Direct enumeration of Borrelia-reactive CD4 T cells ex vivo by using MHC class II tetramers

Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11433-8. doi: 10.1073/pnas.190335897.

Abstract

We characterized antigen-specific CD4(+) T cells in six patients with treatment-resistant Lyme arthritis, using an HLA-DRB1*0401 major histocompatibility complex (MHC) class II tetramer covalently loaded with OspA(164-175), an immunodominant epitope of Borrelia burgdorferi. Direct analysis of OspA-tetramer binding CD4(+) cells in patients expressing the HLA-DRB1*0401 allele revealed frequencies of between <0.005 and 0.1% in peripheral blood (n = 6), and between <0.005 and 3.1% in synovial fluid (n = 3). OspA-tetramer(+)CD4(+) cells were directly cloned at 1 cell per well and expanded by mitogen and IL-2 on allogeneic feeder cells. As measured by [(3)H]thymidine incorporation, 95% of 168 T cell clones from synovial fluid binding the OspA-tetramer were antigen-reactive. Clones generated from peripheral blood revealed a different pattern of responsiveness when compared with clones generated from synovial fluid, as measured by proliferation, IFN-gamma, and IL-13 secretion. These clones, selected on the basis of their peptide binding, also responded to whole protein, but with a different cytokine profile. Our studies demonstrate that MHC class II tetramers can be used in humans to directly identify, isolate, and characterize antigen-reactive T cells from an inflammatory compartment.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, Surface / immunology
  • Bacterial Outer Membrane Proteins / immunology
  • Bacterial Vaccines
  • Biopolymers
  • Borrelia burgdorferi Group / immunology*
  • CD4-Positive T-Lymphocytes / immunology*
  • Clone Cells
  • Cytokines / immunology
  • Dose-Response Relationship, Immunologic
  • HLA-DR Antigens / administration & dosage
  • HLA-DR Antigens / immunology*
  • HLA-DRB1 Chains
  • Humans
  • In Vitro Techniques
  • Lipoproteins*
  • Lyme Disease / immunology
  • Lyme Disease Vaccines / immunology

Substances

  • Antigens, Surface
  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Biopolymers
  • Cytokines
  • HLA-DR Antigens
  • HLA-DRB1 Chains
  • HLA-DRB1*04:01 antigen
  • Lipoproteins
  • Lyme Disease Vaccines
  • OspA protein