Immune profiling of circulating T cells and TILs following neoadjuvant ipilimumab and chemotherapy in non-small cell lung cancer (NSCLC).

26 Background: Ipilimumab (Ipi) is a humanized CTLA-4 antibody that blocks binding of CTLA-4 to B7, permitting T cell activation through CD28. Phased in Ipi added to chemotherapy (C) may enhance efficacy in NSCLC. Methods: Patients with stage 2 or 3A NSCLC received neoadjuvant carboplatin AUC6 plus paclitaxel 200 mg/m2 every 21 days for 3 cycles and Ipi (10 mg/kg) was given on day 1 for cycles 2 and 3. Blood for immune profiling of circulating T cells was collected at baseline, after chemotherapy alone, and after chemotherapy plus Ipi. Tumor infiltrating lymphocytes (TIL) were derived from 7 available tumors. Polychromatic flow cytometry (PFC) analyses were performed on peripheral blood mononuclear cells (PBMC) and TIL. Objective response rates were assessed according to RECIST 1.1 criteria. Results: Of the 24 patients enrolled on this study, objective responses after 3 cycles of neoadjuvant C plus ipi included 2 PD, 8 SD, and 14 PR. Phenotypic analyses revealed that PBMC from all 24 patients were highly activated following two cycles of Ipi (cycle 3) as evidenced by significantly increased frequencies of CD28, ICOS, HLA-DR, PD-1, and CTLA-4 expressing CD4+ cells; and ICOS, HLA-DR, and CTLA-4 expressing CD8+ cells. The frequencies of Tregs were highly variable among the 24 participants. Two of the 24 participants had levels of MDSC cells above 15%. TIL contained far greater frequencies of activated CD4+ and CD8+ cells than found in the PBMC at cycle 3. Tumor associated antigen (TAA)-specific CD4+ or CD8+ cells were detected at baseline in 4 patients (24%), but their relative frequencies remained unaltered by Ipi therapy. No patients developed detectable de novo TAA reactivities while on Ipi therapy. Conclusions: Combined neoadjuvant Ipi plus chemotherapy produced significantly increased frequencies of highly activated CD4+ and CD8+ populations in the peripheral blood and the tumor microenvironment. TAA-specific CD4+ or CD8+ cells were detected in PBMC at baseline in a subset of patients. No TAA-reactive T cells were detected among the 7 TIL samples analyzed. Analysis for predictive or pharmacodynamic biomarkers is ongoing. Clinical trial information: NCT01820754.

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T-cell activation and subset patterns are altered in B-CLL and correlate with the stage of the disease

Blood ◽

10.1182/blood.v74.2.786.786 ◽

1989 ◽

Vol 74 (2) ◽

pp. 786-792 ◽

Cited By ~ 64

Author(s):

TH Totterman ◽

M Carlsson ◽

B Simonsson ◽

M Bengtsson ◽

K Nilsson

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Activation ◽

Healthy Subjects ◽

Lymphocytic Leukemia ◽

Stage Ii ◽

T Cell Subsets ◽

Cd8 Cells ◽

Hla Dr ◽

The Absolute

Abstract Two-color FACS analysis was used to study activated and “functional” T and natural killer (NK) cell subsets of circulating lymphocytes in 23 patients with B-type chronic lymphocytic leukemia (B-CLL) and in 30 healthy subjects. As compared with controls, B-CLL patients had increased absolute numbers of phenotypically activated, HLA-DR+ CD4+ and CD8+ cells and T suppressor/effector (CD11b+CD8+) cells. When patients in Rai stages II through IV (n = 11) were compared with cases in Rai stages O through I (n = 12), the former group of patients had higher numbers of activated CD4+ and CD8+ T cells and decreased levels of suppressor/effector T cells. The absolute numbers of T suppressor/inducer (CD45R+CD4+) cells were elevated in patients with stage O through I disease but within normal range in stage II through IV leukemia. We further showed that the absolute numbers of NK-like (CD16+) cells and their activated counterparts (DR+CD16+) are elevated in B-CLL patients as compared with healthy subjects. The comparison of relative T and NK subsets in the blood of patients and controls showed that the proportions of CD4+, CD8+, and CD16+ cells expressing the activation marker HLA-DR were increased in B-CLL. Furthermore, the percentage of T-suppressor/inducer (CD45R+) cells within the CD4+ population was decreased in the patients. The proportion of T- suppressor/effector (CD11b+) cells within the CD8+ subset was reduced in subjects with stage II-IV disease only. When stimulated in vitro with the T-cell-dependent inducer TPA, B-CLL cells from patients in Rai stages II through IV secreted larger amounts of IgM as compared with cells from stage O through I patients. A positive correlation was observed between the degree of phenotypic activation of CD4+ T-helper cells and their functional capacity to augment IgM secretion by autologous B-CLL cells. Our findings indicate a tumor cell-directed regulatory role of T cells (and possibly NK cells as well) in B-CLL. Furthermore, monitoring of phenotypically activated and functional T- cell subsets may be helpful in the prediction of disease progression and timing of therapy in B-CLL.

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Cyclophosphamide Exerts Significant Immunomodulatory Function in Myeloma Patients Treated with Pomalidomide and Dexamethasone

Blood ◽

10.1182/blood-2018-99-111823 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 4482-4482 ◽

Cited By ~ 1

Author(s):

James Croft ◽

Andrew Hall ◽

Katrina Walker ◽

Amy L Sherborne ◽

Kevin Boyd ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Research Funding ◽

Immune Cell ◽

Cd8 Cells ◽

Hla Dr ◽

Trial Treatment ◽

Equity Ownership ◽

First Time ◽

Immune Profiling

Abstract Background and aims Existing evidence regarding the effect of low-dose cyclophosphamide on immune cells in myeloma patients, in particular in combination with the IMiD® compound pomalidomide is limited. We present here for the first time changes in immune cell sub-group composition associated with the addition of cyclophosphamide to pomalidomide in the randomised MUKseven clinical trial. Material and Methods MUKseven is a randomised phase II study for relapsed or refractory myeloma (RRMM) patients comparing cyclophosphamide (500 mg po d1, 15, 21), pomalidomide and dexamethasone (CPd) versus standard pomalidomide and dex (Pd). Patients with ≥2 prior lines of therapy were randomised 1:1 to CPd or Pd and treated until disease progression. All patients underwent bone marrow sampling and peripheral blood collection, the latter for immune cell immunophenotyping at Cycle 1 Day 1 (C1D1; Baseline), C1D14 (on-treatment), C4D14 (on-treatment) and at disease progression. Peripheral blood (PB) T-cell populations were profiled using multicolor flow cytometry (MFC) designed to assess baseline and pharmacodynamic changes in subpopulations including helper, cytotoxic, naïve, memory, and activated/proliferating phenotypes (CD3, CD4, CD8, CD45RA, CD62L, HLA-DR, Ki67. T-cell sub-populations were defined and their respective % of total lymphocyte population used for downstream analyses. Results In total 102 evaluable RRMM patients were randomised, 51 each to CPd and Pd treatment arms, with comparable clinical baseline characteristics. Patients had received a median of 3 prior lines of treatment. Evaluable PB immune profiling data was available for 93 (91%) patients at Baseline, 83 (81%) at C1D14, 55 (54%) at C4D14 and 26 (25%) at progression. We observed trends for changes in baseline T-cell population composition with increasing numbers of prior lines of therapy. Specifically, mean % CD4+ T-cells decreased from 35% for patients with 2 prior lines (n=18) to 30% with 3 (n=33), 23% with 4 and 20% with ≥5 prior lines of treatment (n=27), whilst the % of CD8+ cells were similar, indicating potential differential cumulative effects of anti-myeloma therapy on T-cell populations. We compared changes in T-cell profiles longitudinally over trial treatment from baseline to C1D14 and C4D14 with summary statistics. Overall, there was a marked increase in activated (HLA-DR+) T-cells with therapy, with a 2-fold increase in mean proportion of activated CD4+ and CD8+ from 3.9% and 10.2% at baseline to 7.8% and 19.9% at C1D14 and 7.2% and 28.2% at C4D14, respectively (Figure 1). Trial treatment was associated with a shift in sub-populations within CD8+ T-cells in particular, with a relative % decrease in naïve (CD45RA+) sub-populations and increase in memory (CD45RA-) populations. To identify differences associated with cyclophosphamide treatment a regression analysis was conducted on the C1D14 time point accounting for the treatment a patient received and incorporating their baseline (C1D1) measurement. The mean% estimates for total T-cells (CD3+) at C1D14 were significantly higher for the CPd arm in comparison to Pd: 72.1% [95% CI: 66.5 - 73.6] vs. 64.2% [58.2 - 66.1] (P=0.004). Estimates for the Pd arm appeared similar to baseline [mean C1D1: 61.7%]. Mean% estimates for CD8+ and CD4+ cells were also significantly higher with CPd treatment at C1D14 compared to Pd: 37.2% [32.6 - 38.0] vs. 33.1% [28.5 - 33.7] (P=0.03) and 26.2% [21.6 - 27.0] vs. 21.8% [21.6 - 27.0] (P=0.016), respectively. Importantly, mean% estimates for activated (HLA-DR+) CD4+ cells were significantly higher for the CPd arm 10.1% [6.9 - 9.4] vs 8.1% [5.1 - 7.2] in the Pd arm (P =0.02). There was a trend for additional increase of activated CD8+ cells by addition of cyclophosphamide to Pd therapy (P=0.06). Discussion We demonstrate for the first time in a randomised trial using systematic longitudinal immune profiling that addition of cyclophosphamide to pomalidomide and dexamethasone is significantly associated with altered T-cell profiles and an increased proportion of activated T-cells. MUKseven clinical endpoint data are reported separately, with improved response rates observed for CPd vs. Pd. Correlation of immune profiles with clinical outcomes and tumour genetics will be presented at the conference when PFS outcome data will be mature. Disclosures Boyd: Novartis: Consultancy, Honoraria; Janssen: Honoraria, Other: Travel and Accommodation expenses; Celgene: Consultancy, Honoraria, Other: Advisory role. Garg:Janssen: Honoraria; Novartis: Other: travel support, Research Funding; Takeda: Other: Travel Grant; Amgen: Honoraria, Other: Travel Support. Pawlyn:Janssen: Honoraria, Other: Travel support; Celgene Corporation: Consultancy, Honoraria, Other: Travel support; Amgen: Consultancy, Honoraria, Other: Travel Support; Takeda Oncology: Consultancy, Other: Travel support. Pierceall:Celgene: Employment, Equity Ownership. Cook:Celgene Corporation: Consultancy, Honoraria, Research Funding, Speakers Bureau; Sanofi: Consultancy, Honoraria, Speakers Bureau; Amgen: Consultancy, Honoraria, Research Funding, Speakers Bureau; Glycomimetics: Consultancy, Honoraria; Bristol-Myers Squibb: Consultancy, Honoraria; Seattle Genetics: Honoraria; Janssen: Consultancy, Honoraria, Research Funding, Speakers Bureau; Takeda: Consultancy, Honoraria, Research Funding, Speakers Bureau; Janssen: Consultancy, Honoraria, Research Funding, Speakers Bureau. Thakurta:Celgene Corporation: Employment, Equity Ownership. Kaiser:Chugai: Consultancy; Takeda: Consultancy, Other: travel support; Celgene: Consultancy, Honoraria, Research Funding; Janssen: Consultancy, Honoraria; Bristol-Myers Squibb: Consultancy, Other: travel support; Amgen: Consultancy, Honoraria.

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The Feature of δRec-ψJα sjTRECs Level and Frequency of 23 TCR Vβ-Dβ1 sjTRECs in Mononuclear Cells, CD4+ and CD8+ T Cells from Cord Blood and Peripheral Blood of Normal Individuals.

Blood ◽

10.1182/blood.v108.11.3873.3873 ◽

2006 ◽

Vol 108 (11) ◽

pp. 3873-3873

Author(s):

Yangqiu Li ◽

Qingsong Yin ◽

Shaohua Chen ◽

Lijian Yang ◽

Grzegorz Przybylski ◽

...

Keyword(s):

T Cells ◽

Quantitative Analysis ◽

Cord Blood ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Output Function ◽

Cd4 Cells ◽

Cd8 Cells ◽

Normal Individuals

Abstract Thymic recent output function is characterized its importance of thymus to T-cell diversity in the periphery of both children and adults. The generation of TCR diversity occurs in the thymus through recombination of gene segments encoding the variable parts of the TCR α and β chains. During these processes, by-products of the rearrangements are generated in the form of signal joint T-cell receptor excision circles (sjTRECs), which is considered as a very valuable tool to estimate thymic function. Quantitative of δRec-ψJα sjTRECs can direct evaluate the recent thymic output function, but it is unable to analyze the particular thymic output function of different TCR Vβ subfamily naive T cells. The complexity of TCR Vβ repertoire is an important factor for immune reconstitution, quantitative analysis of series TCR Vβ-Dβ sjTRECs could be used to evaluate the levels of different Vβ subfamily naive T cells. In the present study, quantitative analysis of δRec-ψJα sjTRECs was performed in mononuclear cells, CD3+, CD4+ and CD8+T cells from peripheral blood of normal individuals and cord blood by real-time PCR(TaqMan). And the analysis of 23 TCR Vβ-Dβ1 sjTRECs was performed by semi-nested PCR. Different amounts of DNA (corresponding to 2*105, 5*104, 1*104 and 1*103 cells respectively) from all samples were amplified to estimate the frequency of TCR Vβ-Dβ sjTRECs. The mean value of δRec-ψJα sjTRECs was detected in 4.10±3.65/1000 PBMCs, 6.37±5.28/1000 CD3+cells, 3.28±1.24/1000 CD4+cells, 4.67±3.63/1000 CD8+cells from normal individuals (n=14) and 35.59±47.56/1000 CBMC, 71.48±86.42/1000 CD3+cells, 41.02±32.9/1000 CD4+ cells, 52.05±52.32/1000 CD8+cells from cord blood (n=9) (p=0.0208, p=0.0096, p=0.0003, p=0.0026, respectively). A part of Vβ subfamily sjTRECs could be detected in all samples from cord blood (Vβ2, 3, 4, 5, 10, 13, 14, 15, 19 and 22) and peripheral blood (Vβ10, 13 and 14) at 5*104 cells level, some of Vβ subfamily sjTRECs could be detected in 1*103 cells level. The frequencies of 23 Vβ-Dβ1 sjTRECs were different at the same cellular concentration. The number of detectable Vβ subfamily sjTRECs was 22.00±0.94/2×105, 18.8±1.87/5×104, 10.40±2.99/1×104 and 0.78±1.39/1×103 CBMCs, as compared with 18.70±2.45/2×105 (p=0.002), 13.7±2.67/5×104 (p<0.001), 5.5±2.07/1×104 (p=0.001) and 0.50±0.71/1×103 (p=0.739) in PBMCs from normal individuals. Similar results were found in CD4+ and CD8+ T cells which were sorted from both CBMCs and PBMCs, the number of detectable Vβ subfamily sjTRECs was 13.90±2.38/1×104 CD4+cells, 11.5±1.96/1×104CD8+cells from cord blood and 5.6±2.68/1×104 CD4+cells (p<0.001) and 8.2±2.57/1×104CD8+cells (p>0.005) from normal individuals. The results indicate that the number of detectable sjTRECs of Vβ subfamilies and the frequencies of most Vβ-Dβ1 sjTRECs in normal PBMCs, CD4+ and CD8+T cells were obviously lower than those in cord blood. In conclusions, the results provide the base data of naïve T cells levels and thymic recent output function in cord blood and peripheral blood of normail individuals in chinese.

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POS0413 COMPREHENSIVE IMMUNE PROFILING OF PERIPHERAL BLOOD IN PSORIATIC ARTHRITIS (PsA) PATIENTS: EXPANSION OF INTERMEDIATE MONOCYTES AND DECREASED T REG AND CD8 T CELLS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.3540 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 436.1-436

Author(s):

A. Grivas ◽

M. Grigoriou ◽

P. Katsimpri ◽

P. Verginis ◽

D. Boumpas

Keyword(s):

T Cells ◽

Psoriatic Arthritis ◽

Disease Activity ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Immune Cell ◽

Psoriatic Skin ◽

Parametric Data ◽

Hla Dr ◽

Intermediate Monocytes

Background:Psoriatic arthritis (PsA) is a heterogeneous inflammatory arthritis that develops in a subset of patients with psoriasis. According to the current paradigm, cells of the innate and adaptive immunity interact with resident tissue fibroblasts mounting an inflammatory response via complex cytokine networks in the skin and joints in which type 1 and type 17 T cells play a dominant role. The abundance and relative contribution of other peripheral blood immune cells to disease pathogenesis as well the molecular signature of peripheral blood mononuclear cells and tissue fibroblasts remain ill defined.Objectives:To comprehensively characterize immune cell subsets driving inflammation in the peripheral blood of patients with active PsA and their impact on psoriatic skin fibroblasts.Methods:Peripheral blood was collected from PsA patients (n=31) and age-/sex-matched healthy individuals (HI) (n=9), after informed consent. Psoriatic skin biopsies were acquired from a subset of 5 patients and 3 HI. All patients fulfilled the CASPAR criteria for the diagnosis and displayed peripheral polyarthritis of moderate- to high-disease activity. Patients’ demographic and clinical data were recorded at time of sampling. Disease activity was assessed using the Disease Activity Index for Psoriatic arthritis (DAPSA) score. Skin psoriasis activity indices, enthesitis and dactylitis were also recorded. Peripheral blood mononuclear cells (PBMCs) were isolated by ficoll density gradient centrifugation. Flow cytometry was performed using a BD FACS-Aria-III and analyzed using FlowJo software. The antibody staining panel utilized aimed at the identification of the following immune cell subsets: Monocyte subsets (HLA-DR+ CD14+/- CD16+/-), Plasmacytoid dendritic cells (HLA- DR+ CD123+), T helper (CD4+), cytotoxic T (CD8+), regulatory T (CD4+ CD25+ CD127-) and B cells (CD19+). Statistical analyses were performed using GraphPad Prism software. Differences between groups were compared using unpaired T test for parametric data; Mann-Whitney and Kruskal Wallis tests for non-parametric data. The level of significance was set at P<0.05.Results:9 males and 22 females PsA patients are included (mean age 50 years and the mean disease duration 19.2 years for skin disease and 5.9 years for arthritis). The mean DAPSA score was 43.4, suggestive of high disease activity, while 8 (26%) patients displayed clinical enthesitis at time of sampling. Flow cytometry analysis revealed aberrancies in peripheral blood immune cell populations. More specifically, PsA patients displayed a significant increase in intermediate monocyte subset (HLA-DR+ CD14+ CD16+) compared to HI with patients with clinical enthesitis demonstrating a more exaggerated expansion of intermediate monocytes compared to patients without enthesitis. A trend towards increased patrolling monocytes (HLA-DR+ CD14- CD16+) was also noted although this did not reach statistical significance. In contrast, both regulatory T cells and cytotoxic CD8+ T cells were significantly decreased probably due to their selective migration at the sites of inflammation. RNA-seq from whole blood and skin fibroblasts from affected skin are in progress.Conclusion:These data demonstrate significant expansion of intermediate monocytes -more pronounced in the enthesitis affected individuals- and decrease in T regulatory cells and T cytotoxic cells in PsA peripheral blood. Increased antigen presentation and co-stimulation mediated via intermediate monocytes in combination with their proangiogenic properties may contribute to disease pathogenesisReferences:[1]Veale, D. J. & Fearon, U. The pathogenesis of psoriatic arthritis. The Lancet (2018) doi:10.1016/S0140-6736(18)30830-4.Disclosure of Interests:None declared

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Distinct cellular immune profiles in the airways and blood of critically ill patients with COVID-19

Thorax ◽

10.1136/thoraxjnl-2020-216256 ◽

2021 ◽

pp. thoraxjnl-2020-216256

Author(s):

Anno Saris ◽

Tom DY Reijnders ◽

Esther J Nossent ◽

Alex R Schuurman ◽

Jan Verhoeff ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

T Cell ◽

T Cell Activation ◽

Inflammatory Mediators ◽

Peripheral Blood ◽

Cell Activation ◽

Mononuclear Cells ◽

Effector Memory ◽

Activated T Cells

BackgroundKnowledge of the pathophysiology of COVID-19 is almost exclusively derived from studies that examined the immune response in blood. We here aimed to analyse the pulmonary immune response during severe COVID-19 and to compare this with blood responses.MethodsThis was an observational study in patients with COVID-19 admitted to the intensive care unit (ICU). Mononuclear cells were purified from bronchoalveolar lavage fluid (BALF) and blood, and analysed by spectral flow cytometry; inflammatory mediators were measured in BALF and plasma.FindingsPaired blood and BALF samples were obtained from 17 patients, four of whom died in the ICU. Macrophages and T cells were the most abundant cells in BALF, with a high percentage of T cells expressing the ƴδ T cell receptor. In the lungs, both CD4 and CD8 T cells were predominantly effector memory cells (87·3% and 83·8%, respectively), and these cells expressed higher levels of the exhaustion marker programmad death-1 than in peripheral blood. Prolonged ICU stay (>14 days) was associated with a reduced proportion of activated T cells in peripheral blood and even more so in BALF. T cell activation in blood, but not in BALF, was higher in fatal COVID-19 cases. Increased levels of inflammatory mediators were more pronounced in BALF than in plasma.InterpretationThe bronchoalveolar immune response in COVID-19 has a unique local profile that strongly differs from the immune profile in peripheral blood. Fully elucidating COVID-19 pathophysiology will require investigation of the pulmonary immune response.

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Quantitative analysis of CD4+CD25+FoxP3+ regulatory T-cells in canine atopic dermatitis in Korea

Veterinární Medicína ◽

10.17221/20/2019-vetmed ◽

2020 ◽

Vol 65 (No. 6) ◽

pp. 250-257

Author(s):

DJ Lee ◽

TH Chung ◽

C Park

Keyword(s):

T Cells ◽

Atopic Dermatitis ◽

Regulatory T Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Canine Atopic Dermatitis ◽

Healthy Dogs ◽

The Difference ◽

Circulating T Cells

Recently, it was suggested that CD4+CD25+FoxP3+ Tregs (Regulatory T-cells) exist in canine skin, although their numbers were not significantly different between healthy and atopic dogs. In this study, we investigated whether Treg frequencies correlate with the clinical features of canine atopic dermatitis (cAD). The goal of this study was to compare and analyse the numbers of the circulating Tregs in atopic and healthy dogs. In the peripheral blood mononuclear cells (PBMC) of healthy dogs, Tregs defined as CD4+CD25+FoxP3+ Tregs in the peripheral blood ranged from 0.3% to 1.5%. By contrast, in atopic dogs, the same population ranged from 0.7% to 8.8%. The percentage of CD4+CD25+FoxP3+ Tregs in gated CD4+ T-cells was significantly higher in the peripheral blood of dogs with atopic dermatitis (n = 9) than in the healthy controls (n = 8). The difference in the Treg levels (CD4+CD25+FoxP3+) (P = 0.012) between the atopic and the healthy groups was statistically significant. The circulating T-cells (phenotype CD4+CD25+FoxP3+ and CD4+FoxP3+) were increased significantly in the atopic dogs. The proportion of CD4+CD25+FoxP3+ Tregs of the atopic dogs decreased with advancing age. These findings suggest that changes in the Tregs may mediate the pathogenesis of CAD.

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Persistent expansions of CD4+ CD8+ peripheral blood T cells

Blood ◽

10.1182/blood.v82.5.1546.1546 ◽

1993 ◽

Vol 82 (5) ◽

pp. 1546-1552 ◽

Cited By ~ 45

Author(s):

P Sala ◽

E Tonutti ◽

C Feruglio ◽

F Florian ◽

A Colombatti

Keyword(s):

T Cells ◽

T Cell ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Blot Hybridization ◽

Southern Blot Hybridization ◽

Cell Receptor ◽

Cd8 Cells ◽

Hla Dr ◽

T Cell Receptor Beta

Abstract CD4+ CD8+ cells are present during T cell differentiation in the thymus. Less than 2% of normal T cells that coexpress CD4 and CD8 also are released in the circulation and are present in the peripheral blood. In this study, nine individuals are described that manifested persistent expansions (11% to 43%) of circulating CD4+ CD8+ T cells that in three cases had large granular lymphocyte (LGL) morphology in the absence of either lymphocytosis or overt lymphoproliferative disorders. Southern blot hybridization of enriched CD4+ CD8+ cells with T-cell receptor beta (TCR beta) and TCR gamma probes showed that most cases had the 12-kb Eco RI germinal band deleted or of decreased intensity. In several individuals new TCR beta-specific bands of different intensity and distinct from case to case suggested either monoclonal or oligoclonal and polyclonal expansions. Immunophenotypic analysis showed that in 7 out of 9 cases the CD4+ CD8+ T cells presented with CD8 dim expression. Furthermore, all the CD4+ CD8+ cells did not express many of the known activation antigens (low or absent CD25, CD38, CD71, HLA-DR), whereas they expressed high levels of CD2, CD29, CD56, and CD57. In addition, the CD4+ CD8+ cells of 5 out of 9 subjects coexpressed CD45RA and CD45RO suggesting that these cells might be “frozen” in an intermediate state between naive and memory T cells. In conclusion, the present CD4+ CD8+ cases fall within a larger spectrum of disorders ranging from apparently normal to reactive or proliferative situations and encompassing cells with LGL morphology or LGL-associated antigens expression either in the presence or in the absence of absolute lymphocytosis that deserve careful follow-up investigations.

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CD28 Monoclonal Antibody Improve Sepsis Mortality by Amending Th17/Treg Balance

10.21203/rs.3.rs-253624/v1 ◽

2021 ◽

Author(s):

Yu Wu ◽

Dai Li ◽

Jian Xie ◽

Han Wang ◽

Yan Meng ◽

...

Keyword(s):

Monoclonal Antibody ◽

T Cells ◽

T Cell ◽

T Cell Activation ◽

Peripheral Blood ◽

Crucial Role ◽

Cell Activation ◽

Mononuclear Cells ◽

Cecal Ligation ◽

Inflammatory Factors

Abstract Background: Sepsis is still developing exorbitantly high mortality. In response to microbial molecules, T cell activation plays a crucial role in sepsis’s initial innate immune reactions. The imbalance between CD4+IL-17+ T helper cells (Th17) and CD4+CD25+ regulatory T cells (Treg) participates in sepsis progression. CD28 signaling pathway was essential for the expression of inflammatory cytokines related to Th17, and play a crucial role in the maintenance of Treg. We investigated the correlations of the balance between Th17 and Treg to prognosis in sepsis patients and influence of anti-CD28 antibody on the ratio of Th17 to Treg in sepsis mice.Methods: 60 sepsis patients' baseline conditions were recorded, and the expressions of inflammatory factors in the peripheral blood and levels of procalcitonin (PCT) were detected. Peripheral blood mononuclear cells (PBMCs) were separated, subtypes of T cells and related biomarkers were measured by Fluorescence-activated cell sorting (FACS). Furthermore, the relationship between the above indicators and patients’ condition scoring (APACHEⅡand SOFA) and ICU hospitalization time were analyzed. To investigate effects of CD28 on the balance of Th17 between Treg, anti-CD28 antibody was intraperitoneal administrated to cecal ligation and puncture (CLP) mice. Results: Compared with septic patients who stay in ICU more than 14 days, the Th17/Treg ratio of patients with ICU hospitalization of fewer than 14 days was significantly lower. The sepsis patients with higher expression of CD28 in peripheral blood lymphocytes have lower APACHE II and SOFA scores. Moreover, the expression of CD28 was significantly higher in sepsis patients than that of healthy donors. After administration of CD28 monoclonal antibody, 7-day mortality and clinical score were significantly improved in septic mice, with splenocyte Th17/Treg ratio decreased. CD28 antibody alleviates the expression of pro-inflammatory factors and spleen injury related to apoptosis. Conclusions: Th17/Treg ratio revealed septic patient severity and can be used as a predictor of ICU stay. CD28 monoclonal antibody could improve 7-day mortality of septic mice by decreasing T cell apoptosis and amending the ratio of Th17/Treg.Trial registration: CHEC2019-133, CTR20191855. Registered 27 August 2019

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CD4+T-bet+, CD4+pSTAT3+ and CD8+T-bet+ T cells accumulate in peripheral blood during NZB treatment

Multiple Sclerosis Journal ◽

10.1177/1352458510392263 ◽

2010 ◽

Vol 17 (5) ◽

pp. 556-566 ◽

Cited By ~ 26

Author(s):

Giovanni Frisullo ◽

Raffaele Iorio ◽

Domenico Plantone ◽

Alessandro Marti ◽

Viviana Nociti ◽

...

Keyword(s):

T Cells ◽

Regulatory T Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Relapsing Remitting ◽

Before And After ◽

Circulating T Cells ◽

Relapsing Remitting Multiple Sclerosis

Circulating T cells and monocytes expressing T-bet, pSTAT1 and pSTAT3 increase in relapsing–remitting multiple sclerosis (RRMS) during relapse. Natalizumab (NZB) is an effective drug in RRMS, but exacerbation of the disease after its discontinuation has been described in some patients. The aim of this research was to study the effect of NZB treatment on circulating lymphomonocyte subpopulations expressing T-bet, pSTAT1, pSTAT3 and CD4+CD25+Foxp3+ regulatory T cells. Flow cytometry was used to evaluate the percentages of circulating CD4+ and CD8+ T cells, CD14+ monocytes and B cells expressing T-bet, pSTAT1, and pSTAT3, and CD4+CD25+Foxp3+ regulatory T cells from RRMS patients before and after 6–12 NZB infusions. In NZB-treated RRMS patients, the percentages of CD4+pSTAT1+ and CD8+pSTAT1+ T cells, CD14+pSTAT1+ monocytes, CD4+T-bet+, CD8+T-bet+ and CD4+pSTAT3+ T cells and CD14+pSTAT3+ monocytes increased after 12 drug infusions and were similar to those observed in untreated relapsing RRMS patients. Otherwise in vitro NZB exposure of peripheral blood mononuclear cells from untreated RRMS patients and controls had no effect. It was concluded that NZB treatment determines an accumulation of CD4+pSTAT1+, CD8+pSTAT1+, CD4+T-bet+, CD8+T-bet+ and CD4+STAT3+ T cells in peripheral blood that may account for the exacerbation of the disease observed in some patients after the discontinuation of the drug.

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Persistent expansions of CD4+ CD8+ peripheral blood T cells

Blood ◽

10.1182/blood.v82.5.1546.bloodjournal8251546 ◽

1993 ◽

Vol 82 (5) ◽

pp. 1546-1552 ◽

Cited By ~ 1

Author(s):

P Sala ◽

E Tonutti ◽

C Feruglio ◽

F Florian ◽

A Colombatti

Keyword(s):

T Cells ◽

T Cell ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Blot Hybridization ◽

Southern Blot Hybridization ◽

Cell Receptor ◽

Cd8 Cells ◽

Hla Dr ◽

T Cell Receptor Beta

CD4+ CD8+ cells are present during T cell differentiation in the thymus. Less than 2% of normal T cells that coexpress CD4 and CD8 also are released in the circulation and are present in the peripheral blood. In this study, nine individuals are described that manifested persistent expansions (11% to 43%) of circulating CD4+ CD8+ T cells that in three cases had large granular lymphocyte (LGL) morphology in the absence of either lymphocytosis or overt lymphoproliferative disorders. Southern blot hybridization of enriched CD4+ CD8+ cells with T-cell receptor beta (TCR beta) and TCR gamma probes showed that most cases had the 12-kb Eco RI germinal band deleted or of decreased intensity. In several individuals new TCR beta-specific bands of different intensity and distinct from case to case suggested either monoclonal or oligoclonal and polyclonal expansions. Immunophenotypic analysis showed that in 7 out of 9 cases the CD4+ CD8+ T cells presented with CD8 dim expression. Furthermore, all the CD4+ CD8+ cells did not express many of the known activation antigens (low or absent CD25, CD38, CD71, HLA-DR), whereas they expressed high levels of CD2, CD29, CD56, and CD57. In addition, the CD4+ CD8+ cells of 5 out of 9 subjects coexpressed CD45RA and CD45RO suggesting that these cells might be “frozen” in an intermediate state between naive and memory T cells. In conclusion, the present CD4+ CD8+ cases fall within a larger spectrum of disorders ranging from apparently normal to reactive or proliferative situations and encompassing cells with LGL morphology or LGL-associated antigens expression either in the presence or in the absence of absolute lymphocytosis that deserve careful follow-up investigations.

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