Neuron-Interacting Satellite Glial Cells in Human Trigeminal Ganglia Have an APC Phenotype This information is current as Monique van Velzen, Jon D. Laman, Alex KleinJan, of September 28, 2021. Angelique Poot, Albert D. M. E. Osterhaus and Georges M. G. M. Verjans J Immunol 2009; 183:2456-2461; Prepublished online 27 July 2009; doi: 10.4049/jimmunol.0900890 Downloaded from http://www.jimmunol.org/content/183/4/2456 References This article cites 44 articles, 14 of which you can access for free at: http://www.jimmunol.org/content/183/4/2456.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 28, 2021 • 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 © 2009 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Neuron-Interacting Satellite Glial Cells in Human Trigeminal Ganglia Have an APC Phenotype1 Monique van Velzen,* Jon D. Laman,†‡ Alex KleinJan,§ Angelique Poot,* Albert D. M. E. Osterhaus,* and Georges M. G. M. Verjans2* Satellite glial cells (SGC) in sensory ganglia tightly envelop the neuronal cell body to form discrete anatomical units. This type of glial cell is considered neuroectoderm-derived and provides physical support to neuron somata. There are scattered hints in the literature suggesting that SGC have an immune-related function within sensory ganglia. In this study, we addressed the hypothesis of human trigeminal ganglia (40 ؍ that SGC are tissue-resident APC. The immune phenotype and function of a large series (n (TG) were assessed by detailed flow cytometry, in situ analyses, and functional in vitro assays. Human TG-resident SGC (TG- SGC) uniformly expressed the common leukocyte marker CD45, albeit at lower levels compared with infiltrating T cells, and the macrophage markers CD14, CD68, and CD11b. In addition, TG-SGC expressed the myeloid dendritic cell (DC) marker CD11c, Downloaded from the T cell costimulatory molecules CD40, CD54, CD80, and CD86 and MHC class II. However, the mature DC marker CD83 was absent on TG-SGC. Functionally, TG-SGC phagocytosed fluorescent bacteria, but were unable to induce an allogeneic MLR. Finally, TG-infiltrating T cells expressed the T cell inhibitory molecules CD94/NKG2A and PD-1, and the interacting TG-SGC expressed the cognate ligands HLA-E and PD-L1, respectively. In conclusion, the data demonstrate that human TG-SGC have a unique leukocyte phenotype, with features of both macrophages and immature myeloid DC, indicating that they have a role as TG-resident APC with potential T cell modulatory properties. The Journal of Immunology, 2009, 183: 2456–2461. http://www.jimmunol.org/ ensory ganglia are part of the peripheral nervous system. geminal ganglion (TG), and reactivates intermittently (6). Re- They contain cell bodies of sensory neurons establishing cent studies in mice and humans emphasized the importance of S the connection between the periphery and CNS. Sensory infiltrating T cells to control latent HSV infections in sensory ganglia lack a blood-nerve barrier and enclose a high number of ganglia (7–9). Virus-specific T cells are directly juxtaposed to satellite glial cells (SGC)3 (1–3). SGC are considered to be neu- latently infected neurons, produce cytokines and cytolytic ef- roectoderm-derived and involved in the maintenance of sensory fector molecules, but do not induce neuronal damage (7, 8, neuron homeostasis by regulating extracellular ion and nutrient 10–12). Current data suggest that the neurons themselves or by guest on September 28, 2021 levels within sensory ganglia (2). In contrast to CNS-resident glial hitherto unrecognized resident cells in latently infected sensory cells, like astrocytes and microglia, SGC have a distinct interaction ganglia induce and coordinate this nonpathogenic chronic T cell with neurons (2, 3). They directly associate with the neuronal response (8, 10–12). soma, so that each neuronal cell body is completely surrounded by In this study, we addressed the hypothesis that SGC are tissue- a sheet of several SGC providing physical support and a protective resident APC. The availability of a series of fresh postmortem barrier (3). The numerous fine invaginations between the neuron human TG specimens enabled us to combine ex vivo and in situ and SGC sheath illustrate their intimate association (2, 3). Upon analyses for the phenotypic and functional characterization of hu- mechanical injury to sensory neurons, SGC undergo morphologi- man TG-resident SGC (TG-SGC). cal changes, proliferate, and up-regulate a variety of growth fac- tors, cytokines, and the glial marker glial fibrillary acidic protein Materials and Methods (2, 4, 5). Clinical specimens Human ␣-herpesviruses, like HSV, are a common threat to Heparinized peripheral blood and TG specimens, i.e., left and right TG, human sensory ganglia. HSV establishes a lifelong latent infec- were obtained from 40 subjects (median age 79 years, range 41–94 tion in neurons within sensory ganglia, predominantly the tri- years) at autopsy with a mean postmortem interval of 6 h (range 2.5– 15.5 h). The TG tissue panel consisted of 34 donors with a CNS disease (mainly Alzheimer’s disease and Parkinson’s disease) and six donors † ‡ without evidence of CNS disease. The cause of death was not related to *Department of Virology, Department of Immunology, MS Center ErasMS, and ␣-herpesvirus infections. No significant differences in the immunolog- §Pulmonary Medicine, Erasmus Medical Center, Rotterdam, The Netherlands ical parameters analyzed were detected between donors with or without Received for publication May 26, 2009. Accepted for publication June 9, 2009. a history of CNS disease (data not shown). Specimens were either snap- The costs of publication of this article were defrayed in part by the payment of page frozen (n ϭ 23) or transferred to tubes (n ϭ 17) containing culture charges. This article must therefore be hereby marked advertisement in accordance medium consisting of RPMI 1640 (Lonza) supplemented with heat- with 18 U.S.C. Section 1734 solely to indicate this fact. inactivated 10% FBS (Greiner) and antibiotics. Written informed con- 1 This study was supported in part by the International Consortium on Anti-Virals (to sent from the donor or next of kin was obtained. The local ethical M.v.V.) and the Dutch MS Research Foundation (to J.D.L.). committees approved the study, which was conducted according to the tenets of the Declaration of Helsinki. 2 Address correspondence and reprint requests to Dr. Georges M.G.M. Verjans, De- partment of Virology, Room Ee1720a, Erasmus Medical Center, s-Gravendijkwal Generation of TG single cell suspensions 230, 3015 CE Rotterdam, the Netherlands. E-mail address: [email protected] 3 Abbreviations used in this paper: SGC, satellite glial cell; DC, dendritic cell; TG, Generation of single cell suspensions from human TG was performed es- trigeminal ganglia; PD, programmed death; PD-L1, PD ligand l. sentially as previously described (12). In brief, the TG were fragmented and subsequently treated with Liberase Blendzyme 3 (0.2 U/ml, Roche) at Copyright © 2009 by The American Association of Immunologists, Inc. 0022-1767/09/$2.00 37°C for 1 h. Dispersed cells were filtered through a 70-␮m pore size cell www.jimmunol.org/cgi/doi/10.4049/jimmunol.0900890 The Journal of Immunology 2457 strainer (BD Biosciences), and the flow-through was collected in PBS 488 nm argon, and 561 nm diode laser to detect DAPI, fluorescein, and containing 1% FBS. From the same donor, PBMC were isolated from Alexa Fluor 610-PE, respectively. heparinized peripheral blood (ϳ4 ml per donor) by density gradient centrifugation on Ficoll-Hypaque (12). Donor PBMC and TG single cell Allogeneic MLR assay suspensions were directly used for phenotypic and functional analyses. CD14-enriched TG-SGC, peripheral blood-derived monocytes and mature Flow cytometry dendritic cells (DC) were used as stimulator cells in allogeneic MLR as- says. Due to the low number of monocytes recovered from TG donors’ Donor-matched PBMC and TG cells were subjected to multicolor flow cyto- PBMC, mature DC were generated from peripheral blood samples of metric analyses using the following fluorochrome-conjugated mAbs: CD3- healthy blood donors (n ϭ 2). To obtain mature DC, CD14-enriched pe- allophycocyanin (UCHT1; DakoCytomation), CD11b-PE (Bear-1; Beckman ripheral blood-derived monocytes were cultured with IL-4 and GM-CSF Coulter), CD11c-allophycocyanin (S-HCL3; BD Biosciences), CD14-FITC for 6 days to generate immature monocyte-derived DC, and subsequently (TU¨ K4; DakoCytomation), CD40-FITC (5C3; BD Biosciences), CD45-PerCP matured with a cytokine mixture as previously described (13, 14). The (2D1; BD Biosciences), CD54-FITC (6.5B5; DakoCytomation), CD68-PE mature DC phenotype, characterized by high CD80, CD83, and CD86 ex- (Y1/82A; BD Biosciences), CD80-FITC (MAB104; Beckman Coulter), pression (15), was confirmed by flow cytometry (data not shown). The CD83-allophycocyanin (HB15e; BD Biosciences), CD86-PE (FUN-1; effector cells, i.e., allogeneic peripheral blood T cells, were labeled with BD Biosciences), HLA-DR-PerCP (L243; BD Biosciences), CD94- CFSE (Invitrogen) at a final concentration of 0.5 ␮M. The stimulator cells FITC (DX22; eBioscience), NKG2A-allophycocyanin (131411; eBio- were cocultured with effector cells at a ratio of 1:10 at 37°C.