The Chemokine Receptor CXCR3 Promotes CD8 + T Cell Accumulation in Uninfected Salivary Glands but Is Not Necessary after Murine Cytomegalovirus Infection This information is current as of September 27, 2021. Sofia Caldeira-Dantas, Thomas Furmanak, Corinne Smith, Michael Quinn, Leyla Y. Teos, Adam Ertel, Drishya Kurup, Mayank Tandon, Ilias Alevizos and Christopher M. Snyder J Immunol published online 29 December 2017 http://www.jimmunol.org/content/early/2017/12/29/jimmun Downloaded from ol.1701272 Supplementary http://www.jimmunol.org/content/suppl/2017/12/29/jimmunol.170127 http://www.jimmunol.org/ Material 2.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! 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Published December 29, 2017, doi:10.4049/jimmunol.1701272 The Journal of Immunology The Chemokine Receptor CXCR3 Promotes CD8+ T Cell Accumulation in Uninfected Salivary Glands but Is Not Necessary after Murine Cytomegalovirus Infection Sofia Caldeira-Dantas,*,†,‡ Thomas Furmanak,* Corinne Smith,* Michael Quinn,* Leyla Y. Teos,x Adam Ertel,{ Drishya Kurup,* Mayank Tandon,x Ilias Alevizos,x and Christopher M. Snyder* Recent work indicates that salivary glands are able to constitutively recruit CD8+ T cells and retain them as tissue-resident memory T cells, independently of local infection, inflammation, or Ag. To understand the mechanisms supporting T cell recruit- ment to the salivary gland, we compared T cell migration to the salivary gland in mice that were infected or not with murine CMV (MCMV), a herpesvirus that infects the salivary gland and promotes the accumulation of salivary gland tissue-resident memory Downloaded from T cells. We found that acute MCMV infection increased rapid T cell recruitment to the salivary gland but that equal numbers of activated CD8+ T cells eventually accumulated in infected and uninfected glands. T cell recruitment to uninfected salivary glands depended on chemokines and the integrin a4. Several chemokines were expressed in the salivary glands of infected and uninfected mice, and many of these could promote the migration of MCMV-specific T cells in vitro. MCMV infection increased the expression of chemokines that interact with the receptors CXCR3 and CCR5, but neither receptor was needed for T cell recruitment to the salivary gland during MCMV infection. Unexpectedly, however, the chemokine receptor CXCR3 was critical for T cell accumu- http://www.jimmunol.org/ lation in uninfected salivary glands. Together, these data suggest that CXCR3 and the integrin a4 mediate T cell recruitment to uninfected salivary glands but that redundant mechanisms mediate T cell recruitment after MCMV infection. The Journal of Immunology, 2018, 200: 000–000. he salivary gland is an exocrine organ composed mostly of infect the salivary gland and use saliva as a major route of transmission acinar cells producing a water-based fluid, which is rich is to new hosts (1–3). How the immune system responds to pathogens in T mucus, ions, and enzymes, that is released in the oral cavity the salivary gland and prevents or limits viral shedding remain poorly via a network of epithelial ducts. Because of this, several viruses, defined. In addition, how pathogens alter the salivary gland envi- by guest on September 27, 2021 including all human betaherpesviruses and gammaherpesviruses, ronment by infection is unknown. CMV is a betaherpesvirus that infects most organs in the body *Department of Immunology and Microbiology, Thomas Jefferson University, Phil- but undergoes prolonged replication in the salivary gland. CMV adelphia, PA 19107; †Life and Health Sciences Research Institute (ICVS), School of infects the acinar and ductal epithelial cells in the salivary gland ‡ Medicine, University of Minho, 4710-057 Braga, Portugal; Life and Health Sciences and, thus, is shed into saliva during replication (4–6). Using the Research Institute (ICVS)/3B’s Associate Laboratory, 4710-057 Braga, Portugal; x Sjo¨gren’s Syndrome and Salivary Gland Dysfunction Unit, National Institute of mouse CMV (MCMV) model, it was shown .40 y ago Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD that lymphocytes enter the salivary gland, resulting in the death of 20892; and {Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107 MCMV-infected epithelial cells (7, 8). MCMV also establishes latency in the salivary gland and readily reactivates in this ORCIDs: 0000-0002-5467-4663 (S.C.-D.); 0000-0002-3804-3861 (T.F.); 0000-0001- 6548-9802 (C.S.); 0000-0002-5854-6730 (L.Y.T.); 0000-0001-7455-1559 (A.E.); tissue during periods of immune suppression, particularly when 0000-0002-3959-7700 (D.K.); 0000-0003-1370-7198 (C.M.S.). CD8+ T cells have been depleted (9). It might be assumed Received for publication September 5, 2017. Accepted for publication November 17, that lymphocytes are drawn to the salivary gland in response to the 2017. CMV infection. Interestingly, however, recent data from several This work was supported by National Institutes of Health Grant AI106810 (to C.M.S.) laboratories, including our own, suggested that Ag-stimulated and Portuguese Foundation for Science and Technology Grant SFRH-BD-52319-2013 + (to S.C.-D.). CD8 T cells could be recruited to the salivary gland constitu- The gene expression data presented in this article have been submitted to the National tively after MCMV infection or even without specific infection of Center for Biotechnology Information Gene Expression Omnibus (https://www.ncbi. the gland (10–12). MCMV-specific T cell populations were nlm.nih.gov/geo/) under accession number GSE107338. abundant in the salivary gland after MCMV infection where they Address correspondence and reprint requests to Dr. Christopher M. Snyder, Thomas Jefferson University, Department of Microbiology and Immunology, Sidney Kimmel adopted an intraepithelial localization and expressed CD69 and Cancer Center, 233 S. 10th Street, BLSB, Room 730, Philadelphia, PA 19107. E-mail CD103 (10, 11), markers of tissue-resident memory T cells (TRM). address: [email protected] TRM are memory T cell subsets that are retained in tissues inde- The online version of this article contains supplemental material. pendently of the circulating pool of T cells, thus providing a rapid Abbreviations used in this article: B6, C57BL/6; FDR, false discovery rate; f.p., defense against reactivation of a latent infection or local reinfec- footpad; GSEA, Gene Set Enrichment Analysis; i.n, intranasal; I.V.+, vasculature localized; I.V.2, parenchyma localized; KO, knockout; LCMV-WE, lymphocytic tion of a previously encountered pathogen (13, 14). Surprisingly, choriomeningitis virus strain WE; MCMV, mouse CMV; PTx, pertussis toxin; even in vitro–activated T cells could migrate to the salivary gland RNA-Seq, RNA sequencing; RT-qPCR, reverse transcriptase quantitative PCR; where they developed into protective TRM in the complete absence T , tissue-resident memory T cell; VACV, vaccinia virus; WT, wild-type. RM of infection or specific inflammation (10–12), leading to the de- + Copyright Ó 2017 by The American Association of Immunologists, Inc. 0022-1767/17/$35.00 scription of the salivary gland as a “sink” for CD8 TRM (10). www.jimmunol.org/cgi/doi/10.4049/jimmunol.1701272 2 CXCR3 PROMOTES T CELL ACCUMULATION IN NAIVE SALIVARY GLANDS It is unknown whether local inflammation or Ag can enhance the establish a mechanism for the surprisingly efficient recruitment of + recruitment or retention of CD8 TRM in the salivary gland. activated T cells to the salivary gland, even after a completely However, this ability of salivary glands to attract and retain pro- irrelevant infection. + tective numbers of CD8 TRM, without a local infection or in- flammation, is quite unexpected. In most other sites in the body, Materials and Methods tissue-localized inflammation and/or Ag is critical for the efficient Mice and infections recruitment of T cells or their retention as T (15–24). The best- RM All mice were purchased from The Jackson Laboratory and bred in-house. studied example of the interplay among Ag, inflammation, and C57BL/6 (B6), B6.SJL-PtprcaPepcb/BoyJ (CD45.1), B6.PL-Thy1a/CyJ tm1Ts TRM formation is the skin, where inflammation alone is sufficient (Thy1.1), and IFN-g–knockout (KO) (B6.129S7-Ifng /J) mice to enable T cell egress from the blood and formation of TRM were used as recipients in the adoptive-transfer experiments and to phenotype cells (22). Interestingly, although infection at one skin assess chemokine expression in the salivary gland. OT-I transgenic mice [B6-Tg(TcraTcrb)1100Mjb/J] were bred with CD45.1 mice, site could lodge T cells at distant skin locations (23, 25), the ef- CXCR3-KO mice (B6.129P2-Cxcr3tm1Dgen/J),