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EBV BMLF1 (280-288) – GLCTLVAML – Epitope peptide of Epstein-Barr virus

EBV BMLF1 (280-288) is an epitope of the early lytic BMLF1 peptide of Epstein-Barr virus. EBV BMLF1 peptide presented by HLA-A*02:01 is one of the most immunogenic T-cell targets from the EBV proteome. EBV BMLF1 (280-288) can be used for the isolation and the stimulation of antigen-specific T cells.

Applications of EBV BMLF1 (280-288)

CD8+ cells have an important role in controlling Epstein-Barr virus infection. Indeed, the expansion of infected B cells is controlled by CD8+ cytotoxic T cells. Therefore, EBV BMLF1 (280-288) is important in study to detect EBV-specific CD8+ T cells by flow cytometry, per example in peripheral blood mononuclear cells of infected patients. EBV BMLF1 (280-288) is also used to analyze and measure the immune response and cytokine production in PBMCs cells to the virus with an application of ELISPOT assay.

 

Technical specification

 EBV BMLF1 (280-288) Sequence : GLCTLVAML
 EBV BMLF1 (280-288) peptide synthesis MW : 920,19 g/mol (C40H73N9O11S2)
 EBV BMLF1 (280-288)   price Purity : > 95%
Peptide Library synthesis Counter-Ion : TFA Salts (see option TFA removal)
Peptide library synthesis EBV BMLF1 (280-288)  Delivery format : Freeze dried in propylene 2mL microtubes
peptide solubility guidelines Peptide Solubility Guideline
buy peptide price Bulk peptide quantities available
 

Price

Product catalog Size Price € HT Price $ HT
SB069-1MG 1 mg 80 100
SB069-5MG 5 mg 280 350
SB069-10MG 10 mg 475 593,75
SB069-50MG 50 mg 1650 2062,5

 

References

1- Falco D. A. et al. Transplantation 74(4):501-510 (2002)
Identification of Epstein-Barr virus-specific CD8+ T lymphocytes in the circulation of pediatric transplant recipients
 

BACKGROUND: Pediatric transplant recipients are at increased risk for Epstein Barr virus (EBV)-related B cell lymphomas. In healthy individuals, the expansion of EBV-infected B cells is controlled by CD8+ cytotoxic T cells. However, immunosuppressive therapy may compromise antiviral immunity. We identified and determined the frequency of EBV-specific T cells in the peripheral blood of pediatric transplant recipients.

METHODS: HLA-B*0801 and HLA-A*0201 tetramers folded with immunodominant EBV peptides were used to detect EBV-specific CD8+ T cells by flow cytometry in peripheral blood mononuclear cells from 24 pediatric liver and kidney transplant recipients. The expression of CD38 and CD45RO on EBV-specific, tetramer-binding cells was also examined in a subset of patients by immunofluorescent staining and flow cytometry.

RESULTS: Tetramer-binding CD8+ T cells were identified in 21 of 24 transplant recipients. EBV-specific CD8+ T cells were detected as early as 4 weeks after transplant in EBV seronegative patients receiving an organ from an EBV seropositive donor. The frequencies (expressed as a percentage of the CD8+ T cells) of the tetramer-binding cells were HLA-B8-RAKFKQLL (BZLF1 lytic antigen peptide) tetramer, range=0.96 to 3.94%; HLA-B8-FLRGRAYGL (EBNA3A latent antigen peptide) tetramer, range=0.03 to 0.59%; and HLA-A2-GLCTLVAML (BMLF1 lytic antigen peptide) tetramer, range=0.06 to 0.76%. The majority of tetramer reactive cells displayed an activated/memory phenotype.

CONCLUSION: Pediatric transplant recipients receiving immunosuppression can generate EBV-specific CD8+ T cells. Phenotypic and functional analysis of tetramer cells may prove useful in defining and monitoring EBV infection in the posttransplant patient.

2- Yang J., Lemas V. M., Flinn I. W., Krone C. and Ambinder R.F. Blood 95(1):241-248 (2000)
Application of the ELISPOT assay to the characterization of CD8+ responses to Epstein-Barr virus antigens
 

CD8+ cells have an important role in controlling Epstein-Barr virus (EBV) infection. We adapted the interferon-γ ELISPOT assay to the quantitative analysis of EBV-specific CD8+ cells. Using peripheral blood mononuclear cells (PBMCs) from healthy donors, we measured both the aggregate response to the virus, using EBV-transformed lymphoblastoid cell lines (LCLs) as stimulators, and the specific responses to 2 A2-restricted peptide epitopes: the subdominant latency membrane protein-2 (LMP2) peptide CLGGLLTMV and the early lytic BMLF1 peptide GLCTLVAML. LCL-responsive CD8+ cells were detected in all EBV-seropositive donors (range 954 to 37 830 spots/106CD8+ cells). LMP2 peptide-responsive CD8+cells were detected in 10 of 11 healthy seropositive A2 donors (range 11 to 83 spots/106 PBMC). BMLF1 peptide-responsive CD8+ cells were detected in all seropositive A2 donors examined (range 13 to 943 spots/106 PBMC). Cytotoxic T-lymphocyte (CTL) lines generated with weekly stimulation of LCLs for therapeutic purposes were also studied. Relative to PBMCs, these CTL lines showed a marked increase in the level of LCL-responsive and LMP2 peptide-responsive CD8+ cells and a lesser degree of expansion of BMLF1 peptide-responsive CD8+ cells. Finally, we applied the ELISPOT assay to monitor adoptive infusion of EBV CTL lines. In 2 patients examined, a transient increase in LCL-responsive CD8+ cells could be detected after infusion. Thus, the ELISPOT assay can be applied to the analysis of CD8+responses to EBV antigens in PBMCs, in ex vivo expanded CTL lines, and in PBMCs from patients treated with ex vivo expanded CTL lines.

3- Nguyen T. O., Bird N. L., Grant E. J., Miles J. J., Thomas P. G., Kotsimbos T. C., Mifsud N. A. and Kedzierska K. Immunol. Cell Biol. 95(1):77-86 (2017)
Maintenance of the EBV-specific CD8+ TCRαβ repertoire in immunosuppressed lung transplant recipients
 

Epstein-Barr virus (EBV) is one of the most common viruses in humans, capable of causing life-threatening infections and cancers in immunocompromised individuals. Although CD8+ T cells provide key protection against EBV, the persistence and dynamics of specific T-cell receptor (TCR) clones during immunosuppression in transplant patients is largely unknown. For the first time, we used a novel single-cell TCRαβ multiplex-nested reverse transcriptase PCR to dissect TCRαβ clonal diversity within GLCTLVAML (GLC)-specific CD8+ T cells in healthy individuals and immunocompromised lung transplant recipients. The GLC peptide presented by HLA-A*02:01 is one of the most immunogenic T-cell targets from the EBV proteome. We found that the GLC-specific TCRαβ repertoire was heavily biased toward TRAV5 and encompassed five classes of public TCRαβs, suggesting that these clonotypes are preferentially utilized following infection. We identified that a common TRAV5 was diversely paired with different TRAJ and TRBV/TRBJ genes, in both immunocompetent and immunocompromised individuals, with an average of 12 different TCRαβ clonotypes/donor. Moreover, pre-transplant GLC-specific TCRαβ repertoires were relatively stable over 1 year post transplant under immunosuppression in the absence or presence of EBV reactivation. In addition, we provide the first evidence of early GLC-specific CD8+ T cells at 87 days post transplant, which preceded clinical EBV detection at 242 days in an EBV-seronegative patient receiving a lung allograft from an EBV-seropositive donor. This was associated with a relatively stable TCRαβ repertoire after CD8+ T-cell expansion. Our findings provide insights into the composition and temporal dynamics of the EBV-specific TCRαβ repertoire in immunocompromised transplant patients and suggest that the early detection of EBV-specific T cells might be a predictor of ensuing EBV blood viremia.

4- Ouyang Q. et al. Mech Ageing Dev. 124(4):477-485 (2003)
An age-related increase in the number of CD8+ T cells carrying receptors for an immunodominant Epstein-Barr virus (EBV) epitope is counteracted by a decreased frequency of their antigen-specific responsiveness
 

The aim of this study was to provide a basis for investigating the effects of one very common environmental factor, Epstein-Barr virus (EBV), on age-related changes in the immune system. To this end, the frequency of CD8(+) T cells carrying receptors for an immunodominant EBV lytic epitope was assessed by direct staining with HLA-peptide tetrameric complexes in 19 very old (>87 years) and 12 young (20-40 years) EBV carriers. The frequency of EBV-tetramer-positive cells within the CD8(+) subset was significantly greater in the old compared to the young group (P=0.001). However, the frequency of EBV antigen-specific IFN-gamma producing T cells, as determined by ELISPOT, was significantly lower in the old (P=0.001). Therefore, the absolute number of functional EBV-specific T cells in the elderly and the young was probably similar. These data suggest CD8 clonal expansions in the elderly, resulting in an accumulation of dysfunctional EBV-specific cells which possibly fill the ‘immunological space’ and could lead to a shrinking of the T cell repertoire for other novel antigens. This may help to explain the increased incidence and case-fatality caused by viruses and intracellular pathogens in the elderly.

5- Tussey L., Speller S., Gallimore A. and Vessey R. Eur J Immunol. 30(7):1823-1829 (2000)
Functionally distinct CD8+ memory T cell subsets in persistent EBV infection are differentiated by migratory receptor expression
 

Human memory T lymphocytes have recently been re-defined as central or effector memory cells (Sallusto, F., Lenig, D., Forster, R., Lipp, M. and Lanzavecchia, A., Nature 1999. 401: 708-712). Effector memory cells (T(em)) are targeted to the peripheral tissues and show rapid effector function in response to antigenic stimulation. Central memory (T(cm)) cells are targeted to the lymph nodes and cannot be immediately activated. In this report HLA-A2-Epstein-Barr virus (EBV) peptide tetramers have been used to characterize the EBV-specific CD8+ T cell subsets in persistent EBV infection. In short-term activation studies two populations of tetramer-positive T cells were identified. One group resembled T(em) cells in that they rapidly produced IFN-gamma and lacked the lymph node homing receptor, CD62L, the second was similar to T(cm) cells since they were CD62L+ but could not be immediately induced to express IFN-gamma.

6- Tan L. C. et al. J Immunol. 162(3):1827-1835 (1999)
A re-evaluation of the frequency of CD8+ T cells specific for EBV in healthy virus carriers
 

EBV is a gammaherpesvirus that can establish both nonproductive (latent) and productive (lytic) infections within the cells of its host. Although T cell responses to EBV latent proteins have been well characterized, little is known about the importance of responses to lytic proteins in long term virus carriers. Here we have compared the frequencies of CD8+ T cells specific for EBV latent and lytic Ags in healthy virus carriers, using three techniques: limiting dilution analysis, enzyme-linked immunospot assay, and FACS staining with tetrameric MHC-peptide complexes. T cells specific for EBV lytic protein epitopes were readily detectable in all donors and were usually more abundant than those specific for latent epitopes. We infer that direct T cell control of viral replicative lesions is maintained in long term carriers of EBV and is an important component of the immune response to this virus. Estimates of CD8+ T cell frequencies varied considerably according to methodology; values obtained from MHC-peptide tetramer staining were, on the average, 4.4-fold higher than those obtained from enzyme-linked immunospot assays, which were, in turn, on the average, 5.3-fold higher than those obtained from limiting dilution analysis. Tetramer staining showed that as many as 5.5% circulating CD8+ T cells in a virus carrier were specific for a single EBV lytic protein epitope. Such values are much greater than previously imagined and illustrate how antigenic challenge from a persistent herpesvirus can influence the composition of the host’s CD8+ T cell pool.