ABSTRACT
PURPOSE: Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8+T cells have been known to play an important role in the pathogenesis of atopic dermatitis (AD). However, the mechanisms underlying the loss of self-tolerance remain unclear. Regulatory T cells (Tregs) play a key role in the development of homeostasis in the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8+CLA+T cells might be underlying mechanism in AD. MATERIALS AND METHODS: CD8+CLA+T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequencies of CD8+CLA+T cells were evaluated. The proliferative responses of CD8+CLA+T cells were assessed by flow cytometry, and the levels of transforming growth factor-beta1 (TGF-beta1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay. RESULTS: Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheral CD8+CLA+T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8+CLA+T cells in AD. Meanwhile, the levels of TGF-beta1 produced by Tregs were significantly lower in AD, and anti-TGF-beta1 abolished such suppression. CONCLUSION: The attenuated inhibitory ability of Tregs on hyper-activated autologous CD8+CLA+T cells, mediated by TGF-beta1, plays an important role in the pathogenesis of AD.
Subject(s)
Adult , Aged , Female , Humans , Male , CD8-Positive T-Lymphocytes/drug effects , Case-Control Studies , Cell Proliferation , Cell Separation , Dermatitis, Atopic/immunology , Granzymes/metabolism , Interleukin-10/metabolism , Lymphocyte Count , Perforin/metabolism , Skin/immunology , T-Lymphocytes, Cytotoxic/drug effects , T-Lymphocytes, Regulatory/drug effects , Transforming Growth Factor beta1/pharmacologyABSTRACT
Background & objectives: Dendritic cells (DCs) are potent antigen presenting cells which proceed from immature to a mature stage during their differentiation. There are several methods of obtaining long lasting mature antigen expressing DCs and different methods show different levels of antigen expressions. We investigated bone marrow derived DCs for the degree of maturation and genetically engineered antigen presentation in the presence of interleukin-4 (IL-4) as a maturity enhancer. Methods: DCs and transfected retrovirus were cultured together in the presence of granulocyte-macrophage colony stimulating factor (GMCSF)-IL4, GMCSF +IL4, lipopolysaccharide (LPS). B 7.1, B7.2 and CD11c were measured by the degree of immune fluorescence using enhanced green fluorescent protein (EGFP) shuttled retrovirus transfected antigen. Degree of MHC class I molecule with antigen presentation of antigen was also evaluated by fluorescence activated cell sorting. The antigen presenting capacity of transfected DCs was investigated. Bone marrow DCs were generated in the presence of GMCSF and IL-4 in vitro. Dividing bone marrow cells were infected with EGFP shuttled retrovirus expressing SSP2 by prolonged centrifugation for three consecutive days from day 5, 6 and 7 and continued to culture in the presence of GMSCF and IL-4 until day 8. Results: IL-4 as a cytokine increased the maturation of retrovirus transfected DCs by high expression of B 7-1 and B 7-2. Also, IL-4 induced DC enhanced by the prolonged centrifugation and it was shown by increased antigen presentation of these dendric cells as antigen presenting cell (APC). Cytolytic effects were significantly higher in cytotoxic T cell response (CTLs) mixed with transfected DCs than CTLs mixed with pulsed DCs. Interpretation & conclusions: There was an enhanced antigen presentation by prolonged expression of antigen loaded MHC class I receptors in DCs in the presence of IL-4 by prolonged centrifugation.
Subject(s)
Antigen Presentation/drug effects , Antigen Presentation/immunology , Antigen-Presenting Cells/immunology , Antigen-Presenting Cells/metabolism , B7 Antigens/genetics , B7 Antigens/metabolism , Bone Marrow/immunology , Bone Marrow/metabolism , Cell Differentiation/genetics , Cell Differentiation/immunology , Centrifugation , Dendritic Cells/drug effects , Dendritic Cells/immunology , Gene Expression Regulation/drug effects , Gene Expression Regulation/immunology , Genes, MHC Class I/immunology , Granulocyte-Macrophage Colony-Stimulating Factor/immunology , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Humans , Interleukin-4/immunology , Interleukin-4/pharmacology , Lipopolysaccharides/pharmacology , Peptides/genetics , Peptides/immunology , Retroviridae/genetics , T-Lymphocytes, Cytotoxic/drug effects , T-Lymphocytes, Cytotoxic/immunology , Transfection/methodsABSTRACT
Trichosanthin, a basic protein purified from the root tuber of Trichosanthes kirilowii, has been used effectively in China to induce midterm abortion in humans. In this paper, we show that trichosanthin at non-cytotoxic concentrations markedly inhibited the mitogen-induced lymphoproliferative response and the generation of a primary alloreactive CTL response in vitro. Similarly, the production of IL-2 by Con A activated splenocytes and the in vitro effector functions of macrophages were also significantly suppressed. In contrast, the cytolytic activity of CTL and NK cells was unimpaired. Moreover, the in vivo activation of NK cells was not significantly altered by a single injection of a non-toxic microgram amount of trichosanthin into mice. However, other immune reactivities such as the induction of a DTH response and the humoral antibody formation to SRBC were markedly depressed. Our data suggest that trichosanthin is a potent immunosuppressive protein that could affect humoral immunity and a variety of cell-mediated processes. In addition, our preliminary results show that this abortifacient protein could also inhibit the growth of a murine malignant tumour (MBL-2), both in vivo and in vitro.