Elsevier

Human Immunology

Volume 64, Issue 2, February 2003, Pages 211-223
Human Immunology

Original contribution
Derivation of HLA-B*0702 transgenic mice: functional CTL repertoire and recognition of human B*0702-restricted CTL epitopes

https://doi.org/10.1016/S0198-8859(02)00786-3Get rights and content

Abstract

Transgenic mice expressing chimeric human leukocyte antigen (HLA)-B*0702 and murine H-2Kb class I molecules were evaluated as a model system to study the immunogenicity of human cytotoxic T lymphocyte epitopes. Immunization of these mice with six known HLA-B*0702–restricted cytotoxic T lymphocyte epitopes emulsified in incomplete Freund’s adjuvant induced significant immune responses specific for all six epitopes. A comparison of the immune responses between HLA-B*0702/Kb and HLA-A*0201/Kb transgenic mice demonstrated that the HLA-B*0702/Kb mice possess a T-cell receptor repertoire capable of recognizing human B*0702 epitopes. However, the magnitude of B*0702-specific responses induced in B*0702/Kb mice were approximately tenfold lower than A*0201-specific responses induced in HLA-A*0201/Kb transgenic mice. A panel of 24 B*0702 motif-bearing peptides was used to examine the relationship between immunogenicity and HLA-B*0702 binding capacity. All seven peptides with high binding affinities of 50% inhibitory concentration ≤50 NM (IC50 50 nM or less) were immunogenic. Similarly, 75% (9 of 12) of the intermediate binders (IC50 nM of 50–500) were also immunogenic. Finally, only two of five peptides with binding capacity > 500 nM were found to have marginal immunogenicity, whereas the other three were completely negative. HLA-B*0702/Kb transgenic mice were found to induce B*0702-specific responses after immunization with whole DNA genes or minigenes, suggesting that, at least to some degree, B*0702 epitopes were generated as a result of natural in vivo processing and presentation.

Section snippets

Abbreviations

    APC

    antigen-presenting cell

    β2m

    β2 microglobulin

    EBNA

    Epstein-Barr virus-encoded nuclear antigen

    HBV

    hepatitis B virus

    HCV

    hepatitis C virus

    IC50

    50% inhibitory concentration

    IFA

    incomplete Freund’s adjuvant

    LPS

    lipopolysaccharide

    MFC

    mean channel fluorescence

    c

    core (e.g., HBVc)

    pol

    polymerase

    env

    envelope

    Pf

    Plasmodium falciparum

    SU

    secretory units (e.g., IFN-γ SU)

Peptides

Peptides were synthesized according to standard t-BOC or F-moc solid-phase synthesis methods [24]. Peptides were purified by analytic reverse-phase high performance liquid chromatography and purity was routinely > 95%. The peptide identify was verified by electrospray mass spectroscopy analysis. Peptides were dissolved in dimethyl sulfoxide at a concentration of 20 mg/ml, stored frozen at 20 °C, and diluted with phosphate-buffered saline (PBS) or PBS 0.05% NP40 before use. The sequence of

Production and characterization of HLA-B*0702/Kb transgenic mice

The B*0702/Kb transgenic mice were derived as described in the Materials and Methods section. The pattern of transgene cell surface expression of these mice as detected by the pan HLA class I mAb, 9.12.1, is shown in Figure 1. The B*0702/Kb chimeric transgene is well-expressed in splenocytes displaying an mean channel fluorescence of approximately 200 (Figure 1A), which is comparable to the level to the murine class I H-2Db antigen detected by mAb 28-14-8S (Figure 1B). This level of expression

Discussion

Previously, we reported on the generation of A*0201/Kb and A*1101/Kb transgenic mice 1, 9. To facilitate the design of epitope-based vaccines that guarantee broadly based and nonethnically biased population coverage, the development of HLA class I transgenic mice other than the well-described A*0201 and A*1101 lines may be of particular importance. In the present study, the derivation and characterization of HLA-B*0702/Kb transgenic mice is described. HLA-B*0702 is one of the most common class

Acknowledgements

This work was supported in part by federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), under contracts NO1-AI-45241 and NO1-A1-95362 to Epimmune Inc. We thank Claire Crimi for excellent technical assistance and Bianca Mothé for critical review of this manuscript.

References (52)

  • D.L. Doolan et al.

    Degenerate cytotoxic T cell epitopes from P. falciparum restricted by multiple HLA-A and HLA-B supertype alleles

    Immunity

    (1997)
  • J. Alexander et al.

    Derivation of HLA-A11/Kb transgenic miceFunctional CTL repertoire and recognition of human A11-restricted CTL epitopes

    J Immunol

    (1997)
  • U. Blum-Tirouvanziam et al.

    Localization of HLA-A21 restricted T cell epitopes in the circumsporozoite protein of Plasmodium falciparum

    J Immunol

    (1995)
  • M. Theobald et al.

    Targeting p53 as a general tumor antigen

    Proc Natl Acad Sci U S A

    (1995)
  • A. Vitiello et al.

    Comparison of cytotoxic T lymphocyte responses induced by peptide or DNA immunizationImplications on immunogenicity and immunodominance

    Eur J Immunol

    (1997)
  • M.J. Palmowski et al.

    Competition between CTL narrows the immune response induced by prime-boost vaccination protocols

    J Immunol

    (2002)
  • A. Vitiello et al.

    Analysis of the HLA-restricted influenza-specific cytotoxic T lymphocyte response in transgenic mice carrying a chimeric human-mouse class I major histocompatibility complex

    J Exp Med

    (1991)
  • P.A. Wentworth et al.

    Differences and similarities in the A2.1-restricted cytotoxic T cell repertoire in humans and human leukocyte antigen-transgenic mice

    Eur J Immunol

    (1997)
  • A. Sette et al.

    The relationship between class I binding affinity and immunogenicity of potential cytotoxic T cell epitopes

    J Immunol

    (1994)
  • M. Shirai et al.

    CTL responses of HLA-A2.1-transgenic mice specific for hepatitis C viral peptides predict epitopes for CTL of humans carrying HLA-A2.1

    J Immunol

    (1995)
  • S. Man et al.

    Definition of a human T cell epitope from influenza A non-structural protein using HLA-A2.1 transgenic mice

    Int Immunol

    (1995)
  • A. Geluk et al.

    Identification of major epitopes of Mycobacterium tuberculosis AG85B that are recognized by HLA-A*0201-restricted CD8+ T cells in HLA-transgenic mice and humans

    J Immunol

    (2000)
  • P. Krausa et al.

    Genetic polymorphism within HLA-A*02Significant allelic variation revealed in different populations

    Tissue Antigens

    (1995)
  • T. Imanishi et al.

    Allele and haplotype frequencies for HLA and complement loci in various ethnic groups

  • E.L. Huczko et al.

    Characteristics of endogenous peptides eluted from the class I MHC molecule HLA-B7 determined by mass spectrometry and computer modeling

    J Immunol

    (1993)
  • J. Sidney et al.

    Specificity and degeneracy in peptide binding to HLA-B7-like class I molecules

    J Immunol

    (1996)
  • Cited by (19)

    • A Distinct Lung-Interstitium-Resident Memory CD8<sup>+</sup> T Cell Subset Confers Enhanced Protection to Lower Respiratory Tract Infection

      2016, Cell Reports
      Citation Excerpt :

      We hope our findings will have implications for the design of safe and effective CD8+ T cell-targeted vaccines against respiratory infections. B6-K0D0;B∗07;02tg (B7tg) transgenic mice have been described elsewhere (Alexander et al., 2003). Mouse breeding, maintenance, and experimentation complied with institutional animal care and use committee (IACUC) regulations.

    • Toxoplasma gondii HLA-B*0702-restricted GRA7 <inf>20-28</inf> peptide with adjuvants and a universal helper T cell epitope elicits CD8 <sup>+</sup> T cells producing interferon-γ and reduces parasite burden in HLA-B*0702 mice

      2012, Human Immunology
      Citation Excerpt :

      The TLR4 agonist was a GLA-SE mimetic that was synthesized by the Infectious Diseases Research Institute (IDRI, Seattle, WA), used as previously described [19]. Female HLA-B*0702 transgenic mice produced at Pharmexa-Epimmune (San Diego, CA) and bred at the University of Chicago express a chimeric HLA-B07/H2-Db major histocompatibility complex (MHC) class I molecule, are on a C57BL/6 × Balb/C background backcrossed through many generations, and have been previously described [11]. HLA-A*1101 mice were also utilized in an experiment.

    • Identification of T. gondii epitopes, adjuvants, and host genetic factors that influence protection of mice and humans

      2010, Vaccine
      Citation Excerpt :

      Wells were washed with sterile PBS and blocked with RPMI-1640 medium containing 10% FCS at room temperature for 2–3 h. PBMCs were then plated in complete RPMI-1640 medium at 2 × 105 cells per well. Peptide or peptide pools were added to each well at 10 μg/ml and plates were incubated at 37 °C and 5% CO2 for 20–24 h. Media containing an equivalent concentration of DMSO was used to measure background response levels, and 5 μg/ml of T. gondii lysate antigen (TLA), 2.5 μg/ml of Concavalin A, or 10 μg/ml of a relevant HLA-B07-restricted Epstein–Barr virus (EBV)-derived peptide [31] was used as a positive control. Plates were successively incubated with 100 μl of 1 μg/ml biotinylated anti-human IFN-γ mAb (7B6-1) for 2 h and streptavidin-conjugated alkaline phosphatase at a 1:1000 dilution for 1 h at room temperature, before spots were developed using 5-bromo-4-chloro-3-indolyl-phosphate/p-nitro blue tetrazolium chloride (BCIP/NBT).

    • TCR Gene Therapy for Cancer

      2022, Methods in Molecular Biology
    View all citing articles on Scopus
    View full text