Glucuronoxylomannan in the Cryptococcus species capsule as a target for Chimeric Antigen Receptor T-cell therapy |
| |
| Authors: | Thiago Aparecido da Silva Paul J Hauser Irfan Bandey Tamara Laskowski Qi Wang Amer M Najjar Pappanaicken R Kumaresan |
| |
| Institution: | 1. Deparment of Pediatric Research, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA;2. Department of Cellular and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil;3. Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA;4. Department of Lymphoma and Myeloma, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA;1. Fundación Institut Guttmann, Institut Universitari de Neurorehabilitació adscrit a la UAB, Hospital de Neurorehabilitació, Barcelona, Spain;2. Universitat Autònoma de Barcelona, Barcelona, Spain;3. Fundació Institut d''Investigació en Ciències de la Salut Germans Trias i Pujol, Barcelona, Spain;4. Research and Education, Banc de Sang i Teixits, Barcelona, Spain;5. Cell Therapy Service, Banc de Sang i Teixits, Barcelona, Spain;6. Cellular Laboratory, Banc de Sang i Teixits, Barcelona, Spain;7. Department of Cell Biology, Physiology and Immunology, Institute of Neurosciences, Universitat Autònoma de Barcelona, Barcelona, Spain;1. Brain Repair Group, School of Biosciences, Cardiff University, Cardiff, UK;2. Department of Anatomy, Faculty of Medical Science, Naresuan University, Phisanulok, Thailand;3. Cardiff School of Sport and Health Sciences, Cardiff Metropolitan University, Cardiff, UK;4. MRC Centre for Neuropsychiatric Genetics and Genomics, School of Medicine, Cardiff University, Cardiff, UK;5. Wales Brain Repair and Intracranial Neurotherapeutics Unit, School of Medicine, Cardiff University, Cardiff, UK;1. Centre for Parasitology and Mycology, Instituto Adolfo Lutz, Avenida Dr. Arnaldo, 355, 01246-000 São Paulo, Brazil;2. Department of Pharmaceutical Sciences, Universidade Federal de São Paulo, Rua São Nicolau 210, 09913-030 Diadema, SP, Brazil;3. Biological Sciences Department, Universidade Federal de São Paulo, Rua São Nicolau 210, 09913-030 Diadema, SP, Brazil;4. Federal University of Mato Grosso do Sul, School of Medicine, MS, Brazil;1. Department of Dermatology, Venereology, and Allergology, University Hospital Würzburg, Würzburg, Germany;2. TICEBA GmbH, Heidelberg, Germany;3. Department of Surgery, University Hospital Würzburg, Würzburg, Germany;4. Renal Division, Brigham and Women''s Hospital, Harvard Medical School, Boston, Massachusetts, USA;5. RHEACELL GmbH & Co. KG, Heidelberg, Germany;6. Department of Dermatology, Brigham and Women''s Hospital, Harvard Medical School, Boston, Massachusetts, USA;7. Division of Plastic Surgery, Brigham and Women''s Hospital, Harvard Medical School, Boston, Massachusetts, USA;8. Department of Medicine, VA Boston Healthcare System, Boston, Massachusetts, USA;9. Division of Genetics, Brigham and Women''s Hospital, Harvard Medical School, Boston, Massachusetts, USA;10. Transplant Research Program, Boston Children''s Hospital, Harvard Medical School, Boston, Massachusetts, USA;11. Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, USA;12. Department of Dermatology and Allergic Diseases, University Hospital, Ulm, Germany;13. School of Medical and Health Sciences, Edith Cowan University, Perth, Australia |
| |
| Abstract: | Background aimsThe genus Cryptococcus comprises two major fungal species that cause clinical infections in humans: Cryptococcus gattii and Cryptococcus neoformans. To establish invasive human disease, inhaled cryptococci must penetrate the lung tissue and reproduce. Each year, about 1 million cases of Cryptococcus infection are reported worldwide, and the infection's mortality rate ranges from 20% to 70%. Many HIV+/AIDS patients are affected by Cryptococcus infections, with 220,000 cases of cryptococcal meningitis reported worldwide in this population every year (C. neoformans infection statistics, via the Centers for Disease Control and Prevention, https://www.cdc.gov/fungal/diseases/cryptococcosis-neoformans/statistics.html). To escape from host immune cell attack, Cryptococcus covers itself in a sugar-based capsule composed primarily of glucuronoxylomannan (GXM). To evade phagocytosis, yeast cells increase to a >45-µm perimeter and become titan, or giant, cells. Cryptococci virulence is directly proportional to the percentage of titan/giant cells present during Cryptococcus infection. To combat cryptococcosis, the authors propose the redirection of CD8+ T cells to target the GXM in the capsule via expression of a GXM-specific chimeric antigen receptor (GXMR-CAR).ResultsGXMR-CAR has an anti-GXM single-chain variable fragment followed by an IgG4 stalk in the extracellular domain, a CD28 transmembrane domain and CD28 and CD3-? signaling domains. After lentiviral transduction of human T cells with the GXMR-CAR construct, flow cytometry demonstrated that 82.4% of the cells expressed GXMR-CAR on their surface. To determine whether the GXMR-CAR+ T cells exhibited GXM-specific recognition, these cells were incubated with GXM for 24 h and examined with the use of brightfield microscopy. Large clusters of proliferating GXMR-CAR+ T cells were observed in GXM-treated cells, whereas no clusters were observed in control cells. Moreover, the interaction of GXM with GXMR-CAR+ T cells was detected via flow cytometry by using a GXM-specific antibody, and the recognition of GXM by GXMR-CAR T cells triggered the secretion of granzyme and interferon gamma (IFN-γ). The ability of GXMR-CAR T cells to bind to the yeast form of C. neoformans was detected by fluorescent microscopy, but no binding was detected in mock-transduced control T cells (NoDNA T cells). Moreover, lung tissue sections were stained with Gomori Methenamine Silver and evaluated by NanoZoomer (Hamamatsu), revealing a significantly lower number of titan cells, with perimeters ranging from 50 to 130 µm and giant cells >130 µm in the CAR T-cell treated group when compared with other groups. Therefore, the authors validated the study's hypothesis by the redirection of GXMR-CAR+ T cells to target GXM, which induces the secretion of cytotoxic granules and IFN-γ that will aid in the control of cryptococcosisConclusionsThus, these findings reveal that GXMR-CAR+ T cells can target C. neoformans. Future studies will be focused on determining the therapeutic efficacy of GXMR-CAR+ T cells in an animal model of cryptococcosis. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
