In one embodiment, an expression system for expressing a UL128 complex is provided herein. The expression system may include a bacterial artificial chromosome (BAC) construct, wherein the BAC construct comprises a viral vector inserted with a set of DNA sequences that encode a UL128 complex. In another embodiment, a vaccine composition for preventing HCMV infection is provided. The vaccine composition may include a viral or bacterial vector capable of expressing a UL128 complex and a pharmaceutically acceptable carrier, adjuvant, additive or combination thereof or additional vector expressing a protein adjuvant. The viral vector may be an MVA and the UL128 complex includes five HCMV proteins or antigenic fragments thereof: UL128, UL130, UL131A, gL, and gH. In some embodiments, the viral vector is further inserted with one or more additional DNA sequences that encode one or more additional HCMVHCMV proteins or antigenic fragments thereof such as pp65, gB or both, or such as gM/gN or gO.

The invention provides a bidirectional hCMV-rhCMV promoter and recombinant vectors and recombinant virus comprising the bidirectional hCMV-rhCMV promoter operably linked to a first transgene in one direction and to a second transgene in the opposite direction. The invention also provides methods of making and using such recombinant vectors and recombinant virus.

The invention is directed to multivalent HCMV immunogenic compositions and their use.

Stealth adapted viruses differ from the conventional viruses from which they are derived in not evoking an inflammatory response. This can occur because of the deletion or mutation of the genes coding for the relatively few virus components, which are normally targeted by the cellular immune system. As part of the stealth adaptation process, exchanges can occur between some and possibly all of the sequences of the initiating virus and sequences of both cellular and bacteria origin. A description is provided on the culturing of stealth adapted viruses. A characteristic feature of cultured stealth adapted virus infected cells is the accumulation of intracellular materials, which will fluoresce under ultraviolet (UV) light in the presence of certain dyes including neutral red and acridine orange.

A hybrid promoter for recombinant expression of proteins of interest is disclosed that combines a mCMV enhancer sequence with a rat EF-1alpha intron sequence. Also disclosed are an expression cassette containing the hybrid promoter and a recombinant expression vector containing the expression cassette. A mammalian host cell, which comprises the recombinant expression vector is also disclosed, as is a method of producing a protein of interest that employs the mammalian host cell, optionally involving tetracycline-inducible expression of the protein.

The present invention relates to the field of immunotherapy, in particular, to adoptive T cell therapy, T cell receptor (TCR) gene therapy and vaccination. The invention provides a method for preparing a nucleic acid encoding the TCR alpha chain construct (TRA) and TCR beta chain construct (TRB) of a TCR construct specific for an epitope from an antigen presented on major histocompatibility complex (MHC), comprising contacting T cells isolated from a donor with a library of artificial antigen presenting cells (APC) comprising cells expressing all MHC I or MHC II alleles present in the donor, preferably, in K562 cells. The TCR construct can be expressed in a T cell, which is useful for adoptive T cell therapy, e.g., of cancer, viral infections or autoimmune diseases. The invention further provides a method for identifying the epitope recognized by said TCR. Immunogenic epitopes recognized by said TCRs can be used to develop vaccine formulations to induce antigen-specific T cell immunity in patients. The invention further provides pairs of two TCR constructs and respective immunogenic epitopes obtained by the method of the invention, wherein the epitopes are from human papillomavirus (HPV) 16 (also designated alphapapillomavirus 9) oncoprotein E5 and human cytomegalovirus (CMV) protein pp65.

A method for treating a patient comprising: (a) providing a composition comprising a population of T cells expressing both a chimeric antigen receptor (CAR) and a T cell receptor specific for a cytomegalovirus (CMV) antigen; (b) administering the composition to the patient; and (c) administering to the patient a viral vector encoding: (i) CMV pp65 and (ii) a fusion protein comprising exon 4 of CMV protein IE I (e4) and exon 5 of CMV protein IE2 (e5) either prior to or subsequent to administering the composition comprising a population of T cells to the patient is described.

The invention provides a bidirectional hCMV-rhCMV promoter and recombinant vectors and recombinant virus comprising the bidirectional hCMV-rhCMV promoter operably linked to a first transgene in one direction and to a second transgene in the opposite direction. The invention also provides methods of making and using such recombinant vectors and recombinant virus.

Compositions and methods to increase the in vivo capacity of CD8+ T cells to kill HIV-infected and reactivated latent HIV-infected T cells (LHITC) to functionally cure HIV infection or improve the clinical course. Compositions and methods to increase the in vivo capacity of CD8+ T cells to kill CMV or CMV-infected cells are also provided.

The technology described herein relates to modified T cells and their use in immunotherapeutic methods. In various examples, the T cells are modified so as to decrease or eliminate OD3ζ, TRAC, and/or TRBC expression.