Effects of Compound Xuanju Capsule on T Lymphocyte Subsets in Patients With Unexplained Recurrent Abortion

Background: Little is known about changes in lymphocyte subsets after SARS-CoV-2 infection. Methods: Clinical data of 580 COVID-19 patients hospitalized in Zhongnan Hospital of Wuhan University from 20 December 2019, to 8 March 2020, were retrospectively analyzed. The relation of lymphocyte subsets and severity or prognosis of disease were analyzed. Results: At 2–3 weeks after the onset of symptoms, lymphocyte subsets decreased to the lowest levels. The levels of lymphocyte subsets in asymptomatic patients were close to healthy persons, except for CD8+ T lymphocyte cells. The levels of lymphocyte subsets in patients with severe illness were lower than that in patients with mild-to-moderate illness (P < 0.01). Similarly, among patients with severe illness, lower levels of lymphocyte subsets were found in dead patients compared to survivors (P < 0.001). Moreover, by comparing the results of the same patients at different stages of the disease, we found levels of lymphocyte subsets were lower in the acute phase compared to that in convalescent-phase (P < 0.001). However, the levels of lymphocyte subsets in patients who had SARS-CoV-2 viral load >5000 copies/ml and 500–5000 copies/ml were at similar levels. Conclusions: Lymphocyte subsets are a good biomarker to assess the severity and prognosis of the disease at 2–3 weeks after the onset of symptoms.

BackgroundThe aim of this study was to investigate the effects and significance of rituximab (RTX) on the levels of T lymphocyte subsets in patients diagnosed with primary membranous nephropathy (PMN).MethodsA total of 58 PMN patients and 25 healthy donors were chosen as the subjects. Among the PMN patients, 40 individuals received RTX treatment and completed at least 6 months of follow-up. All subjects underwent flow cytometry analysis to determine the peripheral blood lymphocyte subsets. The changes in anti-PLA2R antibody titers and 24-hour urinary protein levels were evaluated by ELISA and Biuret method before and after treatment.Results(1) The PMN group exhibited a significantly greater percentage of peripheral blood CD3−CD19+ B cells than the healthy group, which is consistent with the findings of previous reports. Additionally, compared with those in the peripheral blood of healthy individuals, the numbers of CD4+ central memory T cells, CD4+ effector memory T cells, CD4+/CD8+, and CD4+CD25+ T cells in the PMN peripheral blood were markedly greater. However, the number of peripheral blood Treg cells was reduced in the PMN group. (2) After 6 months of RTX treatment, PMN patients exhibited significant decreases in anti-PLA2R antibody titers, 24-hour urinary protein levels, and peripheral blood CD3−CD19+ B cells. Importantly, RTX administration decreased CD4+CD25+ T cells and CD4+/CD8+ in the peripheral blood of PMN patients and improved Treg cell levels. (3) RTX treatment induced alterations in the CD4+ T lymphocyte subsets in PMN patients, which did not correlate with B lymphocyte counts or anti-PLA2R antibody titers.ConclusionsRTX treatment might have a beneficial impact on cellular immunity by effectively restoring the balance of CD4+ T lymphocyte subsets in PMN patients, which is beyond its effects on B cells and antibody production.Trial registrationThe research was registered at the First Affiliated Hospital of Soochow University. Registration Number: MR-32-23-016211. Registration Date: May 31, 2023.

Abnormal Fas/FasL and caspase-3-mediated apoptotic signaling pathways of T lymphocyte subset in patients with systemic lupus erythematosus

Objective: To investigate the characteristics and prognostic value of peripheral blood T lymphocyte subsets in patients with severe influenza. Methods: This was a single-center cross-sectional study in influenza patients admitted to Peking Union Medical College Hospital from August 2017 to April 2018. Peripheral blood lymphocyte subsets were detected by flow cytometry in both patients and 108 healthy controls. Influenza patients were divided into mild group and severe group. Severe patients were further classified into alive and fatal subgroups. Results: A total of 42 influenza patients were recruited in this study, including 24 severe cases (6 deaths). The remaining 18 cases were mild. The peripheral blood lymphocyte counts and lymphocyte subset counts (B, NK, CD4(+)T, CD8(+)T) in either mild patients[795 (571,1 007), 43 (23,144), 70 (47,135), 330 (256,457), 226 (148,366) cells/μl respectively] or severe patients[661 (474,1 151),92 (52,139), 54 (34,134), 373 (235,555), 180 (105,310) cells/μl respectively] were both significantly lower than those of healthy controls [1 963 (1 603,2 394),179 (119,239), 356 (231,496), 663 (531,824), 481 (341,693) cells/μl respectively]. Meanwhile, the T cells and CD8(+)T counts in fatal patients [370 (260,537) cells/μl and 87 (74,105) cells/μl] were significantly lower than those in severe and alive patients [722 (390,990) cells/μl and 222 (154,404) cells/μl]. CD8(+)HLA-DR/CD8(+)and CD8(+)CD38(+)/CD8(+)T cell activating subgroups in mild cases[(53.7±19.2)% and 74.8% (64.1%,83.7%) respectively] were significantly higher than those in severe cases[(38.5±21.7)% and 53.3% (45.3%,67.2%) respectively].Moreover,CD8(+)HLA-DR/CD8(+)count in severe and alive group was higher than that in fatal group [(46.1±19.1)% vs. (18.2±14.6)%, P<0.01]. Logistic regression analysis showed that CD8(+)T cell count (OR=0.952, 95%CI 0.910-0.997, P=0.035) and CD8(+)HLA-DR/CD8(+)T (OR=0.916, 95%CI 0.850-0.987, P=0.022) were both negatively correlated with mortality.Peripheral blood lymphocyte counts in mild cases rapidly decreased within 1 day after diagnosis, and returned to the basic level one week later. Conclusions: All peripheral blood lymphocyte subsets (T,B,NK) in patients with influenza are significantly reduced. These findings are consistent with the immunological characteristics of respiratory viral infections, in which peripheral lymphocytes (especially T cells) migrate to respiratory tract in the early stage and circulate to the peripheral blood after recovery. The activated CD8(+)T cell counts in peripheral blood are negatively correlated with the severity of disease, which could be considered as a prognostic indicator of severe influenza.

IntroductionCellular immune response to cancer is known to be of great importance for tumor control. Moreover, solid tumors influence circulating lymphocytes, which has been shown for several types of cancer. In our prospective study we elucidate changes in lymphocyte subsets in patients with colorectal carcinoma compared to healthy volunteers.MethodsFlow cytometry was performed at diagnosis of colon carcinoma to analyze B cells, T cells and NK cells including various subtypes of each group. Univariate and multivariate analyses including age, gender, tumor stage, sidedness and microsatellite instability status (MSI) were performed.ResultsForty-seven patients and 50 healthy volunteers were included. Median age was 65 years in patients and 43 years in the control group. Univariate analysis revealed lower total lymphocyte counts, lower CD4 + cells, CD8 + cells, B cells and NKs including various of their subsets in patients. In multivariate analysis patients had inferior values of B cells, CD4 + cells and NK cells and various subsets, regardless of age and gender. Naïve, central memory and HLADR + CD8 + cells showed an increase in patients whereas all other altered subsets declined. MSI status had no influence on circulating lymphocytes except for higher effector memory CD8 + cells in MSI-high patients. Localization in the left hemicolon led to higher values of total cytotoxic T cells and various T cell subsets.ConclusionWe found significant changes in circulating lymphocyte subsets in colon carcinoma patients, independent of physiological alterations due to gender or age. For some lymphocyte subsets significant differences according to tumor localization or MSI-status could be seen.

BackgroundSepsis is a life-threatening disease with challenges in clinical management due to delayed diagnosis and immunosuppression. Lymphopenia is a key prognostic indicator in sepsis. microRNAs (miRNAs) are recognized as key immune modulators affecting all stages of inflammation. In this study, we investigated the expression of miR-146b-5p and lymphocyte subsets in sepsis patients and analyzed their correlation with the aim of establishing their combined predictive value for clinical outcomes and advancing personalized treatment strategies.MethodsFrom January 2020 to July 2024, we enrolled 191 patients diagnosed with sepsis at the ICU of the Second Affiliated Hospital of Chongqing Medical University and collected basic clinical data. These patients were categorized into two groups: nonsurvivors (n = 117) and survivors (n = 74). Correlation analysis was employed to analyze the correlation between miR-146b-5p and lymphocyte subsets with disease severity. Binary logistic regression and Cox regression analyses were employed to identify independent risk factors influencing the prognosis of sepsis. The predictive value of miR-146b-5p and lymphocyte subsets for sepsis prognosis was assessed using receiver operating characteristic (ROC) curves.ResultsmiR-146b-5p expression was significantly lower in sepsis patients compared to HD group, with levels in the nonsurvivor group being lower than those in the survivor group. Survival curves for miR-146b-5p indicated that lower levels of miR-146b-5p (<0.272) were associated with a higher mortality rate (HR 3.063). The absolute counts of lymphocytes (Lym), CD3+ T cells, CD4+ T cells, and CD8+ T cells were significantly lower in the sepsis group compared to the HD group. Testing lymphocyte counts at different time points revealed that absolute counts of CD3+ T cells, CD4+ T cells, and CD8+ T cells were consistently lower in the sepsis group across all time intervals. miR-146b-5p levels were predictive of patient prognosis, with the combination of miR-146b-5p and APACHE II scores yielding the highest AUC.ConclusionThe early-stage sepsis-associated downregulation of miR-146b-5p serves as a promising biomarker for severity stratification and prognostic evaluation. The combination of miR-146b-5p with APACHE II scores enhances diagnostic accuracy. Additionally, dynamic monitoring of lymphocyte subsets may facilitate the evaluation of immune status and guide personalized treatment strategies.

Background:The CD4+T cell subsets plays an important role in its pathogenesis, and its new research are constantly being published, but its specific changes between SSc and MCTD are still unclear.Objectives:The aim of the present study was to explore the absolute numbers of CD4+T subsets in peripheral blood(PB) of patients with SSc and MCTD using our modified flow cryometric method and investigate the role in the pathogenesis of both.Methods:The PB samples from 54 patients with SSc, 51 patients with MCTD as well as 30 healthy control subjects were analyzed for lymphocyte subsets using flow cytometry. Of these patients, 19 had pulmonary involvement, including 9 patients with SSc and 10 patients with MCTD. Using directly the percentages from flow cytometry combined with internal standard beads calculated absolute number of peripheral lymphocyte subsets from the subjects in each group.Results:Although there were some changes among CD4+T cell subsets in PB from these SSc patients and MCTD patients, the major alteration was the reductions of Treg cells. Compared with the normal controls, the absolute number of CD4+CD25+FOXP3+Treg cells were significantly decreased in SSc patients and MCTD patients, and the absolute number of Th1 cells in MCTD patients is also significantly reduced. Notably, the absolute numbers of Th17 and Th2 cells were not different from those of normal controls, but the ratios of Th17/Treg in SSc patients and MCTD patients were significantly higher, causing by insufficient number of Treg cells (Fig 1). In addition, in patients with pulmonary involvement, we found that the absolute number of Treg cells was significantly reduced in patients with MCTD, while the absolute number of Th2 cells and Th17 cells was significantly reduced in patients with SSc(Fig 2).Fig 1.Comparison of the levels of CD4+T lymphocyte subsets in SSc patients, MCTD patients and healthy controls: (A) The absolute number of peripheral Th1 cells in patients with MCTD was significantly reduced; (B and C) There was no significant difference in the absolute number of Th2 cells in peripheral blood of different subjects; (D and E) The ratio of Th17/Treg cells in PB of patients with SSc and MCTD were significantly higher.*P&lt; 0.05; **P&lt; 0.01; ***P&lt; 0.001.Conclusion:The number of peripheral Treg cells in patients with SSc and MCTD was significantly reduced, suggesting that that SSc and MCTD progression is associated with the imbalances between pro-inflammation cells to anti-inflammation Treg cells. In addition, we also found that the decrease in peripheral numbers of Treg cells may contribute to the development of MCTD-associated lung disease, whereas in SSc patients who had lung involvement, the reduce in peripheral number of Th17 cells may result in a severe imbalance of Th17/Treg cells, thereby promoting disease progression.Fig 2.Comparison of the levels of CD4+T lymphocyte subsets in patients who had pulmonary involvement and healthy controls: (A) There was no significant difference in the absolute number of Th1 cells in peripheral blood of different subjects; (B and C) The absolute number of peripheral Th2 cells and Th17 cells in patients with SSc were significantly reduced; (D and E) The ratio of Th17/Treg cells in PB of patients with MCTD were higher.*P&lt; 0.05; **P&lt; 0.01; ***P&lt; 0.001.

Effects of venlafaxine and fluoxetine on lymphocyte subsets in patients with major depressive disorder: A flow cytometric analysis

Clinical Study of Lymphocyte Subsets in Patients with BCR/ABL Negative Myeloproliferative Neoplasms

Background and objective: The prognostic value of circulating lymphocyte subsets in cervical cancer patients receiving postoperative radiotherapy remains unclear. This study aimed to explore the prognostic significance of these lymphocyte subsets in this patient population. Methods: Peripheral blood samples were collected from 101 cervical cancer patients prior to receiving postoperative radiotherapy. Flow cytometry was utilized to determine the proportions and absolute counts of lymphocyte subsets, including total T cells, CD4+ T cells, CD8+ T cells, natural killer (NK) cells, and B cells. The Kaplan-Meier method and Cox regression analysis were employed to estimate the overall survival (OS) and identify the key prognostic factors. Receiver operating characteristic (ROC) curves were generated to assess the predictive accuracy. Results: The survival analysis indicated that patients with a decreased proportion of NK cells (P = 0.02) or reduced NK cell counts (P = 0.01) exhibited significantly poorer overall survival (OS) compared to those with higher levels of NK cells. In univariate Cox analysis, both the proportion of NK cells (P = 0.025; HR, 0.33; 95% CI, 0.12-0.87) and NK cell counts (P = 0.015; HR = 0.28) significantly influenced OS. In multivariate analysis, the proportion of CD4+ T cells (P = 0.02; HR, 0.08; 95% CI, 0.01-0.72) and NK cell counts (P = 0.08; HR, 0.11; 95% CI, 0.01-1.37) emerged as independent prognostic factors. The areas under the ROC curves (AUCs) for NK cell counts in predicting 1-, 2-, and 3-year survival were 0.66, 0.76, and 0.68, respectively. Patients diagnosed with stage IIIC1 exhibited a significant reduction in both the absolute counts and the proportion of NK cells compared to those diagnosed with earlier stages, specifically IB3 and IIA. Conclusions: Our study found that pre-treatment levels of circulating NK cell counts and proportions serve as promising prognostic biomarkers for cervical cancer patients undergoing postoperative radiotherapy.

Background: Immune checkpoint inhibitor (ICI) treatments are promising therapies for hepatocellular carcinoma (HCC) patients. However, not all HCC patients benefit from immunotherapy. Therefore, it is urgent to explore markers for the clinical efficacy and prognosis of immunotherapy for liver cancer. This study aimed to investigate changes in peripheral blood lymphocyte subsets after immunotherapy and to assess their predictive and prognostic value.Methods: Sixty-one patients with advanced HCC were enrolled. Peripheral blood samples were collected from HCC patients before and after ICI treatment, and lymphocytes were detected by flow cytometry. The rank sum test, chi-square test, Kaplan‒Meier curve, and Cox regression model were used to determine the relationship between the changes in the percentages of peripheral blood lymphocyte subsets and clinicopathological characteristics, clinical efficacy, progression-free survival (PFS) and overall survival (OS).Results: After ICI treatment, the percentage of CD3+CD8+ T cells increased, and the percentage of B cells decreased. The changes in memory T cells percentages varied according to different immune efficacy groups. Age, history of hepatitis B infection, first-line therapy, and distant metastasis influenced the proportion of peripheral blood lymphocyte subsets in patients with advanced HCC. Furthermore, univariate analysis demonstrated that high percentage changes in the natural killer (NK) cells percentage change predicted longer PFS and OS.Conclusions: ICI treatment alters the percentage of peripheral blood lymphocyte subsets in immunotherapy-treated HCC patients. Changes in the proportion of lymphocyte subsets are influenced by variances in immunological response and clinicopathological features. A high degree of NK cells percentage change in HCC patients treated with ICI represents an independent prognostic predictor.

Ankylosing spondylitis is a complicated consequence of genetic predisposition and environmental factors. Enthesitis is believed to be the hallmark of ankylosing spondylitis, and the chronic inflammatory state of this disease is perpetuated by the disturbances of both the innate immune system and the acquired immune system. To clarify the alteration of immune system in patients with AS, we conducted a meta-analysis concerning the proportions of major lymphocyte subsets in the peripheral blood of AS patients. We systematically searched PubMed and China National Knowledge Infrastructure (CNKI) for articles related to this subject. A total of 95 articles involving 4,020 AS patients and 3,065 healthy controls were included in the analysis. This meta-analysis is performed on R platform using R package “meta”, and Egger’s tests were used to determine the presence of publication bias. Results showed that the percentages of T cells, NK cells and NKT cells were not significantly different between AS patients and healthy controls, but B cells were significantly increased. Among the subsets of T cells, the proportions of CD4+ T cells, Th17 cells, Tfh cells as well as Th1/Th2 ratio were significantly increased, while Tregs were significantly decreased. Subgroup analysis showed that the proportions of Th17 among both PBMCs, T cells and CD4+ T cells were significantly elevated, while Tregs were only significantly lower in PBMCs. Subgroup analysis also demonstrated that Tregs defined by “CD4+CD25+FoxP3+”, “CD4+CD25+CD127low”or “CD4+CD25+CD127-”were significantly downregulated, indicating that the selection of markers could be critical. Further study is warranted in order to elucidate the complicated interactions between different lymphocyte subsets in AS patients. This study implied that the disequilibrium between Th17 and Tregs, as well as between Th1 and Th2 could contribute to the pathogenesis of ankylosing spondylitis, further cementing the understanding that ankylosing spondylitis is a consequence of disrupted balance of innate immune system and acquired immune system.

We investigated whether recombinant human erythropoietin (rhEPO) therapy affected the lymphocyte subsets in patients on long-term maintenance hemodialysis (HD) with severe anemia. Before treatment, the numbers of peripheral blood lymphocyte, CD3+, CD4+, CD8+, and CD20+ cells were decreased in HD patients compared to those in healthy subjects, while the number of CD3+HLA-DR+ cells was increased in HD patients compared to that in healthy subjects. Furthermore, the number of CD4+CD45RA+ (naive T) cells was markedly decreased in HD patients (112 +/- 77 vs 241 +/- 146/microliters; P < 0.01). The number of CD8+S6F1+ (cytotoxic T) cells in HD patients was also less than that in healthy subjects (247 +/- 104 vs 122 +/- 83/microliters; NS). During a 6-month period of rhEPO therapy, we found that the low level of CD4+CD45RA+ cells gradually increased (from 112 +/- 18 to 163 +/- 24/microliters; P < 0.01) with the elevation of hematocrit values (from 21.5 +/- 1.7 to 28.2 +/- 3.5%; P < 0.05). The number of CD3+HLA-DR+ cells decreased after 1 month of rhEPO therapy (from 93 +/- 14 to 46 +/- 13/microliters) and gradually declined throughout the 6-month study period. In our in vitro study, we demonstrated that no effects were observed on [3H]thymidine uptake in the T cell subsets at various concentrations of rhEPO. These results suggest that rhEPO-induced immunoregulation is mediated by an indirect stimulatory effect on the immune system, this stimulation being accompanied by an improvement in physical condition.

To investigate the effect of ganciclovir combined with interferon atomization inhalation on T lymphocyte subsets in patients with Epstein-Barr virus (EBV) infection and its efficacy. Fifty patients with EBV infection who received ganciclovir combined with interferon atomization inhalation were selected as the observation group, and 50 healthy people were selected as the control group. The changes of T lymphocyte subsets in peripheral blood were detected by flow cytometry before treatment and at the 1st, 2nd, 3rd and 4th cycle after treatment. Before treatment, the CD3+, CD4+, CD4+/CD8+ indexes of the patients were significantly lower than those of the control group (P < .05), and the CD8+ level was significantly increased (P < .05). After one cycle of treatment, there was no significant difference in the changes of T lymphocyte subsets compared with those before treatment. After 2 and 3 cycles of treatment, CD3+, CD4+, CD4+/CD8+ values were higher than those before treatment (P > .05), and CD8+ index was lower than that before treatment (P < .05). After the 4th cycle of treatment, CD3+, CD4+, CD4+/CD8+ values were significantly improved (P < .05), and CD8+ index was significantly decreased (P < .05). Ganciclovir combined with interferon atomization inhalation can regulate the changes of T lymphocyte subsets in patients with EBV infection, improve the patient's condition, and has no obvious adverse reactions. Monitoring the changes of T lymphocyte subsets during treatment is more meaningful to predict the therapeutic effect of patients with EB virus infection.

This study aimed to investigate the clusters of lymphocyte subset in patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) and their correlation with clinical characteristics. A total of 247 active AAV patients, 70 AAV patients with induced remission, and 252 healthy controls (HCs) were enrolled. Based on lymphocyte subsets, results were visualised by principal component analysis, and subgroups of patients were identified by cluster analysis. The absolute number of total lymphocytes and lymphocyte subsets were lower in patients with AAV than in HCs. All lymphocyte subsets were negatively correlated with BVAS (p<0.01) in patients with AAV, except for B cells. A decrease in lymphocyte subset count was associated with renal damage in patients with AAV. T lymphocyte subset features recovered during remission compared with the active disease state. However, the percentage and absolute number of B cells markedly decreased in the induced remission group (p<0.0001). Cluster analysis classified patients into three distinctive subgroups: Patients in cluster 2 had the highest white blood cell count, serum albumin level, monocyte count, and eGFR, whereas they had the lowest serum creatinine level and ESR (p<0.05). Patients with decreased lymphocyte subsets showed poor disease outcome. The count of CD3+ T lymphocytes had the best predictive power for identifying disease outcome in AAV patients. The counts of lymphocyte are lower in patients with active AAV than HCs. Decreased lymphocyte subsets may serve as a biomarker for assessing disease severity and predict poor outcome in AAV patients.