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Contributions of the lymphocytic choriomeningitis virus glycoprotein and polymerase to strain-specific differences in murine liver pathogenicity, by A. Bergthaler, D. Merkler, E. Horvath, L. Bestmann and D. D. Pinschewer

Journal of General Virology vol. 88, part 2, pp. 592 - 603

Supplementary Methods

mAbs, hyperimmune serum and neutralization assays. mAbs generated by immunizing mice with LCMV-WE (WEN-3), vsV-IND (VI-7) and vsV-NJ (H6B9D5) have been described previously (Kalinke et al., 1996; Seiler et al., 1998). LCMV-ARM and -WE hyperimmune sera were generated as described previously (Pinschewer et al., 2004). Neutralizing antibodies against LCMV and recombinants were detected in an immunofocus reduction assay as described previously for LCMV (Battegay et al., 1991).

References

Battegay, M., Cooper, S., Althage, A., Banziger, J., Hengartner, H. & Zinkernagel, R. M. (1991). Quantification of lymphocytic choriomeningitis virus with an immunological focus assay in 24- or 96-well plates. J Virol Methods 33, 191-198.

Kalinke, U., Bucher, E. M., Ernst, B., Oxenius, A., Roost, H. P., Geley, S., Kofler, R., Zinkernagel, R. M. & Hengartner, H. (1996). The role of somatic mutation in the generation of the protective humoural immune response against vesicular stomatitis virus. Immunity 5, 639-652.

Pinschewer, D. D., Perez, M., Jeetendra, E., Bächi, T., Horvath, E., Hengartner, H., Whitt, M. A., de la Torre, J. C. & Zinkernagel, R. M. (2004). Kinetics of protective antibodies are determined by the viral surface antigen. J Clin Invest 114, 988-993.

Seiler, P., Brundler, M. A., Zimmermann, C., Weibel, D., Bruns, M., Hengartner, H. & Zinkernagel, R. M. (1998). Induction of protective cytotoxic T cell responses in the presence of high titers of virus-neutralizing antibodies: implications for passive and active immunization. J Exp Med 187, 649-654.

Supplementary Table S1. Differential seroreactivity of wild-type and engineered LCMV viruses

Supplementary Table S2. Sequence comparison of the clone 6 and ARM L segments

[Single PDF of Supplementary Tables S1 and S2] (24 KB)

Supplementary Fig. S1. AST (a) and ALT (b) activities in mice infected i.v. with 2x10⁶ p.f.u. of either rARM/ARMGP or wtARM.

Supplementary Fig. S2. Virus titres in BHK-21 cells infected with the indicated viruses.

Supplementary Fig. S3. Assessment of GP33-, NP396- and GP276-specific CTL responses (a) and enumeration of GP33- and NP396-specific MHC class I tetramer-binding CD8⁺ T cells (b) in C57BL/6 mice infected with 2x10⁶ p.f.u. of rCl.6/WEGP or rCl.6/ARMGP or left uninfected.

[Single PDF of Supplementary Figs S1-S3] (29 KB)

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