Enumeration of human lymphocyte subsets by monoclonal antibodies and flow cytometry: a comparative study using whole blood or mononuclear cells separated by density gradient centrifugation

Paolo De Paoli, Michele Reitano, Sandro Battistin, Carla Castiglia, Gianfranco Santini

Research output: Contribution to journalArticlepeer-review

Abstract

Enumeration of lymphocyte subpopulations by combined use of flow cytometry and monoclonal antibodies is influenced by the separation method used. T or B lymphocyte antigen frequencies do not differ in samples of whole blood or after separation on Ficoll-Paque or Percoll, but there is a significant increase of Leu7+ Leu11+ lymphocytes showing strong natural killer activity at the sample-separation medium interface. At the bottom of the tubes a selective loss of OKT8+, Leu11- cells is found; at this level cytotoxicity is very low. Our data suggest that Leu7+ cells could be subdivided into 2 subpopulations differing in reactivity with the monoclonal antibody to Leu11, natural killer activity and density. Differences observed in the percentages of Leu7+, Leu11+ lymphocytes between whole blood and separated cells could be due to an enrichment produced by centrifugation or to the presence in whole blood of a factor interfering with antibody binding.

Original languageEnglish
Pages (from-to)349-353
Number of pages5
JournalJournal of Immunological Methods
Volume72
Issue number2
DOIs
Publication statusPublished - Sep 4 1984

Keywords

  • density gradient centrifugation
  • lymphocytes
  • monoclonal antibodies
  • whole blood

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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