title: Evaluating frequency and quality of pathogen-specific T cells.
abstract: It is generally accepted that enumeration and characterization of antigen-specific T cells provide essential information about potency of the immune response. Here, we report a new technique to determine the frequency and potency of antigen-specific CD8 T cells. The assay measures changes of intracellular Ca2+in real time by fluorescent microscopy in individual CD8 T cells responding to cognate peptides. The T cells form continuous monolayer, enabling the cells to present the peptides to each other. This approach allows us to evaluate the kinetics of intracellular Ca2+signalling that characterizes the quality of T cell response. We demonstrate the usefulness of the assay examining the frequency and quality of cytomegalovirus-specific CD8 T cells from healthy donor and patient after haploidentical stem cell transplantation. The new assay has a potential to provide essential information determining the status of the immune system, disease morbidity, potency of therapeutic intervention and vaccine efficacy.
authors: Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors: Nadia
authors: N
authors: Anikeeva
authors: Department of Medical Oncology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors: Dolores
authors: D
authors: Grosso
authors: Department of Medical Oncology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.|The Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors: Neal
authors: N
authors: Flomenberg
authors: Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.|Department of Medical Oncology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.|The Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors: Yuri
authors: Y
authors: Sykulev
mesh_terms: D000956:Antigens, Viral / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D002118:Calcium / Q000276:immunology / Q000378:metabolism; D003587:Cytomegalovirus / Q000276:immunology* / Q000502:physiology; D003586:Cytomegalovirus Infections / Q000276:immunology* / Q000821:virology; D058501:Enzyme-Linked Immunospot Assay; D054884:Host-Pathogen Interactions / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D033581:Stem Cell Transplantation
chemical_list: D000956:Antigens, Viral; D007371:Interferon-gamma; D002118:Calcium