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Changes in Virus-Specific Precursors Are Due to Quantitative And Defective CD8 T Cell Responses in Aged Mice Are Due to Quantitative and Qualitative Changes in Virus-Specific Precursors This information is current as Vilma Decman, Brian J. Laidlaw, Travis A. Doering, Jin of October 1, 2021. Leng, Hildegund C. J. Ertl, Daniel R. Goldstein and E. John Wherry J Immunol 2012; 188:1933-1941; Prepublished online 13 January 2012; doi: 10.4049/jimmunol.1101098 http://www.jimmunol.org/content/188/4/1933 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2012/01/13/jimmunol.110109 Material 8.DC1 http://www.jimmunol.org/ References This article cites 55 articles, 23 of which you can access for free at: http://www.jimmunol.org/content/188/4/1933.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on October 1, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2012 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Defective CD8 T Cell Responses in Aged Mice Are Due to Quantitative and Qualitative Changes in Virus-Specific Precursors Vilma Decman,*,† Brian J. Laidlaw,*,† Travis A. Doering,*,† Jin Leng,‡,x Hildegund C. J. Ertl,{ Daniel R. Goldstein,‡,x and E. John Wherry*,† Aging is associated with suboptimal CD8 T cell responses to viral infections. It is not clear whether these poor responses are due to environmental influences or quantitative and qualitative changes in the pool of responding CD8 T cells. Our studies demonstrated several deleterious age-related changes in the pool of Ag-specific CD8 T cells that respond to infection. The majority of CD8 T cells from uninfected aged mice was CD44Hi and had increased expression of inhibitory receptors including PD1, LAG3, 2B4, and CD160. These aged CD44Hi CD8 T cells were transcriptionally similar to exhausted CD8 T cells found during chronic infections. In addition, the number of virus-specific precursors in aged mice prior to infection was decreased up to 10-fold, and many of these Downloaded from Ag-specific precursors had high expression of CD44 and PD1. Finally, TCR transgenic studies demonstrated that the CD44Hi Ag- specific CD8 T cells from unimmunized aged and young mice were qualitatively inferior compared with CD44Lo CD8 T cells from aged or young donors. Thus, a decrease in precursor frequency as well as qualitative changes of CD8 T cells during aging are directly related to impaired immunity. The Journal of Immunology, 2012, 188: 1933–1941. ging is associated with a decline in T cell function and crease in homeostatic proliferation of the existing T cell pool. http://www.jimmunol.org/ an increase in susceptibility to infections. Some of the At least in young mice, T cells generated during lymphopenia- immunological changes associated with aging include induced homeostatic proliferation can have increased functional- A Lo reduced thymic output, a decrease in naive T cells, expansion of ity after in vivo Ag stimulation compared with naive CD44 memory phenotype CD8 T cells, cellular senescence, changes in T cells (9), and these homeostatically induced CD44Hi T cells telomerase expression or telomere length, reduced proliferation, provided protective immunity to bacterial infections (10). How- altered signaling, and reduced IL-2 production (1–5). Two prom- ever, two subpopulations of proliferating T cells exist under inent age-associated changes, the involution of the thymus and lymphopenic conditions, a slow proliferative subset, and a subset a global shift of T cell phenotype from naive to memory, lead that rapidly proliferates to CFSE negative (11). Interestingly, the to a decrease in the number of naive T cells (CD44Lo) and an rapidly proliferative subset is thought to be driven by environ- by guest on October 1, 2021 increase in the proportion of T cells with memory phenotype mental and/or self-Ags and upregulates the inhibitory receptor (CD44Hi) (6, 7). Several studies showed that the transfer of programmed death-1 (PD1) (12). It is unclear, however, if changes 2 2 CD44Lo CD8 T cells into RAG-1 / recipients (lymphopenic in the T cell compartment prior to infection, whether driven by environment) results in the transition to a CD44Hi memory phe- lymphopenia or other signals, impact antiviral immunity. notype within 3 wk. Using adoptive transfer of different numbers A diverse repertoire of naive T cells is essential for a vigorous 2 2 of naive (CD44Lo) cells were transferred to syngeneic RAG-1 / response to infection (13, 14). Reduced thymic output can result recipients, the transition to a memory (CD44Hi) phenotype was in a decline in naive Ag-specific precursors, and this decline can demonstrated to be directly proportional to the extent of homeo- impact the generation of an effective immune response (15, 16). static proliferation (8), suggesting that the gradual decline in new Moreover, clonal expansions of memory phenotype T cells have naive T cell production during aging could lead to a gradual in- been observed in mice, primates, and humans (17). These ex- pansions are believed to be driven by chronic infections, altered *Department of Microbiology, University of Pennsylvania Perelman School of Med- cytokine milieu of the host, or both and could further restrict the icine, Philadelphia, PA 19104; †Institute for Immunology, University of Pennsylvania T cell repertoire and/or compete with other T cell responses (18). Perelman School of Medicine, Philadelphia, PA 19104; ‡Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06510; xDepartment Thus, changes in the global T cell pool and/or changes in T cell of Immunobiology, Yale University School of Medicine, New Haven, CT 06510; and competition might account for some changes in T cell immunity { Immunology Program, Wistar Institute, Philadelphia, PA 19104 observed with age. Received for publication April 15, 2011. Accepted for publication December 13, Although less well documented, cell-intrinsic changes of naive 2011. T cells could also contribute to impaired immunity with aging. This work was supported by the American Heart Association (to V.D.), National Institutes of Health Grants HHSN266200500030C (to E.J.W.) and R01AG028082 Recently, several studies observed phenotypic alterations in the (to D.R.G.), and the Ellison Medical Foundation (to E.J.W.). CD4 T cell pool in aged mice including changes in the expression Address correspondence and reprint requests to Dr. E. John Wherry, University of of several molecules such as PD1, ICOS, CTLA4, and KLRG1, Pennsylvania Perelman School of Medicine, 421 Curie Boulevard, Room 312, Phil- although the consequences of these phenotypic changes remain adelphia, PA 19104. E-mail address: [email protected] poorly understood (19–21). Recent studies also demonstrated that The online version of this article contains supplemental material. decreased thymic output with age leads to an accumulation of Abbreviations used in this article: GSEA, Gene Set Enrichment Analysis; MNV, long-lived naive CD4 T cells that express low amounts of Bim and mouse norovirus; PD1, programmed death-1. have functional defects (22). Less information is available re- Copyright Ó 2012 by The American Association of Immunologists, Inc. 0022-1767/12/$16.00 garding the impact of aging on the naive CD8 T cell pool. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1101098 1934 DEFECTIVE ANTIVIRAL CD8 T CELL PRECURSORS DURING AGING In this study, we demonstrated that aging in mice was associated were done as described previously (9, 25). CD8 and CD3 were used as with an increase in the expression of several inhibitory receptors positive gates. B220, CD19, CD4, CD11b, CD11c, and MHC II were used including PD1, LAG3, 2B4, and CD160 on CD8 T cells. These as dump gates. In addition to tetramer staining, samples were stained for CD44 and PD1. changes were especially pronounced on the CD44Hi population of CD8 T cells from uninfected aged mice with changes first be- Flow cytometry, intracellular cytokine staining, and BrdU coming apparent sometime between 8 and 12 mo of age concur- staining Lo Hi rent with a shift from CD44 to predominantly CD44 T cells. Lymphocytes were stained using standard techniques and analyzed by flow We also demonstrated that the precursor frequency of virus- cytometry. Virus-specific CD8 T cells were quantified using MHC class I specific CD8 T cells from uninfected aged mice declines by peptide tetramer staining. MHC class I peptide tetramers were made and $10-fold. Moreover, aging resulted not just in a decreased pre- used as described previously (23). Abs to CD8, CD44, PD1 (RPMI-30), CD3ε (145-2C11), and CD28 (37.51) were purchased from BioLegend cursor frequency but also in changes in the quality of these pre- (San Diego, CA). Abs to LAG3, 2B4, CD160, and IFN-g were purchased cursors. Many Ag-specific CD8 T cell precursors were CD44Hi from eBioscience (San Diego, CA). Abs to CD4, Mip1a, KLRG1, and and PD1 positive. Transcriptional profiling of total CD44Hi CD8 Tbet were purchased from Invitrogen (Carlsbad, CA), R&D Systems T cells from naive aged mice revealed strong similarities to (Minneapolis, MN), Beckman Coulter (Fullerton, CA), or Santa Cruz Biotechnology (Santa Cruz, CA), respectively. All other Abs were pur- exhausted CD8 T cells found during chronic infections. Studies chased from BD Biosciences (San Diego, CA).
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