scholarly journals Dimethyl fumarate induces changes in B- and T-lymphocyte function independent of the effects on absolute lymphocyte count

2017 ◽  
Vol 24 (6) ◽  
pp. 728-738 ◽  
Author(s):  
Erin E Longbrake ◽  
Claudia Cantoni ◽  
Salim Chahin ◽  
Francesca Cignarella ◽  
Anne H Cross ◽  
...  
Keyword(s):  
T Cell ◽  
Lymphocyte Count ◽  
Mononuclear Cells ◽  
Absolute Lymphocyte Count ◽  
Dimethyl Fumarate ◽  
Cell Phenotype ◽  
Lymphocyte Function ◽  
Functional Changes ◽  
Viral Peptide ◽  
Peptide Stimulation

Background: Dimethyl fumarate (DMF) is used to treat relapsing multiple sclerosis and causes lymphopenia in a subpopulation of treated individuals. Much remains to be learned about how the drug affects B- and T-lymphocytes. Objectives: To characterize changes in B- and T-cell phenotype and function induced by DMF and to investigate whether low absolute lymphocyte count (ALC) is associated with unique functional changes. Methods: Peripheral blood mononuclear cells (PBMCs) were collected from DMF-treated patients, untreated patients, and healthy controls. A subset of DMF-treated patients was lymphopenic (ALC < 800). Multiparametric flow cytometry was used to evaluate cellular phenotypes. Functional response to non-specific and viral peptide stimulation was assessed. Results: DMF reduced circulating memory B-cells regardless of ALC. Follicular T-helper cells (CD4+ CXCR5+) and mucosal invariant T-cells (CD8+ CD161+) were also reduced. DMF reduced T-cell production of pro-inflammatory cytokines in response to polyclonal (PMA/ionomycin) and viral peptide stimulation, regardless of ALC. No differences in activation-induced cell death or circulating progenitors were observed between lymphopenic and non-lymphopenic DMF-treated patients. Conclusion: These data implicate DMF-induced changes in lymphocytes as an important component of the drug’s efficacy and expand our understanding of the functional significance of DMF-induced lymphopenia.

scholarly journals Absolute lymphocyte count proliferation kinetics after CAR T-cell infusion impact response and relapse

2021 ◽  
Vol 5 (8) ◽  
pp. 2128-2136
Author(s):  
Sophia Faude ◽  
Jane Wei ◽  
Kavitha Muralidharan ◽  
Xiaoming Xu ◽  
Gerald Wertheim ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cell ◽  
Lymphocyte Count ◽  
Absolute Lymphocyte Count ◽  
Rapid Expansion ◽  
Proliferation Kinetics ◽  
Car T Cells ◽  
Car T Cell ◽  
Car T

Abstract CD19-directed chimeric antigen receptor (CAR) T cells show characteristic proliferation kinetics after infusion that correlate with response. Clearance of circulating disease, B-cell aplasia (BCA), and cytokine release syndrome (CRS) are used to observe CAR T-cell function, given the lack of commercial CAR T-cell measurement assays. We investigated the utility of common hematology laboratory parameters in 166 patients with B-cell acute lymphoblastic leukemia (B-ALL) who were treated with CAR T-cell therapy targeting CD19. CAR T-cell infusion was followed by disappearance of circulating blasts in 86% of patients at a median of 6 days. After a lag phase, there was a rapid expansion in absolute lymphocyte count (ALC) in the second week that coincided with the appearance of atypical lymphocytes. The expansion phase was followed by a contraction phase with a concomitant decrease in atypical lymphocytes. In vitro CAR T-cell studies showed similar kinetics and morphological changes. Peak ALC and overall expansion was greater in sustained responders compared with that in nonresponders. Patients with early loss of BCA and those with eventual CD19+ minimal residual disease/relapse showed lower overall lymphocyte expansion compared with the controls. Pleomorphic lymphocytosis was noted in the cerebrospinal fluid at post-CAR time points. We conclude that lymphocyte counts and differential can also be used to evaluate CAR T-cell expansion after infusion, along with BCA and CRS. This is the first report to characterize the morphology of CAR T cells and determine the utility of lymphocyte kinetics.

scholarly journals 2508. Absolute Lymphocyte Count and Adenovirus-Specific CD8+ T-cell Immunity as Immunological Predictors of Severe Adenovirus Disease After Kidney Transplantation

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S754-S754
Author(s):  
Jackrapong Bruminhent ◽  
Nopporn Apiwattanakul ◽  
Subencha Pinsai ◽  
Charat Thongprayoon ◽  
Suradej Hongeng ◽  
...  
Keyword(s):  
Kidney Transplantation ◽  
T Cell ◽  
Lymphocyte Count ◽  
Absolute Lymphocyte Count ◽  
Cd8 T Cell ◽  
T Cell Immunity ◽  
Cell Immunity

Efficient T-Cell Compartment in HIV-Positive Patients Receiving Orthotopic Liver Transplant and Immunosuppressive Therapy

2020 ◽  
Author(s):  
Erica Franceschini ◽  
Sara De Biasi ◽  
Margherita Digaetano ◽  
Elena Bianchini ◽  
Domenico Lo Tartaro ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Liver Transplant ◽  
T Cell Activation ◽  
Mononuclear Cells ◽  
Immunosuppressive Treatment ◽  
Orthotopic Liver Transplant ◽  
Cell Phenotype ◽  
Cell Mediated Immunity ◽  
Hiv Patients

Abstract Background In patients undergoing orthotopic liver transplant (OLT), immunosuppressive treatment is mandatory and infections are leading causes of morbidity/mortality. Thus, it is essential to understand the functionality of cell-mediated immunity after OLT. The aim of the study was to identify changes in T-cell phenotype and polyfunctionality in human immunodeficiency virus–positive (HIV+) and –negative (HIV–) patients undergoing immunosuppressive treatment after OLT. Methods We studied peripheral blood mononuclear cells from 108 subjects divided into 4 groups of 27: HIV+ transplanted patients, HIV– transplanted patients, HIV+ nontransplanted patients, and healthy subjects. T-cell activation, differentiation, and cytokine production were analyzed by flow cytometry. Results Median age was 55 years (interquartile range, 52–59 years); the median CD4 count in HIV+ patients was 567 cells/mL, and all had undetectable viral load. CD4+ and CD8+ T-cell subpopulations showed different distributions between HIV+ and HIV– OLT patients. A cluster representing effector cells expressing PD1 was abundant in HIV– transplanted patients and they were characterized by higher levels of CD4+ T cells able to produce interferon-γ and tumor necrosis factor–α. Conclusions HIV– transplanted patients have more exhausted or inflammatory T cells compared to HIV+ transplanted patients, suggesting that patients who have already experienced a form of immunosuppression due to HIV infection respond differently to anti-rejection therapy.

scholarly journals Galectin-9 expression defines exhausted T cells and impaired cytotoxic NK cells in patients with virus-associated solid tumors

2020 ◽  
Vol 8 (2) ◽  
pp. e001849
Author(s):  
Isobel Okoye ◽  
Lai Xu ◽  
Melika Motamedi ◽  
Pallavi Parashar ◽  
John W Walker ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Nk Cells ◽  
Solid Tumors ◽  
Mononuclear Cells ◽  
Cell Phenotype ◽  
Peripheral Blood Mononuclear ◽  
Effector Functions ◽  
Cell Functions ◽  
Cell Effector

BackgroundWe have previously reported that the upregulation of galectin-9 (Gal-9) on CD4+ and CD8+ T cells in HIV patients was associated with impaired T cell effector functions. Gal-9 is a ligand for T cell immunoglobulin and mucin domain-3, and its expression on T cells in cancer has not been investigated. Therefore, we aimed to investigate the expression level and effects of Gal-9 on T cell functions in patients with virus-associated solid tumors (VASTs).Methods40 patients with VASTs through a non-randomized and biomarker-driven phase II LATENT trial were investigated. Peripheral blood mononuclear cells and tumor biopsies were obtained and subjected to immunophenotyping. In this trial, the effects of oral valproate and avelumab (anti-PD-L1) was investigated in regards to the expression of Gal-9 on T cells.ResultsWe report the upregulation of Gal-9 expression by peripheral and tumor-infiltrating CD4+ and CD8+ T lymphocytes in patients with VASTs. Our results indicate that Gal-9 expression is associated with dysfunctional T cell effector functions in the periphery and tumor microenvironment (TME). Coexpression of Gal-9 with PD-1 or T cell immunoglobulin and ITIM domain (TIGIT) exhibited a synergistic inhibitory effect and enhanced an exhausted T cell phenotype. Besides, responding patients to treatment had lower Gal-9 mRNA expression in the TME. Translocation of Gal-9 from the cytosol to the cell membrane of T cells following stimulation suggests persistent T cell receptor (TCR) stimulation as a potential contributing factor in Gal-9 upregulation in patients with VASTs. Moreover, partial colocalization of Gal-9 with CD3 on T cells likely impacts the initiation of signal transduction via TCR as shown by the upregulation of ZAP70 in Gal-9+ T cells. Also, we found an expansion of Gal-9+ but not TIGIT+ NK cells in patients with VASTs; however, dichotomous to TIGIT+ NK cells, Gal-9+ NK cells exhibited impaired cytotoxic molecules but higher Interferon gamma (IFN-γ) expression.ConclusionOur data indicate that higher Gal-9-expressing CD8+ T cells were associated with poor prognosis following immunotherapy with anti-Programmed death-ligand 1 (PD-L1) (avelumab) in our patients’ cohort. Therefore, for the very first time to our knowledge, we report Gal-9 as a novel marker of T cell exhaustion and the potential target of immunotherapy in patients with VASTs.

scholarly journals Effect of dimethyl fumarate on lymphocyte subsets in patients with relapsing multiple sclerosis

2020 ◽  
Vol 6 (2) ◽  
pp. 205521732091861 ◽  
Author(s):  
Guy Buckle ◽  
Daniel Bandari ◽  
Jeffrey Greenstein ◽  
Mark Gudesblatt ◽  
Bhupendra Khatri ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
T Cells ◽  
Confidence Interval ◽  
Cd8 T Cells ◽  
Lymphocyte Count ◽  
Absolute Lymphocyte Count ◽  
Dimethyl Fumarate ◽  
T Cell Subsets ◽  
Routine Practice ◽  
Patients At Risk

Background In patients treated with dimethyl fumarate, absolute lymphocyte count decline typically occurs during the first year and then plateaus; early drops have been associated with the development of severe prolonged lymphopenia. Objective We investigated the effect of dimethyl fumarate on absolute lymphocyte counts and CD4+/CD8+ T cells in patients with relapsing–remitting multiple sclerosis treated with dimethyl fumarate in routine practice. Methods Lymphocyte data were collected via medical chart abstraction. Primary endpoint: change from baseline in absolute lymphocyte count and CD4+/CD8+ counts at 6‐month intervals following dimethyl fumarate initiation. Results Charts of 483 patients were abstracted and 476 patients included in the analysis. Mean baseline absolute lymphocyte count (2.23 × 109/l) decreased by ∼39% (95% confidence interval: –41.1 to –37.2) by month 6 and 44% (95% confidence interval: –46.6 to –42.1) by month 12. CD4+ and CD8+ T-cell subsets strongly correlated with absolute lymphocyte count, with greater decreases from baseline to 6 months vs 6–12 months, and in CD8+ vs CD4+ T cells. Prior natalizumab was not a risk factor for lymphopenia. Conclusion Dimethyl fumarate-associated decline in absolute lymphocyte count in the first 12 months correlated with decline in CD4+ and CD8+ T cells and was independent of prior natalizumab. Absolute lymphocyte count monitoring continues to be an effective strategy to identify patients at risk of prolonged lymphopenia.

SIGNIFICANCE of ABSOLUTE LYMPHOCYTE COUNT at RELAPSE as a PROGNOSTIC FACTOR in PATIENTS with T-CELL NON-HODGKIN'S LYMPHOMA.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2939-2939
Author(s):  
Dae Ro Choi ◽  
Dok Hyun Yoon ◽  
Heui June Ahn ◽  
Yoojin Cho ◽  
Eun Kyoung Kim ◽  
...  
Keyword(s):  
T Cell ◽  
Lymphocyte Count ◽  
Cell Lymphoma ◽  
Prognostic Significance ◽  
Absolute Lymphocyte Count ◽  
Complete Response ◽  
T Cell Lymphoma ◽  
Hodgkin's Lymphoma ◽  
First Relapse ◽  
Ecog Ps

Abstract Abstract 2939 Poster Board II-915 Introduction: Peripheral blood absolute lymphocyte count (ALC) at the time of diagnosis is a prognostic indicator in hematologic malignancies. However, no reports have addressed whether ALC at the time of first relapse (ALC-R) has a prognostic significance in the patients with relapsed T-cell Non-Hodgkin's lymphoma (NHL). We retrospectively studied the prognostic significance of ALC-R in these patients. Patients and Methods: We identified 63 patients who had a documented relapsed disease after having reached a complete response, including at least an unconfirmed complete response between 1993 and 2007 and we analyzed their ALC-R and following variables at the time of first relapse; age, gender, the number of extranodal sites, lactate dehydrogenase, ECOG performance status, stage and international prognostic index. Results: Out of the 63 patients, 23 (36.5%) had a peripheral T-cell lymphoma, not otherwise characterized, while 15 (23.8%) had an extranodal NK/T-cell lymphoma, nasal type, 6 (9.5%) anaplastic large-cell lymphoma, 5 (7.9%) angioimmunoblastic T-cell lymphoma, 2 (3.2%) primary cutaneous T cell lymphoma and 12 (19%) other types. Among them, 32 (50.8%) had an ALC-R ≥ 1.25 × 109/L, 47 (74.6%) had an ECOG PS 0 or 1 and by IPI at relapse, 0, 1, 2, 3, 4, and 5 were 20.6%, 25.4%, 23.8%, 23.8%, 4.8%, and 1.6%, respectively. Univariate analyses showed that good ECOG PS (HR, 0.408; 95% CI 0.190-0.876; p=0.022) and high ALC at relapse (HR, 0.394; 95% CI 0.210-0.740; p=0.004) were associated with longer survival from relapse (Table 1). Multivariate analysis also showed that high ALC at relapse (HR, 0.369; 95% CI 0.187-0.726; p=0.004) and good ECOG PS (HR, 0.295; 95% CI 0.131-0.666; p=0.003) were still associated with longer survival outcome (Table 2). Conclusions: The high ALC-R predicted a better prognosis in patients with relapsed T-cell NHL, suggesting that the host immune system might have a crucial role. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Immunologic differentiation of absolute lymphocyte count with an integrated flow cytometric system: A new concept for absolute T cell subset determinations

Cytometry ◽  
1995 ◽  
Vol 22 (1) ◽  
pp. 48-59 ◽  
Author(s):  
Thomas J. Mercolino ◽  
Mark C. Connelly ◽  
Eric J. Meyer ◽  
Marilyn D. Knight ◽  
John W. Parker ◽  
...  
Keyword(s):  
T Cell ◽  
Lymphocyte Count ◽  
Cell Subset ◽  
Absolute Lymphocyte Count ◽  
T Cell Subset ◽  
System A ◽  
Flow Cytometric

scholarly journals A Simplified Method for Transduction and Expansion of T Cells for Clinical Application

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 4555-4555
Author(s):  
Mariam Khalil ◽  
Aaron Foster ◽  
Christine Gagliardi
Keyword(s):  
Cell Culture ◽  
T Cells ◽  
T Cell ◽  
Mononuclear Cells ◽  
Negative Impact ◽  
Retroviral Vectors ◽  
Transduction Efficiency ◽  
Cell Phenotype ◽  
Similar Distribution ◽  
Specialized Equipment

Abstract Introduction: Genetically modified T cells are being investigated to treat a variety of disorders and have been particularly successful in treating B cell cancers. As more effort is poured into new targets, molecular switches, and various other modifications, development of processes to quickly manufacture new products must keep up. Current manufacturing processes often require highly skilled operators and specialized equipment. Here, we demonstrate a simplified, novel method for transduction of T cells, followed by robust expansion in the G-Rex bioreactor with no need for intervention until harvest. A scaled-up, closed-version of the same process, including a closed harvest step with the GatheRex is currently under evaluation. Methods: Frozen peripheral blood mononuclear cells (PBMCs) from healthy donors were used as starting material. PBMCs were thawed, washed, and activated with soluble anti-CD3 and anti-CD28 antibodies either in cell culture bags (32-C, Saint-Gobain Cell Therapy) or in G-Rex bioreactors (Wilson Wolf Corporation). Cells were cultured in TexMACS GMP medium (Miltenyi Biotec) with IL-7 and IL-15 throughout. For the transduction step, activated PMBCs and retroviral supernatant were incubated in cell culture bags coated with Retronectin (Takara) or in G-Rex bioreactors with vectofusin-1 (Miltenyi Biotec). Viral constructs contained either a CD34 or CD19 marker detectable by antibody staining. For transduction in the G-Rex, various cell densities, volumes, constructs, and multiplicity of infection (MOIs) were tested. Where applicable, the GatheRex device (Wilson Wolf Corporation) was used for volume reduction and harvest. Transduction efficiency and T cell phenotype were measured by flow cytometry. Cell count and viability were assessed with the NC-3000 (Chemometic). Glucose and lactate concentrations were checked daily for in-processing monitoring. Results: Overall transduction efficiency ranged from 30-90% depending on the experimental conditions. Incubating 1x107 activated PBMCs in 10 ml of medium in a 10-cm2 G-Rex (1.0 ml/cm2) with retrovirus at an MOI of 1 resulted in 3% transduced cells. Addition of vectofusin-1 to the same condition yielded transduction efficiency of 44%. Increasing the MOI to 10 lead to 86% transduced cells. Decreasing the transduction volume from 1.0 ml/cm2 to 0.4 ml/cm2 increased transduction efficiency from 34% to 55%. Reducing the volume further to 0.2 ml/cm2 did not improve efficiency, and rather had a negative impact compared to the 0.4 ml/cm2 condition (38%). 16-24 hrs after transduction, the volume of medium was increased to 10.0 ml/cm2 without a wash step. The dilution in place of a wash step had no negative impact on cell viability. 10.0 ml/cm2 medium supported high viability (>90%) and expansion (30-50 fold) over an additional 9 days without operator intervention. The phenotype of cells expanded in the G-Rex contained a mixed population of CD45RO+ and CD45RA+ cells, with a similar distribution of naive and memory cell subsets in G-Rex and bag cultures. Harvest of cells with the GatheRex was efficient; a 1L volume was reduced 10-fold in 5 minutes, and 95% of cells were recovered. Summary: T cells can be transduced with retroviral vectors in the G-Rex bioreactor. Clinically relevant levels of transgene expression can be achieved by combining reagents in the G-Rex, without complicated coating steps or time-consuming spinning steps. This simplified procedure reduces the hands-on time of the T cell transduction to minutes rather than hours. Transgenic cells can be expanded 30-50-fold in the G-Rex with limited operator intervention and without specialized equipment. Disclosures Khalil: Bellicum Pharmaceuticals: Employment. Foster:Bellicum: Employment, Equity Ownership. Gagliardi:Bellicum Pharmaceuticals: Employment.

scholarly journals Mesenchymal Stem Cells Induce Regulatory T-cell Population in Human SLE

2020 ◽  
Vol 19 (4) ◽  
pp. 743-748
Author(s):  
Riyadh Ikhsan ◽  
Agung Putra ◽  
Delfitri Munir ◽  
Dewi Masyithah Darlan ◽  
Bantar Suntoko ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
T Cell ◽  
Lupus Erythematosus ◽  
Mononuclear Cells ◽  
Medical Science ◽  
Cell Subset ◽  
Cell Phenotype ◽  
Control Group ◽  
Trypan Blue Exclusion

Background: The mechanisms underlying peripheral disorders during systemic lupus erythematosus (SLE) were found to be shared with tolerance disorders and mediated by T-regulator (T-reg) cells. Mesenchymal stem cells (MSCs) may inhibit T-cell subset differentiation and induce the T-reg cell phenotype. However, the capacity of MSCs to promote functional T-reg cells in SLE patients remains unclear. Objectives: This study aimed to analyze the capacity of MSCs to induce the production of functional CD4+ CD25+ Foxp3+ T-reg cells, in vitro, under co-culture conditions with human SLE cells. Methods: This study used a pre- and post-test control group design. Peripheral blood mononuclear cells (PBMCs) were extracted from SLE patients at the Kariadi Hospital, and MSCs were derived from human umbilical cords (hUCs) The PBMC control group was treated with standard medium, and the treatment group was co-cultured with hUC-MSCs. After 24 hours of co-culture incubation, T-reg cells were removed from the PBMC pool, using magnetic-activated cell sorting (MACS), and the population was assessed using the trypan blue exclusion assay. Results: A significant increase in the population of T-reg cells was observed (P < 0.001) after 24 hours of co-culture incubation with hUC-MSCs. Conclusion: This study concluded that MSCs have the capacity to enhance the T-reg population in human SLE PBMCs. Bangladesh Journal of Medical Science Vol.19(4) 2020 p.743-748

scholarly journals ICOSL+ plasmacytoid dendritic cells as inducer of graft-versus-host disease, responsive to a dual ICOS/CD28 antagonist

2020 ◽  
Vol 12 (564) ◽  
pp. eaay4799
Author(s):  
Djamilatou Adom ◽  
Stacey R. Dillon ◽  
Jinfeng Yang ◽  
Hao Liu ◽  
Abdulraouf Ramadan ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
T Cell ◽  
Graft Versus Host Disease ◽  
Th17 Cells ◽  
Mononuclear Cells ◽  
Cell Phenotype ◽  
Murine Models ◽  
Host Disease ◽  
Graft Versus Host

Acute graft-versus-host disease (aGVHD) remains a major complication of allogeneic hematopoietic cell transplantation (HCT). CD146 and CCR5 are proteins that mark activated T helper 17 (Th17) cells. The Th17 cell phenotype is promoted by the interaction of the receptor ICOS on T cells with ICOS ligand (ICOSL) on dendritic cells (DCs). We performed multiparametric flow cytometry in a cohort of 156 HCT recipients and conducted experiments with aGVHD murine models to understand the role of ICOSL+ DCs. We observed an increased frequency of ICOSL+ plasmacytoid DCs, correlating with CD146+CCR5+ T cell frequencies, in the 64 HCT recipients with gastrointestinal aGVHD. In murine models, donor bone marrow cells from ICOSL-deficient mice compared to those from wild-type mice reduced aGVHD-related mortality. Reduced aGVHD resulted from lower intestinal infiltration of pDCs and pathogenic Th17 cells. We transplanted activated human ICOSL+ pDCs along with human peripheral blood mononuclear cells into immunocompromised mice and observed infiltration of intestinal CD146+CCR5+ T cells. We found that prophylactic administration of a dual human ICOS/CD28 antagonist (ALPN-101) prevented aGVHD in this model better than did the clinically approved belatacept (CTLA-4-Fc), which binds CD80 (B7-1) and CD86 (B7-2) and interferes with the CD28 T cell costimulatory pathway. When started at onset of aGVHD signs, ALPN-101 treatment alleviated symptoms of ongoing aGVHD and improved survival while preserving antitumoral cytotoxicity. Our data identified ICOSL+-pDCs as an aGVHD biomarker and suggest that coinhibition of the ICOSL/ICOS and B7/CD28 axes with one biologic drug may represent a therapeutic opportunity to prevent or treat aGVHD.

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