Comparison of polyclonal expansion methods to improve the recovery of cervical cytobrush-derived T cells from the female genital tract of HIV-infected women

J Immunol Methods. 2010 Mar 31;354(1-2):68-79. doi: 10.1016/j.jim.2010.02.002. Epub 2010 Feb 8.


Cervical cytobrushing is a useful and non-invasive method for obtaining mucosal mononuclear cells from the female genital tract, but yields few cells. The aim of this study was to compare in vitro expansion protocols (anti-CD3, anti-CD3/CD28 or Dynal anti-CD3/CD28 beads) and cytokine combinations (IL-2, IL-7 and IL-15) to improve cervical T cell yields and viability. Eighteen HIV-infected women were included in this study to compare methods for polyclonal expansion of T cells from the female genital tract and blood. Comparison of T cell yields, viability and maturational status (by differential staining with CD45RO, CCR7 and CD27) was determined following 7 days of in vitro expansion. Anti-CD3 and IL-2 resulted in a 4.5-fold (range 3.7-5.3) expansion of cervical CD3+ T cells in 7 days compared to day 0. Inclusion of anti-CD28 or addition of IL-7 and IL-15 to this combination did not improve expansion. Culturing cells with Dynal beads (1:1) and IL-2, IL-7 and IL-15 gave rise to the highest yields after 7 days in both blood (7.1-fold) and cervix (5.6-fold). While expansion with anti-CD3 led to the accumulation of effector memory T cells (CD45RO+CCR7-CD27-), expansion with Dynabeads selected for accumulation of central memory T cells (CD45RO+CCR7+CD27+). We conclude that in vitro expansion with Dynabeads (1:1) in the presence of IL-2, IL-7 and IL-15 resulted in the greatest increase in viable T cells from both blood and cytobrush. Irrespective of the expansion method used, the T cell memory profile was altered following expansion.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Antigens, CD / metabolism*
  • CD28 Antigens / metabolism
  • CD3 Complex / metabolism
  • Cell Culture Techniques*
  • Cell Differentiation
  • Cell Proliferation*
  • Cell Separation
  • Cell Survival
  • Cells, Cultured
  • Cervix Uteri / immunology*
  • Cervix Uteri / virology
  • Female
  • Flow Cytometry
  • HIV Infections / blood
  • HIV Infections / immunology*
  • HIV Infections / virology
  • Humans
  • Immunologic Memory
  • Immunophenotyping
  • Interleukin-15 / metabolism
  • Interleukin-2 / metabolism
  • Interleukin-7 / metabolism
  • Interleukins / metabolism*
  • Leukocyte Common Antigens / metabolism
  • Mucous Membrane / immunology
  • Mucous Membrane / virology
  • Phenotype
  • Receptors, CCR7 / metabolism
  • Recombinant Proteins / metabolism
  • Specimen Handling*
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / virology
  • Time Factors
  • Tumor Necrosis Factor Receptor Superfamily, Member 7 / metabolism


  • Antigens, CD
  • CCR7 protein, human
  • CD28 Antigens
  • CD3 Complex
  • Interleukin-15
  • Interleukin-2
  • Interleukin-7
  • Interleukins
  • Receptors, CCR7
  • Recombinant Proteins
  • Tumor Necrosis Factor Receptor Superfamily, Member 7
  • Leukocyte Common Antigens