Assessment of major histocompatibility complex class I interaction with Epstein‐Barr virus and human immunodeficiency virus peptides by elevation of membrane H‐2 and HLA in peptide loading‐deficient cells

Stuber, György; Modrow, Susanne; Höglund, Petter; Franksson, Lars; Elvin, John; Wolf, Hans; Kärre, Klas; Klein, George

doi:10.1002/eji.1830221033

Cited by 50 publications

(47 citation statements)

References 44 publications

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“…Screening for MHC class I NS5A epitopes was performed by harvesting spleens from NS5A-DNA-immunized wt C57BL/6J mice and restimulating cultures for 36 h in the presence of the 87 individual peptides in two concentrations (10 and 1 mg/ml) and thereafter determining IFN-g production by ELISPOT assay. Identified and predicted peptides were tested for the stabilization of surface expression of MHC class I molecules on the TAP2-deficient RMA-S cell line (46,47). Two NS5A peptides were identified that bound H-2D…”

Section: Cellular Immune Responsesmentioning

confidence: 99%

A Synthetic Codon-Optimized Hepatitis C Virus Nonstructural 5A DNA Vaccine Primes Polyfunctional CD8+ T Cell Responses in Wild-Type and NS5A-Transgenic Mice

Holmström

Pasetto

Nähr

et al. 2013

The Journal of Immunology

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The hepatitis C virus (HCV) nonstructural (NS) 5A protein has been shown to promote viral persistence by interfering with both innate and adaptive immunity. At the same time, the HCV NS5A protein has been suggested as a target for antiviral therapy. In this study, we performed a detailed characterization of HCV NS5A immunogenicity in wild-type (wt) and immune tolerant HCV NS5A-transgenic (Tg) C57BL/6J mice. We evaluated how efficiently HCV NS5A-based genetic vaccines could activate strong T cell responses. Truncated and full-length wt and synthetic codon-optimized NS5A genotype 1b genes were cloned into eukaryotic expression plasmids, and the immunogenicity was determined after i.m. immunization in combination with in vivo electroporation. The NS5A-based genetic vaccines primed high Ab levels, with IgG titers of >104 postimmunization. With respect to CD8+ T cell responses, the coNS5A gene primed more potent IFN-γ–producing and lytic cytotoxic T cells in wt mice compared with NS5A-Tg mice. In addition, high frequencies of NS5A-specific CD8+ T cells were found in wt mice after a single immunization. To test the functionality of the CTL responses, the ability to inhibit growth of NS5A-expressing tumor cells in vivo was analyzed after immunization. A single dose of coNS5A primed tumor-inhibiting responses in both wt and NS5A-Tg mice. Finally, immunization with the coNS5A gene primed polyfunctional NS5A-specific CD8+ T cell responses. Thus, the coNS5A gene is a promising therapeutic vaccine candidate for chronic HCV infections.

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Section: Cellular Immune Responsesmentioning

confidence: 99%

A Synthetic Codon-Optimized Hepatitis C Virus Nonstructural 5A DNA Vaccine Primes Polyfunctional CD8+ T Cell Responses in Wild-Type and NS5A-Transgenic Mice

Holmström

Pasetto

Nähr

et al. 2013

The Journal of Immunology

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“…After 4 h the cells were washed to remove excess peptide and stained with either BB7.2 Ab (0.5 g/ml) to detect the level of HLA-A2 molecules present on the surface or 1B8 TCRm mAb (0.5 g/ml). Pulsing the cells with peptides that have high affinity for HLA-A2 stabilizes the bound peptide complex and leads to an increase in the BB7.2 staining of the cells that received peptide vs those that did not (negative controls) (21,22). Bound Ab was detected using the PE-conjugated goat anti-mouse IgG H chain-specific polyclonal Ab.…”

Section: T2 Binding Assaymentioning

confidence: 99%

Levels of Specific Peptide-HLA Class I Complex Predicts Tumor Cell Susceptibility to CTL Killing

Weidanz

Nguyen²,

Woodburn³

et al. 2006

The Journal of Immunology

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Recognition of tumor-associated Ags (TAAs) on tumor cells by CTLs and the subsequent tumor cell death are assumed to be dependent on TAA protein expression and to correlate directly with the level of peptide displayed in the binding site of the HLA class I molecule. In this study we evaluated whether the levels of Her-2/neu protein expression on human tumor cell lines directly correlate with HLA-A*0201/Her2/neu peptide presentation and CTL recognition. We developed a TCR mimic (TCRm) mAb designated 1B8 that specifically recognizes the HLA-A2.1/Her2/neu peptide (369–377) (Her2(369)-A2) complex. TCRm mAb staining intensity varied for the five human tumor cell lines analyzed, suggesting quantitative differences in levels of the Her2(369)-A2 complex on these cells. Analysis of tumor cell lines pretreated with IFN-γ and TNF-α for Her2/neu protein and HLA-A2 molecule expression did not reveal a direct correlation between the levels of Her2/neu Ag, HLA-A2 molecule, and Her2(369)-A2 complex expression. However, compared with untreated cells, cytokine-treated cell lines showed an increase in Her2(369)-A2 epitope density that directly correlated with enhanced tumor cell death (p = 0.05). Although a trend was observed between tumor cell lysis and the level of the Her2(369)-A2 complex for untreated cells, the association was not significant. These findings suggest that tumor cell susceptibility to CTL-mediated lysis may be predicted based on the level of specific peptide-MHC class I expression rather than on the total level of TAA expression. Further, these studies demonstrate the potential of the TCRm mAb for validation of endogenous HLA-peptide epitopes on tumor cells.

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“…Flow cytometric analysis was performed and results are reported as percent of HLA-B8 reconstitution based on the following calculation: (mean fluorescence channel of acid-treated cells plus peptide Ϫ mean fluorescence of acid-treated cells without peptide)/(mean fluorescence channel of nonacid-treated cells Ϫ mean fluorescence channel of acid-treated cells without peptide). 31 The HLA-B8-binding peptide GGKKKYKL (HIV-1 gag [23][24][25][26][27][28][29][30] 12 served as the positive control; 10,000 events are collected for each peptide concentration.…”

Section: Hla-b8-binding Assaymentioning

confidence: 99%

Naturally processed and HLA‐B8‐presented HPV16 E7 epitope recognized by T cells from patients with cervical cancer

Oerke¹,

Höhn²,

Zehbe³

et al. 2005

Intl Journal of Cancer

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Several major histocompatibility complex (MHC) alleles have been reported to present peptides derived from the HPV16 E7 oncoprotein to T cells. We describe an overrepresentation of the HLA-B8 allele (28.44%) in cervical cancer patients as compared to the MHC class I allele frequency in a local healthy control population (18.80%) and the identification of an HLA-B8-binding peptide TLHEYMLDL (HPV16 E7 7-15 ), which is able to drive HPV16 E7-specific and MHC class I-restricted T-cell responses in peripheral blood lymphocytes from healthy individuals. TLHEYMLDLspecific T cells recognize the naturally processed and presented peptide on HPV16؉ cervical cancer cells transfected with the HLA-B8 gene defined by IFN-␥ production. This peptide epitope is also recognized by freshly harvested tumor-infiltrating T cells or T cells from tumor-draining lymph nodes from patients with cervical cancer determined by flow cytometry as well as by tetramer in situ staining. HLA-B8-restricted HPV E7 7-15 -specific T cells reside predominantly in the CD8 ؉ CD45RA ؉ CCR7 ؉ precursor or in the differentiated CD8 ؉ CD45RA ؉ CCR7 ؊ T-cell population.

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Assessment of major histocompatibility complex class I interaction with Epstein‐Barr virus and human immunodeficiency virus peptides by elevation of membrane H‐2 and HLA in peptide loading‐deficient cells

Cited by 50 publications

References 44 publications

A Synthetic Codon-Optimized Hepatitis C Virus Nonstructural 5A DNA Vaccine Primes Polyfunctional CD8+ T Cell Responses in Wild-Type and NS5A-Transgenic Mice

A Synthetic Codon-Optimized Hepatitis C Virus Nonstructural 5A DNA Vaccine Primes Polyfunctional CD8+ T Cell Responses in Wild-Type and NS5A-Transgenic Mice

Levels of Specific Peptide-HLA Class I Complex Predicts Tumor Cell Susceptibility to CTL Killing

Naturally processed and HLA‐B8‐presented HPV16 E7 epitope recognized by T cells from patients with cervical cancer

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