title: In Situ Staining and Laser Capture Microdissection of Lymph Node Residing SIV Gag-Specific CD8+ T cells--A Tool to Interrogate a Functional Immune Response Ex Vivo.
abstract: While a plethora of data describes the essential role of systemic CD8+ T cells in the control of SIV replication little is known about the local in situ CD8+ T cell immune responses against SIV at the intact tissue level, due to technical limitations. In situ staining, using GagCM9 Qdot 655 multimers, were here combined with laser capture microdissection to detect and collect SIV Gag CM9 specific CD8+ T cells in lymph node tissue from SIV infected rhesus macaques. CD8+ T cells from SIV infected and uninfected rhesus macaques were also collected and compared to the SIV GagCM9 specific CD8+ T cells. Illumina bead array and transcriptional analyses were used to assess the transcriptional profiles and the three different CD8+ T cell populations displayed unique transcriptional patterns. This pilot study demonstrates that rapid and specific immunostaining combined with laser capture microdissection in concert with transcriptional profiling may be used to elucidate phenotypic differences between CD8+ T cells in SIV infection. Such technologies may be useful to determine differences in functional activities of HIV/SIV specific T cells.
authors.affiliation: Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, L8:01, 17176 Stockholm, Sweden.
authors.forename: Annelie
authors.initials: A
authors.lastname: Tjernlund
authors.affiliation: Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, L8:01, 17176 Stockholm, Sweden.|National Laboratory for HIV Immunology, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.|Department of Medical Microbiology, University of Manitoba, 730 William Ave. Winnipeg, MB, Canada.
authors.forename: Adam
authors.initials: A
authors.lastname: Burgener
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, Stockholm, Sweden.
authors.forename: Jessica M
authors.initials: JM
authors.lastname: Lindvall
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States of America.|Department of Medicine, University of Washington, Seattle, WA, United States of America.
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States of America.|Department of Laboratory Medicine, University of Washington, Seattle, WA, United States of America.
authors.forename: Jia
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, L8:01, 17176 Stockholm, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Öhrmalm
authors.affiliation: Department of Pathology, Vaccine and Gene Therapy Institute, and the Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, OR, United States of America.
authors.forename: Louis J
authors.initials: LJ
authors.lastname: Picker
authors.affiliation: Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, L8:01, 17176 Stockholm, Sweden.
authors.forename: Kristina
authors.initials: K
authors.lastname: Broliden
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States of America.
authors.forename: M Juliana
authors.initials: MJ
authors.lastname: McElrath
authors.affiliation: Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States of America.|Department of Medicine, University of Washington, Seattle, WA, United States of America.|Department of Laboratory Medicine, University of Washington, Seattle, WA, United States of America.
authors.forename: Lawrence
authors.initials: L
authors.lastname: Corey
mesh_terms: D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000473:pathology / Q000821:virology*; D015683:Gene Products, gag / Q000032:analysis* / Q000276:immunology; D061346:Laser Capture Microdissection / Q000379:methods*; D008198:Lymph Nodes / Q000276:immunology / Q000473:pathology / Q000821:virology*; D008253:Macaca mulatta / Q000276:immunology / Q000821:virology*; D016097:Simian Acquired Immunodeficiency Syndrome / Q000276:immunology* / Q000473:pathology / Q000821:virology; D015302:Simian Immunodeficiency Virus / Q000276:immunology / Q000302:isolation & purification*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D015683:Gene Products, gag