In Vitro Nickel-Specific T-Lymphocyte Proliferation: Methodological Aspects

  • E. P. Prens
  • K. Benne
  • T. van Joost
  • R. Benner
Conference paper

Summary

The conditions necessary for optimal and reproducible test results in the nickel specific in vitro T-lymphocyte proliferation assay (NiPA) were studied. For this purpose peripheral blood mononuclear cells (PBMCs) of nickel-allergic subjects were isolated by density-gradient centrifugation and, when necessary, further separated into T cells and non-T cells. The influence of basic variables such as the number of cells per well, culture medium, serum-type and the type of culture dishes (round- or flat-bottomed) on the result of this assay, were investigated in parallel experiments. Furthermore, in this study enhancement of Ni-specific T-lymphocyte proliferation was attempted by the addition of epidermal cells from normal or patch-tested skin areas, recombinant IL-2 after antigen priming or pre-treatment of the cells with neuraminidase. Optimal and reproducible proliferation was obtained in cultures set up with 2 × 105 PBMCs or with purified T cells in the presence of 4–8 × 104 epidermal cells from patch-tested skin, using RPMI 1640 with HEPES and 15% pooled human AB serum in round-bottomed wells. Under the established optimal conditions the NiPA showed a 100% sensitivity and a specificity of 93%. Pre-treatment of PBMCs with neuraminidase and the addition of epidermal cells or rIL-2 after nickel priming enhanced the proliferative response in low-grade sensitized individuals.

Key words

In vitro nickel-proliferation assay Nickel-contact dermatitis 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1989

Authors and Affiliations

  • E. P. Prens
    • 1
  • K. Benne
    • 1
  • T. van Joost
    • 2
  • R. Benner
    • 2
  1. 1.Departments of Dermatology and ImmunologyUniversity Hospital Rotterdam-Dijkzigt and Erasmus UniversityRotterdamThe Netherlands
  2. 2.Department of DermatologyUniversity Hospital Rotterdam-Dijkzigt and Erasmus UniversityRotterdamThe Netherlands

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