HIV-1 Env DNA Vaccine plus Protein Boost Delivered by EP Expands B- and T-Cell Responses and Neutralizing Phenotype In Vivo

A prophylactic vaccine eliciting both broad neutralizing antibodies (bNAbs) to the HIV-1 envelope glycoprotein (Env) and strong T cell responses would be optimal for preventing HIV-1 transmissions. Replication incompetent HIV-1 virus-like particles (VLPs) offer the opportunity to present authentic-structured, virion-associated Env to elicit bNAbs, and also stimulate T cell responses. Here, we optimize our DNA vaccine plasmids as VLP expression vectors for efficient Env incorporation and budding. The original vector that was used in human trials inefficiently produced VLPs, but maximized safety by inactivating RNA genome packaging, enzyme functions that are required for integration into the host genome, and deleting accessory proteins Vif, Vpr, and Nef. These original DNA vaccine vectors generated VLPs with incomplete protease-mediated cleavage of Gag and were irregularly sized. Mutations to restore function within the defective genes revealed that several of the reverse transcriptase (RT) deletions mediated this immature phenotype. Here, we made efficient budding, protease-processed, and mature-form VLPs that resembled infectious virions by introducing alternative mutations that completely removed the RT domain, but preserved most other safety mutations. These VLPs, either expressed from DNA vectors in vivo or purified after expression in vitro, are potentially useful immunogens that can be used to elicit antibody responses that target Env on fully infectious HIV-1 virions.

Download Full-text

Use of an In Vivo FTA Assay to Assess the Magnitude, Functional Avidity and Epitope Variant Cross-Reactivity of T Cell Responses Following HIV-1 Recombinant Poxvirus Vaccination

PLoS ONE ◽

10.1371/journal.pone.0105366 ◽

2014 ◽

Vol 9 (8) ◽

pp. e105366 ◽

Cited By ~ 10

Author(s):

Danushka K. Wijesundara ◽

Charani Ranasinghe ◽

Ronald J. Jackson ◽

Brett A. Lidbury ◽

Christopher R. Parish ◽

...

Keyword(s):

T Cell ◽

T Cell Responses ◽

Cross Reactivity ◽

Functional Avidity ◽

Cell Responses ◽

Epitope Variant ◽

Hiv 1

Download Full-text

Broad and Cross-Clade CD4+ T-Cell Responses Elicited by a DNA Vaccine Encoding Highly Conserved and Promiscuous HIV-1 M-Group Consensus Peptides

PLoS ONE ◽

10.1371/journal.pone.0045267 ◽

2012 ◽

Vol 7 (9) ◽

pp. e45267 ◽

Cited By ~ 21

Author(s):

Rafael Ribeiro Almeida ◽

Daniela Santoro Rosa ◽

Susan Pereira Ribeiro ◽

Vinicius Canato Santana ◽

Esper Georges Kallás ◽

...

Keyword(s):

T Cell ◽

Dna Vaccine ◽

T Cell Responses ◽

Cd4 T Cell ◽

Group Consensus ◽

Cell Responses ◽

Hiv 1

Download Full-text

Vaccination with a Shigella DNA Vaccine Vector Induces Antigen-Specific CD8+ T Cells and Antiviral Protective Immunity

Journal of Virology ◽

10.1128/jvi.75.20.9665-9670.2001 ◽

2001 ◽

Vol 75 (20) ◽

pp. 9665-9670 ◽

Cited By ~ 50

Author(s):

Mohamed T. Shata ◽

David M. Hone

Keyword(s):

T Cell ◽

Dna Vaccine ◽

Protective Immunity ◽

Dna Vaccines ◽

T Cell Responses ◽

T Cell Response ◽

Vaccine Vector ◽

Host Cells ◽

Cell Responses ◽

Hiv 1

ABSTRACT A prototype Shigella human immunodeficiency virus type 1 (HIV-1) gp120 DNA vaccine vector was constructed and evaluated for immunogenicity in a murine model. For comparative purposes, mice were also vaccinated with a vaccinia virus-env(vaccinia-env) vector or the gp120 DNA vaccine alone. Enumeration of the CD8+-T-cell responses to gp120 after vaccination using a gamma interferon enzyme-linked spot assay revealed that a single intranasal dose of the Shigella HIV-1 gp120 DNA vaccine vector elicited a CD8+ T-cell response to gp120, the magnitude of which was comparable to the sizes of the analogous responses to gp120 that developed in mice vaccinated intraperitoneally with the vaccinia-env vector or intramuscularly with the gp120 DNA vaccine. In addition, a single dose of the Shigella gp120 DNA vaccine vector afforded significant protection against a vaccinia-env challenge. Moreover, the number of vaccinia-env PFU recovered in mice vaccinated intranasally with the Shigella vector was about fivefold less than the number recovered from mice vaccinated intramuscularly with the gp120 DNA vaccine. Since theShigella vector did not express detectable levels of gp120, this report confirms that Shigella vectors are capable of delivering passenger DNA vaccines to host cells and inducing robust CD8+ T-cell responses to antigens expressed by the DNA vaccines. Furthermore, to our knowledge, this is the first documentation of antiviral protective immunity following vaccination with a live Shigella DNA vaccine vector.

Download Full-text

Potent CD4+T Cell Responses Elicited by a Bicistronic HIV-1 DNA Vaccine Expressing gp120 and GM-CSF

The Journal of Immunology ◽

10.4049/jimmunol.168.2.562 ◽

2002 ◽

Vol 168 (2) ◽

pp. 562-568 ◽

Cited By ~ 112

Author(s):

Dan H. Barouch ◽

Sampa Santra ◽

Klara Tenner-Racz ◽

Paul Racz ◽

Marcelo J. Kuroda ◽

...

Keyword(s):

T Cell ◽

Dna Vaccine ◽

T Cell Responses ◽

Cd4 T Cell ◽

Gm Csf ◽

Cell Responses ◽

Hiv 1

Download Full-text

HIV-1–Specific Immunodominant T-Cell Responses Drive the Dynamics of HIV-1 Recombination Following Superinfection

Frontiers in Immunology ◽

10.3389/fimmu.2021.820628 ◽

2022 ◽

Vol 12 ◽

Author(s):

Hui Zhang ◽

Shuang Cao ◽

Yang Gao ◽

Xiao Sun ◽

Fanming Jiang ◽

...

Keyword(s):

Amino Acids ◽

T Cell ◽

Immune Escape ◽

T Cell Responses ◽

Cell Responses ◽

Immune Pressure ◽

Cellular Immune ◽

Hiv 1 ◽

Recombination Breakpoints

A series of HIV-1 CRF01_AE/CRF07_BC recombinants were previously found to have emerged gradually in a superinfected patient (patient LNA819). However, the extent to which T-cell responses influenced the development of these recombinants after superinfection is unclear. In this study, we undertook a recombination structure analysis of the gag, pol, and nef genes from longitudinal samples of patient LNA819. A total of 9 pol and 5 nef CRF01_AE/CRF07_BC recombinants were detected. The quasispecies makeup and the composition of the pol and nef gene recombinants changed continuously, suggestive of continuous evolution in vivo. T-cell responses targeting peptides of the primary strain and the recombination regions were screened. The results showed that Pol-LY10, Pol-RY9, and Nef-GL9 were the immunodominant epitopes. Pol-LY10 overlapped with the recombination breakpoints in multiple recombinants. For the LY10 epitope, escape from T-cell responses was mediated by both recombination with a CRF07_BC insertion carrying the T467E/T472V variants and T467N/T472V mutations originating in the CRF01_AE strain. In pol recombinants R8 and R9, the recombination breakpoints were located ~23 amino acids upstream of the RY9 epitope. The appearance of new recombination breakpoints harboring a CRF07_BC insertion carrying a R984K variant was associated with escape from RY9-specific T-cell responses. Although the Nef-GL9 epitope was located either within or 10~11 amino acids downstream of the recombination breakpoints, no variant of this epitope was observed in the nef recombinants. Instead, a F85V mutation originating in the CRF01_AE strain was the main immune escape mechanism. Understanding the cellular immune pressure on recombination is critical for monitoring the new circulating recombinant forms of HIV and designing epitope-based vaccines. Vaccines targeting antigens that are less likely to escape immune pressure by recombination and/or mutation are likely to be of benefit to patients with HIV-1.

Download Full-text

Superior Induction of T Cell Responses to Conserved HIV-1 Regions by Electroporated Alphavirus Replicon DNA Compared to That with Conventional Plasmid DNA Vaccine

Journal of Virology ◽

10.1128/jvi.06535-11 ◽

2012 ◽

Vol 86 (8) ◽

pp. 4082-4090 ◽

Cited By ~ 44

Author(s):

Maria L. Knudsen ◽

Alice Mbewe-Mvula ◽

Maximillian Rosario ◽

Daniel X. Johansson ◽

Maria Kakoulidou ◽

...

Keyword(s):

T Cell ◽

Dna Vaccine ◽

Plasmid Dna ◽

T Cell Responses ◽

Cell Responses ◽

Plasmid Dna Vaccine ◽

Hiv 1

Download Full-text

An Enhanced Synthetic Multiclade DNA Prime Induces Improved Cross-Clade-Reactive Functional Antibodies when Combined with an Adjuvanted Protein Boost in Nonhuman Primates

Journal of Virology ◽

10.1128/jvi.00652-15 ◽

2015 ◽

Vol 89 (18) ◽

pp. 9154-9166 ◽

Cited By ~ 14

Author(s):

Megan C. Wise ◽

Natalie A. Hutnick ◽

Justin Pollara ◽

Devin J. F. Myles ◽

Constance Williams ◽

...

Keyword(s):

T Cell ◽

Neutralizing Antibodies ◽

Vaccine Development ◽

Consensus Sequence ◽

Vaccine Trial ◽

T Cell Responses ◽

Antiretroviral Drugs ◽

Cell Responses ◽

Protein Boost ◽

Hiv 1

ABSTRACTThe search for an efficacious human immunodeficiency virus type 1 (HIV-1) vaccine remains a pressing need. The moderate success of the RV144 Thai clinical vaccine trial suggested that vaccine-induced HIV-1-specific antibodies can reduce the risk of HIV-1 infection. We have made several improvements to the DNA platform and have previously shown that improved DNA vaccines alone are capable of inducing both binding and neutralizing antibodies in small-animal models. In this study, we explored how an improved DNA prime and recombinant protein boost would impact HIV-specific vaccine immunogenicity in rhesus macaques (RhM). After DNA immunization with either a single HIV Env consensus sequence or multiple constructs expressing HIV subtype-specific Env consensus sequences, we detected both CD4+and CD8+T-cell responses to all vaccine immunogens. These T-cell responses were further increased after protein boosting to levels exceeding those of DNA-only or protein-only immunization. In addition, we observed antibodies that exhibited robust cross-clade binding and neutralizing and antibody-dependent cellular cytotoxicity (ADCC) activity after immunization with the DNA prime-protein boost regimen, with the multiple-Env formulation inducing a more robust and broader response than the single-Env formulation. The magnitude and functionality of these responses emphasize the strong priming effect improved DNA immunogens can induce, which are further expanded upon protein boost. These results support further study of an improved synthetic DNA prime together with a protein boost for enhancing anti-HIV immune responses.IMPORTANCEEven with effective antiretroviral drugs, HIV remains an enormous global health burden. Vaccine development has been problematic in part due to the high degree of diversity and poor immunogenicity of the HIV Env protein. Studies suggest that a relevant HIV vaccine will likely need to induce broad cellular and humoral responses from a simple vaccine regimen due to the resource-limited setting in which the HIV pandemic is most rampant. DNA vaccination lends itself well to increasing the amount of diversity included in a vaccine due to the ease of manufacturing multiple plasmids and formulating them as a single immunization. By increasing the number of Envs within a formulation, we were able to show an increased breadth of responses as well as improved functionality induced in a nonhuman primate model. This increased breadth could be built upon, leading to better coverage against circulating strains with broader vaccine-induced protection.

Download Full-text