GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION

M MATSUI, RJ MOOTS, RJ WARBURTON, AL PEACEBREWER, LG TUSSEY, Daniel Quinn, AJ MCMICHAEL, JA FRELINGER

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    Abstract

    During the course of extensive mutagenesis of HLA-A2.1, we examined influenza A matrix peptide (FMP)-specific CTL recognition of HMy2.C1R (C1R) cells expressing mutant HLA-A2.1 molecules, sensitized with synthetic peptide, FMP 58-66, (exogenous peptide), or infected with influenza A virus (endogenous peptide). Most mutants showed equivalent presentation of exogenous and endogenous peptides to FMP-specific CTL. However, five of the mutants differed in this property. Two of the five mutants, F9L and T134K, present exogenous peptide to FMP-specific CTL, but fail to present endogenous peptide to CTL. Western blot analysis using anti-matrix protein Ab indicates that the matrix protein is expressed in these mutants after infection with virus. Interestingly, transfection of these two mutants with a minigene encoding FMP 58-66 results in efficient lysis by FMP-specific CTL. Peptide-binding assays demonstrate that the two mutations dramatically decrease the binding of FMP. However, these mutants bind FMP as well as wild type in the presence of exogenously added human beta(2)-m, suggesting that the lower affinity for beta(2)-m leads to the inability to present endogenous peptide. The remaining three mutants, Y27N, Q32K, and S132C, fail to present exogenous peptide, but present endogenous peptide to FMP-specific CTL. Pulse-chase analyses followed by endoglycosidase-H treatment show that the rate of maturation and processing of the five mutant HLA-A2 molecules in C1R cells is identical to that of wild type. Overall, this study suggests that the assembly and subsequent recognition of endogenous peptide differs from that of exogenous peptide.
    LanguageEnglish
    Pages1088-1096
    JournalJOURNAL OF IMMUNOLOGY
    Volume154
    Issue number3
    Publication statusPublished - Feb 1995

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    Human Influenza
    Peptides
    HLA-A2 Antigen
    N-formylmethionylphenylalanine
    Glycoside Hydrolases
    Influenza A virus
    Virus Diseases
    Mutagenesis
    Transfection
    Proteins
    Western Blotting
    Mutation

    Cite this

    MATSUI, M., MOOTS, RJ., WARBURTON, RJ., PEACEBREWER, AL., TUSSEY, LG., Quinn, D., ... FRELINGER, JA. (1995). GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION. JOURNAL OF IMMUNOLOGY, 154(3), 1088-1096.
    MATSUI, M ; MOOTS, RJ ; WARBURTON, RJ ; PEACEBREWER, AL ; TUSSEY, LG ; Quinn, Daniel ; MCMICHAEL, AJ ; FRELINGER, JA. / GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION. In: JOURNAL OF IMMUNOLOGY. 1995 ; Vol. 154, No. 3. pp. 1088-1096.
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    title = "GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION",
    abstract = "During the course of extensive mutagenesis of HLA-A2.1, we examined influenza A matrix peptide (FMP)-specific CTL recognition of HMy2.C1R (C1R) cells expressing mutant HLA-A2.1 molecules, sensitized with synthetic peptide, FMP 58-66, (exogenous peptide), or infected with influenza A virus (endogenous peptide). Most mutants showed equivalent presentation of exogenous and endogenous peptides to FMP-specific CTL. However, five of the mutants differed in this property. Two of the five mutants, F9L and T134K, present exogenous peptide to FMP-specific CTL, but fail to present endogenous peptide to CTL. Western blot analysis using anti-matrix protein Ab indicates that the matrix protein is expressed in these mutants after infection with virus. Interestingly, transfection of these two mutants with a minigene encoding FMP 58-66 results in efficient lysis by FMP-specific CTL. Peptide-binding assays demonstrate that the two mutations dramatically decrease the binding of FMP. However, these mutants bind FMP as well as wild type in the presence of exogenously added human beta(2)-m, suggesting that the lower affinity for beta(2)-m leads to the inability to present endogenous peptide. The remaining three mutants, Y27N, Q32K, and S132C, fail to present exogenous peptide, but present endogenous peptide to FMP-specific CTL. Pulse-chase analyses followed by endoglycosidase-H treatment show that the rate of maturation and processing of the five mutant HLA-A2 molecules in C1R cells is identical to that of wild type. Overall, this study suggests that the assembly and subsequent recognition of endogenous peptide differs from that of exogenous peptide.",
    author = "M MATSUI and RJ MOOTS and RJ WARBURTON and AL PEACEBREWER and LG TUSSEY and Daniel Quinn and AJ MCMICHAEL and JA FRELINGER",
    year = "1995",
    month = "2",
    language = "English",
    volume = "154",
    pages = "1088--1096",
    journal = "J Immunol",
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    MATSUI, M, MOOTS, RJ, WARBURTON, RJ, PEACEBREWER, AL, TUSSEY, LG, Quinn, D, MCMICHAEL, AJ & FRELINGER, JA 1995, 'GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION', JOURNAL OF IMMUNOLOGY, vol. 154, no. 3, pp. 1088-1096.

    GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION. / MATSUI, M; MOOTS, RJ; WARBURTON, RJ; PEACEBREWER, AL; TUSSEY, LG; Quinn, Daniel; MCMICHAEL, AJ; FRELINGER, JA.

    In: JOURNAL OF IMMUNOLOGY, Vol. 154, No. 3, 02.1995, p. 1088-1096.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION

    AU - MATSUI, M

    AU - MOOTS, RJ

    AU - WARBURTON, RJ

    AU - PEACEBREWER, AL

    AU - TUSSEY, LG

    AU - Quinn, Daniel

    AU - MCMICHAEL, AJ

    AU - FRELINGER, JA

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    N2 - During the course of extensive mutagenesis of HLA-A2.1, we examined influenza A matrix peptide (FMP)-specific CTL recognition of HMy2.C1R (C1R) cells expressing mutant HLA-A2.1 molecules, sensitized with synthetic peptide, FMP 58-66, (exogenous peptide), or infected with influenza A virus (endogenous peptide). Most mutants showed equivalent presentation of exogenous and endogenous peptides to FMP-specific CTL. However, five of the mutants differed in this property. Two of the five mutants, F9L and T134K, present exogenous peptide to FMP-specific CTL, but fail to present endogenous peptide to CTL. Western blot analysis using anti-matrix protein Ab indicates that the matrix protein is expressed in these mutants after infection with virus. Interestingly, transfection of these two mutants with a minigene encoding FMP 58-66 results in efficient lysis by FMP-specific CTL. Peptide-binding assays demonstrate that the two mutations dramatically decrease the binding of FMP. However, these mutants bind FMP as well as wild type in the presence of exogenously added human beta(2)-m, suggesting that the lower affinity for beta(2)-m leads to the inability to present endogenous peptide. The remaining three mutants, Y27N, Q32K, and S132C, fail to present exogenous peptide, but present endogenous peptide to FMP-specific CTL. Pulse-chase analyses followed by endoglycosidase-H treatment show that the rate of maturation and processing of the five mutant HLA-A2 molecules in C1R cells is identical to that of wild type. Overall, this study suggests that the assembly and subsequent recognition of endogenous peptide differs from that of exogenous peptide.

    AB - During the course of extensive mutagenesis of HLA-A2.1, we examined influenza A matrix peptide (FMP)-specific CTL recognition of HMy2.C1R (C1R) cells expressing mutant HLA-A2.1 molecules, sensitized with synthetic peptide, FMP 58-66, (exogenous peptide), or infected with influenza A virus (endogenous peptide). Most mutants showed equivalent presentation of exogenous and endogenous peptides to FMP-specific CTL. However, five of the mutants differed in this property. Two of the five mutants, F9L and T134K, present exogenous peptide to FMP-specific CTL, but fail to present endogenous peptide to CTL. Western blot analysis using anti-matrix protein Ab indicates that the matrix protein is expressed in these mutants after infection with virus. Interestingly, transfection of these two mutants with a minigene encoding FMP 58-66 results in efficient lysis by FMP-specific CTL. Peptide-binding assays demonstrate that the two mutations dramatically decrease the binding of FMP. However, these mutants bind FMP as well as wild type in the presence of exogenously added human beta(2)-m, suggesting that the lower affinity for beta(2)-m leads to the inability to present endogenous peptide. The remaining three mutants, Y27N, Q32K, and S132C, fail to present exogenous peptide, but present endogenous peptide to FMP-specific CTL. Pulse-chase analyses followed by endoglycosidase-H treatment show that the rate of maturation and processing of the five mutant HLA-A2 molecules in C1R cells is identical to that of wild type. Overall, this study suggests that the assembly and subsequent recognition of endogenous peptide differs from that of exogenous peptide.

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    VL - 154

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    JO - J Immunol

    T2 - J Immunol

    JF - J Immunol

    SN - 0022-1767

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    ER -

    MATSUI M, MOOTS RJ, WARBURTON RJ, PEACEBREWER AL, TUSSEY LG, Quinn D et al. GENETIC-EVIDENCE FOR DIFFERENCE BETWEEN INTRACELLULAR AND EXTRACELLULAR PEPTIDES IN INFLUENZA-A MATRIX PEPTIDE-SPECIFIC CTL RECOGNITION. JOURNAL OF IMMUNOLOGY. 1995 Feb;154(3):1088-1096.