Document 0084
 DOCN  M9580084
 TI    Quantification of antigen specific CD8+ T cells using an ELISPOT assay.
 DT    9506
 AU    Miyahira Y; Murata K; Rodriguez D; Rodriguez JR; Esteban M; Rodrigues
       MM; Zavala F; Department of Medical and Molecular Parasitology, New
       York; University Medical Center, NY 10010, USA.
 SO    J Immunol Methods. 1995 Apr 12;181(1):45-54. Unique Identifier :
       AIDSLINE MED/95248136
 AB    An ELISPOT assay to detect and determine the number of antigen specific
       CD8+ T cells was standardized using cloned murine CD8+ T cells specific
       for the epitope SYVPSAEQI of a rodent malaria antigen. This assay is
       based on the detection of IFN-gamma secretion by single cells after
       their stimulation with antigen. The interferon secretion is visualized
       as spots revealed by using enzyme labeled anti-IFN-gamma monoclonal
       antibodies. Using known numbers of cloned murine CD8+ T cells it was
       determined that the assay detects 80-95% of these CD8+ T cells. The
       optimal culture conditions for the stimulation of the CD8+ T cells were
       determined and the antigen concentration, number of antigen presenting
       cells and supplement of growth factors required to perform the assay
       were defined. This ELISPOT assay can be performed with spleen cells from
       immunized mice, and provide the precise number of antigen specific CD8+
       T cells present in mixed lymphocyte populations. This method is more
       sensitive than the chromium-51 release assay, and much simpler than the
       conventional precursor frequency analysis, providing the number of
       antigen specific CD8+ T cells in 36-48 h.
 DE    Amino Acid Sequence  Animal  Antigenic Determinants/ANALYSIS  Antigens,
       Protozoan/*ANALYSIS  CD8-Positive T-Lymphocytes/*IMMUNOLOGY
       Enzyme-Linked Immunosorbent Assay/*METHODS  Interferon Type II/ANALYSIS
       Lymphocyte Count  Lymphocyte Transformation  Malaria/*IMMUNOLOGY  Mice
       Mice, Inbred BALB C  Molecular Sequence Data  Plasmodium
       yoelii/*IMMUNOLOGY  Sensitivity and Specificity  Support, U.S. Gov't,
       Non-P.H.S.  Support, U.S. Gov't, P.H.S.  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).