scholarly journals Convergence of TCR and cytokine signaling leads to FOXO3a phosphorylation and drives the survival of CD4+ central memory T cells

2006 ◽  
Vol 204 (1) ◽  
pp. 79-91 ◽  
Author(s):  
Catherine Riou ◽  
Bader Yassine-Diab ◽  
Julien Van grevenynghe ◽  
Roland Somogyi ◽  
Larry D. Greller ◽  
...  
Keyword(s):  
T Cells ◽  
Molecular Mechanisms ◽  
Memory T Cells ◽  
Ex Vivo ◽  
Global Gene Expression ◽  
Central Memory ◽  
Effector Memory ◽  
Cytokine Signaling ◽  
Global Gene Expression Profiling

The molecular events involved in the establishment and maintenance of CD4+ central memory and effector memory T cells (TCM and TEM, respectively) are poorly understood. In this study, we demonstrate that ex vivo isolated TCM are more resistant to both spontaneous and Fas-induced apoptosis than TEM and have an increased capacity to proliferate and persist in vitro. Using global gene expression profiling, single cell proteomics, and functional assays, we show that the survival of CD4+ TCM depends, at least in part, on the activation and phosphorylation of signal transducer and activator of transcription 5a (STAT5a) and forkhead box O3a (FOXO3a). TCM showed a significant increase in the levels of phosphorylation of STAT5a compared with TEM in response to both IL-2 (P < 0.04) and IL-7 (P < 0.002); the latter is well known for its capacity to enhance T cell survival. Moreover, ex vivo TCM express higher levels of the transcriptionally inactive phosphorylated forms of FOXO3a and concomitantly lower levels of the proapoptotic FOXO3a target, Bim. Experiments aimed at blocking FOXO3a phosphorylation confirmed the role of this phosphoprotein in protecting TCM from apoptosis. Our results provide, for the first time in humans, an insight into molecular mechanisms that could be responsible for the longevity and persistence of CD4+ TCM.

scholarly journals P01.24 The selective HDAC6 inhibitor ITF3756 increases the differentiation to central memory T cells with reduced exhaustion phenotype

2020 ◽  
Vol 8 (Suppl 2) ◽  
pp. A19.2-A20
Author(s):  
C Ripamonti ◽  
C Steinkuhler ◽  
G Fossati
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Memory T Cells ◽  
Ex Vivo ◽  
Central Memory ◽  
Effector Memory ◽  
Memory Cells ◽  
T Effector Cells ◽  
Part Time

BackgroundCentral memory T cells show superior persistence and antitumor immunity compared to effector memory and effector T cells. T effector cells respond quickly to tumors, but they are terminally differentiated and undergo apoptosis upon killing activity. T memory differentiate rapidly into T effector cells and maintain a pool of cells that can continuously differentiate thus sustaining a more lasting response. In adoptive cell therapy (ACT), T cells infused into patients may have a limited time of activity if they are terminally differentiated, and may rapidly undergo exhaustion and apoptosis. The development of new strategies based on novel agents able to generate memory T cells ex-vivo is important for a successful clinical application of ACT.We have studied the effect of a potent and selective HDAC6 inhibitor, ITF3756, on CD8 T cells differentiation during an in vitro induced exhaustion process.Materials and MethodsTo induce exhaustion purified human CD8+ cells were stimulated twice with anti-CD3/CD28 beads (1:2) during 5 days, with or without ITF3756 1μM or 2μM added at all times of stimulation. At day 3 and 5 the expression of exhaustion, memory and effector T cells markers were analyzed by flow cytometry. Cells were also collected at day 5 for genes expression analysis. Expression of exhaustion, T phenotype, metabolic pathway and inflammatory cytokines were investigated by qPCR. Paired two-tailed t-tests was used to determine statistical significance between control versus treatment group at day 3 and 5 in 10 different donors. P-values ≤ 0.05 were considered significant.ResultsITF3756 1μM increased significantly the T central memory phenotype (CD45RO+CD62L+CCR7+) and decreased significantly the T effector phenotype (CD45RO+CD62L-CCR7-). The expression of CD62L in T central memory cells was significantly increased in agreement with the high expression of this marker in naïve and memory T cells. ITF3756 treatment decreased significantly the expression of exhaustion markers PD-1 and LAG-3. No effect was observed on TIM-3 expression. In agreement with the data obtained with protein analysis, treatment with ITF3756 reduced the mRNA level of Pd-1 and Lag-3. Gene expression of Tim-3 was also downmodulated, but this effect did not result in reduction of protein expression at the time of detection. ITF3756 reduced the expression of t-bet (Tbx21) driving T effector differentiation and increased genes related to T memory phenotype (Eomes, Lef-1 and albeit slightly, Tcf-7). T cell activation requires a metabolic reprogramming that supports highly proliferative phenotype and T effector differentiation. ITF3756 treatment decreased both Hif-1α and Glut-1 gene expression that are associated with TCR activation during the exhaustion process. T central memory cells produce less cytokines compared to T effector and effector memory cells. ITF3756 treatment decreased the genes expression of Il-2, Ifn-γ and Tnf-α. All these effects resulted dose dependent.ConclusionsThe selective inhibitor of HDAC6 ITF3756 delays the terminal differentiation of CD8 T cells and increases the percentage of memory T cells with a reduced expression of exhaustion markers in vitro. These results are the basis to further explore the possible use of ITF3756 as a safe ex vivo treatment of CD8 T cells for adoptive cell transfer.Disclosure InformationC. Ripamonti: A. Employment (full or part-time); Significant; Italfarmaco SpA. C. Steinkuhler: A. Employment (full or part-time); Significant; Italfarmaco SpA. G. Fossati: A. Employment (full or part-time); Significant; Italfarmaco SpA.

scholarly journals Metabolic reprogramming of human CD8+ memory T cells through loss of SIRT1

2017 ◽  
Vol 215 (1) ◽  
pp. 51-62 ◽  
Author(s):  
Mark Y. Jeng ◽  
Philip A. Hull ◽  
Mingjian Fei ◽  
Hye-Sook Kwon ◽  
Chia-Lin Tsou ◽  
...  
Keyword(s):  
T Cells ◽  
Molecular Mechanisms ◽  
Memory T Cells ◽  
Cell Metabolism ◽  
Global Gene Expression ◽  
Metabolic Reprogramming ◽  
Transcriptional Reprogramming ◽  
Age Related ◽  
Global Gene Expression Profiling ◽  
And Function

The expansion of CD8+CD28– T cells, a population of terminally differentiated memory T cells, is one of the most consistent immunological changes in humans during aging. CD8+CD28– T cells are highly cytotoxic, and their frequency is linked to many age-related diseases. As they do not accumulate in mice, many of the molecular mechanisms regulating their fate and function remain unclear. In this paper, we find that human CD8+CD28– T cells, under resting conditions, have an enhanced capacity to use glycolysis, a function linked to decreased expression of the NAD+-dependent protein deacetylase SIRT1. Global gene expression profiling identified the transcription factor FoxO1 as a SIRT1 target involved in transcriptional reprogramming of CD8+CD28– T cells. FoxO1 is proteasomally degraded in SIRT1-deficient CD8+CD28– T cells, and inhibiting its activity in resting CD8+CD28+ T cells enhanced glycolytic capacity and granzyme B production as in CD8+CD28– T cells. These data identify the evolutionarily conserved SIRT1–FoxO1 axis as a regulator of resting CD8+ memory T cell metabolism and activity in humans.

scholarly journals Evaluation of Bone Marrow CD8+ tissue-Resident Memory T Cells in Multiple Myeloma

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 4389-4389
Author(s):  
Melania Carlisi ◽  
Salvatrice Mancuso ◽  
Marco Pio La Manna ◽  
Valentina Orlando ◽  
Nadia Caccamo ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
T Cells ◽  
Memory T Cells ◽  
Ex Vivo ◽  
Effector Memory ◽  
In Vitro Experiments ◽  
Resident Memory T Cells ◽  
Homeostatic Cytokines

Background: CD8+ T cell responses are an essential component of the adaptive immune system. After resolution of infection a small population of memory cells is formed. In relation to circulatory patterns, different subsets of memory CD8+ T cells can be identified: the central memory (CM) and the effector memory T cells (EM) (Martin MD, et al., Front Immunol. 2018). In addition, it has been described a subset of resident memory T cells (TRM) permanently living in peripheral tissues, including the bone marrow (BM) (Di Rosa F., et al., Nat Rev Immunol. 2016). It is conceivable that these cells can contribute to the defence toward haematological tumours infiltrating the BM. Therefore, we performed a study to evaluate the frequency and the phenotype of BM CD8+ TRM in patients with multiple myeloma (MM). Moreover, to evaluate the contribution that the microenvironment can have on the homeostatic and functional maintenance of these cells, we performed in vitro experiments of BM-derived mononucleate cells of MM patients cultured in the presence of different homeostatic cytokines. Patients and Methods: we prospectively analysed 21 patients, 16 with a new diagnosis of IgA, IgG and light chain multiple myeloma (MM) and 5 with IgA and IgG smoldering myeloma (SM). At the time of the bone marrow assessment, we collected a sample for the flow cytometry analysis and in vitro cell culture. The ex vivo evaluation of CD8+ TRM frequency and phenotype in BM samples was performed using anti-human mAbs to CD3, CD103, CD69, CD45, CD8, CD45RA and CCR7 (CD197). The sequential gating strategy was: gate on lymphocytes population with CD45 vs. SSC, 7AAD negative cells, exclusion of doublets with FSC-H vs. FSC-A, CD8+CD3+ and evaluation of percentage of CD103+CD69+ cells. Was also established the subsets using CCR7 and CD45RA. Moreover, to evaluate the role of the microenvironment on maintenance of these cells, we performed in vitro experiments of BM-derived mononucleate cells of MM patients cultured in the presence of homeostatic cytokines in maintaining these cells for a long time. BM derived mononucleate cells from patients were then cultured in vitro in complete RPMI with 10% of human serum for 4 days with IL15 (25 ng/ml), IL7 (25 ng/ml) and TGF-β (2 ng/ml), in different combination and in RPMI alone. After culture, we analyzed the frequency of CD8+ TRM and the proliferating fraction with intracellular staining with anti human Ki67 APC. Non parametric Mann-Whitney and Kruskall-wallis tests were performed to determine statistical differences in the distribution of the results using GraphPad Prism 7.00. Values of * p<0.05 were considered significant. Results: the ex vivo average frequency of CD8+ TRM in 16 MM patients was of 0.48% and the phenotype was represented mainly by TEM (72,9%) followed by TEMRA (12.3%) and (7,6%) of naïve cells and (7,2%) of TCM (Fig. A). The comparison with the ex vivo frequency of CD8 TRM in SM patients did not show any significant difference between two groups (data not showed). To evaluate factors capable of maintaining or to induce the expansion of these cells in vitro, we maintained BM-derived mononucleate cells from MM patients for 4 days in presence of homeostatic cytokines, IL-15, IL-7 plus IL-15 and IL-7 together with IL-15 and TGF-β. The result showed an increase of the percentage of CD8+ TRM in all conditions tested, especially in presence of all cytokines (Fig. B), with a percentage of CD8 TRM of 2,74%. Regarding the phenotype distribution, we observed an expansion of CM compared to the other subsets (Fig. C). We also analysed the percentage of CD8+ TRM proliferating through the identification of Ki67 positive cells. Data highlight that IL-15 gives the strongest proliferative input, but also other cytokines contribute to the homeostatic maintenance of these cells (Fig. D). Conclusions: we evaluated the frequency and the phenotype of CD8 TRM in BM of MM patients compared to SM patients with the conclusion that these cells do not differ significantly in percentage and phenotypic distribution in both conditions. In MM patients, the increase of CD8+ TRM cells with a CM phenotype after in vitro culture with the three cytokines could have an anti-tumor role in the control of MM. Further studies are needed to investigate the cytotoxic capacity of these cells against myeloma cells, in order to study their functional role, also in the perspective of a possible use in future therapeutic programs. Disclosures No relevant conflicts of interest to declare.

scholarly journals α 4 β 7 + CD4 + Effector/Effector Memory T Cells Differentiate into Productively and Latently Infected Central Memory T Cells by Transforming Growth Factor β1 during HIV-1 Infection

2018 ◽  
Vol 92 (8) ◽  
Author(s):  
Ka-Wai Cheung ◽  
Tongjin Wu ◽  
Sai Fan Ho ◽  
Yik Chun Wong ◽  
Li Liu ◽  
...  
Keyword(s):  
T Cells ◽  
Cd4 T Cells ◽  
Transforming Growth Factor ◽  
Memory T Cells ◽  
Central Memory ◽  
Effector Memory ◽  
Effector Memory T Cells ◽  
Latently Infected ◽  
Hiv 1

ABSTRACT HIV-1 transmission occurs mainly through mucosal tissues. During mucosal transmission, HIV-1 preferentially infects α 4 β 7 + gut-homing CCR7 − CD4 + effector/effector memory T cells (T EM ) and results in massive depletion of these cells and other subsets of T EM in gut-associated lymphoid tissues. However, besides being eliminated by HIV-1, the role of T EM during the early stage of infection remains inconclusive. Here, using in vitro -induced α 4 β 7 + gut-homing T EM (α 4 β 7 + T EM ), we found that α 4 β 7 + T EM differentiated into CCR7 + CD4 + central memory T cells (T CM ). This differentiation was HIV-1 independent but was inhibited by SB431542, a specific transforming growth factor β (TGF-β) receptor I kinase inhibitor. Consistently, T EM -to-T CM differentiation was observed in α 4 β 7 + T EM stimulated with TGF-β1 (TGF-β). The T CM properties of the TGF-β-induced T EM -derived T CM (α 4 β 7 + T CM ) were confirmed by their enhanced CCL19 chemotaxis and the downregulation of surface CCR7 upon T cell activation in vitro . Importantly, the effect of TGF-β on T CM differentiation also held in T EM directly isolated from peripheral blood. To investigate the significance of the TGF-β-dependent T EM -to-T CM differentiation in HIV/AIDS pathogenesis, we observed that both productively and latently infected α 4 β 7 + T CM could differentiate from α 4 β 7 + T EM in the presence of TGF-β during HIV-1 infection. Collectively, this study not only provides a new insight for the plasticity of T EM but also suggests that the TGF-β-dependent T EM -to-T CM differentiation is a previously unrecognized mechanism for the formation of latently infected T CM after HIV-1 infection. IMPORTANCE HIV-1 is the causative agent of HIV/AIDS, which has led to millions of deaths in the past 30 years. Although the implementation of highly active antiretroviral therapy has remarkably reduced the HIV-1-related morbidity and mortality, HIV-1 is not eradicated in treated patients due to the presence of latent reservoirs. Besides, the pathogenesis in CD4 T cells early after infection still remains elusive. Immediately after HIV-1 mucosal infection, CD4 T cells are preferentially infected and depleted. However, in addition to being depleted, the other roles of the CD4 T cells, especially the effector/effector memory T cells (T EM ), in disease progression are not completely understood. The significance of this study is in revealing a novel mechanism for the formation of latently HIV-1-infected central memory CD4 T cells, a major latent reservoir from CD4 T EM after infection. Our findings suggest previously unrecognized roles of CD4 T EM in HIV-1 pathogenesis.

Adoptive Immunotherapy with Memory T Cells.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. sci-25-sci-25 ◽  
Author(s):  
Helen E. Heslop
Keyword(s):  
T Cells ◽  
T Cell ◽  
Memory T Cells ◽  
Ex Vivo ◽  
Central Memory ◽  
Effector Memory ◽  
Memory Phenotype ◽  
Effector Memory T Cells ◽  
Effector Memory Cells

Clinical adoptive cellular immunotherapy of malignancy and viral infection should transfer T cells that expand in vivo on exposure to antigen and can enter the memory compartment to persist long-term. A number of factors, including cellular phenotype, influence the behavior of the infused line. Primate studies have shown that antigen-specific CD8+ T cell clones only persisted long-term in vivo if they were derived from central memory T cells, but not from effector memory T cells, reacquiring phenotypic and functional properties of memory T cells.1 Other studies have suggested that adoptive transfer of ex vivo-expanded effector memory T cells will have poor survival and clinical efficacy, reporting instead that less differentiated T cells with longer telomeres exhibit longer persistence. These data imply that prolonged ex vivo expansion, required, for example, for T cell cloning, adversely affects subsequent in vivo expansion and survival. However, our trials administering ex vivo-expanded, polyclonal EBV-specific T cell lines demonstrated that expanded effector memory T cells, infused into a lymphodepleted host, can expand massively in vivo, enter the memory compartment, and persist for up to seven years after infusion. Furthermore, in a study infusing trivirus-specific CTLs with effector memory phenotype, we saw expansion of CTLs specific for the latent viruses CMV and EBV. By contrast, adenoviral-specific CTL persisted only in patients who were acutely infected with the agent2 We recently compared non-specifically activated T cells (ATC) with EBV-specific CTLs derived from the same initial peripheral blood collection and expressing distinguishable chimeric GD2-specific chimeric antigen receptors (CARATC and CAR-CTL). In this study, ATCs were cultured for 14 to 21 days. Between 0.9% and 6.1% retained a central memory (CCR7+, CD62L+) phenotype, up to 30% had an effector memory phenotype (CCR7−, CD62L+), and the remainder had a terminally/fully differentiated effector phenotype. By contrast, EBV-CTL were cultured for 30 to 44 days and expressed no CCR7, but up to 50% were CD62L+, and contained cells that were terminally/fully differentiated effectors and effector memory cells. These EBV-CTLs also all had a CD45RO memory phenotype, while about 13% to 60% of ATCs expressed CD45RA, a marker of naïve T cells. Despite these differences in memory subsets, it was the CAR-CTLs that had the clearly greater persistence and could be shown to retain functionality, while CAR-ATC rapidly disappeared from the circulation and could not be recovered. Hence, factors other than phenotype, such as antigenic stimulation and costimulation almost certainly influence cell fate after infusion, and determine whether or not effector memory cells can re-access the central memory pool. Ultimately, strategies that combine selection of optimal phenotype with the provision of antigen stimulation and co-stimulation and a cytokine milieu that favors homeostatic expansion will likely lead to the most effective outcomes following adoptive T cell transfer.

scholarly journals Long‐term in vitro expansion ensures increased yield of central memory T cells as perspective for manufacturing challenges

2021 ◽  
Author(s):  
Stefanie Herda ◽  
Andreas Heimann ◽  
Benedikt Obermayer ◽  
Elisa Ciraolo ◽  
Stefanie Althoff ◽  
...  
Keyword(s):  
T Cells ◽  
Memory T Cells ◽  
Central Memory ◽  
Central Memory T Cells ◽  
In Vitro Expansion

Enhanced Infiltration of Central Memory T Cells to the Lung Tissue during Allergic Lung Inflammation

2021 ◽  
pp. 1-15
Author(s):  
Mashael Alabed ◽  
Asma Sultana Shaik ◽  
Narjes Saheb Sharif-Askari ◽  
Fatemeh Saheb Sharif-Askari ◽  
Shirin Hafezi ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lung Tissue ◽  
Lung Inflammation ◽  
Inflammatory Responses ◽  
Memory T Cells ◽  
Central Memory ◽  
Effector Memory ◽  
Differential Distribution ◽  
Allergic Lung Inflammation

Memory T cells play a central role in regulating inflammatory responses during asthma. However, tissue distribution of effector memory (T<sub>EM</sub>) and central memory (T<sub>CM</sub>) T-cell subtypes, their differentiation, and their contribution to the persistence of lung tissue inflammation during asthma are not well understood. Interestingly, an increase in survival and persistence of memory T cells was reported in asthmatic lungs, which may suggest a shift toward the more persistent T<sub>CM</sub> phenotype. In this report, we investigated the differential distribution of memory T-cell subtypes during allergic lung inflammation and the mechanism regulating that. Using an OVA-sensitized asthma mouse model, we observed a significant increase in the frequency of T<sub>CM</sub> cells in inflamed lungs compared to healthy controls. Interestingly, adoptive transfer techniques confirmed substantial infiltration of T<sub>CM</sub> cells to lung tissues during allergic airway inflammation. Expression levels of T<sub>CM</sub> homing receptors, CD34 and GlyCAM-1, were also significantly upregulated in the lung tissues of OVA-sensitized mice, which may facilitate the increased T<sub>CM</sub> infiltration into inflamed lungs. Moreover, a substantial increase in the relative expression of T<sub>CM</sub> profile-associated genes (EOMES, BCL-6, ID3, TCF-7, BCL-2, BIM, and BMI-1) was noted for T<sub>EM</sub> cells during lung inflammation, suggesting a shift for T<sub>EM</sub> into the T<sub>CM</sub> state. To our knowledge, this is the first study to report an increased infiltration of T<sub>CM</sub> cells into inflamed lung tissues and to suggest differentiation of T<sub>EM</sub> to T<sub>CM</sub> cells in these tissues. Therapeutic interference at T<sub>CM</sub> infiltration or differentiations could constitute an alternative treatment approach for lung inflammation.

scholarly journals In vitro maintaining of CD4+ central and effector memory cells in normal and inflammatory conditions

2020 ◽  
Vol 22 (5) ◽  
pp. 837-846
Author(s):  
E. A. Blinova ◽  
A. V. Kolerova ◽  
V. E. Balyasnikov ◽  
V. A. Kozlov
Keyword(s):  
T Cells ◽  
Memory T Cells ◽  
Cell Receptor ◽  
Whole Body ◽  
Effector Memory ◽  
Adherent Cell ◽  
Memory Cells ◽  
Inflammatory Conditions ◽  
Low Sensitivity

IL-7 is a key factor for the survival and maintenance of CD4+ central (Tcm) and effector (Tem) memory cells in the whole body. In many autoimmune diseases, an elevated level of IL-7 is detected in blood serum and at the site of inflammation, thus suggesting participation of this homeostatic factor in the survival of memory T cells, including auto-reactive clones, in inflammatory disorders. The aim of the study was to investigate the mechanisms of maintaining CD4+ memory T cells under normal and inflammatory conditions. We developed an in vitro model of inflammation, based on induction of pro-inflammatory cytokines, and then evaluated the effects of IL-7 upon purified sorted populations of CD4+Tcm and Tem under normal conditions and in vitro inflammatory model. IL-7 treatment promoted maintenance of CD4+Tcm phenotype in all variants of cultures. In the absence of contact with adherent cell fraction, the IL-7-induced proliferation of Tcm and Tem was slightly reduced, both under normal and inflammatory conditions, thus suggesting low sensitivity of memory T cells to contacts with MHC, and, probably, a requirement for additional signals to provide complete stimulation with IL-7. The last suggestion is also supported by data about CD127 and CD132 expression, i.e., in the absence of contact with MHC, the proportion of CD127+CD132+ cells was decreased in both subpopulations of CD4+ memory cells. Upon in vitro cultures, IL-7 contributed to decreased expression of CD127, and increased expression of CD132 on CD4+Tcm and Tem. We have evaluated the CD4+Tcm and Tem populations by affinity of T cell receptor (TCR), using the level of CD5 expression. Т cells with high TCR affinity for self-antigens are known to have higher expression of CD5. In comparison to Tem, the Tcm contained more CD5high cells. In cultures, IL-7 promoted a high level of CD5 expression on Tcm, which was comparable to levels observed in peripheral blood cells. High CD5 expression on Tem was observed after stimulation with IL-7 in the in vitro inflammatory model. In the absence of contact with MHC, the number of CD5high cells decreased among CD4+Tem and Tcm. Thus, CD4+Tcm cells with high affinity for autologous antigens are probably dependent on the presence of homeostatic factors, in particular, IL-7, and contacts with antigen-presenting cells (APCs). Under conditions of inflammation, no changes were revealed in the mechanism of maintaining CD4+Tcm, in contrast to CD4+Tem. Being less dependent on IL-7 under normal conditions, CD4+CD5highTem are accumulated in the presence of IL-7 under in vitro inflammatory conditions.

scholarly journals WFDC1/ps20 Is a Novel Innate Immunomodulatory Signature Protein of Human Immunodeficiency Virus (HIV)-Permissive CD4+ CD45RO+ Memory T Cells That Promotes Infection by Upregulating CD54 Integrin Expression and Is Elevated in HIV Type 1 Infection

2007 ◽  
Vol 82 (1) ◽  
pp. 471-486 ◽  
Author(s):  
R. Alvarez ◽  
J. Reading ◽  
D. F. L. King ◽  
M. Hayes ◽  
P. Easterbrook ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
T Cells ◽  
Cd4 T Cells ◽  
Memory T Cells ◽  
Ex Vivo ◽  
Interleukin 2 ◽  
Integrin Expression ◽  
Small Interference Rna ◽  
Immunodeficiency Virus

ABSTRACT Understanding why human immunodeficiency virus (HIV) preferentially infects some CD4+ CD45RO+ memory T cells has implications for antiviral immunity and pathogenesis. We report that differential expression of a novel secreted factor, ps20, previously implicated in tissue remodeling, may underlie why some CD4 T cells are preferentially targeted. We show that (i) there is a significant positive correlation between endogenous ps20 mRNA in diverse CD4 T-cell populations and in vitro infection, (ii) a ps20+ permissive cell can be made less permissive by antibody blockade- or small-interference RNA-mediated knockdown of endogenous ps20, and (iii) conversely, a ps20low cell can be more permissive by adding ps20 exogenously or engineering stable ps20 expression by retroviral transduction. ps20 expression is normally detectable in CD4 T cells after in vitro activation and interleukin-2 expansion, and such oligoclonal populations comprise ps20positive and ps20low/negative isogenic clones at an early differentiation stage (CD45RO+/CD25+/CD28+/CD57−). This pattern is altered in chronic HIV infection, where ex vivo CD4+ CD45RO+ T cells express elevated ps20. ps20 promoted HIV entry via fusion and augmented CD54 integrin expression; both of these effects were reversed by anti-ps20 antibody. We therefore propose ps20 to be a novel signature of HIV-permissive CD4 T cells that promotes infection in an autocrine and paracrine manner and that HIV has coopted a fundamental role of ps20 in promoting cell adhesion for its benefit. Disrupting the ps20 pathway may therefore provide a novel anti-HIV strategy.

Cytotoxic minor histocompatibility antigen HA-1–specific CD8+ effector memory T cells: artificial APCs pave the way for clinical application by potent primary in vitro induction

Blood ◽  
2005 ◽  
Vol 106 (1) ◽  
pp. 144-149 ◽  
Author(s):  
Karin Schilbach ◽  
Gunter Kerst ◽  
Steffen Walter ◽  
Matthias Eyrich ◽  
Dorothee Wernet ◽  
...  
Keyword(s):  
T Cells ◽  
Ex Vivo ◽  
Cell Receptor ◽  
Histocompatibility Antigen ◽  
Effector Memory ◽  
Minor Histocompatibility Antigen ◽  
Tcr Repertoire ◽  
Long Time ◽  
In Vitro Induction

Induction of cytotoxic T lymphocytes (CTLs) for treatment of relapsed leukemia after allogeneic stem-cell transplantation is hindered by the laborious and time-consuming procedure of generating dendritic cells for antigen presentation. Artificial antigen-presenting cells (aAPCs) offer the advantage of being readily available in sufficient numbers, thus allowing for a highly standardized in vitro induction of CTLs. We generated aAPCs coated with anti-CD28 antibody (Ab) and either high-density (HD) or low-density (LD) major histocompatibility complex (MHC) class I molecules loaded with HA-1H, a nonapeptide derived from the hematopoiesis-restricted minor histocompatibility antigen HA-1. HD- and LD-aAPCs potently induced HA-1H–specific CD8+ CTLs from untouched CD8+ T cells of healthy donors. CTLs were subsequently purified by magnetic-activated cell sorting. HD- as well as LD-aAPC–induced CTLs exerted high HA-1H–specific cytotoxicity, resembled Tc1 effector memory cells, survived a long time in vitro, and were expanded by a factor varying between 8.2 × 104 and 51 × 104. The T-cell receptor (TCR) repertoire of HA-1H tetramer–positive CTLs was oligoclonal with a prominent usage of Vβ6. The TCR repertoire of tetramer-positive CTLs was distinct from and more restricted than that of tetramer-negative cells. These findings indicate that aAPCs are attractive tools for the ex vivo generation of HA-1H–specific CTLs suitable for immunotherapy of relapsed leukemia.

Sign in / Sign up

Export Citation Format

Share Document