scholarly journals PI3K p110δ regulates T-cell cytokine production during primary and secondary immune responses in mice and humans

Blood ◽  
2010 ◽  
Vol 115 (11) ◽  
pp. 2203-2213 ◽  
Author(s):  
Dalya R. Soond ◽  
Elisa Bjørgo ◽  
Kristine Moltu ◽  
Verity Q. Dale ◽  
Daniel T. Patton ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cytokine Production ◽  
Memory T Cells ◽  
Inflammatory Diseases ◽  
Effector Memory ◽  
Naive T Cells ◽  
Naïve T Cells ◽  
Human T Cells ◽  
Phosphoinositide 3 Kinase

Abstract We have previously described critical and nonredundant roles for the phosphoinositide 3-kinase p110δ during the activation and differentiation of naive T cells, and p110δ inhibitors are currently being developed for clinical use. However, to effectively treat established inflammatory or autoimmune diseases, it is important to be able to inhibit previously activated or memory T cells. In this study, using the isoform-selective inhibitor IC87114, we show that sustained p110δ activity is required for interferon-γ production. Moreover, acute inhibition of p110δ inhibits cytokine production and reduces hypersensitivity responses in mice. Whether p110δ played a similar role in human T cells was unknown. Here we show that IC87114 potently blocked T-cell receptor–induced phosphoinositide 3-kinase signaling by both naive and effector/memory human T cells. Importantly, IC87114 reduced cytokine production by memory T cells from healthy and allergic donors and from inflammatory arthritis patients. These studies establish that previously activated memory T cells are at least as sensitive to p110δ inhibition as naive T cells and show that mouse models accurately predict p110δ function in human T cells. There is therefore a strong rationale for p110δ inhibitors to be considered for therapeutic use in T-cell–mediated autoimmune and inflammatory diseases.

scholarly journals Induction of memory-like CD8 + T cells and CD4 + T cells from human naïve T cells in culture

2021 ◽  
Author(s):  
Yasuhito Tokumoto ◽  
Yasuto Araki ◽  
Yusuke Narizuka ◽  
Yosuke Mizuno ◽  
Susumu Ohshima ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cd8 T Cells ◽  
Cd4 T Cells ◽  
Memory T Cells ◽  
Quiescent State ◽  
Naive T Cells ◽  
Naïve T Cells ◽  
Cells In Culture ◽  
Human T Cells

Abstract Memory T cells are crucial players in vertebrate adaptive immunity but their development is incompletely understood. Here we describe a method to produce human memory-like T cells from naïve human T cells in culture. Using commercially available human T cell differentiation kits, both purified naïve CD8 + T cells and purified naïve CD4 + T cells were activated via T cell receptor signaling and appropriate cytokines for several days in culture. All the T cell activators were then removed from the medium and the cultures were continued in hypoxic condition (1% O2 atmosphere) for several more days; during this period, most of the cells died, but some survived in a quiescent state for a month. The survivors had small round cell bodies, expressed differentiation markers characteristic of memory T cells and restarted proliferation when the T cell activators were added back. We could also induce memory-like T cells from naïve human T cells without hypoxia, if we froze the activated T cells or prepared the naïve T cells from chilled filter buffy coats.

scholarly journals Characteristic patterns of HLA presentation and T cell differentiation in adult-onset Still’s disease

2018 ◽  
Vol 32 ◽  
pp. 205873841879128 ◽  
Author(s):  
Ju-Yang Jung ◽  
Bunsoon Choi ◽  
Hasan MD Sayeed ◽  
Chang-Hee Suh ◽  
Ye Won Kim ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Memory T Cells ◽  
Effector Memory ◽  
Adult Onset Still’S Disease ◽  
Naive T Cells ◽  
Naïve T Cells ◽  
Effector Memory T Cells

We examined the expression of human leukocyte antigen (HLA) and composition of differentiated T cells in the peripheral blood to understand the characteristics of the immune changes in patients with adult-onset Still’s disease (AOSD). This study enrolled patients with AOSD (n = 14), patients with rheumatoid arthritis (RA, n = 20), and healthy controls (HC, n = 20). The percentage of surface-stained cells with HLA-DP, DQ, and DR alleles and the composition of differentiated T cells in peripheral blood leukocytes (PBLs) were evaluated by flow cytometry. AOSD patients exhibited significantly higher percentages of lymphocytes presenting HLA-DP and HLA-DR, and lower percentages of cells presenting HLA-DQ, than RA patients or HC. The proportions of CD4+, CD4+CCR7+, CD4+CD62L–, and CD8+CD62L– cells from PBLs were decreased in AOSD patients relative to RA patients or HCs. By contrast, AOSD patients had higher proportions of CD8+naïve T cells in whole blood relative to RA patients or HC. The proportions of CD4+ effector memory T cells, CD8+ naïve T cells, and CD8+ effector memory T cells in whole blood cells and CD4+ effector memory T cell in lymphocytes were significantly associated with the systemic score. While the proportions of CD4+, CD8+, CCR7+, CD4+CCR7+, CD4+CD62L–, and CD8+CD62L– cells were significantly decreased in AOSD patients, and the proportion of CD8+naïve T cells was elevated in AOSD and correlated with the systemic score. Further studies of a large cohort of AOSD patients will be necessary to evaluate these markers in the pathogenesis of AOSD.

Limited expression of R5-tropic HIV-1 in CCR5-positive type 1–polarized T cells explained by their ability to produce RANTES, MIP-1α, and MIP-1β

Blood ◽  
2000 ◽  
Vol 95 (4) ◽  
pp. 1167-1174 ◽  
Author(s):  
Francesco Annunziato ◽  
Grazia Galli ◽  
Filomena Nappi ◽  
Lorenzo Cosmi ◽  
Roberto Manetti ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Memory T Cells ◽  
Receptor Expression ◽  
Th2 Cells ◽  
Th1 Cells ◽  
Naive T Cells ◽  
Naïve T Cells ◽  
Viral Spread ◽  
Hiv 1

Human T helper (Th) cells (Th1- or Th2-oriented memory T cells as well as Th1- or Th2-polarized naive T cells) were infected in vitro with an R5-tropic HIV-1 strain (BaL) and assessed for their profile of cytokine production, CCR5 receptor expression, and HIV-1 p24 antigen (p24 Ag) production. Higher p24 Ag production was found in CCR5-negative Th2-like memory T cells than in CCR5-positive Th1-like memory T cells. By contrast, p24 Ag production was higher in Th1-polarized activated naive T cells in the first 4 days after infection. However, p24 Ag production in Th1-polarized T cells became comparable or even lower than the production in Th2-polarized populations later in infection or when the cells were infected with HIV-1BaL after secondary stimulation. The higher levels of p24 Ag production by Th1-polarized naive T cells soon after infection reflected a higher virus entry, as assessed by the single round infection assay using the HIV–chloramphenicol acetyl transferase (HIV-CAT) R5-tropic virus that contains the envelope protein of HIV-1 YU2 strain. The limitation of viral spread in the Th1-polarized populations, despite the initial higher level of T-cell entry of R5-tropic strains, was due to the ability of Th1 cells to produce greater amounts of β-chemokines than Th2 cells. In fact, an inverse correlation was observed between Th1-polarized naive T cells and Th1-like memory-activated T cells in regards to p24 Ag production and the release of the following CCR5-binding chemokines: regulated on activation normal T expressed and secreted (RANTES), macrophage inflammatory protein–1 (MIP-1), and MIP-1β. Moreover, infection with the HIV-1BaL strain of Th1-polarized T cells in the presence of a mixture of anti-RANTES, anti–MIP-1, and anti–MIP-1β neutralizing antibodies resulted in a significant increase of HIV-1 expression. These findings suggest that Th1-type responses may favor CD4+ T-cell infection by R5-tropic HIV-1 strains, but HIV-1 spread in Th1 cells is limited by their ability to produce CCR5-binding chemokines.

Naïve T Cell-Derived CTL Recognize Atypical Epitopes of CMVpp65 with Higher Avidity Than CMV-Seropositive Donor-Derived CTL – a Basis for Treatment of Post-Transplant Viral Infection by Adoptive Transfer of T Cells From Virus-naïve Donors

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3002-3002
Author(s):  
Patrick J Hanley ◽  
J. Joseph Melenhorst ◽  
Phillip Scheinberg ◽  
Gail J Demmler-Harrison ◽  
Daniele Lilleri ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Adoptive Transfer ◽  
Memory T Cells ◽  
Conflicts Of Interest ◽  
Target Cells ◽  
Naive T Cells ◽  
Naïve T Cells ◽  
Infected Cells

Abstract Abstract 3002 Adoptive transfer of CMV-specific T cells derived from adult CMV-seropositive (CMVpos) donors can effectively restore antiviral immunity after stem cell transplantation. However due to the absence of CMV antigen-specific memory T cells in cord blood (CB) and adult CMV-seronegative (CMVneg) donors, different culture systems are required to generate virus-specific T cells for adoptive transfer. With a novel protocol we have generated CMVpp65-specific T cells from CB and found that 15/15 CB T cell lines recognized atypical epitopes of pp65. We then explored the generation of CMV-specific CTL from CMVneg donors using a GMP-compliant methodology and studied the epitopes recognized. CD45RA+ naive T cells were selected from the peripheral blood of CMVneg donors and stimulated with pp65-Pepmix-pulsed dendritic cells with supplemented with IL-7, IL-12, and IL-15. For subsequent stimulations T cells were stimulated with pp65-Pepmix-pulsed EBV-LCL and IL-15 or IL-2. CMVpp65-specific T cells (CMV-CTL) expanded from 8 of 11 CMVneg donors were primarily CD8+ T cells (mean 71%). Naïve donor CMV-CTL secreted IFN- γ in response to pp65 peptides (mean 224; range: 38–611 SFC/1×105 cells) compared to irrelevant peptides (mean 12;Range 3–37) as measured in Elispot assays and lysed pp65-pulsed target cells (mean :48; range :15–70%) but not negative controls (mean 22; range 4–40%). These CMV-CTL derived from naive (but not memory) T cells recognized only novel and atypical pp65 epitopes (such as the HLA-A2-restricted epitopes LQTGIHVRV and MLNIPSINV) but not the typical HLA-A2-restricted epitope NLVPMVATV as confirmed by ELISPOT and multimer analysis. These results are similar to CB-derived CTL. Analysis of the avidity of naïve donor CTL specific for the atypical CMV epitopes revealed that the 1/2 maximum effective concentration was similar (mean: 600 pM) to CMVpos CTL recognizing typical epitopes (mean: 300 pM), and more avid than CMVpos CTL recognizing atypical epitopes (mean: 4 nM), highlighting the difference between naïve-derived and memory-derived CTL. TCR sequencing performed on T cells specific for typical (CMVpos) and atypical (CMVpos, CMVneg, and CB) epitopes revealed that CMVpos donor CMV-CTL recognizing typical epitopes were markedly more oligoclonal than CTL recognizing the atypical epitopes derived from CB, CMVpos, or CMVneg donors. To address the concern that atypical epitopes might not be naturally presented by CMV-infected cells and therefore not recognized by in vitro generated CTL, we tested whether CMV CTL (from CB, CMVpos, CMVneg) generated using CMV AD169-infected fibroblasts or CMV VR1814-infected DCs would recognize the same epitopes. As before, CMVpos CMV CTL recognized typical epitopes of pp65 while CB and CMVneg CMV CTL recognized only atypical epitopes, suggesting that the epitopes are naturally processed and presented by APCs, and that the atypical epitopes observed are not an artifact of using exogenous antigens like the pp65 Pepmix. Thus, despite their unusual repertoire, T cells derived from CB or CMVneg donors are likely to control CMV infection. These results reveal major differences in the naïve and memory CMV specific T cell repertoire that merits further exploration. Nevertheless, we demonstrated that atypical epitopes are naturally presented by CMV infected cells and we are now evaluating the clinical efficacy of these CTL in recipients of CBT. These studies should determine if naive T cells primed in vitro are able to persist and establish memory and virus protection in vivo. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Calcineurin Inhibitors Suppress Cytokine Production from Memory T Cells and Differentiation of Naïve T Cells into Cytokine-Producing Mature T Cells

PLoS ONE ◽  
2012 ◽  
Vol 7 (2) ◽  
pp. e31465 ◽  
Author(s):  
Kenshiro Tsuda ◽  
Keiichi Yamanaka ◽  
Hiroshi Kitagawa ◽  
Tomoko Akeda ◽  
Masanao Naka ◽  
...  
Keyword(s):  
T Cells ◽  
Cytokine Production ◽  
Memory T Cells ◽  
Calcineurin Inhibitors ◽  
Naive T Cells ◽  
Naïve T Cells

scholarly journals CD43 is a ligand for E-selectin on CLA+ human T cells

Blood ◽  
2006 ◽  
Vol 107 (4) ◽  
pp. 1421-1426 ◽  
Author(s):  
Robert C. Fuhlbrigge ◽  
Sandra L. King ◽  
Robert Sackstein ◽  
Thomas S. Kupper
Keyword(s):  
T Cells ◽  
T Cell ◽  
Memory T Cells ◽  
Inflammatory Diseases ◽  
Central Element ◽  
Human Memory ◽  
Cell Trafficking ◽  
Immune Functions ◽  
Human T Cells ◽  
Selectin Ligand

The recruitment of memory T cells from blood into tissues is a central element of immune surveillance and adaptive immune responses and a key feature of chronic cutaneous inflammatory diseases such as psoriasis and atopic dermatitis. Human memory T cells that infiltrate skin express the carbohydrate epitope cutaneous lymphocyte-associated antigen (CLA). Expression of the CLA epitope on T cells has been described on P-selectin glycoprotein ligand-1 (PSGL-1) and associated with the acquisition of both E-selectin and P-selectin ligand functions. In this report, we show that CD43, a sialomucin expressed constitutively on T cells, can also be decorated with the CLA epitope and serve as an E-selectin ligand. CLA expressed on CD43 was found exclusively on the high-molecular-weight (125 kDa) glycoform bearing core-2-branched O-linked glycans. CLA+ CD43 purified from human T cells supported tethering and rolling in shear flow via E-selectin but did not support binding of P-selectin. The identification and characterization of CD43 as a T-cell E-selectin ligand distinct from PSGL-1 expands the role of CD43 in the regulation of T-cell trafficking and provides new targets for the modulation of immune functions in skin.

scholarly journals 821 Class specific HDAC inhibition differentially affects the function of specific T cell subsets

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A871-A871
Author(s):  
Nisha Holay ◽  
Uma Giri ◽  
Carla Van Den Berg ◽  
Gail Eckhardt ◽  
Todd Triplett
Keyword(s):  
T Cells ◽  
T Cell ◽  
Histone Deacetylase Inhibitors ◽  
Cytokine Production ◽  
T Cell Subsets ◽  
Global Changes ◽  
Functional Responses ◽  
Naive T Cells ◽  
Naïve T Cells ◽  
Tumor Immunogenicity

BackgroundHistone deacetylase inhibitors (HDACi) are currently being used in the clinic to treat a variety of cancer types. Despite their wide use, the mechanism by which they exert anti-tumor effects is largely unknown. Although originally posited to abrogate tumor proliferation via regulating tumor suppressor genes, responses to monotherapies of HDACi have been shown to be dependent on an adaptive immune system and to enhance responses to immunotherapy. However, whether this mechanism is driven by enhancing tumor immunogenicity or enhancing anti-tumor immune responses is unclear. Understanding this could help identify optimal combination regimens for augmenting immunotherapies. Given the role of epigenetics in regulating T cell differentiation upon antigen encounter into discrete subsets, these studies sought to determine whether HDACi differentially impact naïve from memory T cell subsets.MethodsPBMCs from healthy donors were stimulated with either anti-CD3/anti-CD28 or PMA/Ionomycin in the presence or absence of different HDAC inhibitors (OKI-005, 250 nM; Entinostat, 5 uM; and Vorinostat, 1 uM). Cells were evaluated at different time points by flow cytometric analysis to compare responses by T cell subsets for changes in cytokine production, protein acetylation and other functional responses. Supernatant was collected in separate experiments for comprehensive cytokine bead arrays.ResultsCytokine analysis of supernatants showed clear differences in response to HDACi as while most cytokines decreased, others were either unaffected or increased. We next performed ICS with surface markers to determine if these changes in cytokine production levels were subset specific. Comparisons of naïve and memory subsets found decreased IL-2 levels was primarily attributed to loss of production by naïve T cells. Furthermore, gain of TNFa was almost completely restricted to naïve cells. The preferential responses by naïve T cells was further verified during global changes in acetylated protein levels. Lastly, we found differences between inhibitors on their effects on T cells. As these differences remained even after controlling for potency, this suggests the specificity profiles toward individual HDACs was responsible for their unique effects.ConclusionsThese studies demonstrate clear differences in the effect of HDACi on cytokine production by distinct T cell subsets. Ongoing studies are aimed at elucidating the specific HDACs responsible for regulating T cell effector functions and tumor immunogenicity when targeted. Ultimately, understanding this could help identify inhibitors with the desired specificity profile for combining with immunotherapy.

scholarly journals IgG Immune Complexes Inhibit Naïve T Cell Proliferation and Suppress Effector Function in Cytotoxic T Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Wissam Charab ◽  
Matthew G. Rosenberger ◽  
Haridha Shivram ◽  
Justin M. Mirazee ◽  
Moses Donkor ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Immune Complexes ◽  
Cell Function ◽  
Memory T Cells ◽  
Cytotoxic T Cells ◽  
Cell Subset ◽  
Phenotypic Analysis ◽  
Naive T Cells ◽  
Naïve T Cells

Elevated levels of circulating immune complexes are associated with autoimmunity and with worse prognoses in cancer. Here, we examined the effects of well-defined, soluble immune complexes (ICs) on human peripheral T cells. We demonstrate that IgG-ICs inhibit the proliferation and differentiation of a subset of naïve T cells but stimulate the division of another naïve-like T cell subset. Phenotypic analysis by multi-parameter flow cytometry and RNA-Seq were used to characterize the inhibited and stimulated T cells revealing that the inhibited subset presented immature features resembling those of recent thymic emigrants and non-activated naïve T cells, whereas the stimulated subset exhibited transcriptional features indicative of a more differentiated, early memory progenitor with a naïve-like phenotype. Furthermore, we show that while IgG1-ICs do not profoundly inhibit the proliferation of memory T cells, IgG1-ICs suppress the production of granzyme-β and perforin in cytotoxic memory T cells. Our findings reveal how ICs can link humoral immunity and T cell function.

Short-Term Ionomycin Exposure Activates Naive Murine T-Cells and Induces a Rapid Phenotypic Shift to Memory T-Cell Status: Potential for Use as a Method To Reduce GvHD Activity of Allogeneic T-Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2182-2182
Author(s):  
Mohammad Hossain ◽  
Cynthia R. Giver ◽  
Ned Waller
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Cd4 T Cells ◽  
Cell Activation ◽  
Memory T Cells ◽  
Cell Phenotype ◽  
Naive T Cells ◽  
Memory T Cell ◽  
Naïve T Cells

Abstract We are investigating methods to reduce the graft-versus-host disease (GVHD) potential of donor T-cells while retaining graft-versus-leukemia (GVL) activity in allogeneic HSCT. Previous investigations by our group and others in have shown that naive CD4 T-cells induce severe acute GVHD, while memory CD4 T-cells do not induce GVHD but retain GVL activity in murine transplant models. These findings have led to studies for the development of methods to increase the number of memory T-cells available for transplant. The calcium ionophore, ionomycin, is a T-cell activating agent and mitogen. By increasing intracellular Ca2+ levels, ionomycin is induces T-cell activation through signaling mechanisms including phospholipase C activation, hydrolysis of phosphoinositides, and activation of protein kinase C. Differences in memory and naive T-cell responses to ionomycin have been attributed to resistance of memory T-cells to increases in Ca2+. Memory T-cells lack intracellular Ca2+ stores, and are also resistant to influx of Ca2+. Brief low dose ionomycin exposure (20min, 2μM) of T-cells, leading to increased density of naive T-cells, has previously been exploited as a method for separating memory and naive T-cells by Percoll gradient separation. Since ionomycin exposure induces T-cell activation through native Ca2+ dependent signaling mechanisms, we hypothesized that ionomycin-treated T-cells would shift to an activated/memory T-cell phenotype. Murine splenic T-cells were treated with 1.3μM ionomycin for 4hr. Memory and naive T-cell subsets and activation markers were analyzed by flow cytometry. 75% and 85% of untreated CD4 and CD8 T-cells, respectively, had the CD62L+ naive phenotype. These numbers were dramatically reduced to 7% and 17% after ionomycin exposure, representing a shift to the memory T-cell phenotype. Viability of T-cells was not significantly affected. The majority of remaining CD62L+ naive T-cells expressed activation markers CD25 and CD69. The fraction of CD4+CD25+Foxp3+ regulatory T-cells was also determined by intracellular staining of the transcription factor and co-expression of surface markers. CD4+CD25+Foxp3+ regulatory T-cells represented 4% of untreated CD4 T-cells and 3% of ionomycin-treated CD4 T-cells. While ionomycin has been used for many years in studies of T-cell activation, to our knowledge this is the first demonstration of a rapidly-induced shift of naive T-cells to a memory phenotype. A pilot experiment was conducted testing the GVHD activity of ionomycin-treated splenocytes (SP) in B6→ (B6 × Balb/C)CB6F1 recipients. 5 × 106 T-cell depleted bone marrow cells (TCD-BM) were transplanted along with 10 × 106 treated or untreated SP. Mice that received untreated SP all died from acute GvHD by 34 days after transplant, while all recipients of ionomycin-treated SP survived until the experiement was terminated at day 49 (average weight loss was 25%, data not shown). Continuing experients will refine the dose to further reduce GVHD symptoms and also test GVL activity of the treated cells. Treatment of donor T-cells with ionomycin may represent a clinically applicaple method to engineer donor lymphocyte infusions that are safer for HSCT patients. Figure 1. Survival of CB6F1 recipients after transplant with 5 million B6 TCD-BM and 10 million B6 splenocytes that were either untreated or stimulated ex-vivo with a combination of PMA, ionomycin and brefeldin-A for 4 hours. 5 recipient animals per group. The experiment was terminated at day 49. Figure 1. Survival of CB6F1 recipients after transplant with 5 million B6 TCD-BM and 10 million B6 splenocytes that were either untreated or stimulated ex-vivo with a combination of PMA, ionomycin and brefeldin-A for 4 hours. 5 recipient animals per group. The experiment was terminated at day 49.

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