T Cell Enrichment By Nonadherence to Nylon

Karen S. Hathcock1

1 National Cancer Institute, Bethesda, Maryland
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 3.2
DOI:  10.1002/0471142735.im0302s30
Online Posting Date:  May, 2001
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Abstract

Differential adherence properties of T cells, B cells, and accessory cells such as macrophages or other APC (antigen‐presenting cells) can be employed to enrich for T cells. While the basis for this adherence is not understood, either nylon wool or Sephadex G‐10 can be used as column matrices to which B cells and accessory cells will more readily adhere than T cells. Such procedures offer a convenient, although imprecise, means of enriching T cells through removal of accessory and B cells; use of nylon wool is preferred if both of the latter subsets are to be removed, while Sephadex is used when the goal is primarily to remove accessory cells.

     
 
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Table of Contents

  • Basic Protocol 1: T Cell Enrichment Using Nylon Wool Columns
  • Support Protocol 1: Preparation of Sterilized Nylon Wool Column
  • Commentary
  • Literature Cited
  • Tables
     
 
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Materials

Basic Protocol 1: T Cell Enrichment Using Nylon Wool Columns

  Materials
  • Complete RPMI‐5 medium ( appendix 2A)
  • 50‐ml conical tube
  • Sterilized nylon wool column (see protocol 2 and Table 3.2.1)
  • 3‐way disposable stopcock
  • 19‐G and 23‐G needles
  • Sorvall H‐1000B rotor (or equivalent)
  • Additional reagents and equipment for preparing single‐cell suspensions with removal of red blood cells (unit 3.1) and for cell viability test using trypan blue exclusion ( appendix 3A)
    Table 3.2.1   MaterialsNylon Wool Column Specifications for T Cell Enrichment

    Column size (ml) Nylon wool (g) Loading vol. (ml) Cell capacity (108) Collection vol. (ml)
    12 0.8‐1.0 2 1.5 15‐18
    20 1.6‐2.0 4 3.0 15‐18

Support Protocol 1: Preparation of Sterilized Nylon Wool Column

  Additional Materials
  • 1% HCl
  • Scrubbed nylon fiber/wool (Table 3.2.1; Polysciences)
  • Asbestos gloves
  • Canine grooming brushes (2)
  • 12‐ or 20‐ml disposable syringe (see protocol 1basic protocol)
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Figures

Videos

Literature Cited

Literature Cited
   Julius, M.H., Simpson, E., and Herzenberg, L.A. 1973. A rapid method for the isolation of functional thymus‐derived murine lymphocytes. Eur. J. Immunol. 3:645‐649.
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