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