The effects of dimethyl fumarate and fingolimod on T-cell lymphocyte proliferation in patients with multiple sclerosis

Objective The disease-modifying therapies (DMT), dimethyl fumarate (DMF) and fingolimod (FTY) improve the outcomes in multiple sclerosis (MS) by reducing relapses and numbers and volume of lesions. They mediate their effects through reduction of immune reactivation, which may potentially lead to lymphopaenia and increased risk of infections. Previous studies have examined the effects of these therapies on lymphocyte subsets; however, the in vivo effects on circulating lymphocyte proliferation require further elucidation. The aim of this study was to determine the effects of DMF and FTY on T-cell proliferation in patients with MS. Method We examined T-cell lymphocyte proliferation and lymphocyte subsets in ten patients (five on DMF, five on FTY) before starting DMT and again 4 to 11 months after being maintained on DMT. Results In the FTY-treated group, the mean percentage proliferation was significantly lower using both assays (PHA assay mean percentage change − 51.2 ± 25.97, p < 0.05; anti-CD3/CD28 assay mean percentage change − 39.74 ± 27.85, p < 0.05). There was no statistical difference in T-cell lymphocyte proliferation in the DMF-treated group for either assay (PHA, p = 0.316; anti-CD3/CD28, p = 0.373). Conclusions This pilot study suggests that the T-lymphocytes of patients on FTY have an abnormal proliferation response as well as being reduced in the circulation.

Prognostic significance of circulating lymphocyte subsets in patients with nasopharyngeal carcinoma

The full text of this preprint has been withdrawn by the authors while they make corrections to the work. Therefore, the authors do not wish this work to be cited as a reference. Questions should be directed to the corresponding author.Additionally, the authors have provided this withdrawal declaration: "In this preprint, the criteria for inclusion and exclusion of study designs were not rigorous enough. After data verification, we found that there were some missing or unknown people in the follow-up results of the original data. This part of data may cause information bias. we recognize that the results and conclusions obtained based on these data may be unreliable. After careful discussion, all authors have agreed that, based on the need of scientific accuracy and honesty, it is necessary to withdraw the preprint."

Circulating Lymphocyte Subsets are Prognostic Factors in Patients with Nasopharyngeal Carcinoma

Background: Nasopharyngeal carcinoma (NPC) is a geographically and racially variable disease which has a high incidence in Southeast China. According to previous studies on tumor immunity, we compared multiple clinical parameters and blood indexes to find its relationship with prognosis in NPC patients.Methods: According to the load of EBV at diagnosis, 220 NPC patients receiving concurrent chemoradiotherapy (CRT) were divided into two groups. We compared clinical parameters, peripheral blood mononuclear cells, lymphocyte subsets and biochemical indexes between them. We analyzed distant metastases and overall survival rate between them.Results: In most cases, the two groups showed the same trend. Most blood indexes were decreased during CRT, the decrease in absolute count was more significant than in percentage. The younger age showed the higher CD3+ and CD3+CD8+ percentage. Patients whose EBV DNA≥1500 copies/mL at first diagnosis showed higher pN grade. Among them, higher CD3+CD8+ percentage or lower CD3-CD56+ percentage had batter OS rates. Patients whose EBV DNA<1500 copies/mL at first diagnosis had higher survival rate and longer survival time.Conclusions: CRT cause overall decrease of blood cells in NPC patients. The initial EBV load was related to prognosis. Among all the blood indexes, CD3+CD8+ percentage showed correlation with age and OS rates in patients whose EBV DNA≥1500 copies/mL at first diagnosis, which is worthy of further study.

The Model for End-Stage Liver Disease Score and the Follow-Up Period Can Cause the Shift of Circulating Lymphocyte Subsets in Liver Transplant Recipients

Little is known about the shift of lymphocytes under the condition of the model for end-stage liver disease score and the follow-up period. Then, we detected the peripheral blood from liver transplant recipients by flow cytometry and compared the results. The model for end-stage liver disease score affected the percentages of T-cell subsets and B cells during the short-term follow-up period, but failed to influence the lymphocyte subsets during the long-term follow-up period. In contrast, the follow-up period not only affected the absolute counts of T-cell subsets and natural killer (NK) cells in patients with the low model for end-stage liver disease scores, but also influenced the percentages and absolute counts of T-cell subsets in patients with the high model for end-stage liver disease scores. In the two-way ANOVA, we further revealed that the model for end-stage liver disease score was associated with the percentages of T cells and CD4+ T cells and the absolute numbers of T-cell subsets and B cells, while the follow-up period was associated with the percentages of T-cell subsets and the absolute numbers of lymphocyte subsets. Therefore, patients with either the low model for end-stage liver disease scores or the long-term follow-up period are in a relatively activated immune condition.

COPD is associated with increased pro-inflammatory CD28null CD8 T and NKT-like cells in the small airways

We previously showed increased steroid resistant CD28null CD8+ senescent lymphocyte subsets in peripheral blood from COPD patients. These cells expressed decreased levels of the glucocorticoid receptor (GCR), suggesting their contribution to the steroid resistant property of these cells. COPD is a disease of the small airways. We therefore hypothesized that there would be a further increase in these steroid resistant lymphocytes in the lung, particularly in the small airways. We further hypothesized that the pro-inflammatory/cytotoxic potential of these cells could be negated using prednisolone with low-dose cyclosporin A. Blood, bronchoalveolar lavage, large proximal and small distal airway brushings were collected from 11 COPD patients and 10 healthy aged-matched controls. The cytotoxic mediator granzyme b, pro-inflammatory cytokines IFNγ/TNFα, and GCR were determined in lymphocytes subsets before and after their exposure to 1µM prednisolone and/or 2.5ng/mL cyclosporin A. Particularly in the small airways, COPD subjects showed an increased percentage of CD28null CD8 T-cells and NKT-like cells, with increased expression of granzyme b, IFNγ and TNFα and a loss of GCR, compared with controls. Significant negative correlations between small airway GCR expression and IFNγ/TNFα production by T and NKT-like cells (eg, T-cell IFNγ R= -.834, p=.031) and with FEV1 (R= -890) were shown. Cyclosporine A and prednisolone synergistically increased GCR expression and inhibited pro-inflammatory cytokine production by CD28null CD8- T and NKT-like cells. COPD is associated with increased pro-inflammatory CD28null CD8+ T and NKT-like cells in the small airways. Treatments that increase GCR in these lymphocyte subsets may improve efficacy of clinical treatment.

Reference range of naïve T and T memory lymphocyte subsets in peripheral blood of healthy adult

Naïve T and T memory cell subsets are closely related to immune response and can provide important information for the diagnosis and treatment of immunological and hematological disorders. Lymphocyte compartment undergo dramatic changes during adulthood; age-related reference values derived from healthy individuals are crucial. However, extensively detailed reference values of peripheral blood lymphocytes in whole spectrum of adulthood detected by multi-color flow cytometry on a single platform are rare. 309 healthy adult volunteers were recruited from Tianjin in China. The absolute counts and percentages of CD3+CD4+ T cells, CD3+CD8+ T cells, naïve T cells (Tn), T memory stem cells (Tscm), central memory T cells (Tcm), effector memory T cells (Tem), terminal effector T cells (Tte) were detected by flow cytometry with single platform technologies. Reference range of absolute counts and percentage of T lymphocyte subsets were formulated by different age and gender. The results showed that Tn and Tscm cells, which had stem cell properties, decreased with aging; while, Tcm and Tem increased with aging, which increased from 18 to 64 years old but presented no significant change over the 65 years old. Gender had influence on the fluctuation of lymphocyte subsets, absolute count of CD3+CD8+, CD8+ Tcm, CD8+ Tem in male were higher than those in female. The reference values of percentages and absolute numbers of naïve T and T memory cell subsets can help doctors to understand the immune state of patients and evaluate conditions of prognosis then adjust treatment for patients.

Association of Lymphocyte Subsets in Advanced Non-Small Cell Lung Cancer Patients with the Efficacy and Prognosis of Immune Checkpoint Inhibitor Therapy: A Retrospective Study

Background The adaptation of immune checkpoint inhibitors (ICIs) has achieved promising effects in patients with non-small cell lung cancer (NSCLC). However, not all patients with NSCLC benefit from immunotherapy. There is an urgent need to explore some biomarkers to predict the efficacy and survival rate of patients with advanced NSCLC after immunotherapy. The objective of this study is to monitor the changes of peripheral blood lymphocyte subpopulations and analyze their correlation with the efficacy and prognosis of immunotherapy. Methods A total of 153 patients with advanced NSCLC were examined. Peripheral blood lymphocyte subsets including CD4+ T cells, CD8+ T cells, CD4+/CD8+ ratio, NK cells, and Tregs were collected before any treatment, before immunotherapy, and after 4 cycles of immunotherapy. T-test was used to analyze the influencing factors of lymphocyte subtypes and their changes before and after immunotherapy. Logistic regression was used to analyze the relationship between lymphocyte subtypes and efficacy with ROC curve being plotted. Log-rank test and Cox regression model were used to evaluate the relationship between lymphocyte subtypes and progression-free survival (PFS). Results Gender, age, pathology, distant metastasis, and EGFR mutations all affect the proportion of peripheral blood lymphocyte subpopulations in patients with advanced NSCLC. Compared with baseline, the effective group showed that the proportions of CD4+ T cells, CD4+/CD8+ ratio, NK cells and Tregs were significantly higher, and the proportions of CD8+ T cells were significantly lower in peripheral blood after 4 cycles of immunotherapy. On the contrary, the ineffective group showed no signs of significant differences. Baseline CD4+ T cells, NK cells and Tregs were independent predictors of immunotherapy efficacy and PFS. Conclusions Immune checkpoint inhibitors induce changes in the proportion of peripheral blood lymphocyte subsets in patients with effective immunotherapy. The levels of lymphocyte subsets have a good predictive value for efficacy and prognosis of patients with advanced NSCLC.

Integrated Lymphopenia Analysis in Younger and Older Patients With Multiple Sclerosis Treated With Cladribine Tablets

Cladribine tablets (CladT) preferentially reduce B and T lymphocyte levels. As aging is associated with a decline in immune function, the effect of CladT on lymphocyte levels may differ by age. This post hoc analysis combined data from the Phase 3 CLARITY, CLARITY Extension, and ORACLE-MS studies to examine the effect of age (≤50 or &gt;50 years) on lymphopenia following CladT 3.5 mg/kg (CladT3.5; cumulative dose over 2 years) treatment over 96 weeks. Both CladT3.5 and placebo were given over Weeks 1 and 5 (Year 1 treatment) and Weeks 48 and 52 (Year 2 treatment) from the start of the studies. Absolute lymphocyte count (ALC) and levels of lymphocyte subsets were examined in 1564 patients (Age ≤50 [placebo: N=566; CladT3.5: N=813]; Age &gt;50 [placebo: N=75; CladT3.5: N=110]). In both age groups, following CladT3.5 treatment, nadir for ALC occurred at Week 9 (8 weeks following start of Year 1 treatment) and Week 55 (7 weeks following start of Year 2 treatment) of the 96-week period; for CD19+ B lymphocytes, nadir occurred at Week 9 (Year 1) and Week 52 (Year 2). For CD4+ T lymphocytes, nadir occurred at Week 16 (Year 1) in both age groups, and at Weeks 60 and 72 (Year 2) in the Age ≤50 and &gt;50 groups, respectively. Nadir for CD8+ T lymphocytes occurred at Week 16 (Year 1) and Week 72 (Year 2) in the Age ≤50 group and levels remained in the normal range; nadir occurred at Week 9 (Year 1) and Week 96 (Year 2) in the Age &gt;50 group. Lymphocyte recovery began soon after nadir following CladT3.5 treatment and median levels reached normal range by end of the treatment year in both age groups. By Week 96, ~25% of patients treated with CladT3.5 reported ≥1 episode of Grade ≥3 lymphopenia (Gr≥3L). The rate of certain infections was numerically higher in older versus younger patients who experienced Gr≥3L. In conclusion, CladT3.5 had a similar effect on ALC and lymphocyte subsets in both younger and older patient groups.

Comparison of different sets of immunological tests to identify treatable immunodeficiencies in adult bronchiectasis patients

BackgroundReported prevalence of immunodeficiencies in bronchiectasis patients is variable depending on the frequency and extent of immunological tests performed. ERS Guidelines recommend a minimum bundle of tests. Broadening the spectrum of immunological tests could increase the number of patients diagnosed with an immunodeficiency and those who could receive specific therapy. The primary objective of the present study was to assess the performance of different sets of immunological tests in diagnosing any, primary, secondary, or treatable immunodeficiencies in adults with bronchiectasis.MethodsAn observational, cross-sectional study was conducted at the Bronchiectasis Program of the Policlinico University Hospital in Milan, Italy, from September 2016 to June 2019. Adult outpatients with a clinical and radiological diagnosis of bronchiectasis underwent the same immunological screening during the first visit when clinically stable consisting of: complete blood count, IgA, IgG, IgM, IgG subclasses, total IgE, lymphocyte subsets, and HIV antibodies. The primary endpoint was the prevalence of patients with any immunodeficiencies using five different sets of immunological tests.ResultsA total of 401 bronchiectasis patients underwent the immunological screening. A significantly different prevalence of bronchiectasis patients diagnosed with any, primary, or secondary immunodeficiencies was found across different bundles. 44.6% bronchiectasis patients had a diagnosis of immunodeficiency using when IgG subclasses and lymphocyte subset are added to the minimum bundle suggested by the guidelines.ConclusionA 4-fold increase in the diagnosis of immunodeficiencies can be found in adults with bronchiectasis when IgG subclasses and lymphocyte subsets are added to the bundle of tests recommended by guidelines.