Vaccine properties of a novel marker gene-free recombinant modified vaccinia Ankara expressing immunodominant CMV antigens pp65 and IE1

Abstract

CMV tegument protein pp65 and CMV immediate early gene product IE1 are both considered immunodominant targets of cell-mediated immunity (CMI) and potentially capable of controlling CMV infection. To better assess their role in host defense, we have constructed a novel MVA transfer vector named pZWIIA and generated a recombinant MVA (rMVA) expressing both full-length pp65 and exon4 of IE1 (pp65-IE1-MVA) at high levels, followed by the genetic removal of the bacterial marker gene used to distinguish recombinant forms. Immunogenicity evaluation indicates that pp65-IE1-MVA not only can induce robust primary CMI to both antigens in HLA A2.1 Tg mice, but also can stimulate vigorous expansion of memory T lymphocyte responses to pp65 and IE1 in PBMC of CMV-positive donors. These properties make the MVA-based vaccine ideal for the dual role of priming and boosting CMV-specific T cell immunity as a means to control CMV disease in recipients of hematopoietic cell or solid organ transplantation (HCT or SOT). pZWIIA alone or in combination with other MVA transfer vectors can be used to generate MVA based multiple-antigen vaccine which have application in vaccine development for a wide spectrum of infectious diseases and cancer.

Comparison of monovalent glycoprotein B with bivalent gB/pp65 (GP83) vaccine for congenital cytomegalovirus infection in a guinea pig model: Inclusion of GP83 reduces gB antibody response but both vaccine approaches provide equivalent protection against pup mortality

“…Recombinant MVA vaccines were generated using MVA transfer vector pZWIIA[24, 37]. Details of generation of these vaccines have already been described elsewhere[24].…”

Development of a novel, guinea pig-specific IFN-γ ELISPOT assay and characterization of guinea pig cytomegalovirus GP83-specific cellular immune responses following immunization with a modified vaccinia virus Ankara (MVA)-vectored GP83 vaccine

“…Recombinant MVAs were generated using MVA transfer vector ZWIIA as previously described [31]. Plasmids pKTS 404 and pKTS 437, expressing a truncated, secreted form of GPCMV gB and a full-length form of GP83, respectively, were used as templates for PCR-mediated insertion of each ORF into ZWIIA.…”

A Vaccine Based on the Rhesus Cytomegalovirus UL128 Complex Induces Broadly Neutralizing Antibodies in Rhesus Macaques

“…MVA-RhUL128C, MVARhUL128C⌬, or MVA expressing RhUL128-UL131A subunits was generated by the BAC technology as described below. MVA expressing either RhUL128 or RhUL130 alone was generated by the conventional manipulation strategy in eukaryotic cells as described previously (35). The construction of MVA-RhgB having a deleted transmembrane domain (TM) is described elsewhere (36).…”