Persistent viral replication and the development of T-cell responses after intranasal infection by MCMV
Natural transmission of cytomegalovirus (CMV) has been difficult to observe. However, recent work using the mouse model of murine (M)CMV demonstrated that MCMV initially infects the nasal mucosa after transmission from mothers to pups. We found that intranasal (i.n.) inoculation of C57BL/6J mice resulted in reliable recovery of replicating virus from the nasal mucosa as assessed by plaque assay. After i.n. inoculation, CD8+ T-cell priming occurred in the mandibular, deep-cervical, and mediastinal lymph nodes within 3 days of infection. Although i.n. infection induced “memory inflation” of T cells specific for the M38316–323 epitope, there were no detectable CD8+ T-cell responses against the late-appearing IE3416–423 epitope, which contrasts with intraperitoneal (i.p.) infection. MCMV-specific T cells migrated into the nasal mucosa where they developed a tissue-resident memory (TRM) phenotype and this could occur independently of local virus infection or antigen. Strikingly however, virus replication was poorly controlled in the nasal mucosa and MCMV was detectable by plaque assay for at least 4 months after primary infection, making the nasal mucosa a second site for MCMV persistence. Unlike in the salivary glands, the persistence of MCMV in the nasal mucosa was not modulated by IL-10. Taken together, our data characterize the development of local and systemic T-cell responses after intranasal infection by MCMV and define the nasal mucosa, a natural site of viral entry, as a novel site of viral persistence.
KeywordsCytomegalovirus Nasal mucosa Intranasal infection Tissue-resident memory T cells
This work was supported by Grant AI106810 awarded to C.M.S.
Compliance with ethical standards
Conflict of interest
The authors declare no competing interests.
- 5.Schleiss MR (2006) Role of breast milk in acquisition of cytomegalovirus infection: recent advances. Curr Opin Pediatr 18:48–52Google Scholar
- 6.Dworsky M, Yow M, Stagno S, Pass RF, Alford C (1983) Cytomegalovirus infection of breast milk and transmission in infancy. Pediatrics 72:295–299Google Scholar
- 10.Stahl FR, Heller K, Halle S, Keyser KA, Busche A, Marquardt A, Wagner K, Boelter J, Bischoff Y, Kremmer E, Arens R, Messerle M, Forster R (2013) Nodular inflammatory foci are sites of T cell priming and control of murine cytomegalovirus infection in the neonatal lung. PLoS Pathog 9:e1003828CrossRefGoogle Scholar
- 16.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
- 20.Dunston D, Ashby S, Krosnowski K, Ogura T, Lin W (2013) An effective manual deboning method to prepare intact mouse nasal tissue with preserved anatomical organization. J Vis Exp 78:e50538Google Scholar
- 21.Caldeira-Dantas S, Furmanak T, Smith C, Quinn M, Teos LY, Ertel A, Kurup D, Tandon M, Alevizos I, Snyder CM (2018) The chemokine receptor CXCR3 promotes CD8+ T Cell accumulation in uninfected salivary glands but is not necessary after murine cytomegalovirus infection. J Immunol 200:1133–1145CrossRefGoogle Scholar
- 39.Jordan MC (1978) Interstitial pneumonia and subclinical infection after intranasal inoculation of murine cytomegalovirus. Infect Immun 21:275–280Google Scholar
- 46.Reuter S, Lemmermann NAW, Maxeiner J, Podlech J, Beckert H, Freitag K, Teschner D, Ries F, Taube C, Buhl R, Reddehase MJ, Holtappels R (2019) Coincident airway exposure to low-potency allergen and cytomegalovirus sensitizes for allergic airway disease by viral activation of migratory dendritic cells. PLoS Pathog 15:e1007595CrossRefGoogle Scholar
- 49.Mercer JA, Wiley CA, Spector DH (1988) Pathogenesis of murine cytomegalovirus infection: identification of infected cells in the spleen during acute and latent infections. J Virol 62:987–997Google Scholar
- 53.Seckert CK, Schader SI, Ebert S, Thomas D, Freitag K, Renzaho A, Podlech J, Reddehase MJ, Holtappels R (2011) Antigen-presenting cells of haematopoietic origin prime cytomegalovirus-specific CD8 T cells but are not sufficient for driving memory inflation during viral latency. J Gen Virol 92:1994–2005CrossRefGoogle Scholar