Transcripts expressed in cytomegalovirus latency coding for an antigenic IE/E phase peptide that drives “memory inflation”

  • Angelique Renzaho
  • Julia K. Schmiedeke
  • Marion Griessl
  • Birgit Kühnapfel
  • Christof K. Seckert
  • Niels A. W. LemmermannEmail author
Original Investigation


Roizman’s definition of herpesviral latency, which applies also to cytomegaloviruses (CMVs), demands maintenance of reactivation-competent viral genomes after clearance of productive infection. It is more recent understanding that failure to complete the productive viral cycle for virus assembly and release does not imply viral gene silencing at all genetic loci and all the time. It rather appears that CMV latency is transcriptionally “noisy” in that silenced viral genes get desilenced from time to time in a stochastic manner, leading to “transcripts expressed in latency” (TELs). If a TEL happens to code for a protein that contains a CD8 T cell epitope, protein processing can lead to the presentation of the antigenic peptide and restimulation of cognate CD8 T cells during latency. This mechanism is discussed as a potential driver of epitope-selective accumulation of CD8 T cells over time, a phenomenon linked to CMV latency and known as “memory inflation” (MI). So far, expression of an epitope-encoding TEL was shown only for the major immediate-early (MIE) gene m123/ie1 of murine cytomegalovirus (mCMV), which codes for the prototypic MI-driving antigenic peptide YPHFMPTNL that is presented by the MHC class-I molecule Ld. The only known second MI-driving antigenic peptide of mCMV in the murine MHC haplotype H-2d is AGPPRYSRI presented by the MHC-I molecule Dd. This peptide is very special in that it is encoded by the early (E) phase gene m164 and by an overlapping immediate-early (IE) transcript governed by a promoter upstream of m164. If MI is driven by presentation of TEL-derived antigenic peptides, as the hypothesis says, one should find corresponding TELs. We show here that E-phase and IE-phase transcripts that code for the MI-driving antigenic peptide AGPPRYSRI are independently and stochastically expressed in latently infected lungs.


Antigen presentation Antigenic peptide(s) CD8 T cells Gene m164 IE1 peptide Latency Latent infection Memory inflation (MI) RT-qPCR Stochastic gene expression Transcripts expressed in latency (TEL) Viral gene expression 



This work was supported by the Deutsche Forschungsgemeinschaft (DFG), SFB490, individual project E4 “Antigen presentation under the influence of murine cytomegalovirus immune evasion genes” (M.G., B.K, and C.K.S.), and SFB1292, individual project TP11 “Viral evasion of innate and adaptive immune cells and inbetweeners” (A.R, J.K.S., and N.A.W.L.).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standard

Animal experiments were approved according to German federal law §8 Abs. 1 TierSchG by the ethics committee of the Landesuntersuchungsamt Rheinland-Pfalz, permission number 177-07/G 10-1-052.


  1. 1.
    Roizman B, Sears AE (1987) An inquiry into the mechanisms of herpes simplex virus latency. Annu Rev Microbiol 41:543–571. CrossRefGoogle Scholar
  2. 2.
    Kurz S, Steffens HP, Mayer A, Harris JR, Reddehase MJ (1997) Latency versus persistence or intermittent recurrences: evidence for a latent state of murine cytomegalovirus in the lungs. J Virol 71:2980–2987Google Scholar
  3. 3.
    Reddehase MJ, Lemmermann NA (2019) Cellular reservoirs of latent cytomegaloviruses. Med Microbiol Immunol. Google Scholar
  4. 4.
    Elder E, Sinclair J (2019) HCMV latency: what regulates the regulators? Med Microbiol Immunol. Google Scholar
  5. 5.
    Reddehase MJ, Simon CO, Seckert CK, Lemmermann N, Grzimek NK (2008) Murine model of cytomegalovirus latency and reactivation. Curr Top Microbiol Immunol 325:315–331Google Scholar
  6. 6.
    Seckert CK, Griessl M, Büttner JK, Scheller S, Simon CO, Kropp KA, Renzaho A, Kühnapfel B, Grzimek NK, Reddehase MJ (2012) Viral latency drives ‘memory inflation’: a unifying hypothesis linking two hallmarks of cytomegalovirus infection. Med Microbiol Immunol 201:551–566. CrossRefGoogle Scholar
  7. 7.
    Seckert CK, Griessl M, Buttner JK, Freitag K, Lemmermann N, Hummel M, Liu XF, Abecassis M, Angulo A, Messerle M, Cook CH, Reddehase M (2013) Immune surveillance of cytomegalovirus latency and reactivation in murine models: link to memory inflation. In: Reddehase MJ (ed) Cytomegaloviruses, vol 1. Caister Academic Press, Norfolk, pp 374–416Google Scholar
  8. 8.
    Reddehase MJ, Lemmermann NAW (2018) Mouse model of cytomegalovirus disease and immunotherapy in the immunocompromised host: predictions for medical translation that survived the “Test of Time”. Viruses 25:E693. CrossRefGoogle Scholar
  9. 9.
    Keil GM, Ebeling-Keil A, Koszinowski UH (1984) Temporal regulation of murine cytomegalovirus transcription and mapping of viral RNA synthesized at immediate early times after infection. J Virol 50:784–795Google Scholar
  10. 10.
    Keil GM, Ebeling-Keil A, Koszinowski UH (1987) Immediate-early genes of murine cytomegalovirus: location, transcripts, and translation products. J Virol 61:526–533Google Scholar
  11. 11.
    Keil GM, Ebeling-Keil A, Koszinowski UH (1987) Sequence and structural organization of murine cytomegalovirus immediate-early gene 1. J Virol 61:1901–1908Google Scholar
  12. 12.
    Rawlinson WD, Farrell HE, Barrell BG (1996) Analysis of the complete DNA sequence of murine cytomegalovirus. J Virol 70:8833–8849Google Scholar
  13. 13.
    Reddehase MJ, Koszinowski UH (1984) Significance of herpesvirus immediate early gene expression in cellular immunity to cytomegalovirus infection. Nature 312:369–371CrossRefGoogle Scholar
  14. 14.
    Reddehase MJ, Rothbard JB, Koszinowski UH (1989) A pentapeptide as minimal antigenic determinant for MHC class I-restricted T lymphocytes. Nature 337:651–653. CrossRefGoogle Scholar
  15. 15.
    Messerle M, Keil GM, Koszinowski UH (1991) Structure and expression of murine cytomegalovirus immediate-early gene 2. J Virol 65:1638–1643Google Scholar
  16. 16.
    Kurz SK, Rapp M, Steffens HP, Grzimek NKA, Schmalz S, Reddehase MJ (1999) Focal transcriptional activity of murine cytomegalovirus during latency in the lungs. J Virol 73:482–494Google Scholar
  17. 17.
    Grzimek NK, Dreis D, Schmalz S, Reddehase MJ (2001) Random, asynchronous, and asymmetric transcriptional activity of enhancer-flanking major immediate-early genes ie1/3 and ie2 during murine cytomegalovirus latency in the lungs. J Virol 75:2692–2705. CrossRefGoogle Scholar
  18. 18.
    Simon CO, Holtappels R, Tervo HM, Böhm V, Däubner T, Oehrlein-Karpi SA, Kühnapfel B, Renzaho A, Strand D, Podlech J, Reddehase MJ, Grzimek NK (2006) CD8 T cells control cytomegalovirus latency by epitope-specific sensing of transcriptional reactivation. J Virol 80:10436–10456. CrossRefGoogle Scholar
  19. 19.
    Lemmermann NA, Kropp KA, Seckert CK, Grzimek NK, Reddehase MJ (2011) Reverse genetics modification of cytomegalovirus antigenicity and immunogenicity by CD8 T cell epitope deletion and insertion. J Biomed Biotechnol 2011:812742. CrossRefGoogle Scholar
  20. 20.
    Podlech J, Holtappels R, Pahl-Seibert MF, Steffens HP, Reddehase MJ (2000) Murine model of interstitial cytomegalovirus pneumonia in syngeneic bone marrow transplantation: persistence of protective pulmonary CD8 T cell infiltrates after clearance of acute infection. J Virol 74:7496–7507CrossRefGoogle Scholar
  21. 21.
    Holtappels R, Pahl-Seibert MF, Thomas D, Reddehase MJ (2000) Enrichment of immediate-early 1 (m123/pp 89) peptide-specific CD8 T cells in a pulmonary CD62L(lo) memory-effector cell pool during latent murine cytomegalovirus infection of the lungs. J Virol 74:11495–11503CrossRefGoogle Scholar
  22. 22.
    Karrer U, Sierro S, Wagner M, Oxenius A, Hengel H, Koszinowski UH, Phillips RE, Klenerman P (2003) Memory inflation: continuous accumulation of antiviral CD8 + T cells over time. J Immunol 170:2022–2029 (Corrigendum J Immunol 171:3895) CrossRefGoogle Scholar
  23. 23.
    Welten SPM, Baumann NS, Oxenius A (2019) Fuel and brake of memory T cell inflation. Med Microbiol Immunol. Google Scholar
  24. 24.
    Méndez AC, Rodríguez-Rojas C, Del Val M (2019) Vaccine vectors: the bright side of cytomegalovirus. Med Microbiol Immunol. Google Scholar
  25. 25.
    Cicin-Sain L (2019) Cytomegalovirus memory inflation and immune protection. Med Microbiol Immunol. Google Scholar
  26. 26.
    van den Berg SPH, Pardieck IN, Lanfermeijer J, Sauce D, Klenerman K, van Baarle D, Arens R (2019) The hallmarks of CMV-specific CD8 T cell differentiation. Med Microbiol Immunol. Google Scholar
  27. 27.
    Jackson SE, Sedikides GX, Okecha G, Wills MR (2019) Generation, maintenance and tissue distribution of T cell responses to human cytomegalovirus in lytic and latent infection. Med Microbiol Immunol. Google Scholar
  28. 28.
    Däubner T, Fink A, Seitz A, Tenzer S, Müller J, Strand D, Seckert CK, Janssen C, Renzaho A, Grzimek NK, Simon CO, Ebert S, Reddehase MJ, Oehrlein-Karpi SA, Lemmermann NA (2010) A novel transmembrane domain mediating retention of a highly motile herpesvirus glycoprotein in the endoplasmic reticulum. J Gen Virol 91:1524–1534. CrossRefGoogle Scholar
  29. 29.
    Holtappels R, Thomas D, Podlech J, Reddehase MJ (2002) Two antigenic peptides from genes m123 and m164 of murine cytomegalovirus quantitatively dominate CD8 T cell memory in the H-2d haplotype. J Virol 76:151–164CrossRefGoogle Scholar
  30. 30.
    Messerle M, Crnkovic I, Hammerschmidt W, Ziegler H, Koszinowski UH (1997) Cloning and mutagenesis of a herpesvirus genome as an infectious bacterial artificial chromosome. Proc Natl Acad Sci USA 94:14759–14763CrossRefGoogle Scholar
  31. 31.
    Wagner M, Jonjic S, Koszinowski UH, Messerle M (1999) Systematic excision of vector sequences from the BAC-cloned herpesvirus genome during virus reconstitution. J Virol 73:7056–7060Google Scholar
  32. 32.
    Podlech J, Holtappels R, Grzimek NKA, Reddehase MJ (2002) Animal models: murine cytomegalovirus. In: Kaufmann SHE, Kabelitz D (eds) Methods in microbiology, vol 32. Immunology of infection. Academic Press, London, pp 493–525Google Scholar
  33. 33.
    Simon CO, Seckert CK, Dreis D, Reddehase MJ, Grzimek NK (2005) Role for tumor necrosis factor alpha in murine cytomegalovirus transcriptional reactivation in latently infected lungs. J Virol 79:326–340. CrossRefGoogle Scholar
  34. 34.
    Lemmermann NA, Podlech J, Seckert CK, Kropp KA, Grzimek NK, Reddehase MJ, Holtappels R (2010) CD8 T cell immunotherapy of cytomegalovirus disease in the murine model. In: Kabelitz D, Kaufmann SHE (eds) Methods in microbiology, vol 37. Immunology of infection. Academic Press, London, pp 369–420Google Scholar
  35. 35.
    Holtappels R, Grzimek NK, Simon CO, Thomas D, Dreis D, Reddehase MJ (2002) Processing and presentation of murine cytomegalovirus pORFm164-derived peptide in fibroblasts in the face of all viral immunosubversive early gene functions. J Virol 76:6044–6053CrossRefGoogle Scholar
  36. 36.
    Lefkovits I, Waldman H (1979) Limiting dilution analysis of cells in the immune system. Cambridge University Press, Cambridge, pp 38–82Google Scholar
  37. 37.
    de St Fazekas, Groth S (1982) The evaluation of limiting dilution assays. J Immunol Methods 49:R11–R23CrossRefGoogle Scholar
  38. 38.
    Munks MW, Cho KS, Pinto AK, Sierro S, Klenerman P, Hill AB (2006) Four distinct patterns of memory CD8 T cell responses to chronic murine cytomegalovirus infection. J Immunol 177:450–458CrossRefGoogle Scholar
  39. 39.
    Fink A, Büttner JK, Thomas D, Holtappels R, Reddehase MJ, Lemmermann NA (2014) Noncanonical expression of a murine cytomegalovirus early protein CD8 T cell epitope as an immediate early epitope based on transcription from an upstream gene. Viruses 6:808–831. CrossRefGoogle Scholar
  40. 40.
    Kurz SK, Reddehase MJ (1999) Patchwork pattern of transcriptional reactivation in the lungs indicates sequential checkpoints in the transition from murine cytomegalovirus latency to recurrence. J Virol 73:8612–8622Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute for VirologyUniversity Medical Center, Johannes Gutenberg-University Mainz and Research Center for Immunotherapy (FZI)MainzGermany
  2. 2.Morgan Sindall Professional Services AGBaselSwitzerland
  3. 3.Institute for Medical Microbiology and HygieneUniversity Medical Center, Johannes Gutenberg-University MainzMainzGermany

Personalised recommendations