T-cell repertoire in the blood and lungs of atopic asthmatics before and after ragweed challenge

Vladimir V. Yurovsky, Els J M Weersink, Susan S. Meltzer, Wendy C. Moore, Dirkje S. Postma, Eugene R. Bleecker, Barbara White

Research output: Contribution to journalArticle

Abstract

T cells play a pivotal role in initiating and orchestrating allergic responses in asthma. The goal of this work was to learn whether ragweed challenge in the lungs alters the T-cell repertoire expressed in the blood and lungs of atopic asthmatics. Analyses of cell numbers, differentials, and T-cell subsets in bronchoalveolar lavage (BAL) fluids showed that ragweed challenge was associated with preferential recruitment of CD4+ T cells into the lungs. A reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify T-cell receptor (TCR) gene transcripts from unfractionated, CD4+, and CD8+ T cells in blood and BAL fluids. As judged by RT-PCR, the usage of TCR Vα and Vβ gene families in BAL fluids was similar to that in blood. Ragweed challenge did not change the levels of expression of these V gene families. The clonality of T cells was estimated by analyzing the diversity of TCR V-(D)-J junctional region nucleotide lengths associated with each Vα and Vβ gene family, using sequencing gel electrophoresis. Most V gene families in blood and BAL fluids were associated with multiple junctional region lengths before and after ragweed challenge, indicating polydonal expression. Some V gene families were expressed in an oligoclonal manner in unfractionated. CD4+, and CD8+ T cells in BAL fluids before ragweed challenge, as indicated by a few predominant junctional region lengths. The majority of these V gene families became polyclonal after challenge, compatible with polyclonal T-cell influx during inflammation immediately after ragweed challenge. However, some V gene families became oligoclonal or developed a new oligoclonal pattern of junctional region lengths in BAL T cells after ragweed challenge. Surprisingly, this occurred in both CD4+ and CD8+ T cells. In one of these instances, DNA sequencing of Vβ21 junctional regions in CD8+ T cells confirmed a change from polyclonal to oligoclonal expression after ragweed challenge. These findings show that ragweed challenge is associated with polyclonal influx and oligoclonal activation of both CD4+ and CD8+ T cells in the lungs.

Original languageEnglish (US)
Pages (from-to)370-383
Number of pages14
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume18
Issue number3
StatePublished - 1998
Externally publishedYes

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pulmonary and Respiratory Medicine

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  • Cite this

    Yurovsky, V. V., Weersink, E. J. M., Meltzer, S. S., Moore, W. C., Postma, D. S., Bleecker, E. R., & White, B. (1998). T-cell repertoire in the blood and lungs of atopic asthmatics before and after ragweed challenge. American Journal of Respiratory Cell and Molecular Biology, 18(3), 370-383.