This disclosure provides replication-incompetent adenoviral vectors useful in vaccine development and gene therapy. The disclosed vectors comprise a selective deletion of E3 and are particularly useful for preparation of vaccines development and for gene therapy using toxic transgene products that result in vector instability that occurs when the entire E3 domain is deleted.

Provided is a method for preparing an adenovirus vector vaccine by means of a perfusion culture process. The method comprises a step of culturing adenovirus host cells, and in particular a step of adjusting the perfusion rate by means of at least two stages according to cell density. The method increases the single cell yield of a virus after infection and the specific activity of a virus harvest liquid while achieving high-density growth of adenovirus host cells.

A recombinant vector comprises simian adenovirus 43, 45, 46, 47, 48, 49 or 50 sequences and a heterologous gene under the control of regulatory sequences. A cell line which expresses simian adenovirus 43, 45, 46, 47, 48, 49 or 50 gene(s) is also disclosed. Methods of using the vectors and cell lines are provided.

The present disclosure provides, among other things, helper-dependent adenoviral serotype 35 (Ad35) vectors. In various embodiments, helper-dependent Ad35 vectors can be used to deliver a therapeutic payload to a subject in need thereof. Exemplary payloads can encode replacement proteins, antibodies, CARs, TCRs, small RNAs, and genome editing systems. In certain embodiments, a helper-dependent Ad35 vector is engineered for integration of a payload into a host cell genome. The present disclosure further includes methods of gene therapy that include administration of a helper-dependent Ad35 vector to a subject in need thereof.

Provided are compounds that enhance the efficacy of viruses by increasing spread of the virus in cells, increasing the titer of virus in cells, or increasing the antigen expression from a virus, gene or trans-gene expression from a virus, or virus protein expression in cells. Other uses, compositions and methods of using same are also provided.

Disclosed herein a unique cell line system to generate a novel bovine adenovirus vector that provides more gene insertion capabilities and better immunogenicity for inserted antigens. The unique cell line is used for generating and growing of the new adenovirus vectors for gene delivery or recombinant vaccine production.

The present invention relates to a cell line for producing an adenovirus having a limited autoreplication capability, and a method of preparing the same, and more particularly, to a cell line for producing an adenovirus having no autoreplication capability by expressing at least one selected from an adenoviral E1 protein and an E1A or E1B protein, and a method of preparing the same. Also, the present invention relates to the use of the cell line expressing at least one selected from the adenoviral E1 protein and the E1A or E1B protein.

Disclosed herein are compositions that include modified adenoviruses. Also disclosed are nucleotides, cells, and methods associated with the compositions including their use as vaccines. Also disclosed herein are viral vectors using TET promoter system and methods of producing viruses having the same.

A method for preparing an infectious, recombinant virus vector comprises the steps of: (a) infecting host cells with a first virus comprising an encapsidation defective adenovirus (edAd), the edAd comprising a first defective virus genome; (b) incubating the infected host cells in a culture medium for a period of time sufficient for producing infectious virus particles; and (c) recovering infectious virus particles secreted into a culture supernatant, wherein the edAd or the host cells comprise a second defective virus genome engineered to express a target gene of interest, wherein the edAd or the host cells comprise nucleic acid sequences sufficient for expressing adenovirus (Ad) helper genes necessary for replication of the defective virus DNA; and wherein the edAd or the host cells comprise nucleic acid sequences sufficient from expressing helper functions necessary for producing infectious, replication defective virus particles corresponding to the second virus.

Provided herein are adenoviral vectors comprising nucleotide sequences encoding a Zika virus M and Env antigen, wherein the nucleotide sequence encoding the Zika virus M and Env antigen is operably linked to a cytomegalovirus (CMV) promoter comprising at least one tetracycline operator (TetO) motif. Also provided herein are pharmaceutical compositions comprising the adenoviral vectors, methods of producing the adenoviral vectors, methods of preventing Zika virus or the progression of Zika virus in a subject in need thereof, and kits comprising the adenoviral vectors and host cells.