Supplementary Table S1 from Concurrent Induction of Antitumor Immunity and Autoimmune Thyroiditis in CD4+CD25+ Regulatory T Cell–Depleted Mice (original) (raw)
2023
https://doi.org/10.1158/0008-5472.22363505
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Abstract
AI
The study focuses on the concurrent induction of antitumor immunity and autoimmune thyroiditis in CD4+CD25+ regulatory T cell-depleted mice, analyzing lymph node cell populations post-treatment with α-CD25 monoclonal antibody. Data from various experimental conditions illustrate the effects of Treg cell depletion on immune responses, as reflected in the percentage changes in positive cell staining.
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Parameters controlling the programmed death of mature mouse T lymphocytes in high-dose suppression
Cellular Immunology, 1995
induction, increased proliferation, and greater suppression at high antigen doses. Profound loss of cells at high antigen dose was found to require at least 48 to 72 hr to develop. Antigen add-back experiments showed that strong T cell receptor reengagement of activated, cycling cells was essential for proliferative suppression and cell loss. Increasing the ratio of APC:T lymphocytes to 50: 1 augmented cell death. For antigen-induced death of lymph node T cells, fresh T-depleted splenocytes were more effective than splenocytes that had been irradiated or treated with mitomycin C. Thus, T lymphocyte apoptosis at high antigen doses is a function of the activation response of the T lymphocyte as well as the efficiency of antigen presentation by the APC. These results strengthen the theory that apoptosis takes part in a feedback regulatory mechanism that has been called propriocidal regulation, which limits T cell expansion at high antigen doses. o 1995 Academic PWS, IIIC. 3 Abbreviations used: APCs, antigen-presenting cells; IL-SF& interleukin-2 receptor 01 chain; TCR, T cell receptor; dThd, thymidine; PCC, pigeon cytochrome c; LNTC, lymph node T cells; MBP, myelin basic protein.
Transplantation Journal, 2010
Rabbit antithymocyte globulin therapy (rATG) is a potent lymphocyte-depleting agent commonly used following renal transplantation to reduce the risk of acute rejection. Standard doses (7-10 mg/kg) of rATG result in profound lymphopenia and predispose patients to infection and malignancy. The effects of lower doses of rATG (LoD-rATG, 3-5 mg/kg) on peripheral blood lymphocytes (PBL) are as yet unknown. In this prospective clinical trial, PBL subsets were characterized by flow cytometry over 12 months following LoD-rATG therapy. All patients were initially treated with standard doses of tacrolimus, mycophenolic acid, and prednisone. At 3 months, patients were randomized to either lower doses of tacrolimus or sirolimus to examine the effects of maintenance immunosuppression on PBL reemergence. LoD-rATG therapy resulted in prolonged suppression of CD19 ϩ B cells, total CD3 ϩ T cells, as well as naïve and memory CD4 ϩ T cell and CD4/CD25/Foxp3 ϩ T-regulatory subsets irrespective of chronic immunosuppressive therapy. Selective depletion was only noted in the CD4CD45RA ϩ naïve T-cell subset resulting in an altered memory/naïve CD4 ϩ ratio. LoD-rATG failed to deplete CD8 ϩ T cells, which increased their relative contribution to the total CD3 ϩ pool. All other lymphocyte subsets maintained near normal proportions. Thus, LoD-rATG therapy may lessen the adverse effects of full dose rATG while maintaining overall efficacy.
Immunobiology, 2013
CD4 + CD25 + Foxp3 + T regulatory cells (Tregs) and CD1d-restricted invariant natural killer T (iNKT) cells are two cell types that are known to regulate immune reactions. Depletion or inactivation of Tregs using specific anti-CD25 antibodies in combination with immunostimulation is an attractive modality especially in anti-tumour immunotherapy. However, CD25 is not expressed exclusively on Tregs but also on subpopulations of activated lymphocytes. Therefore, the modulatory effects of the specific anti-CD25 antibodies can also be partially attributed to their interactions with the effector cells. Here, the effector functions of iNKT cells were analysed in combination with anti-CD25 mAb PC61. Upon PC61 administration, ␣-galactosylceramide (␣-GalCer)-mediated activation of iNKT cells resulted in decreased IFN-␥ but not IL-4 production. In order to determine whether mutual interactions between Tregs and iNKT cells take place, we compared IFN␥ production after ␣-GalCer administration in anti-CD25-treated and "depletion of regulatory T cell" (DEREG) mice. Since no profound effects on IFN␥ induction were observed in DEREG mice, deficient in FoxP3 + Tregs, our results indicate that the anti-CD25 antibody acts directly on CD25 + effector cells. In vivo experiments demonstrated that although both ␣-GalCer and PC61 administration inhibited TC-1 tumour growth in mice, no additive/synergic effects were observed when these substances were used in combination therapy.
MRL-.lpr/Ipr (lpr) mice develop profound lymphadenopathy resulting from the accumulation of CD4CD8-(double-negative, DN) cells in the peripheral Iymphoid organs. Earlier studies from our laboratory demonstrated an increased proportion of DN cells in the thymus of lpr mice with age. Inasmuch as the DN thymocytes constitute a heterogenous population of cells, in the present study, wt: investigated the TCR phenotype of DN thymocytes and their responsiveness to activation through the TCR. The DN thymocytes of young (1 month of age) Ipr mice contained -65% CD3+ cells of which -60% were ofl-TCR+ and -39% were y&TCR+ as detected by using pan anti-TCR mAbs. In old (4-6 months of age) or young MRL-+/+ mice, similar proportions of CD3+, o/3-or yb-TCR+ DN thymocytes were detected. Interestingly, however, in old (4-6 months of age) lpr mice, the CD3+ T cells increased to -86% and the majority of these (-81%) were a@TCR+ and only -3% were y&TCR+. Also, in old lpr mice, there was a lo-fold increase in the absolute number of c&TCR+ DN cells in the thymus, whereas, the absolute number of y& TCR+ DN cells in the thymus did not alter significantly. Furthermore, a majority (-84%) of the old Ipr DN thymocytes expressed CD45R, similar to the peripheral DN T cells. In contrast, only a small number (-1%) of DN thymocytes from young Ipr or MRL-+/t mice expressed CD45R. The DN thymocytes from young lpr or MRL-+/+ mice demonstrated strong and similar proliferative responsiveness to stimulation with PMA + calcium ionophore or PMA + IL-2, or to immobilized mAb directed against the TCRs (CD3, @3 and 7s). In contrast, the DN thymocytes and the DN peripheral T cells from old lpr mice demonstrated marked defect in responding to the above stimuli. The present study suggests that with the onset of lymphadenopathy, the DN cells in the thymus of old Ipr mice are increasingly skewed toward the &TCR repertoire, the majority of which express CD45R and respond poorly to mitogenic stimuli or when activated through the TCR. It is suggested that migration of such cells continuously to the periphery may result in severe lymphadenopathy seen in old MRL-lpr/lpr mice. 0 1991 Academic press, IIIC. r This work was supported, in part, by NIH Grants CA 45009 and CA 45010 awarded to M.N. and P.S.N.
CD4+CD25lowGITR+ cells: A novel human CD4+ T-cell population with regulatory activity
European Journal of Immunology, 2011
Treg subsets play a role in sustaining peripheral tolerance, are characterized by markers such as forkhead winged-helix transcription factor (FOXP3) and CD25, and produce suppressive cytokines, such as IL-10 and TGF-b. Glucocorticoid-induced TNF receptor family-related (GITR) protein has been suggested to regulate Treg activity in mice. The aim of our study was to investigate GITR expression in human CD4 1 T lymphocytes and its possible role in Treg function. Results indicate that a subset of CD4 1 T cells in the peripheral blood expresses GITR and low levels of CD25 (CD4 1 CD25 low GITR 1 ). These cells show Treg features as they express FOXP3, IL-10, TGF-b and are anergic but, as opposed to natural Tregs, express low levels of CTLA-4 and are CD127 high . CD4 1 CD25 low GITR 1 cells represent a low percentage of the CD4 1 T-cell population (0.32-1.74%) and are mostly memory cells. Functional experiments demonstrated that CD4 1 CD25 low GITR 1 cells have relevant suppressive activity that depends on TGF-b. Moreover, an anti-GITR Ab inhibited their suppressive activity, as observed in CD4 1 CD25 1 murine Tregs. Taken together, these data indicate that human CD4 1 CD25 low GITR 1 cells represent a distinct Treg subpopulation.
GITR Triggering Induces Expansion of Both Effector and Regulatory CD4+ T Cells In Vivo
The Journal of Immunology, 2009
Glucocorticoid-induced TNF receptor family-related protein (GITR) is expressed on activated and regulatory T cells, but its role on these functionally opposing cell types is not fully understood. Here we describe that transgenic expression of GITR's unique ligand (GITRL) induces a prominent increase of both effector and regulatory CD4 ؉ T cells, but not CD8 ؉ T cells. Regulatory T cells from GITRL transgenic mice are phenotypically activated and retain their suppressive capacity. The accumulation of effector and regulatory T cells is not due to enhanced differentiation of naive T cells, but is a direct result of increased proliferation. Functional consequences of increased numbers of both regulatory and effector T cells were tested in an autoimmune model and show that GITR stimulation is protective, as it significantly delays disease induction. These data indicate that GITR regulates the balance between regulatory and effector CD4 ؉ T cells by enhancing proliferation of both populations in parallel. address: m.a.nolte@amc.nl 4 Abbreviations used in this paper: GITR, Glucocorticoid-induced tumor necrosis factor receptor family-related protein; GITRL, GITR ligand; EAE, experimental autoimmune encephalomyelitis; TG, transgenic; WT, wild type.
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Cancer Research, 2005
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American Journal of Transplantation, 2004
CD4 + CD25 + regulatory T cells (Treg) are potent suppressors, and play important roles in autoimmunity and transplantation. Recent reports suggest that CD4 + CD25 + Treg are not a homogeneous cell population, but the differences in phenotype, function, and mechanisms among different subsets are unknown. Here, we demonstrate CD4 + CD25 + Treg cells can be divided into subsets according to cell-surface expression of CD62L. While both subsets express foxp3 and are anergic, the CD62L + population is more potent on a per cell basis, and proliferates and maintains suppressive function far better than the CD62L-population and unseparated CD4 + CD25 + Treg. The CD62L + population preferentially migrates to CCL19, MCP-1 and FTY720. Both CD62L + and CD62L-subsets prevent the development of autoimmune gastritis and colitis induced by CD4 + CD25-CD45RB high cells in severe combined immunodeficiency (SCID) mice. Overall, these results suggest CD4 + CD25 + Treg are not a homogenous cell population, but can be divided into at least two subsets according to CD62L expression. The CD62L + subset is a more potent suppressor than the CD62L-population or unfractionated CD4 + CD25 + Treg cells, can be expanded far more easily in culture, and is more responsive to chemokine-driven migration to secondary lymphoid organs. These properties may have significant implications for the clinical manipulation of the CD4 + CD25 + CD62L + cells.
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Regulatory T (Treg) cells act to suppress activation of the immune system and thereby maintain immunological homeostasis and tolerance to selfantigens. The frequency and suppressing activity of Treg cells in general are high in different malignancies. We wanted to identify the role and regulation of CD4 + CD25 + FoxP3 + Treg cells in B-cell acute lymphoblastic leukaemia (B-ALL). We have included patients at diagnosis (n = 54), patients in clinical remission (n = 32) and normal healthy individuals (n = 35). These diagnosed patients demonstrated a lower number of CD4 + CD25 + cells co-expressing a higher level of FoxP3, interleukin-10, transforming growth factor-b and CD152/CTLA-4 than the normal population. Treg cells from patients showed a higher suppressive capability on CD4 + CD25 À responder T (Tresp) cells than normal. The frequency and immunosuppressive potential of CD4 + CD25 + FoxP3 + Treg cells became high with the progression of malignancy in BALL. Relative distribution of Tresp and Treg cells was only~5 : 1 in BALL but~35 : 1 in normal healthy individuals, further confirming the elevated immunosuppression in patients. A co-culture study at these definite ex vivo ratios, indicated that Treg cells from BALL patients exhibited higher immunosuppression than Treg cells from normal healthy individuals. After chemotherapy using the MCP841 protocol, the frequency of CD4 + CD25 + cells was gradually enhanced with the reduction of FoxP3, interleukin-10 positivity corresponded with disease presentation, indicating reduced immunosuppression. Taken together, our study indicated that the CD4 + CD25 + FoxP3 + Treg cells played an important role in immunosuppression, resulting in a positive disease-correlation in these patients. To the best of our knowledge, this is the first detailed report on the frequency, regulation and functionality of Treg cells in BALL .
CD4+ CD25+ Treg: divide and rule?
Immunology, 2004
Research on CD4 CD25 regulatory T cells (Treg) has gathered momentum over the last ®ve years but many aspects of their fundamental biology remain elusive. Treg have been considered to be`naturally anergic' based on their failure to proliferate in response to T-cell receptor ligation in vitro. Several recent studies challenge this view and demonstrate a robust proliferative capacity for CD25 cells. The signi®cance of this ®nding for Treg homeostasis and function is considered below.
Blood, 2004
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Time-Dependent Inhibition of TR Cells Generation in MLR by Immunosuppressive Drugs
Transplantation Journal, 2004
Aims: Reconstitution of lethally irradiated B10.AKM (H-2K k ) mice with syngeneic bone marrow cells infected with retroviruses carrying the allogeneic MHC class I gene H-2K b resulted in stable and lifelong expression of K b on bone marrow-derived cells. More importantly, these mice were specifically tolerant to H-2K b skin grafts while still retaining the ability to reject third party skin grafts. Using CD8 transgenic mice, we discovered that alloreactive CD8 T cells underwent negative selection in the thymus, leading to the absence of these potentially alloreactive T cells in the periphery. In this study, we utilize Tg361 CD4 transgenic mice (CBA/Ca (H-2 k )) expressing a TCR specific for the MHC class I gene H-2K b to determine the fate of potentially alloreactive CD4 T cells using our gene therapy protocol. Methods: Tg361 CD4 TCR transgenic mice which express an alloreactive TCR on CD4 T cells specific for K b were utilized to determine the mechanism by which genetic engineering of bone marrow induces donor specific CD4 T cell tolerance after bone marrow transplantation. Lethally irradiated B10.AKM mice were reconstituted with a 6:1 ratio of mock or K b -transduced B10.AKM bone marrow to Tg361 bone marrow. Starting at four weeks after bone marrow transplantation, mice were analyzed for H-2K b expression as well as for the development of Tg361 CD4 T cells. Results: Lethally irradiated recipients that received CD4 transgenic bone marrow in combination with autologous bone marrow transduced with virus encoding H-2K b displayed stable and long-term expression of K b on bone marrow-derived cells. Alloreactive CD4 transgenic T cells were readily detectable in the peripheral blood, spleen and thymus of chimeric recipients; although their levels were 2-fold lower than mock-transduced bone marrow recipients. Despite the presence of potentially alloreactive CD4 T cells in the periphery, chimeric recipients were specifically tolerant to H-2K b expressing skin grafts. Interestingly, chimeric mice which received K b -transduced and CD4 transgenic bone marrow expressed CD25 on 20-30% of Tg361-derived CD4 T cells, compared to only 6% of Tg361-derived CD4 T cells in mock transduced controls. Conclusions: We conclude that genetic engineering of bone marrow induces donor specific CD4 T cell tolerance through deletional and non-deletional mechanisms. We further hypothesize that expression of K b on bone marrow derived cells induces CD4 T cells capable of inhibiting alloreactive T cells.
Journal of Experimental Medicine, 1993
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