Murine cytotoxic T cell response specific for human cytomegalovirus glycoprotein B (gB) induced by adenovirus and vaccinia virus recombinants expressing gB

Berencsi, Klára; Rando, Robert F.; deTaisne, Charles; Paoletti, Enzo; Plotkin, Stanley А.; Gönczöl, Éva

doi:10.1099/0022-1317-74-11-2507

Cited by 26 publications

(12 citation statements)

References 24 publications

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“…None of the viral stocks used in the experiments contained detectable replication-competent virus. Ad5dlE3, a replication-competent AdS with deletion of E3 region, was described (15). Virus (1 x 1011 plaque-forming units/ml) in 0.1 ml of phosphate-buffered saline (PBS, pH 7.4) was infused retrograde into the biliary tract offemale CBA and nu/nu mice as described (16).…”

Section: Methodsmentioning

confidence: 99%

Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy.

Yang

Nunes

Berencsi

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

1,500

944

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An important limitation that has emerged in the use of adenoviruses for gene therapy has been loss of recombinant gene expression that occurs concurrent with the development of pathology In the organ exp g the transgene. We have used liver-dircted approaches to gene therapy in mice to study mecanims that underlie the problems with transient expression and pathology that have characterized in vivo applications of first-generation recombinant adenoviruses (i.e., those deleted of Ela and Elb). Our data are consistent with the following hypothesis. Cells harboring the recombinant viral genome express the transgene as desired; however, lowlevel expression of viral genes also occurs. A virus-specific cellular immune response is stimulated that leads to destruction of the genetically modified hepatocytes, massive hepatitis, and repopulation of the liver with nontransgene-containing hepatocytes. These findigs suggest approaches for improving recombinant adenoviruses that are based on further crippling the virus to limit expression of nondeleted viral genes.Human adenoviruses have emerged as a promising technology for in vivo gene therapy. The 36-kb double-stranded

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Section: Methodsmentioning

confidence: 99%

Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy.

Yang

Nunes

Berencsi

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

1,500

944

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“…After the run, the acrylamide gel was placed in a transfer apparatus with a polyvinylidene difluoride membrane (Bio-Rad), and conditions for transfer were according to the manufacturer's protocol. Development of the polyvinylidene difluoride membrane and detection of the gB proteins were done using the primary antibody (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) together with an ECL bioluminescence kit, using a goat anti-rabbit alkaline phosphatase-conjugated secondary antibody according to the manufacturer's conditions (Pharmacia Amersham, Piscataway, N.J.). The dried membrane was exposed to Kodak X-omat autoradiography film and developed in a standard film developer (model M35A; Kodak, Rochester, N.Y.).…”

Section: Methodsmentioning

confidence: 99%

“…gB680-MVA was selected by color screening in the presence of X-glcA (5-bromo-4-chloro-3-indolyl-␤-D-glucuronide) (Sigma-Aldrich, St Louis, Mo.). After each round of color selection, a portion of lysate from each plaque was immunostained using a gB-specific monoclonal antibody (MAb) (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) to distinguish gB680-positive and gus ϩ MVA from gB680-negative gus ϩ recombinant MVA (rMVA). After 8 to 10 rounds of plaque purification, gB680-MVA virus was grown and amplified on BHK-21 or CEF cells.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, CMV gB has been an attractive candidate for CMV vaccine development. CMV gB vaccines using recombinant gB protein expressed from plasmid DNA and gB expressed in several different viral vectors (ALVAC, adenovirus, and vaccinia virus [VV]) have been investigated with animal models (9,13,23,26,31,40,54). Safety and moderate immunogenicity have been demonstrated with these vaccines, but no licensed CMV vaccine is available.…”

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confidence: 99%

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Recombinant Modified Vaccinia Virus Ankara Expressing a Soluble Form of Glycoprotein B Causes Durable Immunity and Neutralizing Antibodies against Multiple Strains of Human Cytomegalovirus

Wang

Rosa

Maas

et al. 2004

J Virol

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Human cytomegalovirus (CMV) is a viral pathogen that infects both genders, who remain asymptomatic unless they receive immunosuppressive drugs or acquire infections that cause reactivation of latent virus. CMV infection also causes serious birth defects following primary maternal infection during gestation. A safe and effective vaccine to limit disease in this population continues to be elusive. A well-studied antigen is glycoprotein B (gB), which is the principal target of neutralizing antibodies (NAb) towards CMV in humans and has been implicated as the viral partner in the receptor-mediated infection by CMV in a variety of cell types. Antibody-mediated virus neutralization has been proposed as a mechanism by which host immunity could modify primary infection. Towards this goal, an attenuated poxvirus, modified vaccinia virus Ankara (MVA), has been constructed to express soluble CMV gB (gB680-MVA) to induce CMV NAb. Very high levels of gB-specific CMV NAb were produced after two doses of the viral vaccine. NAb were durable within a twofold range for up to 6 months. Neutralization titers developed in immunized mice are equivalent to titers found clinically after natural infection. This viral vaccine, expressing gB derived from CMV strain AD169, induced antibodies that neutralized CMV strains of three different genotypes. Remarkably, preexisting MVA and vaccinia virus (poxvirus) immunity did not interfere with subsequent immunizations of gB680-MVA. The safety characteristics of MVA, combined with the robust immune response to CMV gB, suggest that this approach could be rapidly translated into the clinic.Human cytomegalovirus (HCMV) is a member of the herpesvirus family. It is a major cause of congenital disease, resulting in an estimated 4,000 cases of symptomatic congenital cytomegalovirus (CMV) infection per year in the United States (58). An effective CMV vaccine that can prevent or reduce CMV-associated disease is highly desirable. Early studies have indicated that HCMV gB is the major target of NAb that are induced by naturally acquired CMV infection (16,39). It is the most highly conserved envelope glycoprotein of human herpesviruses (38). Thus, CMV gB has been an attractive candidate for CMV vaccine development. CMV gB vaccines using recombinant gB protein expressed from plasmid DNA and gB expressed in several different viral vectors (ALVAC, adenovirus, and vaccinia virus [VV]) have been investigated with animal models (9,13,23,26,31,40,54). Safety and moderate immunogenicity have been demonstrated with these vaccines, but no licensed CMV vaccine is available. A live attenuated Towne strain of CMV, either alone or with a gB subunit vaccine as a prime-boost, have also been evaluated in human subjects (1, 2, 48).Full-length CMV gB is synthesized as a 907-amino acid (aa) precursor in CMV-infected cells with a predicted molecular mass of 105 kDa, but it can be glycosylated to form a 170-kDa modified protein (17). To enable pharmaceutical development, truncated and secretable forms of gB were derived. These in...

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“…Murine models to predict human cytomegalovirus (CMV) vaccine immunogenicity are available to detect the presence of a specific immune response, such as cytolytic T-lymphocyte (CTL) function (2,5,28), CD4…”

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confidence: 99%

Use of Transgenic HLA A*0201/Kb and HHD II Mice To Evaluate Frequency of Cytomegalovirus IE1-Derived Peptide Usage in Eliciting Human CD8 Cytokine Response

Gallez-Hawkins

Villacres

et al. 2003

J Virol

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Unlike the pp65 protein of human cytomegalovirus (CMV), which has an immunodominant peptide, pp65 [495][496][497][498][499][500][501][502][503] , recognized by human CD8؉ cells in the context of HLA A‫,1020ء‬ the fine peptide specificity for CMV IE1 has shown no such immunodominance. With the use of transgenic HLA A‫/1020ء‬Kb and HHD II mice, a selected pool of IE1 peptides, including IE1 p256-264 , IE1 p297-304 , and IE1 p316-324 , were shown to stimulate cytolytic T-lymphocyte lysis in the context of HLA A‫.1020ء‬ Based on an intracellular gamma interferon response, IE1 p297-304 , a previously unrecognized CD8 epitope, triggered a prominent response to CMV IE1 in HLA A‫1020ء‬ subjects.Murine models to predict human cytomegalovirus (CMV) vaccine immunogenicity are available to detect the presence of a specific immune response, such as cytolytic T-lymphocyte (CTL) function (2, 5, 28), CD4ϩ gamma interferon (IFN-␥) helper response (15, 22), or antibody response to specific CMV proteins (18,20,21). Of interest is the use of the transgenic A2Kb mouse containing the human HLA A‫1020ء‬with the murine ␣3 chain (Kb) (9,19,29). The murine model HHD II is also a transgenic HLA A‫1020ء‬ mouse that contains a disrupted murine major histocompatibility complex (MHC) molecule, forcing the mouse to present all its immunological epitopes through the HLA pathway (10). Both have become useful models to investigate peptide recognition of specific proteins that can be extrapolated to human subjects.The immunodominant HLA A‫-1020ء‬restricted peptide (pp65 [495][496][497][498][499][500][501][502][503] ) of CMV pp65 has been well studied (3,6,12,30), whereas there is no immunodominant peptide recognized for the CMV IE1 protein, despite the prevalence of an IE1-specific CTL response (11,14,16,17). Three reports have described the stimulatory effect of peptides IE1 p315-323 , IE1 p316-324 , and IE1 p354-362 from CMV IE1 in the context of HLA A‫1020ء‬ (11, 17, 24) in a cytokine flow cytometry or CTL assay. However, not all CMV-seropositive subjects respond to these peptides (17). This report describes the use of HLA A2 transgenic mice to identify a novel IE1 peptide, IE1-297, that is recognized by murine CTLs as well as human CD8 cells by cytokine flow cytometry.T2 stabilization. T2 cells are defective for endogenous class I presentation, but the presence of peptide binding to the MHC molecule will stabilize its expression on the cell surface. The stabilized MHC molecule can be detected by flow cytometry with a monoclonal antibody to the HLA A‫1020ء‬ molecule (13). The peptide sequences used in this study were collected from two algorithms for HLA peptide predicted motifs: SYFPEITHI (23)

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Murine cytotoxic T cell response specific for human cytomegalovirus glycoprotein B (gB) induced by adenovirus and vaccinia virus recombinants expressing gB

Cited by 26 publications

References 24 publications

Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy.

Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy.

Recombinant Modified Vaccinia Virus Ankara Expressing a Soluble Form of Glycoprotein B Causes Durable Immunity and Neutralizing Antibodies against Multiple Strains of Human Cytomegalovirus

Use of Transgenic HLA A*0201/Kb and HHD II Mice To Evaluate Frequency of Cytomegalovirus IE1-Derived Peptide Usage in Eliciting Human CD8 Cytokine Response

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