|     home    |     search     |     content     |     mail     |     login     |

study information

date of publication1-jul-2000
journalClin Diagn Lab Immunol 2000 Jul;7(4):687-92
titleNonradioactive techniques for measurement of in vitro T-cell proliferation: alternatives to the [(3)H]thymidine incorporation assay.
title in Dutch
resp. centreCLB
designT-cell proliferation is an important in vitro parameter of in vivo immune function and has been used as a prognostic marker of human immunodeficiency virus type 1 (HIV-1) disease progression. The proliferative capacity of T cells in response to various stimuli is commonly determined by a radioactive assay based on incorporation of [(3)H]thymidine ([(3)H]TdR) into newly generated DNA. In order to assess techniques for application in laboratories where radioactive facilities are not present, two alternative methods were tested and compared to the [(3)H]TdR assay as a "gold standard." As an alternative, T-cell proliferation was measured by flow cytometric assessment of CD38 expression on T cells and by an enzyme-linked immunosorbent assay (ELISA) based on bromo-2'-deoxyuridine (BrdU) incorporation. Peripheral blood mononuclear cells (PBMCs), either in whole blood or Ficoll-Isopaque separated, from a total of 26 HIV-1-positive and 18 HIV-1-negative Dutch individuals were stimulated with CD3 monoclonal antibody (MAb) alone, a combination of CD3 and CD28 MAbs, or phytohemagglutinin. BrdU incorporation after 3 days of stimulation with a combination of CD3 and CD28 MAbs correlated excellently with the [(3)H]TdR incorporation in both study groups (HIV-1 positives, r = 0.96; HIV-1 negatives, r = 0.83). A significant correlation of absolute numbers of T cells expressing CD38 with [(3)H]TdR incorporation, both in HIV-1-positive (r = 0.96) and HIV-1-negative (r = 0.84) individuals, was also observed under these conditions. The results of this study indicate that determination of both the number of CD38-positive T cells and BrdU incorporation can be used as alternative techniques to measure the in vitro T-cell proliferative capacity. The measurement of CD38 expression on T cells provides the additional possibility to further characterize the proliferating T-cell subsets for expression of other surface markers.

Use this form to to view general information about a study.
  • click on 'authors' to view the list of authors who contributed
  • click on 'files' to view a list of files, associated with this study: these files can be of any type: f.i. the full text of the article, or code used in the analysis, or a Powerpoint demonstration.
  • click on 'tables' to view a list of the database tables that were used in this study: more is information on these tables is available.
  • for studies related to the Amsterdam Cohort, the option 'progress' is available: it gives information about the process of collecting information about a study: who was asked for information, when was this done, is the information complete, etc.
  • if you have the authority to change data about the study, the option 'edit' is available: clicking this will take you to a form for editing the main information, with options to edit all aspects.
  • clicking on the PubMedID opens a new browser window with PubMed
  • clicking on the 'resp. centre' gives a list of all studies for this centre
  • click the 'back' button to go to the previous document