Cross-Presentation of the Long-Lived Lymphocytic Choriomeningitis Virus Nucleoprotein Does Not Require Neosynthesis and Is Enhanced via Heat Shock Proteins

Basta, Sameh; Stoessel, Ricarda; Basler, Michael; Broek, Maries van den; Groettrup, Marcus

doi:10.4049/jimmunol.175.2.796

Cited by 64 publications

(100 citation statements)

References 51 publications

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“…1C). We evaluated if the infected LyUV-ADC can supply LCMV antigens for cross-presentation when compared with HEK-NP cells [7,8]. By employing NP396-specific CTL, we confirmed that the infected LyUV-ADC supplied sufficient levels of LCMV-NP for cross-presentation to take place (Fig.…”

Section: Characterization Of Lcmv-infected Uv-irradiated Adcmentioning

confidence: 76%

“…In this study, we employed the LCMV infection model to study how crosspresentation of multiple epitopes translates when studying cross-priming. We used LyUV treatment of the ADC because it fully inactivated the virus and allowed for efficient NP396 crosspresentation similar to HEK-NP cells [7,8]. We initially expected that GP33 would fail to cross-present because it is located in the signal peptide of LCMV-GP [12], but it was cross-presented with low efficiency, probably due to its exceptional long half-life of 6 h [15].…”

Section: Discussionmentioning

confidence: 99%

“…The experiments were conducted in accordance to the guidelines of the Canadian Council on Animal Care. HEK293-NP are stably transfected with LCMV-NP [7,8] and the cell lines BMA and DC2.4 were cultured in RPMI 5% FBS (Invitrogen, ON, Canada) [32,33]. Ribonuclease A from bovin pancrease (RNase A, R4875, 10 mg/mL), lactacystin, BFA, leupeptin, pepstatin A, and chloroquine were purchased from Sigma (Oakvilla, ON, Canada).…”

Section: Methodsmentioning

confidence: 99%

“…The contribution of multiple epitopes from viral proteins to the cross-presentation pathway after infections is not well understood. In fact, much of what is known about the cross-presentation of virus antigens is derived from studies relying on antigen donor cells (ADC) transfected with a single virus protein [7][8][9][10]. The ability of antigens to escape cytosolic degradation in ADC is important during cross-presentation [7,[11][12][13].…”

Section: Introductionmentioning

confidence: 99%

“…In fact, much of what is known about the cross-presentation of virus antigens is derived from studies relying on antigen donor cells (ADC) transfected with a single virus protein [7][8][9][10]. The ability of antigens to escape cytosolic degradation in ADC is important during cross-presentation [7,[11][12][13]. Interestingly, it appears that the capacity of an epitope to access cross-priming may support its immunodominance when considering the overall hierarchy [8,10,14].…”

Section: Introductionmentioning

confidence: 99%

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The outcome of cross‐priming during virus infection is not directly linked to the ability of the antigen to be cross‐presented

et al. 2010

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The initiation of CD8 1 T cell (CTL) immune responses can occur via cross-priming. Recent data suggested a relationship between cross-presentation and immunodominance of epitope-specific T cells. To test this association, we evaluated the efficacy of crosspresentation for several virus epitopes in vitro and examined if this can be extrapolated in vivo. Employing lymphocytic choriomeningitis virus (LCMV), we demonstrate that the cross-presentation and cross-priming of LCMV antigens were dominated by NP396, but not NP205 when analyzing the LCMV-NP. Although with LCMV-GP, cross-presentation was dominated by GP276, and cross-priming was dominated by GP33. Importantly, although NP396 was significantly more efficient than GP33 in cross-presentation, cross-priming of their specific CTL was comparable. In a subsequent virus challenge after cross-priming, GP33-specific CTL dominated the response. Accordingly, based on our data, the ability of viral epitopes to be cross-presented in vitro does not entirely reflect what would occur in cross-priming. Thus, weak cross-presenting antigens may still cross-prime an efficient CTL response depending on other in vivo elements such as the naïve T-cell precursor frequencies.

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Section: Characterization Of Lcmv-infected Uv-irradiated Adcmentioning

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The outcome of cross‐priming during virus infection is not directly linked to the ability of the antigen to be cross‐presented

et al. 2010

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CTL induction by cross‐priming is restricted to immunodominant epitopes

et al. 2009

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CTL are induced by two pathways, i.e. direct priming, where tumor cells present tumor antigens to naïve specific CTL, and cross-priming, where professional APC cross-present captured tumor antigens to CTL. Here, we examined direct priming versus cross-priming after immunizing (H-2 b Â H-2 d ) F1 mice with either H-2 b or H-2 d positive tumor cells transfected with the GP or nucleoprotein (NP) of lymphocytic choriomeningitis virus (LCMV). Cross-priming was observed for the immunodominant epitopes LCMV-gp33 andnp118, although direct induction resulted in higher CTL frequencies. In contrast, CTL specific for the subdominant epitopes LCMV-gp283 or -np396 were induced only if epitopes were presented directly on MHC class I molecules of the immunizing cell. The broader repertoire and the higher CTL frequencies induced after vaccination with haplotype-matched tumor cells resulted in more efficient anti-tumor and antiviral protection. Firstly, our results indicate that certain virus and tumor antigens may not be detected by CD8 1 T cells because of impaired cross-priming. Secondly, efficient cross-priming contributes to the immunodominant nature of a tumor-specific CTL epitope. Thirdly, vaccine strategies using autologous or syngenic antigen-expressing cells induce a broader repertoire of tumor-specific CTL and higher CTL frequencies.Key words: CD8 T cells . Cross presentation/priming . Tumor immunology IntroductionProfessional APC (pAPC) capture cells in the periphery, migrate to secondary lymphoid organs, cross-present the specific antigen and activate naïve CTL. Cross-priming enables the immune system to detect and respond to pathogens, tumors and tissue antigens that are exclusively expressed outside of secondary lymphoid organs. Cross-priming may be essential for the immunosurveillance of tumors, since most tumor cells lack costimulatory molecules and thus cannot efficiently stimulate protective anti-tumoral CTL immunity [1][2][3]. Alternatively, tumor cells directly present the internally synthesized tumor antigen via MHC class I molecules and activate naïve tumorspecific CTL [2,4]. This direct priming pathway requires that tumor cells reach secondary lymphoid organs. Different cellular pathways in the cross-presentation of exogenous antigen have been characterized. They can be separated into TAP-dependent and TAP-independent mechanisms [5,6]. Current literature favors the view that TAP-dependent mechanisms are dominating in vivo [6].Several studies have shown that stable cellular proteins in particulate form are the major source of cross-presented antigens in vivo, whereas soluble proteins, peptides, HSP-peptide complexes, RNA and DNA seem to play only a minor role in crosspresentation [7][8][9]. Although different cell types, including B cells, macrophages and endothelial cells, have been reported to have the capacity to cross-present antigens, CD81 DC are the predominant cell population capturing and cross-presenting tumor antigens 704 [10][11][12][13]. The maturation of DC and the efficiency of cross-priming c...

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Treg suppress CTL responses upon immunization with HSP gp96

Liu

Qiu

et al. 2009

Eur J Immunol

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HSP gp96-based vaccines have been trialled in rodent models and, more recently, in humans. Better understanding of gp96's immunomodulatory role will help with the design of more effective strategies for treatment of cancer and infectious diseases. In this study, we monitored the activities of T cells and activation of Treg in BABL/c mice after immunization using different doses of gp96 as adjuvant. We found that co-injection of gp96 simultaneously stimulated both CTL and Treg activity. Activation of CTL at low dose was far more pronounced than Treg activation. Treg population and suppression increased with gp96 dose, eventually abrogating the T-cell response induced by immunization. Lowdose cyclophosphamide treatment could restore the T-cell responses lost after high-dose gp96 adjuvant injection by suppression of Treg activation. We further examined the effect of different doses of gp96 or N355 peptide administration on tumor rejection. Our results provide new insights into the mechanisms of gp96-mediated balance between regulatory and responder T cells, which may facilitate future development of an effective gp96-based therapeutic vaccine.Key words: CTL . gp96 . Hepatitis B virus . Treg Introduction HSP gp96, a counterpart of the cytosolic HSP90 family residing in the endoplasmic reticulum, plays important roles in the activation of innate and adaptive immunity. This molecule has the unique ability to associate with antigenic peptides from tumor, virus and intracellular bacteria. Peptides bound by gp96 are taken up by APC through cell surface receptor CD91 and can be presented by both MHC class I and MHC class II molecules [1][2][3]. In mouse models, gp96-peptide complexes have been shown to prime and induce MHC class I-restricted CTL responses by crosspresentation of peptides to MHC class I molecules. Additionally, gp96 may act as co-stimulator and activator of CD4 1 and CD8 1T cells [4], increasing cell proliferation and secretion of cytokines. The gp96 protein also induces innate immune responses. Interaction of gp96 with APC (e.g. dendritic cells) leads to maturation or enhanced function of dendritic cells [5]. Recently gp96 was demonstrated to function in the expression of both intracellular and cell surface TLR [6]. In rodent models, autologous tumor-derived gp96-peptide complexes are highly effective in the elimination of tumor burden [7,8]. Immunization of mice with gp96 complexed with specific CTL epitopes derived from various viruses has been shown to elicit virus-specific CTL or protective immunity [1,9,10]. Since gp96 has been demonstrated to act as a powerful adjuvant, clinical trials of gp96-based therapeutic vaccines have been initiated for treatment of cancer [8,11].Previous study in our lab found that gp96 and its N-terminal fragment, N355, have the ability to augment CTL responses against to hepatitis B virus (HBV). However, higher amounts of Ã These authors contributed equally to this work. 3110gp96 were detrimental to the adjuvant effect and decreased the capability of mice to generate an i...

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Cross-Presentation of the Long-Lived Lymphocytic Choriomeningitis Virus Nucleoprotein Does Not Require Neosynthesis and Is Enhanced via Heat Shock Proteins

Cited by 64 publications

References 51 publications

The outcome of cross‐priming during virus infection is not directly linked to the ability of the antigen to be cross‐presented

The outcome of cross‐priming during virus infection is not directly linked to the ability of the antigen to be cross‐presented

CTL induction by cross‐priming is restricted to immunodominant epitopes

Treg suppress CTL responses upon immunization with HSP gp96

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