Identification of functional subsets by flow cytometry: Intracellular detection of cytokine expression

Vernon C. Maino, Louis Picker

Research output: Contribution to journalArticle

188 Citations (Scopus)

Abstract

Methods for analysis of T cell function have traditionally relied upon measurements of proliferation or cytokine expression in bulk cultures of PBMC in long term incubations with polyclonal mitogens or putative antigen. These techniques suffer from the drawback that they do not enable analysis of single cell responses in the context of unselected cellular backgrounds. In addition these methods are not sensitive enough to rapidly assess rare event responses characteristic of cognate memory T cell responses. This review discusses recently developed flow cytometric methods designed to rapidly assess leukocyte subset cytokine responses to polyclonal activators and specific antigen in PBMC and whole blood samples. These procedures determine the percentages of activated cells and the identification of leucocyte subsets capable of expressing various cytokines and cell surface antigens. The ability to assess key intracellular functional markers by multiparameter flow cytometry offers some unique advantages in a number of clinical applications. The technical simplicity and rapidity of the flow cytometric intracellular cytokine detection techniques described in this report, as well as the widespread availability of appropriate flow cytometers and cell surface directed antibodies in clinical laboratories, suggests the possibility that this technique could be broadly applicable to the clinical evaluation of immune status. Since any cell type can be identified with this approach, responses to a variety of clinically relevant stimuli in virtually any leukocyte subset can be evaluated including monocyte responses to LPS, and T cell responses to mitogens and a variety of bacterial and viral antigens. The significance of measuring low frequency antigen-specific responses with respect to clinical significance in assessing immune status in a variety of clinical conditions and determining efficacy or immunotoxicity of drugs and vaccine antigens is discussed.

Original languageEnglish (US)
Pages (from-to)207-215
Number of pages9
JournalCommunications in Clinical Cytometry
Volume34
Issue number5
DOIs
StatePublished - Oct 15 1998
Externally publishedYes

Fingerprint

Flow Cytometry
Cytokines
Antigens
Leukocytes
T-Lymphocytes
Mitogens
Single-Cell Analysis
Bacterial Antigens
Viral Antigens
Surface Antigens
Monocytes
Vaccines
Antibodies
Pharmaceutical Preparations

Keywords

  • Activation
  • Antigen
  • Cytokines
  • Flow cytometry
  • Intracellular

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Identification of functional subsets by flow cytometry : Intracellular detection of cytokine expression. / Maino, Vernon C.; Picker, Louis.

In: Communications in Clinical Cytometry, Vol. 34, No. 5, 15.10.1998, p. 207-215.

Research output: Contribution to journalArticle

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