scholarly journals AB0041 CD8+ T CELLS HAVE AN ELEVATED PROLIFERATIVE CAPACITY IN GIANT CELL ARTERITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1323.1-1323
Author(s):  
R. Reitsema ◽  
R. Hid Cadena ◽  
W. Abdulahad ◽  
A. Boots ◽  
P. Heeringa ◽  
...  
Keyword(s):  
T Cells ◽  
Giant Cell Arteritis ◽  
Giant Cell ◽  
Cd8 T Cells ◽  
Mononuclear Cells ◽  
Effector Memory ◽  
Newly Diagnosed ◽  
Proliferative Capacity ◽  
Ki 67 ◽  
Age And Sex

Background:Giant cell arteritis (GCA) is the most frequent form of systemic vasculitis affecting the large- and medium-sized vessels. The involvement of innate immune cells and CD4+ T cells in the pathogenesis of GCA has been extensively studied. Interestingly, recent findings suggest a role for CD8+ T cells in disease development (1). However, CD8+ subsets and their functional capacities have not yet been studied in detail.Objectives:This study aims to characterize the phenotype and proliferative capacity of CD8+ T cells in newly diagnosed GCA patients and GCA patients in remission compared to healthy age- and sex- matched controls.Methods:To determine the phenotype of CD8+ T cells in GCA, newly diagnosed, untreated GCA patients (baseline, n=14), GCA patients in stable glucocorticoid-free remission (GC-FR, n=10) and age- and sex-matched healthy controls (HCs, n=18) were enrolled. Peripheral blood mononuclear cells (PBMCs) were stained with fluorochrome-conjugated antibodies directed against CD3, CD4, CD8, CCR7, CD45RO, Ki-67, CD69 and CD25 and analyzed by flow cytometry. The following differentiation subsets were defined: CD8+ T naive (CD45RO-CCR7+), central memory (TCM, CD45RO+CCR7+), effector memory (TEM, CD45RO+CCR7-) and effector memory re-expressing CD45RA (TEMRA, CD45RO-CCR7-) cells. Secondly, the proliferative capacity of CD8+ T cells was determined in isolated CD3+ T cells of 10 GCA baseline, 10 GCA GC-FR patients and 19 HCs after 5 days of stimulation with plate-bound anti-CD3 or anti-CD3 plus soluble anti-CD28 using a dye-based proliferation assay.Results:A reduced frequency of CD8+ TEMcells was found in GCA baseline patients compared to HCs (p=0.025). Furthermore, a higher frequency of Ki-67+ cells was detected among CD8+ TEMcells in GCA baseline patients than in HCs (p=0.0007), suggesting a higher proliferative activityin vivo.In addition,in vitrostimulation with anti-CD3 and anti-CD3+anti-CD28 led to higher percentages of divided CD8+ T cells in GCA baseline and GC-FR patients than in HCs (p<0.05). Moreover, the frequencies of CD8+ TEMRAcells and the percentage of divided CD8+ T cells upon CD3 stimulation strongly correlated in GCA baseline patients (R=0.79, p=0.009) and GCA GC-FR patients (R=0.67, p=0.039) but not in HCs (R=0.31, p=0.25).Conclusion:GCA baseline patients demonstrate a higher frequency of proliferating circulating CD8+ TEMcells, defined by Ki-67 expression, than HCs. In addition, functional data on induced proliferative capacity suggest that CD8+ T cells from GCA baseline patients are more rapidly activated by crosslinking CD3 and CD3+CD28, suggesting either reduced regulation in these patients or more intrinsic threshold changes. Furthermore, the induced proliferative capacity is also elevated in patients in stable glucocorticoid-free remission. Whether the increased proliferative capacity of total CD8+ T cells in GCA patients is causally linked to the increased frequencies of CD8+ TEMRAcells in these patients requires further investigation.References:[1]Samson M, Ly KH, Tournier B, Janikashvili N, Trad M, Ciudad M, et al. Involvement and prognosis value of CD8+ T cells in giant cell arteritis. J Autoimmun. 2016;72:73–83.Disclosure of Interests:Rosanne Reitsema: None declared, Rebeca Hid Cadena: None declared, Wayel Abdulahad: None declared, Annemieke Boots Consultant of: Grünenthal Gmbh until 2017, Peter Heeringa: None declared, Elisabeth Brouwer Consultant of: Roche (consultancy fee 2017 and 2018 paid to the UMCG), Speakers bureau: Roche (2017 and 2018 paid to the UMCG)

scholarly journals IMMU-30. UTILIZING A NOVEL MASS CYTOMETRY-BASED IMMUNOMONITORING PLATFORM FOR THE CHARACTERIZATION OF VACCINE-REACTIVE, EPITOPE-SPECIFIC CD8+ T-CELLS IN HLA-A*0201+ PATIENTS WITH K27M+ DIFFUSE MIDLINE GLIOMAS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi125-vi125
Author(s):  
Jared Taitt ◽  
Payal Watchmaker ◽  
Takahide Nejo ◽  
Neil Almeida ◽  
Kaori Okada ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cd8 T Cells ◽  
Time Course ◽  
Mononuclear Cells ◽  
Expression Profiles ◽  
Peptide Vaccine ◽  
Effector Memory ◽  
Mass Cytometry ◽  
Peripheral Blood Mononuclear

Abstract Diffuse midline glioma (DMG), including diffuse intrinsic pontine glioma (DIPG) constitutes up to 20% of pediatric brain cancer and has a median survival of less than one year. We have identified a novel HLA-A*02:01-restricted neoantigen epitope encompassing the H3.3K27M mutation and implemented a pilot clinical trial through the Pacific Pediatric Neuro-Oncology Consortium (PNOC007). Newly diagnosed DIPG patients who are HLA-A2+ and H3.3K27M+ underwent radiation therapy, and then received the H3.3K27M peptide vaccine and tetanus toxoid (TT) peptide emulsified in Montanide in combination with poly-ICLC every 3 weeks for a total of 24 weeks. Our objective is to characterize vaccine-induced H3.3K27M-specific T-cell subpopulations in peripheral blood mononuclear cells through the evaluation of surface markers correlated with activation, memory, and exhaustion phenotypes utilizing a novel H3.3K27M-specific dextramer-based mass cytometry method. Through this approach, the temporal expansion of vaccine-reactive CD8+ T-cells was observed in all of patients (n = 4) who completed a minimum of 18 weeks on the study. These T-cells were subsequently stratified into discrete clusters on a tSNE plot using canonical CD8+ T-cell markers. Resultant clusters were further classified by their expression profiles, revealing distinct effector memory and exhausted subpopulations. Chronological monitoring of these groups indicates the time course-dependent development and persistence of vaccine-reactive exhausted and effector memory CD8+ T-cells in 75% of patients analyzed. Furthermore, a comparative analysis of myeloid subpopulations revealed an inverse correlation between the expansion of monocytic myeloid-derived suppressor cells (M-MDSCs) and length of enrollment in the trial. Future plans include the analysis of regulatory T-cells (Tregs) and MDSCs of all enrolled patients to solidify the relationship between the length of stay on the study and prevalence of immunosuppressive populations. This methodology offers insight into the progression of vaccine-induced patient immune responses and exhibits promise as a platform that may be extrapolated to other immunotherapies.

Circulating CD8+ T cells in polymyalgia rheumatica and giant cell arteritis: A review

2001 ◽  
Vol 30 (4) ◽  
pp. 257-271 ◽  
Author(s):  
Victor M. Martinez-Taboada ◽  
Ricardo Blanco ◽  
Concepción Fito ◽  
Maria Jose Bartolome Pacheco ◽  
Miguel Delgado-Rodriguez ◽  
...  
Keyword(s):  
T Cells ◽  
Giant Cell Arteritis ◽  
Giant Cell ◽  
Cd8 T Cells ◽  
Polymyalgia Rheumatica

Clonally Expanded CD8 T Cells in Patients with Polymyalgia Rheumatica and Giant Cell Arteritis

1996 ◽  
Vol 79 (3) ◽  
pp. 263-270 ◽  
Author(s):  
Victor M. Martinez-Taboada ◽  
Jörg J. Goronzy ◽  
Cornelia M. Weyand
Keyword(s):  
T Cells ◽  
Giant Cell Arteritis ◽  
Giant Cell ◽  
Cd8 T Cells ◽  
Polymyalgia Rheumatica

Activation of central/effector memory T cells in advanced gastric cancer patients treated with antiprogrammed death-1 antibody.

2019 ◽  
Vol 37 (4_suppl) ◽  
pp. 54-54 ◽  
Author(s):  
Hirofumi Ohmura ◽  
Kyoko Yamaguchi ◽  
Fumiyasu Hanamura ◽  
Mamoru Ito ◽  
Akitaka Makiyama ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
T Cells ◽  
T Cell ◽  
Advanced Gastric Cancer ◽  
Cd8 T Cells ◽  
Mononuclear Cells ◽  
Effector Memory ◽  
Cytotoxic T Cell ◽  
Programmed Death ◽  
Programmed Death 1

54 Background: Anti-programmed death-1 (PD-1) monoclonal antibody, nivolumab, enhances antitumor activity by inhibiting the interaction of PD-1 and programmed death-1 ligand 1 (PD-L1) and has shown efficacy for advanced gastric cancer (AGC) in the salvage line. However, specific subsets of immune cells predominantly activated during the period of anti-PD-1 therapy for AGC have not been clarified. Methods: Peripheral blood mononuclear cells of 20 AGC patients treated with nivolumab were prospectively obtained before the initial and second administrations of nivolumab, and at the time of progressive disease (PD). The proportion of immune cell subsets were systematically analyzed by flow cytometry, including the expression of costimulatory and coinhibitory molecules such as T-cell immunoglobulin and mucin domain 3 (TIM-3), Lymphocyte-activation gene 3 (LAG-3), T-cell immunoreceptor with Ig and ITIM domains (TIGIT), cytotoxic T cell antigen-4 (CTLA-4), CD28, OX40, and inducible T cell costimulator (ICOS). Association between changes in the proportion of the subsets and therapeutic effect were analyzed. Results: Median progression free survival (PFS) of the whole patients was 51 days (95% CI 35–83). After a single course of nivolumab, patients showed a significant increase in activated effector memory and activated effector subsets of CD4+/CD8+ T cells (p = 0.018, 0.018, 0.032, 0.024). At the time of PD, proportions of myeloid dendritic cell, IgM memory B cell and Tfh-Th1/17 cell subsets decreased (p = 0.024, 0.013, 0.0039). On the other hand, LAG3 positive CD4+/CD8+ T cells, TIM-3 positive CD4+/CD8 T cells increased at the time of PD (p = 0.013, 0.032, 0.042, 0.042). Significant positive correlations were found between PFS and the proportion of LAG3 positive CD4+/CD8+ T cells (p = 0.0056, 0.0054), OX 40 positive CD4+/CD8+ T cells (p = 0.0034, 0.0006) prior to the initial nivolumab therapy. Conclusions: Nivolumab therapy enhances activation of effector memory and effector subsets of CD4+/CD8+ T cells. The expression level of LAG3 and OX40 on T cells might be correlated with efficacy of nivolumab therapy.

Involvement and prognosis value of CD8 + T cells in giant cell arteritis

2016 ◽  
Vol 72 ◽  
pp. 73-83 ◽  
Author(s):  
Maxime Samson ◽  
Kim Heang Ly ◽  
Benjamin Tournier ◽  
Nona Janikashvili ◽  
Malika Trad ◽  
...  
Keyword(s):  
T Cells ◽  
Giant Cell Arteritis ◽  
Giant Cell ◽  
Cd8 T Cells

scholarly journals IMMU-23. A NOVEL MASS CYTOMETRY-BASED MULTI-PARAMETER CHARACTERIZATION OF NEOANTIGEN-REACTIVE CD8+ T-CELLS IN PATIENTS PARTICIPATING IN PNOC007 H3.3K27M PEPTIDE VACCINE CLINICAL TRIAL

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii364-iii365
Author(s):  
Jared Taitt ◽  
Takahide Nejo ◽  
Payal Watchmaker ◽  
Neil Almeida ◽  
Kaori Okada ◽  
...  
Keyword(s):  
Clinical Trial ◽  
T Cells ◽  
Clinical Outcomes ◽  
Cd8 T Cells ◽  
Mononuclear Cells ◽  
Expression Profiles ◽  
Peptide Vaccine ◽  
Effector Memory ◽  
Mass Cytometry ◽  
Peripheral Blood Mononuclear

Abstract BACKGROUND We have identified an HLA-A*02:01-restricted neoantigen epitope encompassing the H3.3K27M mutation and implemented a multi-center clinical trial of the peptide vaccine through the Pacific Pediatric Neuro-Oncology Consortium (PNOC007) for patients with diffuse midline glioma (DMG), including diffuse intrinsic pontine glioma (DIPG). We sought to characterize vaccine-reactive CD8+T-cells subpopulations using their precise activation and developmental status to find their associations with clinical outcomes. METHODS Mass cytometry (CyTOF) analysis was performed on patient-derived peripheral blood mononuclear cells collected at baseline as well as pre-specified time points throughout the study. Each cell subtype was characterized via tSNE-clustering based on their expression profiles and quantified as a fraction of total CD45+cells. H3.3K27M-reactive CD8+T-cells were evaluated using an H3.3K27M-HLA-A2 dextramer along with a panel of T-cell and myeloid markers. RESULTS Among all 29 patients enrolled, we analyzed samples from all 19 DIPG and 9 of 10 non-brainstem DMG cases, of which 18 had longitudinal samples available (range: 2–5). Utilizing a novel CyTOF-based immuno-monitoring platform, the expansion of H3.3K27M-reactive CD8+T-cells, defined as a 25% increase at any time-point relative to baseline, was observed in 7 of these 18 patients. Survival analyses indicated that the expansion of H3.3K27M-reactive CD8+T-cells, particularly the effector-memory phenotype, positively correlated with longer overall survival (OS) (median: 16.1 vs 9.7 months, p=0.03), whereas an abundance of early and monocytic myeloid-derived suppressor cells at baseline correlated with shorter OS among DIPG patients (9.5 vs 14.3 months, p=0.002). CONCLUSION Our novel immuno-monitoring approach offers insight into how vaccine-induced immune responses impact clinical outcomes.

scholarly journals Effects of storage time and temperature on highly multiparametric flow analysis of peripheral blood samples; implications for clinical trial samples

2021 ◽  
Vol 41 (2) ◽  
Author(s):  
Amelia Jerram ◽  
Thomas V. Guy ◽  
Lucinda Beutler ◽  
Bavani Gunasegaran ◽  
Ronald Sluyter ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Gradient Centrifugation ◽  
Effector Memory ◽  
Low Density ◽  
Blood Draw ◽  
Blood Samples

Abstract We sought to determine the effect of time and temperature of blood sample storage before preparation of human peripheral blood mononuclear cells (PBMCs) by Ficoll-hypaque density gradient centrifugation. Blood samples from healthy donors were stored at room temperature (RT) or refrigerated at 4°C before preparation of PBMCs. Cell yield and viability, and proportions of major cell populations within PBMCs, as determined by fluorescence flow cytometry, were assessed for both fresh and cryopreserved samples. Highly multiparametric mass cytometry was performed on cryopreserved PBMCs. We found that refrigeration had marked negative effects on subsequent PBMC yield. Storage at RT led to co-purification of low density neutrophils with PBMCs, but had no detectable effects on the proportions of multiple cell subsets including, but not limited to, monocytes, NK cells, B cells, Treg cells, and naïve, central memory and effector memory CD4+ and CD8+ T cells and CD45RA-positive terminal effector CD8+ T cells. Expression of a number of cell surface receptors, including CXCR5, CCR6, CXCR3 and TIGIT, but not CD247 was reduced after RT storage before PBMC preparation, and this effect correlated with the degree of low density neutrophil contamination. As such, when PBMC preparation cannot be undertaken immediately after blood draw, storage at RT is far superior to refrigeration. RT storage leads to neutrophil activation, but does not compromise measurement of PBMC subset distribution. However caution must be applied to interpretation of cytometric measurements of surface molecules such as chemokine receptors.

scholarly journals Impaired Anti-Tumor T cell Response in Hepatocellular Carcinoma

Cancers ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 627 ◽  
Author(s):  
Nada Chaoul ◽  
Serena Mancarella ◽  
Luigi Lupo ◽  
Gianluigi Giannelli ◽  
Francesco Dituri
Keyword(s):  
Hepatocellular Carcinoma ◽  
T Cells ◽  
Cd8 T Cells ◽  
Mononuclear Cells ◽  
Immune Cell ◽  
T Cell Response ◽  
Effector Memory ◽  
Peripheral Blood Mononuclear ◽  
Immune Cell Subsets ◽  
Memory Cd4 T Cells

Different subsets of lymphocytes have the capacity to promote or counteract the progression of solid cancers, including hepatocellular carcinoma (HCC). Therefore, to determine the infiltrative ability and functional status of major immune cell subtypes into tumor may lead to novel insights from the perspective of immunotherapy. After obtaining single cell suspensions from freshly collected specimens of HCC tumor, along with paired peritumor tissues and peripheral blood mononuclear cells (PBMCs) from 14 patients, we flow-cytometrically identified and quantified the relative frequencies of lymphocyte subsets within the tissues of origin. We found that the recruitment in the tumor of cytotoxic cells, namely the terminally differentiated CD4+ and CD8+ T cells (TEFF), is impaired, whereas the effector memory CD4+ T cells (TEM) are more attracted in this site. Concerning the other subsets, the frequency of NK CD56hi and NKT CD56hi cells infiltration in the tumor is increased, whereas that of NKT CD56low is reduced. Although CD4+ and CD8+ T cells settled in the tumor show a higher degree of activation than the circulating counterpart, they occur with a more exhausted phenotype. Overall, these data demonstrate the prevalently immunosuppressive nature of HCC microenvironment, and prompt us to search for strategies to enhance the activity of anti-tumor immune cell subsets.

507 High dimensional flow cytometry analysis in newly diagnosed acute myeloid leukemia predicts patients outcomes

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A543-A543
Author(s):  
Francesco Mazziotta ◽  
Rupkatha Mukhopadhyay ◽  
Hanna A Knaus ◽  
Anish Chowdhury ◽  
Amanda Blackford ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Cd8 T Cells ◽  
Myeloid Leukemia ◽  
Cd8 T Cell ◽  
Effector Memory ◽  
Newly Diagnosed ◽  
Response To Chemotherapy ◽  
Acute Myeloid

BackgroundWe have previously characterized phenotypic and transcriptional profile of CD8+ T cells in acute myeloid leukemia (AML) and their differences between responders vs. nonresponders to chemotherapy.1 Goal of ongoing work was to further probe uniqueness of AML in sculpting CD8+ T cell responses and the plasticity of their signatures upon chemotherapy response.MethodsWe first examined the cumulative expression of multiple inhibitory receptors (IRs) (detected by 2 different panels) on CD8+ T cells and created an IR-score which summarizes the relative amount of PD-1, Tim3, KLRG1, 2B4, CD160, CD57, and BTLA-positive CD8+ T-cells in relation to the well-characterized maturation states of CD8+ T cells. Serial bone marrow samples from 33 newly diagnosed AML patients with well-annotated clinical data (21 complete responders (CR) and 12 nonresponders (NR) to chemotherapy) and 11 healthy controls (HC) were analyzed. FInally, using custom made R code, we performed dimensionality reduction, clustering, and pseudotime analysis.ResultsThe IR-score discriminated NR and CR (p = 3e-02, AUC 0.84) after treatment with CD57 and KLRG1 accounting for most of this difference (p = 2e-02, AUC = 0.79). Next we investigated CD8+ T cell populations that best correlated with response to chemotherapy. FlowSOM revealed seven major clusters: naive and naive-like, CD28+KLRG1+ activated-effector, CD28+KLRG1+PD1+ dysfunctional, PD1+CD57+ senescent effector-memory and two clusters of terminally differentiated CD45RA+KLRG1+ cells. Since the activation and differentiation states accounted for most of the subpopulation variability, we grouped the clusters into resting (naive, naive-like), activated (activated-effector, dysfunctional), and terminally differentiated cells (senescent effector-memory, terminally differentiated). UMAP, developmental trajectories and differential abundance testing showed increased frequency of activated cells at diagnosis (p-adj = 2.9e-05) and of resting cells after treatment (p-adj = 1.3e-02) in CR, while terminally differentiated T cells prevailed in NR (p-adj = 5.3e-08) after treatment (figures 1 and 2).Abstract 507 Figure 1UMAP embedding of T cells in CR, NR, at diagnosis (BM_DG) and after chemotherapy (BM_post), HC colored by T cell state (resting, activated, terminal differentiated), overlaid with a contour plotAbstract 507 Figure 2Boxplots showing the differential cluster abundance and adjusted p-values for CR, NR, at diagnosis (BM_DG) and after chemotherapy (BM_post), HC in the three different T cell states (resting, activated, terminal differentiated)ConclusionsThe increased number of functional activated T cells at diagnosis and the persistence of a naive/naive-like reservoir at the time of response is a signature associated with achievement of CR. Lack of response (NR) correlates with accumulation of the terminally differentiated and senescent cells in the bone marrow. These results uncover an intertwined relationship between skewing of T cell differentiation and clinical response to chemotherapy. The data provide rationale to either remove senescent or augment activity of naïve/naïve-like T cells as a strategy to reinforce antileukemia immunity.ReferenceKnaus HA, Berglund S, Hackl H, et al. Signatures of CD8+ T cell dysfunction in AML patients and their reversibility with response to chemotherapy. JCI Insight 2018; 3(21).

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