Humoral and CD4+ T helper (Th) cell responses to the hepatitis C virus non-structural 3 (NS3) protein: NS3 primes Th1-like responses more effectively as a DNA-based immunogen than as a recombinant protein

Lazdina, Una; Hultgren, Catharina; Frelin, Lars; Chen, Margaret; Lodin, Karin; Weiland, Ola; Leroux‐Roels, Geert; Quiroga, Juan Antonio; Milich, David R.; Sällberg, Matti

doi:10.1099/0022-1317-82-6-1299

Cited by 32 publications

(36 citation statements)

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“…With respect to the NS3 protein of HCV, which has been evaluated in many laboratories as a therapeutic vaccine candidate, 7,8,13,23,34,35 some important findings have recently been made. The NS3 protein has three enzymatic functions in the viral life cycle.…”

Section: Discussionmentioning

confidence: 99%

“…23 Staining of coNS3/4A plasmid-transfected, and SFVinfected, BHK cells revealed a similar perinuclear and cytoplasmic distribution of the NS3 as previously observed, confirming an unchanged subcellular localization ( Figure 1). 13,23 Effect of codon optimization and mRNA amplification on humoral responses…”

Section: Characterization Of the Codon-optimized Ns3/4a Gene And Sfv mentioning

confidence: 99%

“…13,23 To compare indirectly the T helper 1 (Th1) and Th2 skewing of the T-cell response primed by wtNS3/4A, coNS3/4A, and wtNS3/4A-SFV immunizations, the levels of NS3-specific IgG1 (Th2) and IgG2a (Th1) antibodies were analyzed ( Figure 2). The IgG2a/IgG1 ratio in mice immunized with rNS3 with or without adjuvant was always o1 regardless of the murine haplotype, 27 signaling a Th2-dominated response.…”

Section: Characterization Of the Codon-optimized Ns3/4a Gene And Sfv mentioning

confidence: 99%

“…[17][18][19][20] We noted that when using a genetic immunogen containing the complete NS3/4A protease, the humoral responses were surprisingly strong. 13 Previous reports had shown that the presence of the cofactor NS4A increases the intracellular stability of NS3 and targets NS3 to intracellular membranes, factors that possibly may affect immunogenicity. 21,22 However, we could recently show that the presence of NS4A, by some mechanism, increases the expression levels of NS3, which explains the increased immunogenicity conferred by NS4A.…”

Section: Introductionmentioning

confidence: 99%

“…It has been shown that DNA immunizations can induce both specific antibodies and cell-mediated responses against the structural and nonstructural (NS) HCV proteins in mice. [6][7][8][9][10][11][12][13][14][15][16] The NS protein 3 of HCV has been considered as a possible vaccine target. The reasons for this are that the NS3 protein shows a limited genetic variability, it performs multiple enzymatic functions, and it is a relatively large protein.…”

Section: Introductionmentioning

confidence: 99%

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Codon optimization and mRNA amplification effectively enhances the immunogenicity of the hepatitis C virus nonstructural 3/4A gene

et al. 2004

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We have recently shown that the NS3-based genetic immunogens should contain also hepatitis C virus (HCV) nonstructural (NS) 4A to utilize fully the immunogenicity of NS3. The next step was to try to enhance immunogenicity by modifying translation or mRNA synthesis. To enhance translation efficiency, a synthetic NS3/4A-based DNA (coNS3/4A-DNA) vaccine was generated in which the codon usage was optimized (co) for human cells. In a second approach, expression of the wild-type (wt) NS3/4A gene was enhanced by mRNA amplification using the Semliki forest virus (SFV) replicon (wtNS3/4A-SFV). Transient tranfections of human HepG2 cells showed that the coNS3/4A gene gave 11-fold higher levels of NS3 as compared to the wtNS3/4A gene when using the CMV promoter. We have previously shown that the presence of NS4A enhances the expression by SFV. Both codon optimization and mRNA amplification resulted in an improved immunogenicity as evidenced by higher levels of NS3-specific antibodies. This improved immunogenicity also resulted in a more rapid priming of cytotoxic T lymphocytes (CTLs). Since HCV is a noncytolytic virus, the functionality of the primed CTL responses was evaluated by an in vivo challenge with NS3/4A-expressing syngeneic tumor cells. The priming of a tumor protective immunity required an endogenous production of the immunogen and CD8 þ CTLs, but was independent of B and CD4 þ T cells. This model confirmed the more rapid in vivo activation of an NS3/4A-specific tumor-inhibiting immunity by codon optimization and mRNA amplification. Finally, therapeutic vaccination with the coNS3/4A gene using gene gun 6-12 days after injection of tumors significantly reduced the tumor growth in vivo. Codon optimization and mRNA amplification effectively enhances the overall immunogenicity of NS3/4A. Thus, either, or both, of these approaches should be utilized in an NS3/4A-based HCV genetic vaccine.

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Section: Discussionmentioning

confidence: 99%

Section: Characterization Of the Codon-optimized Ns3/4a Gene And Sfv mentioning

confidence: 99%

Section: Characterization Of the Codon-optimized Ns3/4a Gene And Sfv mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Codon optimization and mRNA amplification effectively enhances the immunogenicity of the hepatitis C virus nonstructural 3/4A gene

et al. 2004

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Hepatitis C Virus-Mediated Modulation of Cellular Immunity

Brenndörfer

Sällberg

2012

Arch. Immunol. Ther. Exp.

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The hepatitis C virus (HCV) is a major cause of chronic liver disease globally. A chronic infection can result in liver fibrosis, liver cirrhosis, hepatocellular carcinoma and liver failure in a significant ratio of the patients. About 170 million people are currently infected with HCV. Since 80 % of the infected patients develop a chronic infection, HCV has evolved sophisticated escape strategies to evade both the innate and the adaptive immune system. Thus, chronic hepatitis C is characterized by perturbations in the number, subset composition and/or functionality of natural killer cells, natural killer T cells, dendritic cells, macrophages and T cells. The balance between HCV-induced immune evasion and the antiviral immune response results in chronic liver inflammation and consequent immune-mediated liver injury. This review summarizes our current understanding of the HCV-mediated interference with cellular immunity and of the factors resulting in HCV persistence. A profound knowledge about the intrinsic properties of HCV and its effects on intrahepatic immunity is essential to be able to design effective immunotherapies against HCV such as therapeutic HCV vaccines.

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