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Biotin-FluM1 (58-66) – Epitope peptide of Influenza virus

Biotin-FluM1 (58-66) is the N-ter biotinylated version of FluM1 (58-66). Biotin-FluM1 (58-66) can be used in the analysis of antigen-specific T cells.

FluM1 (58-66) peptide

FluM1 (58-66) is a short part of the matrix protein of Influenza A virus which is the most abundant component of this enveloped virus localized under the viral lipid envelope. The GILGFVFTL epitope is highly conserved in Influenza A virus strains at a rate of 93% for 69 strains tested. Human Influenza epitopes may bind MHC molecules and then may be recognized by CD8+ cytotoxic T cells.

Applications of FluM1 (58-66)

FluM1 (58-66) is used to stimulate CTL responses in peripheral blood mononuclear cells (PBMCs). Then, ELISPOT assay is used to quantify peptide epitope specificity and IFN-γ releasing effector cells.

FluM1 (58-66) has shown CTL responses qualified of immunodominant with restriction by HLA-A*02:01. It has also been detected CTL responses when FluM1 (58-66) is restricted by all HLA-C. Therefore, FluM1 (58-66) may help to understand the reaction of immune system against Influenza virus of each populations having different HLA type.

FluM1 (58-66) has indeed prompted research to develop T-cell vaccine strategies capable of inducing specific CTL responses in patients upon immunization with Influenza M1 antigenic epitope. MVA-NP+M1 vaccine use GILGFVFTL epitope and is tested in clinical trial.

Potential cross-reactivity with HIV-1 p17 Gag (77-85)

Moreover, FluM1 (58-66) share similarities with HIV-1 p17 Gag (77-85) which can potentially show a cross-reactivity between these epitopes. It has been demonstrated a cross-reactivity and results suggest that immunity following infection by Influenza virus causes specific immune response to HIV-1 p17 Gag (77-85).

 

Technical specification

Biotin-FluM1 peptide (58-66) buy Sequence : Biotin-GILGFVFTL
Biotin-FluM1 peptide (58-66) synthesis MW : 1192,47 g/mol (C59H89N11O13S)
Biotin-FluM1 peptide (58-66) price Purity : > 95%
Peptide Library synthesis Counter-Ion : TFA Salts (see option TFA removal)
Peptide library synthesis Biotin FluM1 peptide (58-66) 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
SB074-1MG 1 mg 156 195
SB074-5MG 5 mg 546 682,5

 

References

1- Houser K., Subbarao K. Cell Host Microbe. 17(3):295-300
Influenza vaccines: challenges and solutions
 

Vaccination is the best method for the prevention and control of influenza. Vaccination can reduce illness and lessen severity of infection. This review focuses on how currently licensed influenza vaccines are generated in the U.S., why the biology of influenza poses vaccine challenges, and vaccine approaches on the horizon that address these challenges.

2- Bednarek M. A., Sauma S. Y., Gammon M. C., Porter G., Tamhankar S., Williamson A. R. and Zweerink H. J. J Immunol. 147(12):4047-4053 (1991)
The minimum peptide epitope from the influenza virus matrix protein. Extra and intracellular loading of HLA-A2
 

Influenza virus matrix protein-derived peptides were synthesized based on the amino acid motifs for HLA-A2 bound self peptides. Among these peptides a nonamer (amino acids 58 through 66: G I L G F V F T L) was found to be 100 to 1000 times more effective than the commonly used peptide 57-68 (K G I L G F V F T L T V) in sensitizing HLA-A2+ target cells to lysis by influenza virus specific cytotoxic T lymphocytes. The sensitizing activity of the 12-mer 57-68 was not due to contamination with shorter and more active peptides. Intracellular expression of peptide 58-66 (mediated by a stable expression plasmid with DNA coding for this peptide) also sensitized HLA-A2+ cells to lysis. Peptide 58-66 stimulated human PBMC to generate CTL that recognized peptides 58-66 and 57-68 in association with HLA-A2.

3- Antrobus R. D. et al. PLoS One. 7(10):e48322 (2012)
style= »color: #000000; »>A T cell-inducing influenza vaccine for the elderly: safety and immunogenicity of MVA-NP+M1 in adults aged over 50 years  

BACKGROUND: Current influenza vaccines have reduced immunogenicity and are of uncertain efficacy in older adults. We assessed the safety and immunogenicity of MVA-NP+M1, a viral-vectored influenza vaccine designed to boost memory T cell responses, in a group of older adults.

METHODS: Thirty volunteers (aged 50-85) received a single intramuscular injection of MVA-NP+M1 at a dose of 1·5×10(8) plaque forming units (pfu). Safety and immunogenicity were assessed over a period of one year. The frequency of T cells specific for nucleoprotein (NP) and matrix protein 1 (M1) was determined by interferon-gamma (IFN-γ) ELISpot, and their phenotypic and functional properties were characterized by polychromatic flow cytometry. In a subset of M1-specific CD8(+) T cells, T cell receptor (TCR) gene expression was evaluated using an unbiased molecular approach.

RESULTS: Vaccination with MVA-NP+M1 was well tolerated. ELISpot responses were boosted significantly above baseline following vaccination. Increases were detected in both CD4(+) and CD8(+) T cell subsets. Clonality studies indicated that MVA-NP+M1 expanded pre-existing memory CD8(+) T cells, which displayed a predominant CD27(+)CD45RO(+)CD57(-)CCR7(-) phenotype both before and after vaccination.

CONCLUSION: MVA-NP+M1 is safe and immunogenic in older adults. Unlike seasonal influenza vaccination, the immune responses generated by MVA-NP+M1 are similar between younger and older individuals. A T cell-inducing vaccine such as MVA-NP+M1 may therefore provide a way to circumvent the immunosenescence that impairs routine influenza vaccination.

4- Schmittel A., Keilholz U., Bauer S., Kuhne U., Stevanovic S., Thiel E. and Scheibenbogen C. J Immunol Methods. 247(1-2):17-24 (2001)
Application of the IFN-γ ELISPOT assay to quantify T cell responses against proteins
 

The ELISPOT assay has been established for the direct ex vivo quantification of peptide-reactive T lymphocytes from peripheral blood mononuclear cells (PBMC). In this report we studied the ELISPOT assay conditions for the detection of T cell responses against protein antigens including Tetanus toxoid (TT), purified-protein-derivative (PPD) and a synthetic 30 mer peptide derived from influenza matrix protein (IMP) containing the HLA-A*0201-restricted 9 mer peptide epitope GILGFVFTL as a model for a viral protein. We found several aspects to be crucial for a sensitive detection of T cell responses against proteins including a pellet preincubation step and the monocyte concentration. Using optimized assay conditions specific CD4+ T cell responses against TT and PPD as well as CD8+ T cell responses against IMP can be quantified directly ex vivo from peripheral blood mononuclear cells.

5- Choo J. A. L., Liu J., Toh X., Grotenberg G. M. and Ren E. C. J. Virol. 88(18):10613-10623 (2014)
The Immunodominant Influenza A Virus M158–66 Cytotoxic T Lymphocyte Epitope Exhibits Degenerate Class I Major Histocompatibility Complex Restriction in Humans
 

Cytotoxic T lymphocytes recognizing conserved peptide epitopes are crucial for protection against influenza A virus (IAV) infection. The CD8 T cell response against the M158–66 (GILGFVFTL) matrix protein epitope is immunodominant when restricted by HLA-A*02, a major histocompatibility complex (MHC) molecule expressed by approximately half of the human population. Here we report that the GILGFVFTL peptide is restricted by multiple HLA-C*08 alleles as well. We observed that M158–66 was able to elicit cytotoxic T lymphocyte (CTL) responses in both HLA-A*02- and HLA-C*08-positive individuals and that GILGFVFTL-specific CTLs in individuals expressing both restriction elements were distinct and not cross-reactive. The crystal structure of GILGFVFTL–HLA-C*08:01 was solved at 1.84 Å, and comparison with the known GILGFVFTL–HLA-A*02:01 structure revealed that the antigen bound both complexes in near-identical conformations, accommodated by binding pockets shaped from shared as well as unique residues. This discovery of degenerate peptide presentation by both HLA-A and HLA-C allelic variants eliciting unique CTL responses to IAV infection contributes fundamental knowledge with important implications for vaccine development strategies.

6- Acierno P. M., Newton D. A., Brown E. A., Maes L. A., Baatz J. E. and Gattoni-Celli S. J. Transl. Med. 1(3) (2003)
Cross-reactivity between HLA-A2-restricted FLU-M1:58–66 and HIV p17 GAG:77–85 epitopes in HIV-infected and uninfected individuals
 

BACKGROUND: The matrix protein of the influenza A virus and the matrix and capsid proteins of the human immunodeficiency virus (HIV) share striking structural similarities which may have evolutionary and biological significance. These similarities led us to hypothesize the existence of cross-reactivity between HLA-A2-restricted FLU-M1:58–66 and HIV-1 p17 GAG:77–85 epitopes.

METHODS: The hypothesis that these two epitopes are cross-reactive was tested by determining the presence and extent of FLU/GAG immune cross-reactivity in lymphocytes from HIV-seropositive and seronegative HLA-A2+ donors by cytotoxicity assays and tetramer analyses. Moreover, the molecular basis for FLU/GAG cross-reactivity in HIV-seropositive and seronegative donors was studied by comparing lymphocyte-derived cDNA sequences corresponding to the TCR-β variable regions, in order to determine whether stimulation of lymphocytes with either peptide results in the expansion of identical T-cell clonotypes.

RESULTS: Here, we report evidence of cross-reactivity between FLU-M1:58–66 and HIV-1 p17 GAG:77–85 epitopes following in vitro stimulation of PBMC derived from either HIV-seropositive or seronegative HLA-A2+ donors as determined by cytotoxicity assays, tetramer analyses, and molecular clonotyping.

CONCLUSION: These results suggest that immunity to the matrix protein of the influenza virus may drive a specific immune response to an HLA-A2-restricted HIV gag epitope in HIV-infected and uninfected donors vaccinated against influenza.