Induction of neutralizing antibodies and T cell responses by dengue virus type 2 envelope domain III encoded by plasmid and adenoviral vectors

Khanam, Saima; Khanna, Navin; Swaminathan, Sathyamangalam

doi:10.1016/j.vaccine.2006.06.031

Cited by 40 publications

(31 citation statements)

References 38 publications

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“…However the critical question that we addressed and showed is that our neutralizing antibody titers were protective in both vaccines, even without IL-4 detection. These results are in agreement with another study where the levels of IFN- were higher than IL-4, obtained from splenocytes in proliferative response after immunization with a plasmid enconding the domain III of the dengue 2 E protein [32].…”

Section: Discussionsupporting

confidence: 93%

“…In our vaccinal strategy, the response to stimulus in both vaccine candidates (pVAC1WDEN3 and pVAC3DEN3) cultivated spleen cells, produced a high quantity of IFN- in comparison of IL-4 production. IL-4 is a B-cell stimulatory cytokine and contributes markedly to the generation of a dengue virus-neutralizing antibody response [32]. However the critical question that we addressed and showed is that our neutralizing antibody titers were protective in both vaccines, even without IL-4 detection.…”

Section: Discussionmentioning

confidence: 79%

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Evaluation of immunogenicity elicited from two DNA vaccine candidates that expresses the prM and E genes of the dengue-3 virus

Paula¹,

Franca²,

Lima³

et al. 2010

Health

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In this work, we report the evaluation of two DNA vaccines against dengue-3 virus (DENV-3). The first construction, called pVAC3DEN3, was engineered inserting the pre-membrane (prM) and envelope (E) gene of DENV-3 truncated with a restriction site between them, as previously described. The second construction was developed cloning the full gene sequence of prM and E from DENV-3 virus in pCI plasmid for mammalian expression and was denominated pVAC1WDEN3. The results showed that both constructions were capable of expressing the prM and E proteins, as demonstrated by ELISA and immunoblotting detection in cell culture transfected with the plasmids. After positive "in vitro" results, the vaccine candidates were used to immunize BALB/c mice and the elicited response was investigated. After immunization by intramuscular inoculation with three doses of each vaccinal clone the animals were sacrificed, the cytokine levels and T cell response were analyzed in the spleens, after three days of culture with stimulus, our analysis showed that the two constructions elicited T cell responses measured by BrdU incorporation assay and high levels of IFN-γ, detected in the supernatant of the cultures. Moreover, both constructions induced detectable titers of neutralizing antibodies in mice. And finally the survival rate of the immunized animals after intracerebral challenge was analyzed, showing a better result in the pVAC3DEN3 group with an 80% survival compared with a 50% survival of the pVAC1 WDEN3.Thus, these data showed that our two constructions were able to induce specific immune response and protects mice against a lethal challenge with DENV-3, and these vaccine candidates can be employed to develop a viable dengue vaccine.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 79%

Evaluation of immunogenicity elicited from two DNA vaccine candidates that expresses the prM and E genes of the dengue-3 virus

Paula¹,

Franca²,

Lima³

et al. 2010

Health

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show abstract

“…Although there are a number of studies describing DNA vaccines against dengue Azevedo et al, 2011;Galula et al, 2014;Khanam et al, 2006;Konishi et al, 2006;Ocazionez Jimenez & Lopes da Fonseca, 2000;Prompetchara et al, 2014;Ramanathan et al, 2009;Raviprakash et al, 2001Raviprakash et al, , 2006, to date there has been only one human clinical trial for a dengue DNA vaccine involving a Phase 1 study of a plasmid expressing the PrM and E proteins of DENV1 . In all cases, these vaccines have been proved to be safe and well-tolerated in humans, although low immunogenicity is still a concern associated with genetic vaccines in general (Coban et al, 2011;Danko et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

Secretion of dengue virus envelope protein ectodomain from mammalian cells is dependent on domain II serotype and affects the immune response upon DNA vaccination

Slon-Campos

Poggianella

Marchese

et al. 2015

Journal of General Virology

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Dengue virus (DENV) is currently among the most important human pathogens and affects millions of people throughout the tropical and subtropical regions of the world. Although it has been a World Health Organization priority for several years, there is still no efficient vaccine available to prevent infection. The envelope glycoprotein (E), exposed on the surface on infective viral particles, is the main target of neutralizing antibodies. For this reason it has been used as the antigen of choice for vaccine development efforts. Here we show a detailed analysis of factors involved in the expression, secretion and folding of E ectodomain from all four DENV serotypes in mammalian cells, and how this affects their ability to induce neutralizing antibody responses in DNA-vaccinated mice. Proper folding of E domain II (DII) is essential for efficient E ectodomain secretion, with DIII playing a significant role in stabilizing soluble dimers. We also show that the level of protein secreted from transfected cells determines the strength and efficiency of antibody responses in the context of DNA vaccination and should be considered a pivotal feature for the development of E-based DNA vaccines against DENV.

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“…This includes the use of fusion proteins (1,22,35,64), combinations with strong adjuvants such as Freund's adjuvant (1,2,22,35), strings of DIII (7,27), expression by adenovirus vectors (28), or coupling to large carriers such as a bacteriophage (54) to mimic its presentation at the surface of the virus. However, despite its low immunogenicity when it is used alone, DIII proved to be an excellent boosting antigen and strongly enhanced the neutralizing antibody response of mice preimmunized with inactivated virion or sE.…”

Section: Discussionmentioning

confidence: 99%

Immunodominance and Functional Activities of Antibody Responses to Inactivated West Nile Virus and Recombinant Subunit Vaccines in Mice

Zlatkovic

Stiasny

Heinz

2011

J Virol

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Factors controlling the dominance of antibody responses to specific sites in viruses and/or protein antigens are ill defined but can be of great importance for the induction of potent immune responses to vaccines. West Nile virus and other related important human-pathogenic flaviviruses display the major target of neutralizing antibodies, the E protein, in an icosahedral shell at the virion surface. Potent neutralizing antibodies were shown to react with the upper surface of domain III (DIII) of this protein. Using the West Nile virus system, we conducted a study on the immunodominance and functional quality of E-specific antibody responses after immunization of mice with soluble protein E (sE) and isolated DIII in comparison to those after immunization with inactivated whole virions. With both virion and sE, the neutralizing response was dominated by DIIIspecific antibodies, but the functionality of these antibodies was almost four times higher after virion immunization. Antibodies induced by the isolated DIII had an at least 15-fold lower specific neutralizing activity than those induced by the virion, and only 50% of these antibodies were able to bind to virus particles. Our results suggest that immunization with the tightly packed E in virions focuses the DIII antibody response to the externally exposed sites of this domain which are the primary targets for virus neutralization, different from sE and isolated DIII, which also display protein surfaces that are cryptic in the virion. Despite its low potency for priming, DIII was an excellent boosting antigen, suggesting novel vaccination strategies that strengthen and focus the antibody response to critical neutralizing sites in DIII.The availability of high-resolution structures is a prerequisite for understanding structural determinants of immunogenicity and immunodominance. Knowledge of factors that control and/or influence these properties of antigens can lead to an improvement of existing vaccines and the rational design of new vaccine antigens and regimens (13). Flaviviruses are among those human-pathogenic viruses for which detailed structural information is available through the combined use of X-ray crystallography and cryo-electron microscopy (cryo-EM) (25, 30, 36-38, 40, 49, 65). The most important representatives are the mosquito-borne yellow fever (YF), dengue (DEN), Japanese encephalitis (JE), and West Nile (WN) viruses, as well as tick-borne encephalitis (TBE) virus (14). These viruses have a significant impact on public health and the potential for emergence in new geographic regions, as exemplified by the expansion of WN virus in the Americas since its first introduction into the United States in 1999 (17). Human vaccines are in general use against YF virus (live attenuated), JE virus (live attenuated and inactivated whole virus), and TBE virus (inactivated whole virus) but not yet against DEN and WN viruses (48).Mature flavivirions are composed of an isometric capsid containing the positive-stranded RNA genome and a lipid envelope with two me...

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Induction of neutralizing antibodies and T cell responses by dengue virus type 2 envelope domain III encoded by plasmid and adenoviral vectors

Cited by 40 publications

References 38 publications

Evaluation of immunogenicity elicited from two DNA vaccine candidates that expresses the prM and E genes of the dengue-3 virus

Evaluation of immunogenicity elicited from two DNA vaccine candidates that expresses the prM and E genes of the dengue-3 virus

Secretion of dengue virus envelope protein ectodomain from mammalian cells is dependent on domain II serotype and affects the immune response upon DNA vaccination

Immunodominance and Functional Activities of Antibody Responses to Inactivated West Nile Virus and Recombinant Subunit Vaccines in Mice

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