1075), setrusumab, sevilumab, sibrotuzumab, sifalimumab (V H -SEQ ID NO:825,V L -SEQ ID NO:1076), siltuximab (V H -SEQ ID NO:826,V L -SEQ ID NO:1077), simtuzumab (V H -SEQ ID NO:827, V L -SEQ ID NO:1078), siplizumab, siltrazumab, sirukumab (V H -SEQ ID NO:828,V L -SEQ ID NO:1079), sofituzumab (V H -SEQ ID NO:829,V L -SEQ ID NO:1080), solanezumab (V H -SEQ ID NO:830,V L-SEQ ID NO:1081), solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab, sulesomab, sputumumab, stimulimab, suvizumab (V H -SEQ ID NO:831, V L -SEQ ID NO:1082), subratoxumab, tabalumab (V H -SEQ ID NO:832, V L -SEQ ID NO:1083), tacatuzumab, tadocizumab, talacutuzumab, talizumab, tamtubetumab, tanezumab (V H -SEQ ID NO:833, V L -SEQ ID NO:1084), taplitumomab, tarexuzumab (V H -SEQ ID NO:834, V L -SEQ ID NO:1085), taborimab, tefibazumab, terimomab, telisotuzumab, tenatumomab (V H -SEQ ID NO:835, V L -SEQ ID NO:1086), teneliximab, teplizumab (V H -SEQ ID NO:836,V L -SEQ ID NO:1087), tepositamab, teprotumumab (V H -SEQ ID NO:837, V L -SEQ ID NO:1088), tesidolumab (V H -SEQ ID NO:838,V L -SEQ ID NO:1089), tetulomab, tezepelumab (V H -SEQ ID NO:839,V L -SEQ ID NO:1090), tiburizumab, tildrakizumab (V H -SEQ ID NO:364, V L -SEQ ID NO:429), tigatuzumab (V H -SEQ ID NO:840,V L -SEQ ID NO:1091), timigituzumab, timolumab (V H -SEQ ID NO:841, V L-SEQ ID NO:1092), tiragotumab, tislelizumab, tisotumab (V H -SEQ ID NO:842, V L -SEQ ID NO:1093), tocilizumab (V H -SEQ ID NO:367, V L -SEQ ID NO:432), tomzotuximab, toralizumab, tosatoxumab (V H -SEQ ID NO:843, V L -SEQ ID NO:1094), tositumomab, tobetumab (V H -SEQ ID NO:844, V L -SEQ ID NO:1095), tralokinumab (V H -SEQ ID NO:845, V L -SEQ ID NO:1096), trastuzumab (V H -SEQ ID NO:846,V L -SEQ ID NO:1097), tregalizumab (V H -SEQ ID NO:847, V L -SEQ ID NO:1098), tremelimumab (V H -SEQ ID NO:848,V L -SEQ ID NO:1099), trevoglumab (V H -SEQ ID NO:849,V L -SEQ ID NO:1100), tucotuzumab, tuvilumab, ublituximab (V H -SEQ ID NO:381, V L -SEQ ID NO:446), urocupulumab (V H -SEQ ID NO:850,V L -SEQ ID NO:1101), urelumab (V H -SEQ ID NO:851, V L -SEQ ID NO:1102), urtoxazumab, ustekinumab (V H -SEQ ID NO:358,V L -SEQ ID NO:423), utomilumab, vadastuximab (V H -SEQ ID NO:852, VL -SEQ ID NO:1103), banalimab, bundletuzumab (V H -SEQ ID NO:853, V L -SEQ ID NO:1104), vanticutumab (V H -SEQ ID NO:854, V L -SEQ ID NO:1105), vanucizumab (bispecific:V H1 -SEQ ID NO:855,V L1 -SEQ ID NO:1106, V H2 -SEQ ID NO:357, V L2 -SEQ ID NO:422), bapaliximab, valisacumab, varlilumab (V H -SEQ ID NO:856,V L -SEQ ID NO:1107), batelizumab (V H -SEQ ID NO:857, V L -SEQ ID NO:1108), vedolizumab (V H -SEQ ID NO:858,V L -SEQ ID NO:1109), veltuzumab (V H -SEQ ID NO:859,V L -SEQ ID NO:1110), bepalimomab, besencumab (V H -SEQ ID NO:860,V L -SEQ ID NO:1111), visilizumab (V H -SEQ ID NO:861, V L -SEQ ID NO:1112), bovalilizumab, volociximab, bonlerolizumab, bopratelimab, borsetuzumab (V H -SEQ ID NO:862, V L -SEQ ID NO:1113), votumumab, bunakizumab, xentuzumab (V H -SEQ ID NO:863, V L -SEQ ID NO:1114), zalutumumab (V H -SEQ ID NO:864,V L -SEQ ID NO:1115), zanolimumab (V H-SEQ ID NO:865,V L -SEQ ID NO:1116), zatuximab, zenoctuzumab, dilarimumab, zolbetuximab, and zolimomab. In some embodiments, the antibody is not any one or more of the above-mentioned antibodies (e.g., it does not comprise any one or more heavy chain variable regions and / or light chain variable regions of any one or more of the above-mentioned antibodies). In some embodiments, the antibody is not an anti-CTLA4 antibody and / or an anti-PD-L1 antibody.

[0105] In some embodiments, an antibody of the disclosure has at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of the light chain variable region of any of the antibodies described herein or known in the art. (e.g., an antibody of the present disclosure comprises a light chain variable region having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any of the light chain variable regions disclosed in the preceding paragraph). In some embodiments, an antibody of the disclosure has at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of the heavy chain variable region of any of the antibodies described herein or known in the art. (e.g., an antibody of the present disclosure comprises a heavy chain variable region having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the sequence of any of the heavy chain variable regions disclosed in the preceding paragraph).In some embodiments, an antibody of the disclosure comprises a light chain variable region and a heavy chain variable region that have at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the light chain variable region and heavy chain variable region sequences of any of the antibodies described herein or known in the art. and heavy chain variable regions (e.g., antibodies of the present disclosure comprise light chain variable regions and heavy chain variable regions that have at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity, respectively, to the sequences of any of the light chain variable regions and heavy chain variable regions disclosed in the preceding paragraph).

[0106] Exemplary Antibody Sequences In some embodiments, one or more polynucleotides of the present disclosure encode an antibody (e.g., a full-length antibody, an antibody fragment, etc.) comprising a heavy chain variable region comprising HVR-H1, HVR-H2, and / or HVR-H3 of any of the antibodies described herein or known in the art. Methods for identifying HVR-H1, HVR-H2, and / or HVR-H3 in a given heavy chain variable region are generally known to those of skill in the art (see, e.g., abysis.org; methods used in Al-Lazikani et al., (1997) JMB 273, 927-948; Martin, ACR (1996) Proteins 25(1):130-3, etc.). In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising one, two, or three HVRs selected from HVR-H1 comprising a sequence selected from SEQ ID NOs: 1-59, HVR-H2 comprising a sequence selected from SEQ ID NOs: 60-122, and / or HVR-H3 comprising a sequence selected from SEQ ID NOs: 123-185. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3 of any of the antibodies shown in Table 1. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3 of any of the heavy chain variable regions shown in Table 2, or set forth in SEQ ID NOs: 355-419 or SEQ ID NOs: 614-865.

[0107] In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a light chain variable region comprising the HVR-L1, HVR-L2, and / or HVR-L3 of any of the antibodies described herein or known in the art. Methods for identifying the HVR-L1, HVR-L2, and / or HVR-L3 in a given light chain variable region are generally known to those of skill in the art (see, e.g., abysis.org; Al-Lazikani et al., (1997) JMB 273, 927-948; Martin, ACR (1996) Proteins 25(1):130-3, etc.). In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a light chain variable region comprising one, two, or three HVRs selected from HVR-L1 comprising a sequence selected from SEQ ID NOs:186-242, HVR-L2 comprising a sequence selected from SEQ ID NOs:243-294, and / or HVR-L3 comprising a sequence selected from SEQ ID NOs:295-354. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3 of any of the antibodies shown in Table 1. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3 of any of the light chain variable regions shown in Table 2, or set forth in SEQ ID NOs:420-482 or SEQ ID NOs:866-1116.

[0108] In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region comprising HVR-H1, HVR-H2, and / or HVR-H3 of any of the antibodies described herein or known in the art, and a light chain variable region comprising HVR-L1, HVR-L2, and / or HVR-L3 of any of the antibodies described herein or known in the art. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising one, two, or three HVRs selected from HVR-H1 comprising a sequence selected from SEQ ID NOs: 1-59, HVR-H2 comprising a sequence selected from SEQ ID NOs: 60-122, and / or HVR-H3 comprising a sequence selected from SEQ ID NOs: 123-185, and a light chain variable region comprising one, two, or three HVRs selected from HVR-L1 comprising a sequence selected from SEQ ID NOs: 186-242, HVR-L2 comprising a sequence selected from SEQ ID NOs: 243-294, and / or HVR-L3 comprising a sequence selected from SEQ ID NOs: 295-354. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of any of the antibodies set forth in Table 1. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of any of the heavy chain and light chain variable regions shown in Table 2, or set forth in SEQ ID NOs:355-419, SEQ ID NOs:420-482, SEQ ID NOs:614-865, or SEQ ID NOs:86-1116.

[0109] Table 1. HVRs of exemplary antibodies TIFF2025186440000002.tif221152TIFF2025186440000003.tif218152TIFF2025186440000004.tif218152

[0110] In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising HVR-H1 comprising the sequence of SEQ ID NO:1, HVR-H2 comprising the sequence of SEQ ID NO:60, and HVR-H3 comprising the sequence of SEQ ID NO:123. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising HVR-H1 comprising the sequence of SEQ ID NO:2, HVR-H2 comprising the sequence of SEQ ID NO:61, and HVR-H3 comprising the sequence of SEQ ID NO:124. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising HVR-H1 comprising the sequence of SEQ ID NO:3, HVR-H2 comprising the sequence of SEQ ID NO:62, and HVR-H3 comprising the sequence of SEQ ID NO:125.

[0111] In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a light chain variable region comprising an HVR-L1 comprising the sequence of SEQ ID NO: 186, an HVR-L2 comprising the sequence of SEQ ID NO: 243, and an HVR-L3 comprising the sequence of SEQ ID NO: 295. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a light chain variable region comprising an HVR-L1 comprising the sequence of SEQ ID NO: 187, an HVR-L2 comprising the sequence of SEQ ID NO: 244, and an HVR-L3 comprising the sequence of SEQ ID NO: 296. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a light chain variable region comprising an HVR-L1 comprising the sequence of SEQ ID NO: 188, an HVR-L2 comprising the sequence of SEQ ID NO: 245, and an HVR-L3 comprising the sequence of SEQ ID NO: 2297.

[0112] In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising an HVR-H1 comprising the sequence of SEQ ID NO: 1, an HVR-H2 comprising the sequence of SEQ ID NO: 60, an HVR-H3 comprising the sequence of SEQ ID NO: 123, an HVR-L1 comprising the sequence of SEQ ID NO: 186, an HVR-L2 comprising the sequence of SEQ ID NO: 243, and an HVR-L3 comprising the sequence of SEQ ID NO: 295. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising an HVR-H1 comprising the sequence of SEQ ID NO: 2, an HVR-H2 comprising the sequence of SEQ ID NO: 61, an HVR-H3 comprising the sequence of SEQ ID NO: 124, an HVR-L1 comprising the sequence of SEQ ID NO: 187, an HVR-L2 comprising the sequence of SEQ ID NO: 244, and an HVR-L3 comprising the sequence of SEQ ID NO: 296. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising an HVR-H1 comprising the sequence of SEQ ID NO:3, an HVR-H2 comprising the sequence of SEQ ID NO:62, an HVR-H3 comprising the sequence of SEQ ID NO:125, an HVR-L1 comprising the sequence of SEQ ID NO:188, an HVR-L2 comprising the sequence of SEQ ID NO:245, and an HVR-L3 comprising the sequence of SEQ ID NO:297.

[0113] In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region of any of the antibodies described herein or known in the art. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region comprising a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence selected from SEQ ID NOs:355-419 or SEQ ID NOs:614-865. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region comprising a sequence selected from SEQ ID NOs:355-419 or SEQ ID NOs:614-865. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising the heavy chain variable region of any of the antibodies shown in Table 2.

[0114] In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a light chain variable region of any of the antibodies described herein or known in the art. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a light chain variable region comprising a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence selected from SEQ ID NOs:420-482 or SEQ ID NOs:866-1116. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a light chain variable region comprising a sequence selected from SEQ ID NOs:420-482 or SEQ ID NOs:866-1116. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising the light chain variable region of any of the antibodies shown in Table 2.

[0115] In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising the heavy chain variable region and the light chain variable region of any of the antibodies described herein or known in the art. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising a sequence having at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence selected from SEQ ID NOs: 355-419 or SEQ ID NOs: 420-482 or SEQ ID NOs: 420-483. In some embodiments, one or more polynucleotides of the disclosure encode an antibody comprising a heavy chain variable region comprising a sequence selected from SEQ ID NOs:355-419 or SEQ ID NOs:614-865, and a light chain variable region comprising a sequence selected from SEQ ID NOs:420-482 or SEQ ID NOs:866-1116.In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region and a light chain variable region that each independently have at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the heavy chain variable region and light chain variable region of any of the antibodies set forth in Table 2. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising the heavy chain variable region and light chain variable region of any of the antibodies set forth in Table 2.

[0116] Table 2: Heavy and light chain variable regions of exemplary antibodies TIFF2025186440000005.tif25152TIFF2025186440000006.tif217152TIFF20251864400 00007.tif217152TIFF2025186440000008.tif217152TIFF2025186440000009.tif187152

[0117] In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region comprising the sequence of SEQ ID NO:355 and a light chain variable region comprising the sequence of SEQ ID NO:420. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region comprising the sequence of SEQ ID NO:356 and a light chain variable region comprising the sequence of SEQ ID NO:421. In some embodiments, one or more polynucleotides of the present disclosure encode an antibody comprising a heavy chain variable region comprising the sequence of SEQ ID NO:357 and a light chain variable region comprising the sequence of SEQ ID NO:422.

[0118] A polynucleotide of the present disclosure that encodes an antibody may further encode additional coding and non-coding sequences, including, but not limited to, sequences encoding additional polypeptide tags (e.g., polypeptide tags encoded in-frame with an antibody to produce a fusion protein), introns (e.g., natural, modified, or heterologous introns), 5'UTRs and / or 3'UTRs (e.g., natural, modified, or heterologous 5'UTRs and / or 3'UTRs), etc. Examples of suitable polypeptide tags may include, but are not limited to, purification tags (e.g., his-tags, flag-tags, maltose-binding protein, and glutathione-S-transferase tags), detection tags such as tags that can be detected photometrically (e.g., green fluorescent protein or red fluorescent protein), and tags with detectable enzymatic activity (e.g., alkaline phosphatase), tags containing secretory sequences, signal sequences, leader sequences, and / or stabilizing sequences, any combination of protease cleavage sites (e.g., furin cleavage sites, TEV cleavage sites, thrombin cleavage sites, etc.). In some embodiments, the 5' UTR and / or 3' UTR improves the stability, localization, and / or translation efficiency of the polynucleotide. In some embodiments, the 5' UTR and / or 3' UTR improves the level and / or duration of protein expression. In some embodiments, the 5'UTR and / or 3'UTR contain elements (e.g., one or more miRNA binding sites) that can prevent or reduce off-target expression (e.g., inhibit expression in particular cell types (e.g., neuronal cells), at particular times in the cell cycle, at particular developmental stages, etc.). In some embodiments, the 5'UTR and / or 3'UTR contain elements (e.g., one or more miRNA binding sites) that can enhance antibody expression in particular cell types.

[0119] In some embodiments, the polynucleotides of the disclosure encode (in frame) a leader, signal and / or secretory sequence at the N-terminus of the encoded antibody. Any leader, signal and / or secretory sequence known in the art can be encoded by the polynucleotides of the present disclosure, including, for example, a naturally occurring antibody signal sequence (e.g., any of the antibody leader sequences described in Retter I et al. VBASE2, an integrative V gene database. Nucleic Acids Res. 2005 Jan 1;33:D671-4), or a heterologous or synthetic signal sequence (e.g., those described in von Heijne G. (1983) Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem 133(1)17-21; Martoglio B. and Dobberstein B. (1998) Signal sequences: More than just greasy peptides. Trends Cell Biol 8(10),410-5; Hegde RS and Bernstein HD (2006) The surprising complexity of signal sequences. Trends Biochem Sci 31(10),563-71; Kapp K., Schrempf S., Lemberg MK and Dobberstein B. (2009) Post-Targeting Functions of Signal Peptides. Chapter in: Protein Transport into the Endoplasmic Reticulum, Landes Bioscience, and see sequences disclosed at www.signalpeptide.de. Exemplary secretory sequences include the human CD33 leader sequence TIFF2025186440000010.tif4128, human IL2 leader sequence TIFF2025186440000011.tif4128, human tissue plasminogen activator leader sequence TIFF2025186440000012.tif4128, human antibody leader sequences (Tables 3A-3C) and synthetic secrecon leader sequences Includes TIFF2025186440000013.tif4128.

[0120] Table 3A: Human antibody heavy chain leader sequences TIFF2025186440000014.tif59146

[0121] Table 3B: Human antibody light chain (kappa) leader sequences TIFF2025186440000015.tif59152

[0122] Table 3C: Human antibody light chain (lambda) leader sequences TIFF2025186440000016.tif63146

[0123] In some embodiments, a polynucleotide of the present disclosure encoding an antibody is operably linked to one or more (e.g., one or more, two or more, three or more, four or more, five or more, ten or more, etc.) regulatory sequences. The term "regulatory sequence" can include enhancers, insulators, promoters, and other expression control elements (e.g., polyadenylation signals). Any suitable enhancer(s) known in the art can be used, including, for example, enhancer sequences derived from mammalian genes (e.g., globin, elastase, albumin, alpha-fetoprotein, insulin, etc.), enhancer sequences derived from eukaryotic viruses (e.g., the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, adenovirus enhancers, etc.), and any combination thereof. Any suitable insulator(s) known in the art can be used, including, for example, the HSV chromatin boundary (CTRL / CTCF binding / insulator) elements CTRL1 and / or CTRL2, the chicken hypersensitive site 4 insulator (cHS4), the human HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE), the scaffold / matrix attachment region (S / MAR) from the human interferon beta gene (IFNB1), and any combination thereof.Any suitable promoter known in the art (e.g., a promoter suitable for transcription in a mammalian host cell) can be used, including, for example, promoters obtained from the genomes of viruses (e.g., polyomavirus, fowlpox virus, adenovirus (such as adenovirus 2), bovine papillomavirus, avian sarcoma virus, cytomegalovirus, retroviruses, hepatitis B virus, simian virus 40 (SV40), etc.), promoters derived from heterologous mammalian genes (e.g., from an actin promoter (e.g., a β-actin promoter), a ubiquitin promoter (e.g., a ubiquitin C (UbC) promoter), a phosphoglycerate kinase (PG) promoter, an immunoglobulin promoter, a heat shock promoter, etc.), promoters derived from homologous mammalian genes (e.g., a native human immunoglobulin promoter), synthetic promoters (such as the CAGG promoter), and any combination thereof (provided such promoters are compatible with the host cell). Regulatory sequences can include those that direct constitutive expression of the nucleic acid, as well as tissue-specific regulatory sequences and / or inducible or repressible sequences.

[0124] In some embodiments, the polynucleotides of the present disclosure are operably linked to one or more heterologous promoters. In some embodiments, the one or more heterologous promoters are one or more of a constitutive promoter, a tissue-specific promoter, a temporal promoter, a spatial promoter, an inducible promoter, and a repressible promoter. In some embodiments, the one or more heterologous promoters are one or more of a human cytomegalovirus (HCMV) immediate early promoter, a human elongation factor-1 (EF1) promoter, a human β-actin promoter, a human UbC promoter, a human PGK promoter, a human immunoglobulin promoter, a synthetic CAGG promoter, and any combination thereof. In some embodiments, the polynucleotides of the present disclosure encoding an antibody are operably linked to an HCMV promoter.

[0125] In some embodiments, a polynucleotide of the present disclosure does not include a coding sequence for (e.g., a transgene encoding) a collagen alpha-1 (VII) chain polypeptide (COL7). In some embodiments, a polynucleotide of the present disclosure does not include a coding sequence for (e.g., a transgene encoding) a lysyl hydroxylase 3 polypeptide (LH3). In some embodiments, a polynucleotide of the present disclosure does not include a coding sequence for (e.g., a transgene encoding) a keratin type I cytoskeletal 17 polypeptide (KRT17). In some embodiments, a polynucleotide of the present disclosure does not include a coding sequence for (e.g., a transgene encoding) a transglutaminase (TGM) polypeptide (e.g., a human transglutaminase polypeptide such as a human TGM1 polypeptide). In some embodiments, the polynucleotides of the present disclosure do not include coding sequences (e.g., coding sequences of transgenes encoding) for cosmetic proteins (e.g., collagen proteins, fibronectin, elastin, lumican, vitronectin / vitronectin receptor, laminin, neuromodulators, fibrillin, additional skin extracellular matrix proteins, etc.). In some embodiments, the polynucleotides of the present disclosure do not include coding sequences (e.g., coding sequences of transgenes encoding) for collagen alpha-1(VII) chain polypeptides, lysyl hydroxylase 3 polypeptides, keratin type I cytoskeletal 17 polypeptides, and / or any chimeric polypeptides thereof. In some embodiments, the polynucleotides of the present disclosure do not include coding sequences (e.g., coding sequences of transgenes encoding) for collagen alpha-1(VII) chain polypeptides, lysyl hydroxylase 3 polypeptides, keratin type I cytoskeletal 17 polypeptides, transglutaminase (TGM) polypeptides (e.g., human transglutaminase polypeptides such as human TGM1 polypeptide), cosmetic proteins, and / or any chimeric polypeptides thereof.

[0126] Polynucleotides encoding single-chain antibodies In some embodiments, a recombinant nucleic acid (e.g., a recombinant herpesvirus genome) of the present disclosure comprises one or more polynucleotides encoding an antibody, wherein the antibody is a single-chain antibody. Any form of single-chain antibody known in the art can be encoded by a polynucleotide of the present disclosure. In some embodiments, a single-chain antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, a single-chain antibody comprises a heavy chain variable region and a light chain variable region separated by a linker polypeptide. In some embodiments, a single-chain antibody comprises, from N-terminus to C-terminus, 1) a heavy chain variable region, 2) a linker polypeptide, and 3) a light chain variable region. In some embodiments, a single-chain antibody comprises, from N-terminus to C-terminus, 1) a heavy chain variable region, 2) a linker polypeptide, and 3) a heavy chain variable region. In some embodiments, a single-chain antibody further comprises an antibody hinge region (e.g., an IgG1 hinge region). An exemplary IgG1 hinge region is set forth as SEQ ID NO:603. In some embodiments, the single chain antibody further comprises an antibody Fc region (e.g., an IgG1 Fc region). An exemplary human IgG1 Fc region is shown as SEQ ID NO:604. In some embodiments, the single chain antibody is an scFv-Fc antibody (e.g., an scFv fused to the hinge region and Fc region of an IgG1 antibody heavy chain).

[0127] Any suitable linker polypeptide known in the art can be used in the single chain antibodies of the present disclosure, including, for example, Contains TIFF2025186440000017.tif57151.

[0128] Exemplary polypeptides encoding single-chain antibodies comprising a leader sequence, an antibody heavy chain variable region, a linker polypeptide, an antibody light chain variable region, and an antibody hinge region and Fc region are set forth as SEQ ID NOs:578-583.

[0129] Polynucleotides containing multiple expression cassettes In some embodiments, a recombinant nucleic acid (e.g., a recombinant herpesvirus genome) of the disclosure comprises one or more polynucleotides encoding an antibody, wherein at least one of the polynucleotides comprises two or more expression cassettes. In some embodiments, the polynucleotide comprises, from 5' to 3', a first expression cassette encoding a polypeptide comprising an antibody heavy chain variable region (e.g., a full-length antibody heavy chain) and a second expression cassette encoding a polypeptide comprising an antibody light chain variable region (e.g., a full-length antibody light chain). In some embodiments, the polynucleotide comprises, from 5' to 3', a first expression cassette encoding a polypeptide comprising an antibody light chain variable region (e.g., a full-length antibody light chain) and a second expression cassette encoding a polypeptide comprising an antibody heavy chain variable region (e.g., a full-length antibody heavy chain). In some embodiments, the first and second expression cassettes have separate regulatory sequences (e.g., promoters, enhancers, polyadenylation signals, etc.).

[0130] In some embodiments, the first and second expression cassettes are present in the same orientation in the DNA. In some embodiments, the first and second expression cassettes are present in the DNA facing each other. Without wishing to be bound by theory, integrating the two expression cassettes in the antisense orientation (opposite DNA strands) can help avoid read-through and ensure proper expression of each cassette.

[0131] Polynucleotides encoding polycistronic mRNAs In some embodiments, a recombinant nucleic acid (e.g., a recombinant herpesvirus genome) of the present disclosure comprises one or more polynucleotides encoding an antibody, wherein at least one of the polynucleotides encodes a polycistronic mRNA. In some embodiments, the polycistronic mRNA comprises 1) a first open reading frame (ORF) encoding a polypeptide comprising an antibody heavy chain variable region (e.g., an antibody heavy chain), and 2) a second open reading frame (ORF) encoding a polypeptide comprising an antibody light chain variable region (e.g., an antibody light chain). In some embodiments, the polycistronic mRNA comprises 1) a first open reading frame (ORF) encoding a polypeptide comprising an antibody light chain variable region (e.g., an antibody light chain), and 2) a second open reading frame (ORF) encoding a polypeptide comprising an antibody heavy chain variable region (e.g., an antibody heavy chain). In some embodiments, the polycistronic mRNA further comprises an internal ribosome entry site (IRES) separating the first and second ORFs. In some embodiments, the polycistronic mRNA comprises, from 5' to 3', a first ORF encoding a polypeptide comprising an antibody heavy chain variable region-IRES-a second ORF encoding a polypeptide comprising an antibody light chain variable region. In some embodiments, the polycistronic mRNA comprises, from 5' to 3', a first ORF encoding a polypeptide comprising an antibody light chain variable region-IRES-a second ORF encoding a polypeptide comprising an antibody heavy chain variable region.

[0132] Any suitable IRES known in the art can be used in the polycistronic mRNAs of the present disclosure, including, for example, IRES derived from viruses (e.g., IRES derived from poliovirus, rhinovirus, encephalomyocarditis virus (EMCV), foot-and-mouth disease virus, hepatitis C virus, swine fever virus, Rous sarcoma virus, human immunodeficiency virus, cricket paralysis virus, Kaposi's sarcoma-associated herpesvirus, etc.), IRES derived from cellular mRNAs (e.g., IRES derived from growth factor mRNAs such as fibroblast growth factor 2, platelet-derived growth factor B, and vascular endothelial growth factor; IRES derived from transcription factor mRNAs such as antennapedia, ultrabithorax, and NF-κB inhibitor; c-myc, pim-1, and protein kinase p58). PITSLRE such as IRESs derived from oncogene mRNAs, synthetic IRES (e.g., CP148 IRES), as well as others (see, e.g., Mokrejs et al. (2007) A Bioinformatical Approach to the Analysis of Viral and Cellular Internal Ribosome Entry Sites. Columbus F editors. New Messenger RNA Research Communications. Hauppauge, NY: Nova Science Publishers; pp. 133-166; see also Mokrejs et al. (2006) Nucleic Acids Res 1;34(Database issue):D125-30).

[0133] In some embodiments, the IRES is the CP148 IRES. An exemplary nucleic acid sequence encoding the CP148 IRES is set forth as SEQ ID NO:584. In some embodiments, the IRES is the EMCV IRES. An exemplary nucleic acid sequence encoding the EMCV IRES is set forth as SEQ ID NO:585.

[0134] In some embodiments, the nucleic acid sequence encoding the IRES has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleic acid sequence selected from SEQ ID NO: 584 or SEQ ID NO: 585. In some embodiments, the nucleic acid sequence encoding the IRES has the sequence of SEQ ID NO: 584 or SEQ ID NO: 585.

[0135] Exemplary polynucleotides encoding a polycistronic mRNA comprising 1) a first ORF encoding a leader sequence and an antibody light chain (kappa), 2) an IRES, and 3) a second ORF encoding a leader sequence and an antibody heavy chain (IgG1) are set forth as SEQ ID NOs:586-588.

[0136] Polynucleotides encoding chimeric polypeptides In some embodiments, a recombinant nucleic acid (e.g., a recombinant herpesvirus genome) of the present disclosure comprises one or more polynucleotides encoding a chimeric polypeptide comprising an antibody heavy chain variable region and an antibody light chain variable region, separated by a cleavable linker polypeptide. In some embodiments, the chimeric polypeptide comprises a first amino acid sequence comprising an antibody heavy chain and a second amino acid sequence comprising an antibody light chain, separated by a cleavable linker polypeptide. In some embodiments, the chimeric polypeptide comprises, from N-terminus to C-terminus, 1) a first amino acid sequence comprising an antibody light chain variable region (e.g., antibody light chain), 2) a cleavable linker, and 3) a second amino acid sequence comprising an antibody heavy chain variable region (e.g., antibody heavy chain). In some embodiments, the chimeric polypeptide comprises, from N-terminus to C-terminus, 1) a first amino acid sequence comprising an antibody heavy chain variable region (e.g., antibody heavy chain), 2) a cleavable linker, and 3) a second amino acid sequence comprising an antibody light chain variable region (e.g., antibody light chain).

[0137] Any cleavable linker polypeptide known in the art can be used in the chimeric polypeptides of the present disclosure, including, for example, Contains TIFF2025186440000018.tif31128.

[0138] In some embodiments, the linker polypeptide comprises a sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence selected from SEQ ID NOs: 589-592. In some embodiments, the linker polypeptide comprises a sequence selected from SEQ ID NOs: 589-592.

[0139] Exemplary polynucleotides encoding chimeric polypeptides comprising a leader sequence, an antibody light chain, a linker polypeptide, a leader sequence, and an antibody heavy chain are set forth as SEQ ID NOs:593-595.

[0140] two or more polynucleotides In some embodiments, a recombinant nucleic acid (e.g., a recombinant herpesvirus genome) of the present disclosure comprises two or more polynucleotides encoding an antibody. In some embodiments, the recombinant nucleic acid comprises a first polynucleotide encoding a polypeptide comprising an antibody heavy chain variable region (e.g., a full-length antibody heavy chain) and a second polynucleotide encoding a polypeptide comprising an antibody light chain variable region (e.g., a full-length antibody light chain). In some embodiments, the recombinant nucleic acid comprises two copies of the first polynucleotide encoding a polypeptide comprising an antibody heavy chain variable region (e.g., a full-length antibody heavy chain) and two copies of the second polynucleotide encoding a polypeptide comprising an antibody light chain variable region (e.g., a full-length antibody light chain). In some embodiments, the recombinant nucleic acid comprises two copies of the first polynucleotide encoding a polypeptide comprising an antibody heavy chain variable region (e.g., a full-length antibody heavy chain) and one copy of the second polynucleotide encoding a polycistronic mRNA comprising first and second ORFs, each encoding a polypeptide comprising an antibody light chain variable region (e.g., a full-length antibody light chain), separated by an IRES (e.g., any of the IRESs described herein). In some embodiments, the recombinant nucleic acid comprises one copy of a first polynucleotide encoding a polycistronic mRNA comprising first and second ORFs, each encoding a polypeptide comprising an antibody heavy chain variable region (e.g., a full-length antibody heavy chain) separated by an IRES (e.g., any of the IRESs described herein), and two copies of a second polynucleotide encoding a polypeptide comprising an antibody light chain variable region (e.g., a full-length antibody light chain).

[0141] In some embodiments, the recombinant nucleic acid comprises a first polynucleotide encoding a first antibody and a second polynucleotide encoding a second antibody. In some embodiments, the first and second antibodies are the same. In some embodiments, the first and second antibodies are different.

[0142] Recombinant nucleic acids In some embodiments, the present disclosure relates to a recombinant nucleic acid comprising any one or more of the polynucleotides described herein. In some embodiments, the recombinant nucleic acid is a vector (e.g., an expression vector, a display vector, etc.). In some embodiments, the vector is a DNA vector or an RNA vector. Generally, any vector suitable for maintaining, propagating, and / or expressing a polynucleotide to produce one or more polypeptides in a subject can be used. Examples of suitable vectors can include, for example, plasmids, cosmids, episomes, transposons, and viral vectors (e.g., adenoviral vectors, adeno-associated viral vectors, vaccinia viral vectors, Sindbis viral vectors, measles vectors, herpes viral vectors, lentiviral vectors, retroviral vectors, etc.). In some embodiments, the vector is a herpes viral vector. In some embodiments, the vector is capable of autonomous replication in a host cell. In some embodiments, the vector is not capable of autonomous replication in a host cell. In some embodiments, the vector is capable of integration into host DNA. In some embodiments, the vector is not capable of integration into host DNA (e.g., is episomal). Methods for producing vectors containing one or more polynucleotides of interest are well known to those of skill in the art and include, for example, by chemical synthesis or by the artificial manipulation of isolated segments of nucleic acid (e.g., by genetic engineering techniques).

[0143] In some embodiments, the recombinant nucleic acid of the present disclosure is a herpes simplex virus (HSV) amplicon. Herpes virus amplicons, including their structural characteristics and methods of production, are generally known to those of skill in the art (see, e.g., de Silva S. and Bowers W. "Herpes Virus Amplicon Vectors", Viruses 2009, 1, 594-629). In some embodiments, the herpes simplex virus amplicon is an HSV-1 amplicon. In some embodiments, the herpes simplex virus amplicon is an HSV-1 hybrid amplicon. Examples of HSV-1 hybrid amplicons include, but are not limited to, HSV / AAV hybrid amplicons, HSV / EBV hybrid amplicons, HSV / EBV / RV hybrid amplicons, and / or HSV / Sleeping Beauty hybrid amplicons. In some embodiments, the amplicon is an HSV / AAV hybrid amplicon. In some embodiments, the amplicon is an HSV / Sleeping Beauty hybrid amplicon.

[0144] In some embodiments, the recombinant nucleic acid of the present disclosure is a recombinant herpesvirus genome. The recombinant herpesvirus genome can be a recombinant genome derived from any member of the DNA virus family Herpesviridae known in the art, including, for example, a recombinant herpes simplex virus genome, a recombinant varicella-zoster virus genome, a recombinant human cytomegalovirus genome, a recombinant herpesvirus 6A genome, a recombinant herpesvirus 6B genome, a recombinant herpesvirus 7 genome, a recombinant Kaposi's sarcoma-associated herpesvirus genome, and any combination thereof or any derivative thereof. In some embodiments, the recombinant herpesvirus genome contains a greater number of inactivating mutations (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.). In some embodiments, the one or more inactivating mutations are present in one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) herpesvirus genes. In some embodiments, the recombinant herpesvirus genome is attenuated (e.g., compared to a corresponding wild-type herpesvirus genome). In some embodiments, the recombinant herpesvirus genome is replication-competent. In some embodiments, the recombinant herpesvirus genome is replication-deficient.

[0145] In some embodiments, the recombinant nucleic acid is a recombinant herpes simplex virus (HSV) genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome, a recombinant herpes simplex virus type 2 (HSV-2) genome, or any derivative thereof. In some embodiments, the recombinant herpes simplex virus genome is a recombinant HSV-1 genome. In some embodiments, the recombinant herpes simplex virus genome is replication-competent. In some embodiments, the recombinant herpes simplex virus genome is replication-deficient. In some embodiments, the recombinant herpes simplex virus genome comprises one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) inactivating mutations. In some embodiments, one or more inactivating mutations are present in one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) herpes simplex virus genes. As used herein, "inactivating mutation" may refer to any mutation that results in a gene product (RNA or protein) that is reduced in amount and / or function, undetectable, or eliminated (e.g., compared to the corresponding sequence lacking the inactivating mutation). Examples of inactivating mutations may include, but are not limited to, deletions, insertions, point mutations, and rearrangements in transcriptional control sequences (promoters, enhancers, insulators, etc.) and / or coding sequences of a given gene or regulon. Any suitable method for measuring the amount of a gene product or regulon product known in the art can be used, including, for example, qPCR, Northern blot, RNAseq, Western blot, ELISA, etc.

[0146] In some embodiments, the recombinant herpes simplex virus genome comprises inactivating mutations in at least one, at least two, at least three, at least four, at least five, at least six, at least seven, or all eight of the infected cell protein (or infected cell polypeptide) (ICP)0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL)41, and / or UL55 herpes simplex virus genes. In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivating mutation in the ICP34.5 herpes simplex virus gene (one or both copies) and / or the ICP47 herpes simplex virus gene (e.g., to avoid production of immunostimulatory virus). In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivating mutation in the ICP34.5 herpes simplex virus gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivating mutation in the ICP47 herpes simplex virus gene. In some embodiments, the recombinant herpes simplex virus genome does not contain inactivating mutations in the ICP34.5 herpes simplex virus gene (one or both copies) and the ICP47 herpes simplex virus gene, hi some embodiments, the recombinant herpes simplex virus genome is not oncolytic.

[0147] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies) and further comprises an initiating mutation in the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, the ICP47 gene, the UL41 gene, and / or the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies) and an inactivating mutation in the ICP4 gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies) and an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies) and an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies), an inactivating mutation in the ICP4 gene (one or both copies), and an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies), an inactivating mutation in the ICP4 gene (one or both copies), and an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies), an inactivating mutation in the ICP22 gene, and an inactivating mutation in the UL41 gene.In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies), an inactivating mutation in the ICP4 gene (one or both copies), an inactivating mutation in the ICP22 gene, and an inactivating mutation in the UL41 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, and / or the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome further comprises an inactivating mutation in the ICP27 gene, the ICP47 gene, and / or the UL55 gene.

[0148] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or both copies). In some embodiments, the recombinant herpes complex virus genome comprises an inactivating mutation in the ICP4 gene (one or both copies) and further comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP22 gene, the ICP27 gene, the ICP47 gene, the UL41 gene, and / or the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or both copies) and an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or both copies) and an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or both copies), an inactivating mutation in the ICP22 gene, and an inactivating mutation in the UL41 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the ICP4 gene (one or both copies), the ICP22 gene, and / or the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome further comprises an inactivating mutation in the ICP0 gene, the ICP27 gene, the ICP47 gene, and / or the UL55 gene.

[0149] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene and further comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP27 gene, the ICP47 gene, the UL41 gene, and / or the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene and an inactivating mutation in the UL41 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the ICP22 gene and / or the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome further comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP27 gene, the ICP47 gene, and / or the UL55 gene.

[0150] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP27 gene and further comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, the ICP47 gene, the UL41 gene, and / or the UL55 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the ICP27 gene.

[0151] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP47 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP47 gene and further comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, the UL41 gene, and / or the UL55 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the ICP47 gene.

[0152] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL41 gene and further comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, the ICP47 gene, and / or the UL55 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the UL41 gene.

[0153] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene and further comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, the ICP47 gene, and / or the UL41 gene. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the UL55 gene.

[0154] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation (e.g., deletion) in the internal repeat (joint) region, which includes the internal repeat long (IRL) region and the internal repeat short (IRS) region. In some embodiments, the inactivation (e.g., deletion) of the joint region removes one copy each of the ICP4 gene and the ICP0 gene. In some embodiments, the inactivation (e.g., deletion) of the joint region further inactivates (e.g., deletes) the promoters of the ICP22 gene and the ICP47 gene. If necessary, expression of one or both of these genes can be restored by inserting an immediate-early promoter into the recombinant herpes simplex virus genome (see, e.g., Hill et al. (1995). Nature 375(6530):411-415; Goldsmith et al. (1998). J Exp Med 187(3):341-348). Without wishing to be bound by theory, it is believed that inactivating (e.g., deleting) the joint region may contribute to the stability of the recombinant herpes simplex virus genome and / or allow the recombinant herpes simplex virus genome to accommodate more and / or larger transgenes.

[0155] In some embodiments, the recombinant herpes simplex virus genome comprises inactivating mutations in the ICP4 gene (one or both copies), the ICP22 gene, and the ICP27 gene. In some embodiments, the recombinant herpes simplex virus genome comprises inactivating mutations in the ICP4 gene (one or both copies), the ICP27 gene, and the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome comprises inactivating mutations in the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, the ICP47 gene, and the UL55 gene. In some embodiments, the inactivating mutations in the ICP4 gene (one or both copies), the ICP27 gene, and / or the UL55 gene are deletions of the coding sequence of the ICP4 gene (one or both copies), the ICP27 gene, and / or the UL55 gene. In some embodiments, the inactivating mutations in the ICP22 and ICP47 genes are deletions in the promoter regions of the ICP22 and ICP47 genes (e.g., the coding sequences of ICP22 and ICP47 are intact but not transcriptionally active). In some embodiments, the recombinant herpes simplex virus genome comprises deletions in the coding sequences of the ICP4 gene (one or both copies), the ICP27 gene, and the UL55 gene, and deletions in the promoter regions of the ICP22 and ICP47 genes. In some embodiments, the recombinant herpes simplex virus genome further comprises an inactivating mutation in the ICP0 gene (one or both copies) and / or the UL41 gene.

[0156] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies) and the ICP4 gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), and the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, and the ICP27 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, and the UL55 gene. In some embodiments, the inactivating mutation in the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, and / or the UL55 gene comprises a deletion of the coding sequence of the ICP0 gene (one or both copies), the ICP4 gene (one or both copies), the ICP22 gene, the ICP27 gene, and / or the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome further comprises an inactivating mutation in the ICP47 gene and / or the UL41 gene.

[0157] In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one, two, three, four, five, six, seven, or more viral loci. Examples of suitable viral loci may include, but are not limited to, the ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, ICP47, tk, UL41, and UL55 herpes simplex virus loci. In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one or both ICP4 viral loci (e.g., a recombinant virus harboring a polynucleotide encoding an antibody (or portion thereof) within one or both ICP4 loci). In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within the ICP22 viral locus (e.g., a recombinant virus harboring a polynucleotide encoding an antibody (or portion thereof) within the ICP22 locus). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of this disclosure within the UL41 viral locus (e.g., a recombinant virus harboring a polynucleotide encoding an antibody (or portion thereof) within the UL41 locus). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of this disclosure within the ICP47 viral locus (e.g., a recombinant virus harboring a polynucleotide encoding an antibody (or portion thereof) within the ICP47 locus). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of this disclosure within one or both ICP4 viral loci and one or more polynucleotides of this disclosure within the ICP22 viral locus (e.g., a recombinant virus harboring a polynucleotide encoding an antibody heavy chain within one or both ICP4 loci and a polynucleotide encoding an antibody light chain within the ICP22 locus; a recombinant virus harboring a polynucleotide encoding an antibody heavy chain within one or both ICP4 loci and a polynucleotide encoding a polycistronic mRNA encoding two copies of an antibody light chain within the ICP22 locus, etc.).In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one or both ICP4 viral loci and one or more polynucleotides of the present disclosure within the UL41 viral locus (e.g., a recombinant virus harboring a polynucleotide encoding an antibody heavy chain at one or both ICP4 loci and a polynucleotide encoding an antibody light chain at the UL41 locus; a recombinant virus harboring a polynucleotide encoding an antibody heavy chain at one or both ICP4 loci and a polynucleotide encoding a polycistronic mRNA encoding two copies of an antibody light chain at the UL41 locus, etc.). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within the UL41 viral locus and one or more polynucleotides of the present disclosure within the ICP22 viral locus (e.g., a recombinant virus harboring a polynucleotide encoding an antibody heavy chain at the UL41 locus and a polynucleotide encoding an antibody light chain at the ICP22 locus; a recombinant virus harboring a polynucleotide encoding an antibody light chain at the UL41 locus and a polynucleotide encoding an antibody heavy chain at the ICP22 locus, etc.). In some embodiments, the recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one or more ICP4 viral loci, one or more polynucleotides of the present disclosure within the ICP22 viral locus, and one or more polynucleotides of the present disclosure within the UL41 viral locus (e.g., a recombinant virus carrying a polynucleotide encoding an antibody heavy chain at one or both ICP4 loci, a polynucleotide encoding an antibody light chain at the ICP22 locus, and a polynucleotide encoding an antibody light chain at the UL41 locus; a recombinant virus carrying a polynucleotide encoding an antibody light chain at one or both ICP4 loci, a polynucleotide encoding an antibody heavy chain at the ICP22 locus, and a polynucleotide encoding an antibody heavy chain at the UL41 locus, etc.).

[0158] In some embodiments, the recombinant herpesvirus genome (e.g., recombinant herpes simplex virus genome) has been engineered to reduce or eliminate expression of one or more toxic herpesvirus genes (such as one or both copies of the HSV ICP4 gene, the ICP22 gene, the UL41 gene, and / or the ICP27 gene). In some embodiments, the recombinant herpesvirus genome (e.g., recombinant herpes simplex virus genome) has been engineered to reduce the cytotoxicity of the recombinant genome (e.g., when introduced into a target cell) compared to a corresponding wild-type herpesvirus genome (e.g., a wild-type herpes simplex virus genome). In some embodiments, the cytotoxicity (e.g., in human keratinocytes and / or fibroblasts) of the recombinant viral genome (e.g., recombinant herpes simplex virus genome) is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 95%, at least about 100%, at least about 105 ... %, at least about 85%, at least about 90%, at least about 95%, or at least about 99% (e.g., measuring the relative cytotoxicity of a recombinant ΔICP4 (one or both copies) herpes simplex virus genome versus a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines); measuring the relative cytotoxicity of a recombinant ΔICP4 (one or both copies) / ΔICP22 herpes simplex virus genome versus a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines), etc.).In some embodiments, the cytotoxicity (e.g., in human keratinocytes and / or fibroblasts) of the recombinant herpesvirus genome (e.g., recombinant herpes simplex virus genome) is at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 50-fold, at least about 75-fold, at least about 100-fold, at least about or by about 250-fold, at least about 500-fold, at least about 750-fold, at least about 1000-fold, or more (e.g., measuring the relative cytotoxicity of a recombinant ΔICP4 (one or both copies) herpes simplex virus genome versus a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines); measuring the relative cytotoxicity of a recombinant ΔICP4 (one or both copies) / ΔICP22 herpes simplex virus genome versus a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines), etc.). Methods for measuring cytotoxicity are known to those of skill in the art and include, for example, the use of vital dyes (formazan dyes), protease biomarkers, MTT assays (or assays using related tetrazolium salts, such as XTT, MTS, water-soluble tetrazolium salts), measuring ATP content, etc.

[0159] In some embodiments, the recombinant herpesvirus genome (e.g., recombinant herpes simplex virus genome) is engineered to have a reduced effect on host cell proliferation following exposure of a target cell to the recombinant genome, compared to a corresponding wild-type herpesvirus genome (e.g., a wild-type herpes simplex virus genome). In some embodiments, the target cell is a human cell. In some embodiments, the target cell is a cell of the epidermis and / or dermis. In some embodiments, the target cell is a cell of the eye. In some embodiments, the target cell is a cell of a joint. In some embodiments, the target cell is a cell of the lung. In some embodiments, host cell proliferation (e.g., of human keratinocytes and / or fibroblasts) after exposure to the recombinant genome is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 100%, at least about 110%, at least about 120%, at least about 130%, at least about 140%, at least about 150%, at least about 160%, at least about 170%, at least about 180%, at least about 190%, at least about 210%, at least about 220%, at least about 230%, at least about 240%, at least about 250%, at least about 260%, at least about 270%, at least about 280%, at least about 290%, at least about 300%, at least about 310%, at least about 320%, at least about 330%, at least about 340%, at least about 350%, at least about 360%, at least about 370%, at least about 380%, at least about 390%, at least about 400%, at least about 410%, at least about 420%, at least about 430%, at least about 440%, at least about 450%, at least about 460%, at least about 470%, at least about 480%, at least about 490%, at least about 500%, at least about 550%, at least about 550%, at least about 600%, at least about 65 0%, at least about 95%, or at least about 99% faster (e.g., measuring the relative cell proliferation following exposure to a recombinant ΔICP4 (one or both copies) herpes simplex virus genome relative to the cell proliferation following exposure to a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell line); measuring the relative cell proliferation following exposure to a recombinant ΔICP4 (one or both copies) / ΔICP22 herpes simplex virus genome relative to the cell proliferation following exposure to a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell line), etc.).In some embodiments, host cell proliferation (e.g., of human keratinocytes and / or fibroblasts) following exposure to the recombinant genome is at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 50-fold, at least about 75-fold, at least about 100-fold, at least about 250-fold, at least about 500-fold, at least about 10 ... at least about 750 times or at least about 1000 times faster (e.g., measuring the relative cell proliferation after exposure to a recombinant ΔICP4 (one or both copies) herpes simplex virus genome relative to the cell proliferation after exposure to a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines); measuring the relative cell proliferation after exposure to a recombinant ΔICP4 (one or both copies) / ΔICP22 herpes simplex virus genome relative to the cell proliferation after exposure to a wild-type herpes simplex virus genome in human keratinocytes or fibroblasts (primary cells or cell lines), etc.). Methods for measuring cell proliferation are known to those skilled in the art and include, for example, by using a Ki67 cell proliferation assay, a BrdU cell proliferation assay, etc.

[0160] A vector (e.g., a herpes virus vector) can contain one or more polynucleotides of the present disclosure in a form suitable for expression of the polynucleotide in a host cell. The vector can include one or more regulatory sequences operably linked (e.g., as described above) to the polynucleotide to be expressed.

[0161] In some embodiments, a recombinant nucleic acid (e.g., a recombinant herpes simplex virus genome) of the present disclosure comprises one or more of the polynucleotides described herein inserted into the recombinant nucleic acid in any orientation. When a recombinant nucleic acid comprises two or more polynucleotides described herein (e.g., two or more, three or more, etc.), the polynucleotides can be inserted in the same or opposite orientation to each other. Without wishing to be bound by theory, incorporating two polynucleotides (e.g., two transgenes) into a recombinant nucleic acid (e.g., a vector) in an antisense orientation can be useful to avoid read-through and ensure proper expression of each polynucleotide.

[0162] IV. Viruses Certain aspects of the present disclosure relate to viruses comprising any of the polynucleotides and / or recombinant nucleic acids described herein. In some embodiments, the viruses are capable of infecting one or more target cells of a subject (e.g., a human). In some embodiments, the viruses are suitable for delivering polynucleotides and / or recombinant nucleic acids to one or more target cells of a subject (e.g., a human subject). In some embodiments, the one or more target cells are one or more cells of a mucosa or skin (e.g., one or more cells of the epidermis, dermis, and / or subcutaneous tissue). In some embodiments, the one or more cells are selected from keratinocytes, melanocytes, Langerhans cells, Merkel cells, mast cells, fibroblasts, and / or adipocytes. In some embodiments, the one or more cells are keratinocytes. In some embodiments, the one or more cells are present in the stratum corneum, stratum granulosum, stratum spinosum, stratum basale, and / or basement membrane. In some embodiments, the one or more target cells are one or more epidermal cells. In some embodiments, the one or more target cells are one or more dermal cells. In some embodiments, the one or more target cells are one or more cells of a joint. In some embodiments, the one or more target cells are one or more cells of the eye, hi some embodiments, the one or more target cells are one or more cells of the airways and / or lungs.

[0163] Any suitable virus known in the art can be used, including, for example, adenovirus, adeno-associated virus, retrovirus, lentivirus, Sendai virus, herpes virus (e.g., herpes simplex virus), vaccinia virus, and / or any hybrid or derivative thereof. In some embodiments, the virus is attenuated. In some embodiments, the virus is replication-deficient. In some embodiments, the virus is replication-competent. In some embodiments, the virus has been modified to alter its tissue tropism compared to the tissue tropism of the corresponding unmodified wild-type virus. In some embodiments, the virus has reduced cytotoxicity compared to the corresponding wild-type virus. Methods for producing viruses containing recombinant nucleic acids are well known to those of skill in the art.

[0164] In some embodiments, the virus is a member of the DNA virus family Herpesviridae, which includes, for example, herpes simplex virus, varicella-zoster virus, human cytomegalovirus, herpesvirus 6A, herpesvirus 6B, herpesvirus 7, and Kaposi's sarcoma-associated herpesvirus. In some embodiments, the herpesvirus is attenuated. In some embodiments, the herpesvirus is replication-deficient. In some embodiments, the herpesvirus is replication-competent. In some embodiments, the herpesvirus has reduced cytotoxicity compared to the corresponding wild-type herpesvirus. In some embodiments, the herpesvirus is not oncolytic.

[0165] In some embodiments, the virus is a herpes simplex virus. The herpes simplex virus comprising the recombinant nucleic acid may be produced by the processes disclosed in, for example, WO2015 / 009952 and / or WO2017 / 176336. In some embodiments, the herpes simplex virus is attenuated. In some embodiments, the herpes simplex virus is replication-deficient. In some embodiments, the herpes simplex virus is replication-competent. In some embodiments, the herpes simplex virus is herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), or any derivative thereof. In some embodiments, the herpes simplex virus is herpes simplex virus type 1 (HSV-1). In some embodiments, the HSV-1 is attenuated. In some embodiments, the HSV-1 is replication-deficient. In some embodiments, the HSV-1 is replication-competent. In some embodiments, the HSV-1 has reduced cytotoxicity compared to the corresponding wild-type HSV-1. In some embodiments, the HSV-1 is not oncolytic.

[0166] In some embodiments, the herpes simplex virus is modified to alter its tissue tropism compared to the tissue tropism of the unmodified wild-type herpes simplex virus. In some embodiments, the herpes simplex virus comprises a modified envelope. In some embodiments, the modified envelope comprises one or more (e.g., one or more, two or more, three or more, four or more, etc.) mutated herpes simplex virus glycoproteins. Examples of herpes simplex virus glycoproteins can include, but are not limited to, glycoproteins gB, gC, gD, gH, and gL. In some embodiments, the modified envelope alters the tissue tropism of the herpes simplex virus compared to the wild-type herpes simplex virus.

[0167] In some embodiments, the transduction efficiency (in vitro and / or in vivo) of a virus of the present disclosure (e.g., a herpes virus, such as herpes simplex virus) of one or more target cells (e.g., one or more human keratinocytes and / or fibroblasts) is at least about 25%. For example, the transduction efficiency of the virus of one or more target cells can be at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5%, or more. In some embodiments, the virus is a herpes simplex virus, and the transduction efficiency of the virus of one or more target cells (e.g., one or more human keratinocytes and / or fibroblasts) is between about 85% and about 100%. In some embodiments, the virus is a herpes simplex virus, and the viral transduction efficiency of one or more target cells (e.g., one or more human keratinocytes and / or fibroblasts) is at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%. Methods for measuring viral transduction efficiency in vitro or in vivo are well known to those of skill in the art and include, for example, qPCR analysis, deep sequencing, Western blotting, fluorescence analysis (e.g., fluorescent in situ hybridization (FISH), fluorescent reporter gene expression, immunofluorescence, FACS), and the like.

[0168] V. Pharmaceutical Compositions and Formulations Certain aspects of the present disclosure relate to pharmaceutical compositions and formulations comprising any of the recombinant nucleic acids (e.g., recombinant herpesvirus genomes) and / or viruses (e.g., herpesviruses comprising recombinant genomes) described herein and a pharmaceutically acceptable excipient or carrier.

[0169] In some embodiments, the pharmaceutical composition or formulation comprises any one or more of the viruses (e.g., herpes viruses) described herein. In some embodiments, the pharmaceutical composition or formulation comprises about 10 4 ~about 10 12 For example, the pharmaceutical composition or formulation may contain about 10 plaque-forming units (PFU) / mL of virus. 4 ~about 10 12 , about 10 5 ~about 10 12 , about 10 6 ~about 10 12 , about 10 7 ~about 10 12 , about 10 8 ~about 10 12 , about 10 9 ~about 10 12 , about 10 10 ~about 10 12 , about 10 11 ~about 10 12 , about 10 4 ~about 10 11 , about 10 5 ~about 10 11 , about 10 6 ~about 10 11 , about 10 7 ~about 10 11 , about 10 8 ~about 10 11 , about 10 9 ~about 10 11 , about 10 10 ~about 10 11 , about 10 4 ~about 10 10 , about 10 5 ~about 10 10 , about 10 6 ~about 10 10 , about 10 7 ~about 10 10 , about 10 8 ~about 10 10, about 10 9 ~about 10 10 , about 10 4 ~about 10 9 , about 10 5 ~about 10 9 , about 10 6 ~about 10 9 , about 10 7 ~about 10 9 , about 10 8 ~about 10 9 , about 10 4 ~about 10 8 , about 10 5 ~about 10 8 , about 10 6 ~about 10 8 , about 10 7 ~about 10 8 , about 10 4 ~about 10 7 , about 10 5 ~about 10 7 , about 10 6 ~about 10 7 , about 10 4 ~about 10 6 , about 10 5 ~about 10 6 or about 10 4 ~about 10 5 In some embodiments, the pharmaceutical composition or formulation may contain about 10 PFU / mL of virus. 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 11 or about 10 12 containing PFU / mL of virus.

[0170] Pharmaceutical compositions and formulations can be prepared by mixing active ingredient(s) (such as recombinant nucleic acids and / or viruses) having a desired degree of purity with one or more pharmaceutically acceptable carriers or excipients. Pharmaceutically acceptable carriers or excipients are generally non-toxic to recipients at the dosages and concentrations employed, and include, but are not limited to, buffers (e.g., phosphates, citrates, acetates, and other organic acids); antioxidants (e.g., ascorbic acid and methionine); preservatives (e.g., octadecyldimethylbenzyl ammonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); amino acids (e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine). low molecular weight (less than about 10 residues) polypeptides; proteins (e.g., serum albumin, gelatin, or immunoglobulins); polyols (e.g., glycerol (e.g., formulations containing 10% glycerol)); hydrophilic polymers (e.g., polyvinylpyrrolidone); monosaccharides, disaccharides, and other carbohydrates (including glucose, mannose, or dextrins); chelating agents (e.g., EDTA); sugars (e.g., sucrose, mannitol, trehalose, or sorbitol); salt-forming counterions (e.g., sodium); metal complexes (e.g., Zn-protein complexes); and / or non-ionic surfactants (e.g., polyethylene glycol (PEG)). A complete discussion of pharmaceutically acceptable carriers is available in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., NJ 1991).

[0171] In some embodiments, the pharmaceutical composition or formulation comprises one or more lipid (e.g., cationic lipid) carriers. In some embodiments, the pharmaceutical composition or formulation comprises one or more nanoparticle carriers. Nanoparticles are submicron (less than about 1000 nm) sized drug delivery vehicles that can carry encapsulated drugs (such as synthetic small molecules, proteins, peptides, cells, viruses, and nucleic acid-based biologicals) for immediate or controlled release. Various molecules (e.g., proteins, peptides, recombinant nucleic acids, etc.) can be efficiently encapsulated in nanoparticles using processes well known in the art. In some embodiments, a molecule "encapsulated" in a nanoparticle can refer to a molecule (such as a virus) that is contained within the nanoparticle, or attached and / or associated with the surface of the nanoparticle, or any combination thereof. Nanoparticles for use in the compositions or formulations described herein can be any type of biocompatible nanoparticle known in the art, including, for example, nanoparticles comprising poly(lactic acid), poly(glycolic acid), PLGA, PLA, PGA, and any combination thereof (see, e.g., Vauthier et al. Adv Drug Del Rev. (2003) 55:519-48, US2007 / 0148074, US2007 / 0092575, US2006 / 0246139, US5753234, US7081483, and WO2006 / 052285).

[0172] In some embodiments, the pharmaceutically acceptable carrier or excipient may be compatible or suitable for any route of administration known in the art, including, for example, intravenous, intramuscular, subcutaneous, dermal, oral, intratracheal, sublingual, buccal, topical, transdermal, intradermal, intraperitoneal, intraorbital, intravitreal, subretinal, transmucosal, intraarticular, implant, inhalation, intrathecal, intracerebroventricular, and / or intranasal administration. In some embodiments, the pharmaceutically acceptable carrier or excipient is compatible or suitable for topical, transdermal, subcutaneous, and / or intradermal administration. In some embodiments, the pharmaceutical composition or formulation is compatible or suitable for topical, transdermal, subcutaneous, and / or intradermal administration. In some embodiments, the pharmaceutically acceptable carrier or excipient is compatible or suitable for topical, transdermal, and / or intradermal administration. In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for topical, transdermal, and / or intradermal administration. In some embodiments, the pharmaceutically acceptable carrier or excipient is adapted or suitable for oral, sublingual, nasal, intranasal, intratracheal, or buccal administration, or administration by inhalation. In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for oral, sublingual, nasal, intranasal, intratracheal, or buccal administration, or administration by inhalation. In some embodiments, the pharmaceutically acceptable carrier or excipient is adapted or suitable for topical (ophthalmic), intravitreal, subretinal, and / or intraorbital administration. In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for topical (ophthalmic), intravitreal, subretinal, and / or intraorbital administration. In some embodiments, the pharmaceutically acceptable carrier or excipient is adapted or suitable for intra-articular administration. In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for intra-articular administration.

[0173] Examples of carriers or excipients compatible or suitable for use in the pharmaceutical compositions or formulations of the present disclosure may include, but are not limited to, ointments, oils, pastes, creams, aerosols, suspensions, emulsions, fatty ointments, gels, powders, liquids, lotions, solutions, sprays, patches (e.g., transdermal patches or microneedle patches), adhesive strips, microneedles or microneedle arrays, and inhalants. In some embodiments, the carrier or excipient (e.g., a pharmaceutically acceptable carrier or excipient) comprises one or more (e.g., one or more, two or more, three or more, four or more, five or more, etc.) ointments, oils, pastes, creams, aerosols, suspensions, emulsions, fatty ointments, gels, powders, liquids, lotions, solutions, sprays, adhesive strips, and inhalants. In some embodiments, the carrier comprises a patch (e.g., a patch that adheres to the skin), such as a transdermal patch or a microneedle patch. In some embodiments, the carrier comprises a microneedle or a microneedle array. Methods for making and using microneedle arrays suitable for composition delivery are generally known in the art (Kim Y. et al. "Microneedles for drug and vaccine delivery". Advanced Drug Delivery Reviews 2012, 64(14):1547-68).

[0174] In some embodiments, the pharmaceutical composition or formulation further comprises one or more additional ingredients. Examples of additional ingredients include binders (such as pregelatinized maize starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose), fillers (such as lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethylcellulose, polyacrylates, or calcium hydrogen phosphate), lubricants (such as magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, maize starch, polyethylene glycol, sodium benzoate, acetic acid, etc.). The polynucleotide may include, but is not limited to, an ester or a mixture of hydroxypropyl methylcellulose, ...

[0175] Compositions and formulations to be used for in vivo administration (e.g., pharmaceutical compositions and formulations) are generally sterile. Sterility is readily accomplished, for example, by filtration through sterile filtration membranes.

[0176] In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein can be used to deliver one or more polynucleotides encoding an antibody (e.g., a therapeutic antibody) to one or more cells of a subject and / or to express the antibody (e.g., a therapeutic antibody) in one or more tissues of a subject. In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein can be used for therapy. In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations can be used to treat a disease or condition that would benefit from administration of an antibody (e.g., a therapeutic antibody). In some embodiments, any of the recombinant nucleic acids, viruses and / or pharmaceutical compositions or formulations described herein are administered to treat or prevent any of the following conditions: psoriasis (e.g., chronic plaque psoriasis), atopic dermatitis, pyoderma gangrenosum, bullous dermatitis, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behcet's disease, cancer (e.g., skin cancer, breast cancer, lymphoma, colorectal cancer, head and neck cancer, etc.), hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, psoriatic arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, rheumatoid arthritis, psoriatic arthritis, rheumatoid ... The compositions may be used to treat one or more of arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, autoimmune diseases (e.g., multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, celiac disease, etc.), asthma, uveal melanoma, thyroid eye disease, infectious diseases, transplant / tissue / organ rejection, and / or neurological diseases (e.g., Alzheimer's disease).

[0177] In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein can be used in the preparation or manufacture of a medicament. In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein can be used in the preparation or manufacture of a medicament useful for delivering one or more polynucleotides encoding an antibody (e.g., a therapeutic antibody) to one or more cells of a subject and / or for expressing an antibody (e.g., a therapeutic antibody) in one or more tissues of a subject. In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein can be used in the preparation or manufacture of a medicament useful for treating a disease or condition that would benefit from administration of an antibody (e.g., a therapeutic antibody). In some embodiments, any of the recombinant nucleic acids, viruses and / or pharmaceutical compositions or formulations described herein are administered to treat or prevent any of the following conditions: psoriasis (e.g., chronic plaque psoriasis), atopic dermatitis, pyoderma gangrenosum, bullous dermatitis, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behcet's disease, cancer (e.g., skin cancer, breast cancer, lymphoma, colorectal cancer, head and neck cancer, etc.), hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing leukemia, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing leukemia, rheumatoid arthritis, psoriatic arthritis, rheumatoid ... The composition may be used in the preparation or manufacture of a medicament useful for treating one or more of spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, autoimmune diseases (e.g., multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, celiac disease, etc.), asthma, uveal melanoma, thyroid eye disease, infectious diseases, transplant / tissue / organ rejection, and / or neurological diseases (e.g., Alzheimer's disease).

[0178] VI. Method Certain aspects of the present disclosure relate to methods of delivering an antibody to a subject, the method comprising administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs, and / or compositions or formulations described herein. In some embodiments, the present disclosure relates to methods of locally delivering an antibody to one or more specific target tissues in a subject (e.g., eye tissue, joint tissue, skin tissue, lung tissue, etc.), the method comprising administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs, and / or compositions or formulations described herein. In some embodiments, the subject is a human.

[0179] In some embodiments, local delivery of an antibody using a recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation described herein reduces or eliminates systemic exposure to the antibody in a subject, for example, compared to a subject treated with a purified antibody delivered via a conventional route of antibody administration (e.g., intravenous or subcutaneous administration). Methods of measuring systemic exposure to an antibody are well known to those of skill in the art and include, for example, by measuring the amount of antibody present in the subject's blood or serum by ELISA. In some embodiments, use of a recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation described herein to locally deliver an antibody to a subject reduces or eliminates one or more side effects of the antibody compared to side effects observed following systemic exposure of the subject to the same antibody (e.g., a comparison of one or more side effects of antibody expression in a subject following delivery (e.g., topical, intra-articular, intravitreal, etc.) of a recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation described herein to side effects following systemic (e.g., intravenous or subcutaneous) administration of a purified antibody). Examples of side effects resulting from systemic exposure to therapeutic antibodies include, for example, allergic reactions, chills, weakness, diarrhea, nausea, vomiting, rash, itching, high blood sugar levels, cough, constipation, shortness of breath, peripheral edema, headache, fever, muscle aches and pains, loss of appetite, elevated triglyceride levels, insomnia, abdominal pain, back pain, dizziness, hypotension, anaphylaxis, infections, cancer, serum sickness, autoimmune thyroiditis, arteriovenous thrombosis, congestive heart failure, bleeding, interstitial lung disease, hepatitis, gastrointestinal perforation, enteritis, mucositis, stomatitis, anemia, decreased white blood cell count, and / or hypothyroidism. Methods for assessing side effects of antibodies are well known to those of skill in the art.

[0180] In some embodiments, the use of a recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation described herein for local delivery of an antibody to a subject improves one or more pharmacokinetic properties of the antibody at the site of interest (e.g., intradermal, intraarticular, intraocular, etc.) compared to the pharmacokinetic properties of the antibody at the site of interest following systemic (e.g., intravenous) administration of the purified antibody. For a review of the pharmacokinetic properties of antibodies, see, e.g., Ryman and Meibohm "Pharmacokinetics of Monoclonal Antibodies," CPT Pharmacometrics Syst Pharmacol. 2017 Sep;6(9):576-588.

[0181] In some embodiments, the use of a recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation described herein for local delivery of an antibody to a subject improves the tissue accessibility and / or penetration of the antibody at a site of interest (e.g., intradermal, intra-articular, intraocular, intra-respiratory, or intrapulmonary, etc.) compared to the tissue accessibility and / or penetration of the antibody at the site of interest following systemic (e.g., intravenous or subcutaneous) administration of the purified antibody. In some embodiments, the use of a recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation described herein for local delivery of an antibody to a subject improves the antibody concentration at a site of interest (e.g., intradermal, intra-articular, intraocular, intra-respiratory, or intrapulmonary, etc.) compared to the antibody concentration at the site of interest following systemic (e.g., intravenous or subcutaneous) administration of the purified antibody. For example, in some embodiments, the use of a recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation described herein for local delivery of an antibody increases the antibody concentration at a site of interest (e.g., intradermal, intra-articular, intraocular, respiratory tract, or pulmonary, etc.) by at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 99%, or more, compared to the antibody concentration at the site of interest following systemic (e.g., intravenous or subcutaneous) administration of the purified antibody. In some embodiments, the use of a recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation described herein for local delivery of an antibody increases the antibody concentration at a site of interest (e.g., intradermal, intra-articular, intraocular, respiratory tract, or lung, etc.) by at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 50-fold, at least about 75-fold, at least about 100-fold, at least about 250-fold, at least about 500-fold, at least about 750-fold, or at least about 1000-fold compared to the antibody concentration at the site of interest following systemic (e.g., intravenous or subcutaneous) administration of the purified antibody.Methods for measuring antibody concentrations in tissue samples are readily available to those of skill in the art and include, for example, by Western blot, ELISA, immunofluorescence, mass spectrometry, and the like.

[0182] Other aspects of the present disclosure relate to methods for providing prophylactic, palliative, and / or therapeutic relief of one or more signs or symptoms of a disease in a subject, the method comprising administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. The disease can be any disease known in the art that can benefit from treatment with a therapeutic antibody, including, for example, psoriasis (e.g., chronic plaque psoriasis), atopic dermatitis, pyoderma gangrenosum, bullous diseases, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behçet's disease, cancer (e.g., skin cancer, breast cancer, lymphoma, colorectal cancer, head and neck cancer, etc.), hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, and the like. In some embodiments, the disease includes, but is not limited to, arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, autoimmune diseases (e.g., multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, celiac disease, etc.), asthma, uveal melanoma, thyroid eye disease, infectious diseases, transplant / tissue / organ rejection, neurological diseases (e.g., Alzheimer's disease), etc. In some embodiments, the disease is not cancer.

[0183] The recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein can be administered by any suitable method or route known in the art, including, but not limited to, oral administration, sublingual administration, buccal administration, intranasal administration, intratracheal administration, topical administration, rectal administration, by inhalation, transdermal administration, subcutaneous injection, intradermal injection, intravenous (IV) injection, intraarterial injection, intramuscular injection, intracardiac injection, intraosseous injection, intraperitoneal injection, transmucosal administration, intravaginal administration, intravitreal administration, intraorbital administration, subconjunctival administration (e.g., using a subconjunctival depot), suprachoroidal administration, subretinal administration, intraarticular administration, periarticular administration, local administration, epicutaneous administration, or any combination thereof. Accordingly, the present disclosure encompasses methods of delivering any of the recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein to an individual (or a specific site or tissue thereof).

[0184] In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation used in the methods of the disclosure is administered to a subject dermally, topically, transdermally, subcutaneously, intradermally, transmucosally, sublingually, nasally, bucally, intranasally, intratracheally, intravitreally, subconjunctivally, suprachoroidally, subretinally, intraarticularly, or by inhalation. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to a subject topically. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to a subject intradermally. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to a subject orally, sublingually, bucally, intranasally, intranasally, intratracheally, or by inhalation. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to a subject intraarticularly. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered intraorbitally, intravitreally, subconjunctivally, suprachoroidally, subretinally or topically (ophthalmically) to the subject.

[0185] In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to the subject once, hi some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to the subject at least twice (e.g., at least twice, at least three times, at least four times, at least five times, at least ten times, etc.). In some embodiments, at least about 1 hour (e.g., at least about 1 hour, at least about 6 hours, at least about 12 hours, at least about 18 hours, at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 15 days, at least about 20 days, at least about 30 days, at least about 40 days, at least about 50 days, at least about 60 days, at least about 70 days, at least about 80 days, at least about 90 days, at least about 100 days, at least about 120 days, etc.) elapses between administrations (e.g., between a first administration and a second administration, between a second administration and a third administration, etc.). In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to a subject once, twice, three times, four times, five times, or more times daily. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered to one or more affected or non-affected areas of the subject.

[0186] Another aspect of the present disclosure relates to a method of administering an antibody to the epidermis and / or dermis of a subject, the method comprising topically, transdermally, and / or intradermally administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is not systemically exposed to the antibody (e.g., the antibody is not detectable in the serum). In some embodiments, the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation is administered topically. In some embodiments, the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation is administered intradermally. In some embodiments, the subject is human. In some embodiments, the subject is afflicted with a skin disease or disorder. In some embodiments, the subject is afflicted with one or more of psoriasis, atopic dermatitis, pyoderma gangrenosum, bullous disease, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorder, bullous pemphigoid, Behçet's disease, skin cancer, and / or hidradenitis suppurativa. In some embodiments, the disease or disorder is not cancer (eg, not skin cancer).

[0187] In some embodiments, one or more portions of a subject's skin are abraded or permeabilized before treatment with the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation described herein. Any suitable method known in the art for abrading skin or increasing skin permeability can be used, including, for example, using a skin roller, repeatedly using adhesive strips to remove layers of skin cells (tape stripping), rubbing with a scalpel or blade, using sandpaper, using chemical permeation enhancers (e.g., cell-penetrating polypeptides) or electrical energy, using sonic or ultrasonic energy, using light (e.g., laser) energy, using a micron-sized needle or blade that is long enough to pierce but not completely penetrate the epidermis, etc.

[0188] Another aspect of the present disclosure relates to a method of administering an antibody to the mucosa of a subject, the method comprising administering an effective amount of any of the recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein to the subject topically, mucosally, orally, sublingually, nasally, intranasally, intratracheally, by inhalation, or bucally. In some embodiments, the subject is not systemically exposed to the antibody (e.g., the antibody is not detectable in the serum). In some embodiments, the recombinant nucleic acid, virus, drug, and / or composition is administered sublingually. In some embodiments, the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation is administered topically. In some embodiments, the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation is administered bucally. In some embodiments, the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation is administered intranasally. In some embodiments, the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation is administered by inhalation. In some embodiments, the subject is a human.

[0189] Another aspect of the present disclosure relates to a method of administering an antibody to the airways or lungs of a subject, the method comprising administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein orally, sublingually, nasally, intranasally, intratracheally, by inhalation, or bucally. In some embodiments, the subject has not been systemically exposed to the antibody (e.g., the antibody is not detectable in the serum). In some embodiments, the recombinant nucleic acid, virus, drug and / or composition is administered sublingually. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered bucally. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered intranasally. In some embodiments, the recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation is administered by inhalation. In some embodiments, the subject is human. In some embodiments, the subject is suffering from a disease or disorder of the airways or lungs (e.g., a respiratory disease such as asthma, lung cancer, a respiratory infection, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, etc.).

[0190] Another aspect of the present disclosure relates to a method of administering an antibody to one or more joints of a subject, the method comprising intra-articularly administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject has not been systemically exposed to the antibody (e.g., the antibody is not detectable in the serum). In some embodiments, the subject is suffering from a disease of the joint. In some embodiments, the subject is suffering from one or more of arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, and / or enteropathic arthritis. In some embodiments, the subject is human.

[0191] Other aspects of the present disclosure relate to methods of administering an antibody to one or both eyes of a subject, the method comprising topically, intravitreally, intraorbitally, subconjunctivally, subretinally, or suprachoroidally administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject has not been systemically exposed to the antibody (e.g., the antibody is not detectable in the serum). In some embodiments, the subject is afflicted with an ocular disease. In some embodiments, the subject is afflicted with an autoimmune disease affecting the eye. In some embodiments, the subject is afflicted with uveal melanoma or thyroid eye disease. In some embodiments, the subject is human.

[0192] VII.Host cells Certain aspects of the present disclosure relate to one or more host cells comprising any of the recombinant nucleic acids described herein. Any suitable host cell (prokaryotic or eukaryotic) known in the art can be used, including, for example, eubacteria, such as gram-negative or gram-positive microorganisms, for example, Escherichia (e.g., E. coli), Enterobacter, Erminia, Klebsiella, Proteus, Salmonella (e.g., S. typhimurium), and the like. Prokaryotic cells, including Enterobacteriaceae, such as Serratia (e.g., S. marcescans) and Shigella, and Bacilli, such as B. subtilis and B. licheniformis; fungal cells (e.g., S. cerevisiae); insect cells (e.g., S2 cells); and SV40-transformed monkey kidney strain CV1 (COS-7, ATCC CRL 1651), human embryonic kidney line (293 cells or 293 cells subcloned for growth in suspension culture), baby hamster kidney cells (BHK, ATCC CCL 10), mouse Sertoli cells (TM4), monkey kidney cells (CV1 ATCC CCL 70), African green monkey kidney cells (VERO-76, ATCC CRL-1587), human cervical carcinoma cells (HELA, ATCC CCL 2), canine kidney cells (MDCK, ATCC CCL 34), buffalo rat hepatocytes (BRL 3A, ATCC CRL 1442), human lung cells (W138, ATCC CCL 75), human hepatocytes (Hep G2, HB 8065), mouse mammary carcinoma (MMT 060562, ATCC CCL51), TRI cells, MRC 5 cells, FS4 cells, human hepatoma line (Hep G2), Chinese hamster ovary (CHO) cells, including DHFR-CHO cells, and mammalian cells, including myeloma cell lines such as NS0 and Sp2 / 0.In some embodiments, the host cell is a human cell or a non-human primate cell. In some embodiments, the host cell is a cell derived from a cell line. Examples of suitable host cells or cell lines may include, but are not limited to, 293 cells, HeLa cells, SH-Sy5y cells, Hep G2 cells, CACO-2 cells, A549 cells, L929 cells, 3T3 cells, K562 cells, CHO-K1 cells, MDCK cells, HUVEC cells, Vero cells, N20 cells, COS-7 cells, PSN1 cells, VCaP cells, CHO cells, etc.

[0193] In some embodiments, the recombinant nucleic acid is a herpes simplex virus vector. In some embodiments, the recombinant nucleic acid is a herpes simplex virus amplicon. In some embodiments, the recombinant nucleic acid is an HSV-1 amplicon or an HSV-1 hybrid amplicon. In some embodiments, a virus comprising one or more recombinant nucleic acids described herein is produced by contacting a host cell containing a helper virus with an HSV-1 amplicon or an HSV-1 hybrid amplicon described herein. In some embodiments, the virus is recovered from the supernatant of the contacted host cells. Methods for producing a virus by contacting a host cell containing a helper virus with an HSV-1 amplicon or an HSV-1 hybrid amplicon are known in the art.

[0194] In some embodiments, the host cell is a complementing host cell. In some embodiments, the complementing host cell expresses one or more genes that are inactivated in any of the viral vectors described herein. In some embodiments, the complementing host cell is contacted with a recombinant herpesvirus genome described herein (e.g., a recombinant herpes simplex virus genome). In some embodiments, contacting the complementing host cell with the recombinant herpesvirus genome produces a herpesvirus comprising one or more recombinant nucleic acids described herein. In some embodiments, the virus is recovered from the supernatant of the contacted host cell. Methods for producing virus by contacting a complementing host cell with a recombinant herpes simplex virus are generally described in WO2015 / 009952 and / or WO2017 / 176336.

[0195] VIII. Products or Kits Certain aspects of the present disclosure relate to articles of manufacture or kits that include any of the recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations described herein. In some embodiments, the articles of manufacture or kits include a package insert containing instructions for administering the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation (e.g., to provide a method of locally delivering an antibody to one or more tissues of a subject in need thereof (e.g., the epidermis and / or dermis of a subject) by administering the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation).

[0196] Suitable containers for the recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation may include, for example, bottles, vials, bags, tubes, and syringes. Containers may be formed from a variety of materials, such as glass, plastic (such as polyvinyl chloride or polyolefin), or alloys (such as stainless steel or Hastelloy). In some embodiments, the container comprises a label on or associated with the container, the label indicating directions for use. The article of manufacture or kit may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, package inserts, etc.

[0197] IX. Enumerated Embodiments Embodiment 1: A recombinant herpes simplex virus (HSV) genome comprising one or more polynucleotides encoding an antibody.

[0198] Embodiment 2: The recombinant genome of embodiment 1, wherein the antibody is an antibody fragment.

[0199] Embodiment 3: The recombinant genome of embodiment 2, wherein the antibody fragment is a Fab, Fab', Fab'-SF, F(ab')2, Fv, scFv, or scFv-Fc fragment.

[0200] Embodiment 4: The recombinant genome of embodiment 1, wherein the antibody is a full-length antibody.

[0201] Embodiment 5: The recombinant genome of any one of embodiments 1 to 4, wherein the antibody is a murine antibody, a chimeric antibody, a humanized antibody, a human antibody, a monoclonal antibody, or a multispecific antibody.

[0202] Embodiment 6: The recombinant genome of any one of embodiments 1 to 5, wherein the antibody is an IgA antibody, an IgD antibody, an IgE antibody, an IgG antibody, or an IgM antibody.

[0203] Embodiment 7: The recombinant genome of any one of embodiments 1 to 6, wherein the antibody is an IgG antibody.

[0204] Embodiment 8: The recombinant genome of embodiment 7, wherein the IgG antibody is an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody.

[0205] Embodiment 9: The recombinant genome of any one of embodiments 1 to 8, wherein the antibody is an agonist antibody.

[0206] Embodiment 10: The recombinant genome of any one of embodiments 1 to 8, wherein the antibody is an antagonist antibody.

[0207] Embodiment 11: The recombinant genome of any one of embodiments 1 to 10, wherein the antibody comprises a heavy chain variable region comprising HVR-H1, HVR-H2, and HVR-H3, wherein the HVR-H1 comprises a sequence selected from the group consisting of SEQ ID NOs: 1-59, the HVR-H2 comprises a sequence selected from the group consisting of SEQ ID NOs: 60-122, and / or the HVR-H3 comprises a sequence selected from the group consisting of SEQ ID NOs: 123-185.

[0208] Embodiment 12: The recombinant genome of embodiment 11, wherein the heavy chain variable region comprises a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 355-419.

[0209] Embodiment 13: The recombinant genome of any one of embodiments 1 to 12, wherein the antibody comprises a light chain variable region comprising HVR-L1, HVR-L2, and HVR-L3, wherein the HVR-L1 comprises a sequence selected from the group consisting of SEQ ID NOs: 186 to 242, the HVR-L2 comprises a sequence selected from the group consisting of SEQ ID NOs: 243 to 294, and / or the HVR-L3 comprises a sequence selected from the group consisting of SEQ ID NOs: 295 to 354.

[0210] Embodiment 14: The recombinant genome of embodiment 13, wherein the light chain variable region comprises a sequence having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence selected from SEQ ID NOs: 420-482.

[0211] Embodiment 15: The recombinant genome of any one of embodiments 1 to 14, wherein said recombinant genome is a recombinant HSV-1 genome, a recombinant HSV-2 genome, or any derivative thereof.

[0212] Embodiment 16: The recombinant genome of any one of embodiments 1 to 15, wherein the recombinant genome comprises an inactivating mutation in a herpes simplex virus gene.

[0213] Embodiment 17: The recombinant genome of embodiment 16, wherein the herpes simplex virus gene is selected from the group consisting of infected cell protein (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), long unique region (UL) 41, and UL55.

[0214] Embodiment 18: The recombinant genome of embodiment 17, wherein said recombinant genome comprises an inactivating mutation in one or both copies of said ICP4 gene.

[0215] Embodiment 19: The recombinant genome of embodiment 17 or 18, wherein the recombinant genome comprises an inactivating mutation in the ICP22 gene.

[0216] Embodiment 20: The recombinant genome of any one of embodiments 17 to 19, wherein the recombinant genome comprises an inactivating mutation in the UL41 gene.

[0217] Embodiment 21: The recombinant genome of any one of embodiments 17 to 20, wherein the recombinant genome comprises an inactivating mutation in the ICP0 gene.

[0218] Embodiment 22: The recombinant genome of any one of embodiments 17 to 21, wherein the recombinant genome comprises an inactivating mutation in the ICP27 gene.

[0219] Embodiment 23: The recombinant genome of any one of embodiments 16 to 22, wherein the inactivating mutation is a deletion of the coding sequence of the gene(s).

[0220] Embodiment 24: The recombinant genome of any one of embodiments 1 to 23, wherein the recombinant genome has reduced cytotoxicity when introduced into a target cell compared to a wild-type herpes simplex virus genome.

[0221] Embodiment 25: The recombinant genome of embodiment 24, wherein the target cell is a human cell.

[0222] Embodiment 26: The recombinant genome of embodiment 24 or 25, wherein the target cell is a keratinocyte or a fibroblast.

[0223] Embodiment 27: The recombinant genome of any one of embodiments 1 to 2, wherein the recombinant genome comprises the one or more polynucleotides within one or more viral loci.

[0224] Embodiment 28: The recombinant genome of embodiment 27, wherein said recombinant genome comprises one or more polynucleotides within one or both ICP4 viral loci.

[0225] Embodiment 29: The recombinant genome of embodiment 27 or 28, wherein said recombinant genome comprises said one or more polynucleotides within an ICP22 viral locus.

[0226] Embodiment 30: The recombinant genome of any one of embodiments 27 to 29, wherein said recombinant genome comprises said one or more polynucleotides within the UL41 viral locus.

[0227] Embodiment 31: A herpes simplex virus (HSV) comprising a recombinant genome of any one of embodiments 1 to 30.

[0228] Embodiment 32: The virus of embodiment 31, wherein the HSV is replication-competent.

[0229] Embodiment 33: The virus of embodiment 31, wherein the HSV is replication-deficient.

[0230] Embodiment 34: The virus of any one of embodiments 31 to 33, wherein the HSV has reduced cytotoxicity compared to wild-type herpes simplex virus.

[0231] Embodiment 35: The virus of any one of embodiments 31 to 34, wherein the HSV is herpes simplex virus type 1, herpes simplex virus type 2, or any derivative thereof.

[0232] Embodiment 36: A pharmaceutical composition comprising the recombinant genome of any one of embodiments 1 to 30 or the virus of any one of embodiments 31 to 35, and a pharmaceutically acceptable excipient.

[0233] Embodiment 37: The pharmaceutical composition of embodiment 36, wherein the pharmaceutically acceptable excipient is suitable for topical, transdermal, subcutaneous, intradermal, transmucosal, sublingual, nasal, buccal, intraorbital, intravitreal, subconjunctival, suprachoroidal, intraarticular, and / or inhaled administration.

[0234] Embodiment 38: The pharmaceutical composition of embodiment 36 or 37, wherein the pharmaceutically acceptable excipient is suitable for topical administration.

[0235] Embodiment 39: The pharmaceutical composition of any one of embodiments 36 to 38, wherein the pharmaceutically acceptable excipient comprises hydroxypropyl methylcellulose gel.

[0236] Embodiment 40: The pharmaceutical composition of any one of embodiments 36 to 39, wherein the pharmaceutically acceptable excipient comprises a phosphate buffer.

[0237] Embodiment 41: The pharmaceutical composition of any one of embodiments 36 to 40, wherein the pharmaceutically acceptable excipient comprises glycerol.

[0238] Embodiment 42: The pharmaceutical composition of any one of embodiments 36 to 41, wherein the pharmaceutically acceptable excipient comprises a lipid carrier.

[0239] Embodiment 43: The pharmaceutical composition of any one of embodiments 36-42, wherein the pharmaceutically acceptable excipient comprises a nanoparticle carrier.

[0240] Embodiment 44: A method of administering an antibody to a subject, comprising administering to the subject an effective amount of the virus of any one of embodiments 31 to 35 or the pharmaceutical composition of any one of embodiments 36 to 43.

[0241] Embodiment 45: A method for providing prophylactic, palliative and / or therapeutic alleviation of one or more signs or symptoms of a disease in a subject, comprising administering to the subject an effective amount of the virus of any one of embodiments 31 to 35 or the pharmaceutical composition of any one of embodiments 36 to 43.

[0242] Embodiment 46: The method of embodiment 44 or 45, wherein the virus or composition is administered topically, transdermally, subcutaneously, intradermally, transmucosally, sublingually, intranasally, bucally, intravitreally, subconjunctivally, suprachoroidally, intraarticularly, or by inhalation.

[0243] Embodiment 47: The method of embodiment 45 or 46, wherein the disease is selected from the group consisting of psoriasis, atopic dermatitis, pyoderma gangrenosum, bullous disease, pemphigus, pemphigus vulgaris, pemphigus foliaceus, autoimmune bullous skin disorders, bullous pemphigoid, Behcet's disease, cancer, hidradenitis suppurativa, arthritis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, juvenile idiopathic arthritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, enteropathic arthritis, autoimmune diseases, melanoma, uveal melanoma, and thyroid eye disease.

[0244] Embodiment 48: A method of administering an antibody to the epidermis and / or dermis of a subject, comprising topically, transdermally, or intradermally administering to the subject an effective amount of the virus of any one of embodiments 31 to 35 or the pharmaceutical composition of any one of embodiments 36 to 43.

[0245] Embodiment 49: The method of embodiment 48, wherein the subject's skin is abraded prior to administration.

[0246] Embodiment 50: A method of administering an antibody to the mucosa of a subject, comprising administering to the subject topically, mucosally, sublingually, nasally, or bucally an effective amount of the virus of any one of embodiments 31 to 35 or the pharmaceutical composition of any one of embodiments 36 to 43.

[0247] Embodiment 51: A method of administering an antibody to one or more joints of a subject, comprising intra-articularly administering to the subject an effective amount of the virus of any one of embodiments 31 to 35 or the pharmaceutical composition of any one of embodiments 36 to 43.

[0248] Embodiment 52: A method of administering an antibody to one or both eyes of a subject, comprising topically, intraorbitally, intravitreally, subconjunctivally, or suprachoroidally administering to the subject an effective amount of the virus of any one of embodiments 31 to 35 or the pharmaceutical composition of any one of embodiments 36 to 43.

[0249] Embodiment 53: The method of any one of embodiments 44 to 52, wherein the subject is a human.

[0250] Embodiment 54: The method of any one of embodiments 44 to 53, wherein the subject has not been systemically exposed to the antibody.

[0251] The specification is considered to be sufficient to enable one skilled in the art to practice the disclosure. Various modifications of the disclosure, in addition to those shown and described herein, will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. [Example]

[0252] The present disclosure will be more fully understood by reference to the following examples. However, they should not be construed as limiting the scope of the present disclosure. It is understood that the examples and embodiments described herein are for illustrative purposes only, and that various modifications or changes in light thereof will be suggested to those skilled in the art and are within the spirit and scope of the present application and the scope of the appended claims.

[0253] Example 1: Modified herpes simplex virus vectors encoding antibodies To create a modified herpes simplex virus genome vector capable of expressing an antibody in target mammalian cells (such as human keratinocytes or fibroblasts), the herpes simplex virus genome (Figure 1A) is first modified to inactivate one or more herpes simplex virus genes. Such modifications can reduce the toxicity of the genome in mammalian cells. Next, variants of these modified / attenuated recombinant virus constructs are created so that they carry one or more polynucleotides encoding the desired antibody. These variants include: (1) a recombinant ΔICP4 / ΔICP22 modified HSV-1 genome comprising an expression cassette containing a coding sequence for an antibody fragment (e.g., scFv-Fc) under the control of a heterologous promoter integrated into each ICP4 locus (Figure 1B); (2) a recombinant ΔICP4 modified HSV-1 genome comprising an expression cassette containing a coding sequence for an antibody fragment (e.g., scFv-Fc) under the control of a heterologous promoter integrated into each ICP4 locus (Figure 1C); (3) a recombinant ΔICP4 / ΔICP22 modified HSV-1 genome comprising an expression cassette containing a coding sequence for an antibody heavy chain under the control of a first heterologous promoter and a coding sequence for an antibody light chain under the control of a second heterologous promoter on the same DNA strand integrated into each ICP4 locus (Figure 1D); and (4) a recombinant ΔICP4 / ΔICP22 modified HSV-1 genome comprising an expression cassette containing a coding sequence for an antibody heavy chain under the control of a first heterologous promoter and a coding sequence for an antibody light chain under the control of a second heterologous promoter on the same DNA strand integrated into each ICP4 locus (Figure 1D). (5) a recombinant ΔICP4 / ΔICP22 modified HSV-1 genome (Figure 1F) comprising an expression cassette containing an antibody heavy chain coding sequence under the control of a first heterologous promoter on one DNA strand and an antibody light chain coding sequence under the control of a second heterologous promoter on the other DNA strand, integrated into each ICP4 locus; (6) a recombinant ΔICP4 modified HSV-1 genome (Figure 1G) comprising an expression cassette containing an antibody heavy chain coding sequence under the control of a first heterologous promoter on one DNA strand and an antibody light chain coding sequence under the control of a second heterologous promoter on the other DNA strand, integrated into each ICP4 locus;(7) a recombinant ΔICP4 / ΔICP22 modified HSV-1 genome (Figure 1H) comprising an expression cassette encoding a polycistronic mRNA under the control of a heterologous promoter integrated into each ICP4 locus, wherein the polycistronic mRNA contains coding sequences for antibody heavy and light chains separated by an internal ribosome entry site (IRES); (8) a recombinant ΔICP4 / ΔICP22 modified HSV-1 genome comprising an expression cassette encoding a polycistronic mRNA under the control of a heterologous promoter integrated into each ICP4 locus, wherein the polycistronic mRNA is separated by an internal ribosome entry site (IRES). (9) a recombinant ΔICP4 / ΔICP22 / ΔUL41 modified HSV-1 genome (Figure 1J) containing an expression cassette containing an antibody heavy chain coding sequence under the control of a heterologous promoter integrated into each ICP4 locus, and an expression cassette containing an antibody light chain coding sequence under the control of a heterologous promoter integrated into the UL41 locus and the ICP22 locus; (10) a recombinant ΔICP4 / ΔICP22 / ΔUL41 modified HSV-1 genome containing an expression cassette containing an antibody heavy chain coding sequence under the control of a heterologous promoter integrated into each ICP4 locus; a recombinant ΔICP4 / ΔICP22 / ΔUL41 modified HSV-1 genome (Figure 1K) containing an expression cassette containing a coding sequence for an antibody light chain under the control of a promoter and an expression cassette containing a coding sequence for an antibody heavy chain under the control of a heterologous promoter integrated into the UL41 locus and the ICP22 locus; (11) an expression cassette containing a coding sequence for an antibody heavy chain under the control of a heterologous promoter integrated into each ICP4 locus and a coding sequence for a polycistronic mRNA under the control of a heterologous promoter integrated into the UL41 locus; a recombinant ΔICP4 / ΔUL41 modified HSV-1 genome (Figure 1L), in which the polycistronic mRNA contains two copies of the coding sequence of an antibody light chain separated by an internal ribosome entry site (IRES); (12) an expression cassette containing the coding sequence of an antibody heavy chain under the control of a heterologous promoter integrated into each ICP4 locus, and an expression cassette containing the coding sequence of a polycistronic mRNA under the control of a heterologous promoter integrated into the ICP22 locus, in which the polycistronic mRNAIncluded are a recombinant ΔICP4 / ΔICP22 modified HSV-1 genome (Figure 1M) containing two copies of an antibody light chain coding sequence separated by an internal ribosome entry site (IRES), and a recombinant ΔICP4 / ΔICP22 / ΔUL41 modified HSV-1 genome (Figure 1N) containing an expression cassette containing an antibody light chain coding sequence under the control of a heterologous promoter integrated into the UL41 locus and an expression cassette containing an antibody heavy chain coding sequence under the control of a heterologous promoter integrated into the ICP22 locus. The modified herpes simplex virus genome can be engineered as described above, except that the coding sequence for the full-length antibody heavy chain is replaced with coding sequences for the antibody heavy chain variable region and constant region 1 (CH1) so that the recombinant herpes simplex virus produces Fab fragments.

[0254] These modified herpes simplex virus genome vectors are transfected into engineered Vero cells that are modified to express one or more herpes virus genes. These engineered Vero cells secrete the replication-defective herpes simplex virus with the modified genome packaged inside into the supernatant of the cell culture. The supernatant is then collected, concentrated, and sterilized through a 5 μm filter.

[0255] Example 2: Construction and validation of recombinant herpesviruses encoding antibodies First, a recombinant attenuated HSV vector was designed to express a full-length antibody. A polynucleotide construct was created (e.g., as shown in Figures 1H and 1I) that encoded, from 5' to 3', a first ORF encoding the antibody light chain (including the leader sequence), a synthetic IRES, and a second ORF encoding the antibody heavy chain (including the leader sequence). Next, an IRES-based construct (encoded in an expression cassette further comprising a heterologous promoter and polyA sequence) was inserted into both copies of the HSV1 ICP4 locus. This first viral construct (HSV-flAb2) was engineered to express an anti-human CD20 IgG1 chimeric antibody containing a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:356 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:421.

[0256] Plaques identified in the transgenic cell lawn were picked and screened to identify vectors with correctly inserted IRES constructs. Briefly, Vero cells were infected with putative IRES virus isolates, infection was allowed to proceed for 5 days, and cell supernatants were collected and tested for the presence of secreted full-length antibodies by ELISA according to the manufacturer's instructions (Abcam, catalog no. ab195215). All isolates tested expressed and secreted the encoded antibodies into the cell supernatant; however, the antibodies were produced at relatively low levels. The most productive isolate of HSV-flAb2 secreted 0.844 ng / mL of the encoded full-length chimeric antibody into the supernatant of infected Vero cells.

[0257] Instead of expressing full-length antibodies from engineered vectors, which require the use of multiple ORFs encoding antibody light and heavy chains, recombinant HSV1 vectors were constructed to express single-chain antibodies from a single ORF (see, e.g., Figures 1B and 1C). Here, single-chain antibodies were constructed to contain a leader sequence, a heavy chain variable region sequence, a light chain variable region sequence, a linker polypeptide connecting the heavy and light chain variable regions, and an Fc region (i.e., scFv-Fc antibodies). Two different variants of each single-chain antibody were designed to encompass the possible relative orientations of both the light chain variable region sequence and the heavy chain variable region sequence: the "Fc1" variant of each antibody contained, from n- to c-terminus, leader sequence-heavy chain variable region-linker sequence-light chain variable region-and Fc region, and the "Fc2" variant of each antibody contained, from n- to c-terminus, leader sequence-light chain variable region-linker sequence-heavy chain variable region-and Fc region. These single-chain antibody constructs (encoded in expression cassettes further comprising a heterologous promoter and polyA sequence) were then inserted into both copies of the HSV1 ICP4 locus. A summary of the sequences used for each scFv-Fc antibody tested is shown in Table 4 below.

[0258] Table 4. HSV-encoded scFv-Fc antibodies TIFF2025186440000019.tif103152

[0259] Viral plaques were identified and harvested from the lawn of engineered cells, and isolates were individually screened in Vero cells to identify vectors producing the encoded antibodies as described above. Cell supernatants were harvested from infected Vero cells and tested for the presence of secreted antibodies by ELISA according to the manufacturer's instructions (Abcam, catalog number ab195215 for single-chain antibodies containing human Fc sequences; Abcam, catalog number ab151276 for single-chain antibodies containing mouse Fc sequences). Table 5 below shows the calculated average concentrations of each antibody secreted into the supernatant from infected cells.

[0260] Table 5. Concentration of single-chain antibodies secreted from HSV-infected cells TIFF2025186440000020.tif55128

[0261] Surprisingly, all single-chain antibodies were secreted into the cell supernatant at much higher concentrations (at least 800-fold higher) than the full-length HSV-flAb2 antibody. In direct comparison, the Fc2 single-chain variant of HSV-Ab2 was detected at a concentration 19,114-fold higher than the full-length variant of HSV-Ab2.

[0262] Next, a dose-ranging experiment was performed to measure antibody secretion from human cells infected with engineered HSV1 vectors encoding single-chain antibodies. Immortalized human keratinocytes (HaCaT) were either mock-infected (MOI 0) or infected with HSV vectors at various multiplicities of infection (MOIs) for 48 hours. The antibody-encoding vectors used in this experiment were HSV-Ab1Fc1 (single-chain human anti-human TNFα antibody), HSV-Ab1Fc2 (single-chain human anti-human TNFα antibody), HSV-Ab2Fc2 (single-chain anti-human CD20 chimeric antibody), HSV-Ab66Fc1 (single-chain mouse anti-mouse IL-4Ra antibody), and HSV-Ab66Fc2 (single-chain mouse anti-mouse IL-4Ra antibody). After infection, supernatants were collected from infected cells and centrifuged at 11,000 × g for 5 minutes. The antibodies secreted into the clarified supernatants were then quantified using an anti-human IgG (Figure 2A) ELISA kit (Abcam, catalog no. ab195215) or an anti-mouse IgG (Figure 2B) ELISA kit (Abcam, catalog no. ab151276). While little or no antibodies were detected in mock-infected cells, secreted human and chimeric antibodies were reliably detected in a dose-dependent manner in infected human keratinocytes (Figure 2A). Single-chain mouse antibodies were also detected in secreted human keratinocytes, but their secretion did not appear to be dose-dependent (Figure 2B).

[0263] Finally, to confirm the functionality of the HSV-encoded antibody, we investigated the ability of the single-chain human anti-human TNFα antagonist antibody, HSV-Ab1Fc1, to inhibit ELISA-based TNFα detection. Immortalized human keratinocytes (HaCaT) were left uninfected or infected with HSV-Ab1Fc1 at MOIs of 0.3, 1, and 3 for 48 hours. After infection, cell supernatants were collected and clarified by centrifugation at 11,000 xg for 5 minutes six times. Recombinant human TNFα (1000 pg / mL) was added to the clarified cell supernatant and incubated at 37°C for 2 hours. Next, an anti-TNFα ELISA (Abcam, catalog no. ab181421) was performed to determine whether the single-chain Ab1Fc1 produced by the engineered virus could mediate inhibition of TNFα detection. Detectable levels of TNFα were significantly reduced in cell supernatants harvested from infected human cells compared with mock-infected cells (Fig. 3 ), confirming that the secreted single-chain antibody was indeed functional.

[0264] Taken together, the data presented in this example demonstrate that (1) recombinant HSV-1 vectors were successfully constructed to encode full-length antibodies and antibody fragments, (2) the engineered vectors expressed / secreted single-chain antibodies at levels hundreds to thousands of times higher than full-length antibodies, (3) the vectors were able to reliably express fully human, chimeric, and murine antibodies (containing Fc regions from multiple IgG isotypes) at comparable levels, (4) HSV-mediated antibody secretion from human cells was dose-dependent, and (5) the secreted antibodies were functional. Without wishing to be bound by theory, it is believed that engineered herpesvirus vectors provide a novel method for administering multiple types of antibodies to humans.

[0265] Example 3: Herpesvirus-encoded antibodies in a murine atopic dermatitis (AD) model Atopic dermatitis (AD) is a chronic, relapsing, and often intensely pruritic inflammatory disorder of the skin. The primary cause of this disease appears to be a defective epithelial barrier resulting from a combination of genetic predisposition, immune system function, and environmental factors. Treatment for moderate to severe AD has remained largely unchanged for decades and relies on broad-spectrum immunosuppressants. However, in recent years, several therapeutic approaches using systemic administration of antibodies targeting various aspects of the complex immune activation pathway in atopic dermatitis have been explored in clinical settings. These include antibodies targeting the IL-4 pathway (e.g., dupilumab), the IL-12 / 23p40 pathway (e.g., ustekinumab), the IL-13 pathway (e.g., tralokinumab, lebrikizumab), the IL-17 pathway (e.g., secukinumab), and the IL-31 pathway (e.g., nemolizumab). Indeed, systemic therapy (biweekly subcutaneous injections) using the monoclonal antibody dupilumab (anti-IL-4Ra) was recently approved by the FDA for the treatment of atopic dermatitis. However, systemic administration of therapeutic antibodies that target immune pathways has been shown to suppress the immune system and place patients at significant risk for infections and other complications.

[0266] Topical application of vitamin D3 or its synthetic analogues induces AD-like inflammation in mouse skin. A skin inflammation model induced by the vitamin D analogue calcipotriol (MC903) showed that topical application of MC903 increased levels of TSLP and group 2 (IL-5) cytokines. + and IL-13 + MC903 has recently attracted attention because it induces innate lymphocyte infiltration into the skin, thereby mimicking certain immune disturbances observed in skin lesions in humans with AD. Topical administration of MC903 has also been shown to induce increased IL-4 signaling in treated animals (Martel et al., Yale J Biol Med (2017)).

[0267] The goals of the in vivo experiments described here were (1) to establish a murine MC903-induced model of atopic dermatitis and demonstrate the potential for HSV-encoded antibody expression in these animals when the virus is administered topically to AD-like skin, and (2) to evaluate whether the murine surrogate single-chain antibody of dupilumab (anti-IL-4Ra) alleviates one or more AD symptoms in this animal model.

[0268] Materials and Methods For nucleic acid analysis, skin biopsies were lysed in buffer RLT (AllPrep DNA / RNA Mini Kit, Qiagen, catalog number 80204) containing DTT (G-Biosciences, catalog number 3483-12-3) using sonication at 25% amplitude with a QSONICA ultrasonicator (125W, 20Hz). After sonication, any insoluble material was pelleted using high-speed centrifugation. DNA and RNA were isolated using the AllPrep DNA / RNA Mini kit (Qiagen) according to the manufacturer's protocol. For qPCR / qRT-PCR analysis, 50 ng of DNA or RNA was used per reaction in a total reaction volume of 25 μL. DNA quantification was measured by qPCR analysis using TaqMan® Fast Advanced Master Mix (Applied Biosystems), and RNA quantification was measured by qRT-PCR analysis using Quantabio 1-Step RT-qPCR ToughMix. All samples were performed in duplicate or triplicate.

[0269] For immunofluorescence, 5 μm sections were taken from OCT-frozen tissues, mounted on slides, and air-dried for up to 1 hour. Slides were then immersed in 100% methanol (MeOH) at -20°C for 10 minutes and air-dried. Methanol-fixed sections were rehydrated by washing three times (5 minutes each) in PBS at room temperature, then incubating in 3% H2O2 at room temperature for 10 minutes, followed by three washes with PBS. Next, the samples were incubated with blocking solution (Power Block) in a humidified chamber for 1 hour at room temperature. Excess blocking solution was removed, and the sections were stained with a small volume of primary antibody (Ab) solution (30–50 μL primary Ab solution / section) prepared in antibody dilution buffer. Sections were incubated with primary antibody (mouse anti-human IgG antibody; Abcam, catalog no. ab200699) for 16 hours at 4°C or 1 hour at room temperature, washed three times with TBST (TBS + 0.025% Triton X-100) for 5 minutes at room temperature, and then secondary Ab (Alexa Fluor® 488-conjugated goat anti-mouse antibody; ThermoFisher, catalog no. A-11029) was applied at a dilution of 1:200 in antibody dilution buffer for 30 minutes at room temperature in a humidified chamber. Slides were again washed three times with TBST, and stained sections were mounted with mounting medium (ProLong™ Gold Antifade Mountant with DAPI, ThermoFisher, catalog no. P36931) and covered with a coverslip. After dehydration (approximately 24 hours), sections were imaged using an ECHO fluorescence microscope.

[0270] For hematoxylin and eosin (H&E) staining, 5 μm sections were taken from cryopreserved tissue, mounted on slides, and air-dried for up to 1 hour. Dried slides were rehydrated by immersion in double-distilled water for 2 minutes at room temperature. Next, sections were incubated in 2x Gill's hematoxylin (VWR) for 2 minutes at room temperature, then immersed in acid alcohol 2-3 times, immersed in ammonia blue 3-4 times, and incubated in eosin (eosin Y solution 1%, VWR) for 2 minutes. Between each step, samples were rinsed 3-4 times with tap water. Stained and rinsed sections were gradually dehydrated using ethanol (EtOH), first with 95% EtOH twice for 2 minutes each, then with 100% EtOH twice for 2 minutes each. Sections were then cleared by rinsing three times for 2 minutes in Histo-Clear, mounted with mounting medium (Permount™ Mounting Medium, @P15-100), and capped with a coverslip. After approximately 24 hours of dehydration, sections were imaged using a bright-field microscope.

[0271] result Establishment of AD animal models An in vivo study was initiated to demonstrate that MC903 induces atopic dermatitis-like symptoms in treated animals. Fourteen C57BL / 6J mice were used in this study (two mice per group). MC903 was prepared at a concentration of 100 μM in ethanol (EtOH). On day 1, mice were anesthetized with isoflurane, their backs were shaved and treated with a chemical depilatory, and 25 μL of EtOH or MC903 / EtOH was applied to the left and right ears (both sides) and four marked dorsal sites (approximately 2 cm each). 2) were applied to the affected area. Mice were then re-treated with EtOH or MC903 / EtOH at the same sites on days 2, 3, 4, and 5. Next, on days 5 or 7, select cohorts of animals received topical treatment with HSV-Ab1Fc1 (or vehicle control) formulated in a gel vehicle at the left and right ears and two MC903-treated dorsal sites, and intradermal injections of HSV-Ab1Fc1 (or vehicle control) at the remaining two MC903-treated dorsal sites. These mice were then euthanized on days 7 and 9, respectively. The HSV-AbFc1 vector (fully human anti-TNFα single-chain antibody) was used in this experiment as a proof-of-concept for evaluating antibody expression in AD-like lesions, as this virus was adequately purified and characterized at the start of the study. The experimental design is outlined in Table 6.

[0272] Table 6. Overview of study design to establish MC903 model TIFF2025186440000021.tif96146

[0273] At the designated termination date, animals were euthanized and the dorsal treatment site was harvested using an 8 mm punch biopsy, while the entire ear was excised. Half of each biopsy / ear was flash-frozen in liquid nitrogen for nucleic acid analysis, while the other half was processed for immunofluorescence analysis (as described above).

[0274] To confirm the appearance of atopic dermatitis-like lesions in mouse skin after topical MC903 treatment (days 1–5), qRT-PCR analysis was performed on harvested tissues to quantify changes in the expression of specific markers of atopic dermatitis previously shown by others to be upregulated in MC903-exposed skin. Specifically, the mean fold change in TSLP or IL-4 transcripts in GAPDH-corrected mouse ear or dorsal tissues treated with MC903 was calculated using the delta-delta Ct method compared with the corresponding EtOH-treated ear or dorsal tissues. As previously observed by other groups, repeated topical MC903 exposure significantly induced the expression of TSLP (Figure 4A) and IL-4 (Figure 4B) in treated mouse skin, confirming the establishment of an AD-like mouse model. Interestingly, the kinetics of TSLP and IL-4 expression were different, with TSLP levels peaking on day 5 in both ear and dorsal skin, whereas IL-4 expression had a more delayed kinetics, with highest levels of transcript expression on day 7 in dorsal skin and day 9 in ear skin.

[0275] Concurrent with increased expression of TSLP and IL-4 was thickening of the ear skin in the MC903-treated sites compared with the ethanol control-treated sites (Figure 5), further confirming that MC903 induced atopic dermatitis-like symptoms in these mice.

[0276] Next, we investigated the potential for HSV-encoded antibody expression following topical administration to atopic dermatitis-like lesions using an engineered virus (HSV-Ab1Fc1) encoding an exemplary fully human single-chain antibody (Ab1Fc1). For immunofluorescence analysis, ear and dorsal skin tissue sections were prepared from animals exposed to MC903 on days 1–5 and topically treated with HSV-Ab1Fc1 (formulated in a methylcellulose gel carrier) or a solvent control (formulated in a methylcellulose gel carrier) on days 5 or 7. Figure 6A shows representative immunofluorescence images of ear and dorsal skin harvested on day 7 after topical treatment with HSV-Ab1Fc1 or a solvent control on day 5. Figure 6B shows representative immunofluorescence images of ear and dorsal skin harvested on day 9 after topical treatment with HSV-Ab1Fc1 or a solvent control on day 7. Strong human IgG protein expression was detected in both ear and dorsal tissues infected with HSV-Ab1Fc1 but not in the vehicle control, suggesting that the virus was able to deliver its virally encoded antibody cargo to atopic dermatitis-like lesions after topical exposure.

[0277] Taken together, this data demonstrates the successful establishment of an AD-like phenotype after MC903 treatment and the feasibility of potent local delivery of virally encoded antibodies to AD-like lesions after topical HSV administration.

[0278] Evaluation of antibody efficacy after local delivery of herpesvirus Having confirmed the atopic dermatitis-like nature of the lesions induced by MC903 therapy (see, e.g., Figure 4), we next tested the efficacy of a topical HSV-encoded antibody in reducing one or more signs or symptoms of atopic dermatitis in this animal model. As mentioned above, dupilumab (an anti-human IL-4Ra antibody) is currently the only FDA-approved antibody for atopic dermatitis and is administered systemically by biweekly subcutaneous injection. Here, we engineered a recombinant herpesvirus (HSV-Ab66Fc1) to express a dupilumab surrogate antibody (Ab66Fc1, a mouse anti-mouse IL-4Ra single-chain antibody) targeting mouse IL-4Ra to test the antibody's ability to reduce MC903-mediated ear thickening. Furthermore, we chose to use an antibody targeting the IL-4 pathway, in part due to the observation that MC903 significantly induced IL-4 signaling in C57BL / 6J mice (Figure 4B).

[0279] Eight C57BL / 6J mice were used in this study (two mice per group). MC903 was prepared at a concentration of 100 μM in ethanol (EtOH). The backs of mice were shaved with electric clippers and treated with a chemical depilatory compound on day -2. On day 1, mice were anesthetized with isoflurane, and 25 μL of EtOH or MC903 / EtOH was applied to the left and right ears (both sides) and four marked dorsal sites (approximately 2 cm each). 2 ) on the left and right ears. Mice were re-treated with EtOH or MC903 / EtOH at the same sites on study days 2, 3, 4, and 5. Next, per study schedule, selected cohorts of animals received five topical treatments of HSV-Ab66Fc1 formulated in a gel vehicle or vehicle control at the left and right ears and two dorsal sites treated with EtOH or MC903 / EtOH, and five intradermal injections of HSV-Ab66Fc1 or vehicle control at the remaining two dorsal sites treated with EtOH or EtOH / MC903. The experimental design is outlined in Table 6.

[0280] Table 6. Overview of study design to evaluate efficacy of HSV-encoded antibodies TIFF2025186440000022.tif69152TOP=Topical treatment;ID=Intradermal injection

[0281] On day 10, 8 mm punch biopsies were taken from the dorsal skin, and the entire ear was simultaneously excised, and the tissues were processed for nucleic acid analysis. qPCR data from the tissue samples showed that the engineered HSV genome encoding the Ab66Fc1 transgene efficiently transduced the MC903-treated ear skin (Figure 7A) and, to a lesser extent, the MC903-treated dorsal skin (Figure 7B) of immunocompetent animals. Significant viral transduction was also observed in the ethanol-treated ear skin, likely due to the thinness and fragility of this tissue type. Not only did the viral genome efficiently transduce the ear skin, but mouse anti-IL-4Ra single-chain antibodies were also robustly expressed after infection, as assessed by qRT-PCR analysis (Figure 7C). The lowest Ab66Fc1 transcripts were detected in MC903-treated dorsal skin, consistent with the significantly lower transduction efficiency observed in this tissue, potentially due to the less skin damage induced by MC903 in this tissue type. No Ab66Fc1 DNA or RNA was detected in tissue treated with the solvent control, demonstrating the specificity of the single-chain antibody assay. Unfortunately, because single-chain antibodies contain mouse IgG Fc, specific immunofluorescence detection of the expressed protein in these samples was not possible due to endogenous mouse IgG.

[0282] To evaluate the efficacy of topical therapy, we measured ear thickness in both ears of each mouse daily from day 1 to day 10 using digital calipers (Figure 8A). As expected, MC903 exposure induced significant ear thickening in treated mice compared with ethanol exposure. The mean ear thickness of ethanol / vehicle-treated animals was 0.19 mm at day 10, while the mean thickness of MC903 / vehicle-treated animals at this same time point was 0.6125 mm. However, by day 5, a significant reduction in ear thickness (>22% thinning) was observed in MC903 / Ab66Fc1-treated ears compared with MC903 / vehicle-treated ears, which was consistently maintained throughout the experiment. The antibody did not cause any apparent change in ear thickness in ethanol-exposed ears (compare EtOH / vehicle with EtOH / Ab66 in Figure 8A), confirming the specificity of the antibody for reducing this atopic dermatitis-like phenotype.

[0283] Interestingly, the improvement in MC903-induced ear thickening on day 5 mediated by topical treatment with HSV-Ab66Fc1 was comparable to the improvement observed on day 5 in a similar MC903 mouse model following administration of an anti-IL17c antibody (the pathway targeted by AD antibodies currently in human clinical trials, e.g., secukinumab), as previously reported (see Figure 5 in WO2017060289). However, despite the similarity between the positive ear thickness results observed in this study and those described in WO2017060289, the therapeutic approaches differed significantly. The study described in WO2017060289 required systemic administration of a therapeutic antibody via intraperitoneal injection at a high dose and included an antibody pretreatment step in which the first dose of antibody was administered 3 days before initiating MC903 treatment. This example used local (rather than systemic) administration of the formulation and did not use any antibody pretreatment (the first HSV-Ab66Fc1 application was given after the first MC903 application).

[0284] On day 10, harvested ears from each mouse were weighed prior to tissue manipulation. Consistent with the ear thickness measurements, MC903 treatment significantly increased mean ear weight compared to ethanol controls, an increase that was partially restored by HSV-Ab66Fc1 therapy (Figure 8B). It was also noted that at harvest on day 10, MC903 / vehicle-treated ears felt much stiffer / less flexible than MC903 / Ab66-treated ears, providing a qualitative measure of treatment efficacy.

[0285] Taken together, this data demonstrates that engineered recombinant herpesviruses can successfully express their encoded cargo in atopic dermatitis-like lesions following topical treatment, and further demonstrates that expression of antagonistic antibodies targeting pathways known to play a role in human AD disease results in quantitative and qualitative improvement of the disease phenotype. Without wishing to be bound by theory, it is believed that topical administration of antibody-encoding herpesvirus vectors offers a novel and unique method for delivering effective immunotherapy (e.g., for treating inflammatory diseases / conditions such as atopic dermatitis) while limiting systemic exposure to the antibody.

[0286] Sequence information SEQUENCE LISTING <110> Krystal Biotech, Inc. <120> COMPOSITIONS AND METHODS FOR ANTIBODY DELIVERY <150> US 62 / 692,514 <151> 2018-06-29 <150> US 62 / 713,066 <151> 2018-08-01 <160> 1122 <170> FastSEQ for Windows Version 4.0 <210> 1 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 1 Asp Tyr Ala Met His 1 5 <210> 2 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 2 Ser Tyr Asn Met His 1 5 <210> 3 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 3 Asn Tyr Gly Met Asn 1 5 <210> 4 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 4 Thr Tyr Trp Leu Gly 1 5 <210> 5 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 5 Asn Tyr Trp Met Asn 1 5 <210> 6 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 6 Asp Tyr His Ile His 1 5 <210> 7 <211> 5 <212> PRT <213> Artificial Sequence 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Artificial Sequence <220> <223> Synthetic Construct <400> 41 Asp Ser...

Claims

[Claim 1] The invention described in the specification of this application.

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