Clonal Analysis of Human T-Cell Responses to Fractionated House Dust Mite Antigens (<i>Dermatophagoides pteronyssinus</i>)

Abstract

In the present study, we investigated the cellular basis of house dust mite-driven immune responsiveness in an atopic individual with perennial rhinitis. We established 40 human T-cell clones (CD3+, 4+, 8-) reactive to Dermatophagoides pteronyssinus (Dp) antigen under the restriction of HLA-DR. By using the crude Dp antigen and its 14 molecular weight (MW) fractions, we analyzed the distribution of T-cell-recognizing sites in the whole Dp extract. We tested T-cell-mediated immunity through two parameters; the identification of Dp fractions inducing T-cell proliferation, and the ability of T-cell clones to secrete IL-2, IL-4, and IFN-gamma. According to a prominent peak among fraction-driven T-cell proliferation, we observed that T-cell clones that recognized 45,000- to 95,000-MW fractions were common, while clones reactive to 15,000- to 25,000-MW fractions were less frequent. Several clones were also reactive to antigens of Dermatophagoides farinae or other insects. Based on the responses of cloned T cells, we observed at least 9 distinct T epitopes in crude Dp antigen. These T-cell clones had a heterogenous secretory pattern of cytokines. T-cell clones showed no association between their ability to produce regulatory cytokine and their recognition of particular Dp fractions.