Summary
Pichinde virus (PV) infection of mice results in induction of a strong H-2 restricted, virus-specific cytotoxic T lymphocyte (CTL) response and rapid clearance of the virus. To define the specificities of CTL induced by PV infection, we constructed vaccinia virus recombinants containing cloned cDNAs corresponding to full-length (VVNP) and a truncated form (VVNP51–561) of the nucleoprotein (NP) gene of PV. Radioimmunoprecipitation analysis of infected cell lysates indicated that VVNP expressed a PV-specific product identical in size to that of authentic NP, while vaccinia virus recombinants containing truncated NP produced a polypeptide consistent with the synthesis of amino acids 51–561 of Pichinde virus NP. Interestingly, cells infected with VVNP synthesized easily detectable, but much lower levels of nucleoprotein relative to both PV and VVNP51–561. Primary virus-specific CTL induced in three different strains of inbred mice following intravenous infection with PV were able to lyse syngeneic target cells infected with PV but did not markedly lyse syngeneic targets expressing full-length or truncated NP following recombinant vaccinia virus infection. Similarly, secondary anti-PV specific CTL generated following in vitro restimulation by PV or selectively restimulated with vaccinia recombinants did not significantly lyse target cells expressing NP. Further, infection of mice with VVNP and VVNP51–561 did not induce CTLs specific for PV and did not prime mice for the generation of memory anti-PV CTL in vivo. These results suggest that PV gene products other than NP, such as the GPC or L protein, contain the major target epitope(s) recognized by PV-specific CTL.
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References
Auperin D, Dimock K, Cash P, Rawls WE, Leung WC, Bishop DHL (1982) Analyses of the genomes of prototype Pichinde arenavirus and a virulent derivative of Pichinde Munchique: evidence for sequence conservation of the 3′ termini of their viral RNA species. Virology 116: 363–367
Auperin DD, Galinski M, Bishop DHL (1984) The sequences of the N protein gene and intergenic region of the S RNA of Pichinde arenavirus. Virology 134: 208–219
Auperin DD, Romanowski V, Galinski M, Bishop DHL (1984) Sequencing studies of Pichinde arenavirus S RNA indicate a novel coding strategy, and ambisense viral S RNA. J Virol 52: 897–904
Auperin DD, Esposito JJ, Lange JV, Bauer SP, Knight J, McCormick JB, Sasso SD (1988) Construction of a recombinant vaccinia virus expressing the Lassa virus glycoprotein gene and protection of guinea pigs from a lethal Lassa virus infection. Virus Res 9: 233–248
Bangham CRM, Openshaw PJM, Ball LA, King AMQ, Wertz GW, Askonas BA (1986) Human and murine cytotoxic T cells specific to respiratory syncytial virus recognize the viral nucleoprotein (N), but not the major glycoprotein (G), expressed by vaccinia virus recombinants. J Immunol 137: 3973–3977
Bennink JR, Yewdell JW, Smith GL, Moss B (1986) Recognition of cloned influenza virus hemagglutinin gene products by cytotoxic T lymphocytes. J Virol 57: 786–791
Buchmeier MJ, Rawls WE (1977) Variation between strains of hamsters in the lethality of Pichinde virus infections. Infect Immun 16: 413–421
Buchmeier MJ, Lewicki HA, Tomori O, Oldstone MBA (1981) Monoclonal antibodies to lymphocytic choriomeningitis and Pichinde viruses: generation, characterization, and cross-reactivity with other arenaviruses. Virology 113: 73–85
Chakrabarti S, Brechling K, Moss B (1985) Vaccinia virus expression vector: coexpression of β-galactosidase provides visual screening of recombinant virus plaques. Mol Cell Biol 5: 3403–3409
Clegg JCS, Lloyd G (1987) Vaccinia recombinant expressing Lassa-virus internal nucleocapsid protein protects guinea pigs against Lassa fever. Lancet 2: 186–188
Cochran MA, Puckett C, Moss B (1985) In vitro mutagenesis of the promoter region for a vaccinia virus gene: evidence for tandem early and late regulatory signals. J Virol 54: 30–37
Fisher-Hoch SP, McCormick JB, Auperin D, Brown BG, Castor M, Perez G, Ruo S, Conaty A, Brammer L, Bauer S (1989) Protection of rhesus monkeys from fatal Lassa fever by vaccination with a recombinant vaccinia virus containing the Lassa virus glycoprotein gene. Proc Natl Acad Sci USA 86: 317–321
Geballe AP, Mocarski ES (1988) Translational control of cytomegalovirus gene expression is mediated by upstream AUG codons. J Virol 62: 3334–3340
Gould K, Cossins J, Bastin J, Brownlee GG, Townsend A (1989) A 15 amino acid fragment of influenza nucleoprotein synthesized in the cytoplasm is presented to class I-restricted cytotoxic T lymphocytes. J Exp Med 170: 1051–1056
Hany M, Oehen S, Schulz M, Hengartner H, Mackett M, Bishop DHL, Overton H, Zinkernagel RM (1989) Anti-viral protection and prevention of lymphocytic choriomeningitis or of the local footpad swelling reaction in mice by immunization with vaccinia-recombinant virus expressing LCMV-WE nucleoprotein or glycoprotein. Eur J Immunol 19: 417–424
Harnish DG, Leung WC, Rawls WE (1981) Characterization of polypeptides immunoprecipitable from Pichinde virus-infected BHK-21 cells. J Virol 38: 840–848
Harnish DG, Dimock K, Bishop DHL, Rawls WE (1983) Gene mapping in Pichinde virus: assignment of viral polypeptides to genomic L and S RNAs. J Virol 46: 638–641
King AMQ, Stott EJ, Langer SJ, Young KK-Y, Ball LA, Wertz GW (1987) Recombinant vaccinia viruses carrying the N gene of human respiratory syncytial virus: studies of gene expression in cell culture and immune response in mice. J Virol 61: 2885–2890
Kit S, Dubbs D (1977) Regulation of herpesvirus thymidine kinase acitvity in LM (tk —) cells transformed by ultra-violet irradiated herpes simplex virus. Virology 76: 331–340
Klavinskis LS, Whitton JL, Oldstone MBA (1989) Molecularly engineered vaccine which expresses an immunodominant T-cell epitope induces cytotoxic T lymphocytes that confer protection form lethal virus infection. J Virol 63: 4311–4316
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Leung W-C, Harnish D, Ramsingh A, Dimock K, Rawls WE (1981) Gene mapping in Pichinde virus. In: Bishop DHL, Compans RW (eds) Replication of negative strand viruses. Elsevier, Amsterdam, pp 51–57
Mackett M, Smith GL, Moss B (1984) General method of production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol 49: 857–864
Mifune K, Carter M, Rawls WE (1973) Characterization studies of the Pichinde virus — a member of the Arenavirus group. Proc Soc Exp Biol Med 136: 637–644
Morrison HG, Bauer SP, Lange JV, Esposito JJ, McCormick JB, Auperin DD (1989) Protection of guinea pigs from Lassa fever by vaccinia virus recombinants expressing the nucleoprotein or the envelope glycoproteins of Lassa virus. Virology 171: 179–188
Oldstone MBA, Whitton JL, Lewicki H, Tishon A (1989) Fine dissection of a nine amino acid glycoprotein epitope, a major determinant recognized by lymphocytic choriomeningitis virus-specific class I-restricted H-2Db cytotoxic T lymphocytes. J Exp Med 168: 559–570
Puddington L, Bevan MJ, Rose JK, Lefrancois L (1986) N protein is the predominant antigen recognized by vesicular stomatitis virus-specific cytotoxic T cells. J Virol 60: 708–717
Romanowski V, Matsuura Y, Bishop DHL (1985) Complete sequence of the S RNA of lymphocytic choriomeningitis virus (WE strain) compared to that of Pichinde arenavirus. Virus Res 3: 101–114
Rosenthal KL (1988) Specificity of antiviral cytotoxic T lymphocytes. In: Lopez C (ed) Immunobiology and pathogenesis of persistent virus infections. American Society for Microbiology, Washington, DC, pp 135–145
Schulz M, Aichele P, Vollenweider M, Bobe FW, Cardinaux F, Hengartner H, Zinkernagel RM (1989) Major histocompatibility complex-dependent T cell epitopes of lymphocytic choriomeningitis virus nucleoprotein and their protective capacity against viral disease. Eur J Immunol 19: 1657–1667
Shivaprakash M, Harnish D, Rawls W (1988) Characterization of temperature-sensitive mutants of Pichinde virus. J Virol 62: 4037–4043
Townsend ARM, McMicheal AJ, Carter NP, Huddleston JA, Brownlee GG (1984) Cytotoxic T cell recognition of the influenza nucleoprotein and hemagglutinin expressed in transfected mouse L cells. Cell 39: 13–25
Townsend ARM, Gotch FM, Davey J (1985) Cytotoxic T cells recognize fragments of the influenza nucleoprotein. Cell 42: 457–467
Townsend ARM, Rothbard J, Gotch FM, Bahadur G, Wraith D, McMicheal AJ (1986) The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell 44: 959–968
Townsend ARM, Bastin J, Gould K, Brownlee G, Andrew M, Coupar B, Boyle D, Chan S, Smith G (1988) Defective presentation to class I-restricted cytotoxic T lymphocytes in vaccinia-infected cells is overcome by enhanced degradation of antigen. J Exp Med 168: 1211–1224
Trapido H, Sanmartin C (1971) Pichinde virus: a new virus of the Tacaribe group from Columbia. Am J Trop Med Hyg 36: 631–641
Venkatesan S, Baroudy BM, Moss B (1981) Distinctive nucleotide sequences adjacent to multiple initiation and termination sites of an early vaccinia virus gene. Cell 125: 805–813
Volkmer H, Bertholet C, Jonjic S, Wittek R, Koszinowski UH (1987) Cytolytic T lymphocyte recognition of the murine cytomegalovirus nonstructural immediate-early protein pp 89 expressed by recombinant vaccinia virus. J Exp Med 166: 668–677
Walker CM, Rawls WE, Rosenthal KL (1984) Generation of memory cell-mediated immune responses after secondary infection of mice with Pichinde virus. J Immunol 132: 469–474
Whitton JL, Oldstone MBA (1989) Class I MHC can present an endogenous peptide to cytotoxic T lymphocytes. J Exp Med 170: 1033–1038
Whitton JL, Gebhard JR, Lewicki H, Tishon A, Oldstone MBA (1988) Molecular definition of a major cytotoxic T lymphocyte epitope in the glycoprotein of lymphocytic choriomeningitis virus. J Virol 62: 687–695
Whitton JL, Southern PJ, Oldstone MBA (1988) Analysis of the cytotoxic T lymphocyte responses to glycoprotein and nucleoprotein components of lymphocytic choriomeningitis virus. Virology 162: 321–327
Whitton JL, Tishon A, Lewicki H, Gebhard J, Cook T, Salvato M, Joly E, Oldstone MBA (1989) Molecular analysis of a five-amino-acid cytotoxic T-lymphocyte (CTL) epitope: an immunodominant region which induces nonreciprocal CTL cross-reactivity. J Virol 63: 4303–4310
Yanish-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vector and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33: 103–119
Yewdell JW, Bennink JR, Smith GL, Moss B (1985) Influenza A virus nucleoprotein is a major target antigen for cross-reactive anti-influenza A virus cytotoxic T lymphocytes. Proc Natl Acad Sci USA 82: 1785–1789
Yewdell JW, Bennink JR, Mackett M, Lefrancois L, Lyles DS, Moss B (1986) Recognition of cloned vesicular stomatitis virus internal and external gene products by cytotoxic lymphocytes. J Exp Med 163: 1529–1538
Young RA, Davis RW (1983) Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci USA 80: 1194–1198
Young RA, Davis RW (1983) Yeast RNA polymerase II genes: isolation with antibody probes. Science 222: 778–782
Zinkernagel RM, Rosenthal KL (1980) Experiments and speculation on antiviral specificity of T and B cells. Immunol Rev 58: 131–155
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Ozols, D.Y., Harnish, D.G., Rawls, W.E. et al. Assessment of the specificity of cytotoxic T lymphocytes for the nucleoprotein of Pichinde virus using recombinant vaccinia viruses. Archives of Virology 115, 209–225 (1990). https://doi.org/10.1007/BF01310531
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DOI: https://doi.org/10.1007/BF01310531