Peripheral Blood Lymphocyte Subsets Research Articles

High Expression of Plasma IL-1β Levels and Transition of Regulatory T-Cell Subsets Correlate with Disease Progression in Myelodysplastic Syndrome

Objective: Myelodysplastic syndrome (MDS) is a group of heterogeneous hematopoietic clonal diseases. Recent studies have confirmed that there are essential immune differences in the progression of MDS. However, the detailed mechanisms remain unclear. In order to explore the changes of immune disorders during the occurrence and progression of MDS, and identify critical differentially expressed factors as well as prognostic relevance. What's more, to probe the possible immunological mechanism in the progression of MDS. Methods: The study had a prospective design. A total of 86 patients with newly diagnosed MDS were enrolled in this study, where patients were classified as low risk group and high risk group based on the revised international prognostic scoring system (IPSS-R) score. The expression profiles of multiple cytokines in patients’ plasma were determined using Luminex microarray assay, and flow cytometry were used to measure lymphocyte subsets in peripheral blood, especially T regulatory cell (Treg) and its subsets: induced regulatory Treg (iTreg: CD45RO+CD3+CD4+CD25+CD127low+) and naïve regulated Treg (nTreg: CD45RA+CD3+CD4+CD25+CD127low+), and the percentage of Monocytic-myeloid derived suppressor cells (M-MDSCs) in bone marrow. The mRNA expression of NLRP3 inflammasome related molecules was detected by real-time quantitative PCR. P <0 .05 was considered statistically significant. Results: In the high risk group, the plasma levels of some cytokines or chemokines with promoting or amplifying inflammation signals in directly or indirectly way, including IL-9, MIP-1β,GRO-α and TNF-β, and two growth factors influencing the immune microenvironment by regulating the growth and differentiation of corresponding cells, including β-NGF, PDGF-bb, were significantly lower (P<0.05).The expression level of IL-1β was significantly higher in the high risk group (P<0.05) and its expression was significantly correlated with the percentage of M-MDSCs (P<0.0001). What's more, the percentage of bone marrow M-MDSCs facilitated immune tolerance was significantly higher. Although there is no significant difference of Treg between the two groups, both the percentages and absolute counts of nTreg/iTreg was decreased significantly in high-risk group of MDS patients (P<0.05) (Figure). After adjusting the Cox model, nTreg/iTreg was an independent protective factor for OS (P=0.038, HR=0.081, 95% CI: 0.008-0.869), and elevated IL-1β level was an independent prognostic factor for PFS (P=0.018, HR=2.554, 95% CI: 1.175-5.552). Then, we found that the mRNA expression of NLRP3, ASC, Caspase 1 and IL-1β were significantly increased in the high risk group(P<0.05). Conclusion: All above results demonstrated that the expression of pro-inflammatory cytokines and chemokines was increased in low risk MDS, while the amplification of bone marrow M-MDSCs and the transformation of nTreg to iTreg may be the key immune mechanisms leading to disease progression. The activation of NLRP3/IL-1β pathway may play an important role in the progression of disease. The serum levels of IL-1β and the percentage of nTreg /iTreg ratio may be an effective indicator to predict the prognosis of MDS. It provided a basis for clinical prediction and early intervention to improve the prognosis. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Characterizing Immunodeficiency in Patients with Telomere Biology Disorders

Telomere biology disorders (TBD) are caused by germline mutations in telomere related genes that lead to excessive telomere shortening. Affected genes include TERT and TERC, which encode two components of the telomerase complex. TBDs are multisystem disorders that present with a constellation of findings including bone marrow failure and lung and/or liver disease. Immunodeficiency has been described but is not well characterized in a large cohort of TBD patients. In the present study, we report the clinical incidence and severity of infections, as well as laboratory evidence of immunodeficiency in 83 patients with TBD. The electronic medical record was queried for the clinical history of patients with TBD who were evaluated at the National Institutes of Health Clinical Center from 2002 to present. Laboratory data included immunoglobulins (IgG, IgA, IgM) and peripheral blood lymphocyte subsets (CD4, CD19). Fifteen patients did not have immunoglobulin levels and 29 patients did not have lymphocyte subsets available. A significant infection history was defined as occurrence of infections that were opportunistic, recurrent, and/or required hospitalization. Recurrent infections were defined as two or more severe infections in one year, three or more respiratory infections (e.g. sinusitis, otitis, bronchitis) in one year, or the need for antibiotics for two months per year. Diminished immune function was defined by CD4 T cell counts: patients with CD4 < 200 cells x 106/L were classified as immunodeficient, and CD4 >200 cells x 106/L but < 500 cells x 106/L as immune impaired. Reference ranges for female and male patients were used to determine whether levels are below or within normal limits. Development of lung and/or liver fibrosis were also assessed. Chi-square tests were used to test the association between the two categorical variables. Kaplan-Meier curves and log rank tests were used to estimate overall survival (OS) and test the differences of survival time distributions among infection groups and immune status groups. Patient characteristics are in Table 1. TERT and TERC were the most commonly mutated genes, and others included DKC1, PARN, TINF2, CTC1, RTEL1, and NAF1. Of 49 patients with laboratory data available, the majority (65%) had depressed CD4 counts with 11 immunodeficient patients, 21 immune impaired patients, and 17 patients with normal counts. Median age was similar in the 3 groups. Three (27%) immunodeficient patients and 2 (10%) immune impaired patients had an opportunistic infection. Two (18%) immunodeficient patients and 6 (29%) immune impaired patients reported recurrent infections. Nine (43%) immune impaired patients required hospitalization from infection. No patients with normal CD4 count had opportunistic or recurrent infections, but 4 had required hospitalization for infection. Lower CD4 count was associated with having a significant infection history, and specifically recurrent infections, and being hospitalized for infection (p=0.0417, p=0.0471, p=0.0154 respectively). We observed varied associations between other clinical factors and immune dysfunction or infection. Low levels of CD19 B cells were observed in 14 patients, and associated with development of recurrent infections (P = 0.0003) but not opportunistic or infections requiring hospitalization. Two patients had deficient IgG and IgA levels while 8 had deficient IgM. However, immunoglobulins were not associated with any significant infection history. All immunodeficient and immune impaired patients had concurrent bone marrow failure. Lung fibrosis was recorded in 6 (54%) immunodeficient and 6 (27%) immune impaired patients, and liver fibrosis in 4 (36%) immunodeficient and 4 (19%) immune impaired patients. Of 11 patients with both lung and liver fibrosis, 4 (36%) were immunodeficient, 2 (18%) were immune impaired, and 5 (45%) did not have CD4 counts available. There was no association between gene mutation and development of low CD4 count or clinically significant infections. OS was lower in TBD patients with clinically significant infections (p=0.00011) or low CD4 counts (p=0.016) (Figure 1). Immune dysfunction is common in TBD and patients are at increased risk of clinically significant infections. Low CD4 counts are associated with worse OS. TBD patients should be routinely screened for immunodeficiency to allow appropriate clinical monitoring, antimicrobial prophylaxis, and education. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The predictive value of peripheral blood cells and lymphocyte subsets in oesophageal squamous cell cancer patients with neoadjuvant chemoradiotherapy.

Neoadjuvant chemoradiotherapy (nCRT) is a standard treatment option for patients with stage III oesophageal cancer. Approximately 30% of oesophageal cancer patients will have a pathological complete response (pCR) after nCRT. However, available clinical methods cannot accurately predict pCR for patients. We aimed to find more indicators that could be used to predict the pathological response to nCRT. A total of 84 patients with stage III oesophageal squamous cell cancer were enrolled in this study. Ten patients failed to have surgery as a result of progressive disease (PD). Among the patients who underwent surgery, 32 patients had a pathologic complete response (pCR), whereas 42 patients showed no or partial response (npCR) after nCRT. Routine blood test results and lymphocyte subset assessments before and after nCRT were retrospectively analysed. Univariate and multivariate analyses were used to identify independent predictors of the clinical curative effect of nCRT. Eventually, nomograms were established for predicting the PD and pCR rates. The numbers of lymphocytes, B lymphocytes, T lymphocytes, Th lymphocytes, Ts lymphocytes, and NK cells and the percentages of B lymphocytes and NK cells were decreased significantly after nCRT (P < 0.0001), whereas the percentages of T lymphocytes and Ts lymphocytes increased (P < 0.0001). Univariate analysis showed that age, the length of the lesion, the level of haemoglobin before nCRT, and the amount of change in haemoglobin were related to PD, and the percentage of NK cells after nCRT was related to pCR. Multivariate logistic analysis demonstrated that the length of the lesion, the neutrophil-to-lymphocyte ratio (NLR) before nCRT, and the amount of change in haemoglobin were independent predictors of PD, whereas the percentage of NK cells after nCRT was an independent predictor of pCR. Lymphocyte subsets changed dramatically during nCRT, and these changes together with baseline and posttreatment lymphocyte subsets have predictive value in determining the response to nCRT for oesophageal cancer.

Remifentanil combined with dexmedetomidine on the analgesic effect of breast cancer patients undergoing modified radical mastectomy and the influence of perioperative T lymphocyte subsets.

To study the analgesic effect of breast cancer patients undergoing modified radical mastectomy (MRM) and the influence of perioperative T lymphocyte subsets by remifentanil combined with dexmedetomidine. 80 breast patients were divided into control group and research group based on the anesthesia protocol. Patients in control group was given remifentanil for anesthesia induction and maintenance, and patients in research group was given remifentanil and dexmedetomidine for anesthesia induction and maintenance. We compared the anesthesia time, operation time, surgical blood loss, postoperative wake-up time, extubation time, incidence of adverse reactions, VAS score and T lymphocyte subsets in peripheral blood in the two groups of patients. The baseline data including age, height, weight and BMI, ASA classification, stage of breast cancer, frequency of neoadjuvant therapy, and surgical characteristics including anesthesia time, operation time and bleeding volume all have no significant difference between two groups (P > 0.05). Compared to control group, the time of wake up and extubation in patients of research group were all significantly decreased (P < 0.05), and significantly decreased MBP and HR after loading dose of dexmedetomidine in research group (P < 0.05). The VAS scores of patients at 4, 8, 12, 16, 20 and 24 h after surgery in the research group are all significantly lower than those in the control group (P < 0.05). Before induction of anesthesia, there was no significant difference in the ratio of CD4+, CD8+ and CD4+/CD8+ T lymphocytes in peripheral blood between the two groups (P > 0.05). At 1 h during operation and 24 h after operation, the ratio of CD4+ and CD4+/CD8+ cells in the research group was significantly higher than these of the control group (P < 0.05), while the ratio of CD8+ cells was lower than that of the control group (P < 0.05). For breast cancer patients undergoing MRM, the use of remifentanil combined with dexmedetomidine can enhance postoperative analgesia and reduce postoperative immunosuppression.

POLYMYALGIA RHEUMATICA AFTER MRNA COVID-19 VACCINATION

<h3>Introduction</h3> Polymyalgia rheumatica (PMR) is an inflammatory disorder characterized by muscle pain and stiffness. We present a 67-year-old man who developed PMR after immunization with the mRNA COVID-19 vaccine. <h3>Case Description</h3> A 67-year-old male with a history of viral pericarditis, non-ischemic cardiomyopathy, Raynaud's phenomenon, and prostate cancer presented for evaluation of acute-onset proximal muscle pain and weakness, extreme fatigue, malaise, and weight loss. Symptoms developed 10 days after the first dose of the mRNA COVID-19 vaccine. No associated fever or skin rashes. No history of natural COVID-19 infection. A workup for malignancy was negative. CRP and ESR were elevated. ANA was positive (1:80, speckled pattern). ENAs were negative. CK and aldolase were normal. He had low IgG at 328 mg/dL with normal IgA and IgM. However, repeat IgG levels a week later were normal (1130 mg/dL), raising the possibility of a laboratory error. Lymphocyte subsets in peripheral blood were normal. IgG titers to the SARS-COV2 spike and nucleocapsid proteins were consistent with COVID-19 immunity from prior immunization. He was treated with low-dose oral prednisone with resolution of his symptoms and has been slowly tapering prednisone as tolerated. <h3>Discussion</h3> This case report raises the possibility that PMR may be triggered by an immune response to the mRNA COVID-19 vaccine in susceptible individuals. However, further studies involving a larger population are required to assess whether this association is causative or rather coincidental.

Peripheral Blood Lymphocyte Subsets as a Risk Predictor of Patients with Endometrioid Endometrial Cancer.

This study aimed to explore lymphocyte subsets for the personalized prediction of endometrioid endometrial cancer (EEC) risk and evaluated the correlation between immune cells and International Federation of Gynecology and Obstetrics (FIGO) staging in patients with EEC. A case-control study was conducted to analyze the clinical data of 421 patients admitted to Fujian Maternity and Child Health Hospital from October 2017 to December 2021. t-tests or Mann-Whitney U-tests were used to analyze the percentages and absolute counts of peripheral blood lymphocyte subsets in patients with EEC and patients without cancer. The independent risk factors for ECC and FIGO stage were analyzed via multivariate binary logistic regression. The receiver operating characteristic curve was calculated to evaluate the prediction efficacy of risk factors on ECC. The CD4+ T% in the 121 patients with EEC was lower than in the 300 patients without cancer (P = 0.013). The absolute counts of peripheral CD4+ T (P = 0.002) and T cells (P = 0.007) in 37 patients with EEC were lower than in 51 patients without cancer. Multivariate binary logistic regression analysis showed that CD4+ T% and natural killer cell (NK)% were independent risk factors for FIGO staging in patients with EEC. NK% was significantly higher in patients with advanced stage (FIGO III and IV) than those with early EEC (FIGO I and II) (P = 0.004). To determine the early and advance FIGO stage of EEC, the cutoff value of NK% was calculated as 19.94%. With the decrease of CD4+ T counts, the immune status of patients with EEC is impaired. NK cells may help in the evaluation of the prognosis of patients with EEC and are likely to be an independent risk factor for FIGO staging in patients with EEC.

Protective Effect of the Polysaccharides from Taraxacum mongolicum Leaf by Modulating the p53 Signaling Pathway in H22 Tumor-Bearing Mice.

Dandelion is an edible plant with a variety of bioactive components. This paper mainly reports the antitumor activity of dandelion polysaccharide DLP120 on H22 tumor-bearing mice. DLP120 is an acidic polysaccharide composed of pectin and arabinogalactan. The results indicate that DLP120 markedly inhibited tumor growth in a dose-dependent manner and attenuated and regulated negative effects on organs. In addition, DLP120 not only increased the viability of spleen lymphocytes and natural killer (NK) cells, but also increased the proportion of lymphocyte subsets in peripheral blood. Furthermore, Hematoxylin-Eosin (HE) staining showed that tumor tissues and cells exhibited typical pathology features. Annexin V FITC/PI staining and cell cycle distribution results further confirmed apoptosis and cell cycle arrest in S and G2 phases. Notably, there was a significant accumulation of reactive oxygen species. Western blotting results demonstrated that the expression of p53 was up-regulated in the DLP120 group. Moreover, the pro-apoptotic protein Bax was up-regulated while the inhibitory-apoptotic protein Bcl-2 was down-regulated. In addition, the expression of Fas and FasL, associated with the death receptor pathway, were also up-regulated. Overall, administration of DLP120 in H22 tumor-bearing mice can not only enhance immunity but also directly induce tumor cell apoptosis.

Changes of T Lymphocyte Subsets in Peripheral Blood of Patients with Intermediate and Advanced Cervical Cancer before and after Nimotuzumab Combined with Chemoradiotherapy

Introduction: Cervical cancer (CC) is the main malignant tumor of gynecology with high mortality. This study aimed to explore the changes of T lymphocyte subsets in the peripheral blood of patients with intermediate and advanced CC (IACC) treated with nimotuzumab (Nimo) combined with chemoradiotherapy (CRT). Methods: Peripheral blood was extracted before and after treatment, and patients were randomly divided into the CRT group (N = 68) or the Nimo + CRT group (N = 68). The levels of tumor markers squamous cell carcinoma antigen (SCCA), carbohydrate antigen 125 (CA125), and carcinoembryonic antigen (CEA), tumor indexes leptin and insulin-like growth factor-2 (IGF2), and key secretory factors (interferon γ/tumor necrosis factor α/interleukin (IL)-4/IL-13) of Th1 and Th2 cells in the serum were detected. The levels of T lymphocyte subsets CD3+, CD4+, CD8+, and CD4+/CD8+ and the levels of Th1/Th2 and Th17/Treg in CD4+ cell subsets were determined by flow cytometry. The objective remission rate, progression-free survival (PFS), and overall survival (OS) of patients were analyzed, and Kaplan-Meier curves were drawn to show the prognosis of patients. Results: After treatment, the levels of SCCA, CA125, CEA, leptin, and IGF2 in the serum of patients with IACC were decreased, the level of Th1/Th2 was increased, and the Th17/Treg level was decreased. The treatment effect of Nimo + CRT was more significant than that of CRT alone. Survival curve analysis showed that Nimo + CRT could prolong PFS and OS in patients with IACC. In addition, the follow-up results of patients showed that Nimo did not increase the incidence and severity of adverse reactions caused by CRT. Conclusion: Nimo combined with CRT can protect the immune function of patients, improve the effective rate, and prolong the survival rate of patients with IACC.

Peripheral blood lymphocyte subsets are associated with the clinical outcomes of prostate cancer patients

BackgroundMany studies have sought associations between the total peripheral blood lymphocyte count and prostate cancer (PCa) prognosis, but not peripheral lymphocyte subsets. We explored the associations between the absolute counts of peripheral lymphocyte subsets and PCa patient survival. Methods135 PCa patients were included in this study. The log-rank test and Kaplan-Meier method were employed to compare overall survival (OS) and progression-free survival (PFS) rates. Univariate and multivariate Cox’s regression analyses were employed to identify prognostic factors. Flow cytometry analysis was used to determine the numbers of peripheral lymphocyte subsets. ResultsPCa patients with lower absolute counts of certain lymphocyte subsets showed poorer PFS and OS than those with higher absolute counts of these cells. The numbers of CD4+ T cells, CD3+ T cells, and natural killer (NK) cells were significantly higher in PCa patients of tumor node metastasis (TNM) Ⅰ-Ⅱ stages than those of TNM ⅠIII-IV stages. Univariate and multivariate Cox’s regression analyses of OS and PFS indicated that neutrophil numbers > 4.81*109/L, CD4+ T cells ≤ 254 /μL, and NK cells ≤ 136 /μL were unfavorably prognostic for patients with PCa. ConclusionsLower absolute counts of certain peripheral lymphocyte subsets (NK cells ≤ 136/μL and CD4+ T cells ≤ 254/μL) are prognostically unfavorable for PCa patients.

Dynamic analysis of immune status in patients with intracranial germ cell tumor and establishment of an immune risk prognostic model.

IntroductionImmune status was evaluated by means of lymphocyte subset counts and immune factors in cancer. This study analyzed the peripheral blood immune index and survival outcomes in intracranial germ cell tumor (iGCT) patients.MethodsPeripheral blood lymphocyte subset counts and levels of interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor (TNF), and interferon-γ (IFN) from 133 iGCT patients were collected and retrospectively analyzed. Their clinical information was extracted from the hospital database, and prognosis was confirmed by telephone visit. Patients (n=11) underwent prospective review and their samples of peripheral blood lymphocytes were verified.ResultsA total of 113 (84.2%) patients received comprehensive treatments, including 96 standard therapy (combination of full course chemotherapy and radiology with or without surgery) and 17 comprehensive but non-standard therapy (either without full course chemotherapy or with non-standard radiotherapy) and 98 (73.7%) reached complete or partial response. T lymphocytes (CD3+), cytotoxic T cells (CD3+CD8+ or Tc), and B lymphocytes (CD19+) decreased (p=0.047, p=0.004, and p<0.001, respectively), while activated cytotoxic T lymphocytes (CD8+CD25+) and IFN increased (p<0.001 and p=0.002, respectively) after treatment. Median survival was 45.33 months, and patients with increased Tc cells and activated Tc cells as well as IFN presented encouraging outcomes (p=0.039, p=0.041, and p=0.017 respectively). Regression analysis showed that non-increased Tc cells and non-increased activated Tc cells were independent factors of poor prognosis (p=0.016, HR=3.96, 95%CI=1.288-12.20; p=0.002, HR=4.37 95%CI= 1.738-10.97). Standard chemo-radiotherapy was independently related to reduced risk of death(p=0.022, HR=0.19, 95%CI=0.044-0.79). Consistence was seen in a nomogram established through retro and prospective studies. An immune risk model indicated the activated group (with both increased activated T cells and IFN levels) had the best prognosis, the mildly activated type with elevated IFN levels had intermediate outcome, and patients with the silent immune status had the worst outcomes (Log rank test, p=0.011).ConclusionImplementation of standard comprehensive treatments led to positive responses. Dynamic monitoring of peripheral blood lymphocyte subsets can be used as an auxiliary indicator for prognosis judgment.

Serum 3-phenyllactic acid level is reduced in benign multiple sclerosis and is associated with effector B cell ratios

Background3-phenyllactic acid (PLA) is produced by both intestinal bacteria and the human host. PLA exists in its D- and L- chiral forms. It modulates human immune functions, thereby acting as a mediator of bacterial-host interactions. We aim to determine the amount and potential influence of PLA on clinical and immunological features of MS. MethodsWe measured D- and L-PLA levels in bacterial supernatants and in sera of 60 MS patients and 25 healthy controls. We investigated potential associations between PLA levels, clinical features of MS, serum cytokine levels and ratios of peripheral blood lymphocyte subsets. ResultsMultiple gut commensal bacteria possessed the capacity to generate D- and L-PLA. MS patients with benign phenotype showed markedly lower PLA levels than healthy controls or other MS patients. Fingolimod resistant patients had higher PLA levels at baseline. Furthermore, MS patients with higher PLA levels tended to display increased memory B and plasma cell ratios, elevated IL-4 levels and increased ratios of IL-4 and IL-10 producing T cell subsets. ConclusionCollectively, our work indicates that reduced serum levels of PLA could be associated with a favorable clinical course in MS and possibly be used as a biomarker.

Studies of CD45+ and CD46+ expression on the peripheral blood lymphocyte subsets of the post-COVID patients

The SARS-CoV-2 virus can enter the cells using S1 viral spike (S) protein, not only by binding to ACE2, but also through other cellular receptors. These candidate receptors include CD46, which, like CD45, belongs to pan-leukocyte receptors and is expressed on all types of lymphocytes. In turn, SARS-CoV-2 infection is accompanied by damage to almost all compartments of the immune system, mainly T lymphocytes. The purpose of the study was to evaluate the expression levels of CD45+ and CD46+ in various subpopulations of lymphocytes in patients who had undergone SARS-CoV-2 infection.&#x0D; 72 patients who had undergone SARS-CoV-2 infection were examined. Using flow cytometry technique, we determined CD45+ and CD46+ (panleukocyte marker for lymphocyte gating), CD45+ and CD46+, CD3+ (T lymphocytes), CD45+ and CD46+, CD3+, CD4+ (helper inducers), CD45+ and CD46+, CD3+, CD8+ (cytotoxic T-lymphocytes), CD45+ and CD46+, CD3+, CD56+ (TNK cells) CD45+ and CD46+, CD3-, CD56+ (natural killers), CD45+ and CD46+, CD3-, CD19+ (B lymphocytes), CD45+ and CD46+, CD3+, CD4+, CD25+ (activated helpers, early activation of lymphocytes), CD45+ and CD46+, CD3+, HLA-DR (activated T lymphocytes late activation of lymphocytes). Our studies have shown that a decrease in CD46+ expression in T lymphocytes (CD3+) is accompanied by similar decrease of its expression in cytotoxic T lymphocytes (CD3+, CD8+), TNK (CD3+, CD56+), as well as in helpers T carrying markers of early activation (CD3+, CD4+, CD25+). At the same time, the most pronounced decrease was observed both among total T lymphocytes and cytotoxic T cells. In these patients, the expression level of CD46+ in B lymphocytes was slightly increased. Recent data suggest that there is no involvement of CD46 receptor on B lymphocytes. Our data suggest that the SARS-CoV-2 virus may affect the CD46 receptor. Such exposure may lead to promotion of the long-COVID (post-COVID) symptoms in such patients, thus requiring new approaches to correction of these disorders.

Reference long-read isoform-aware transcriptomes of 4 human peripheral blood lymphocyte subsets.

Long-read sequencing technologies such as isoform sequencing can generate highly accurate sequences of full-length mRNA transcript isoforms. Such long-read transcriptomics may be especially useful in investigations of lymphocyte functional plasticity as it relates to human health and disease. However, no long-read isoform-aware reference transcriptomes of human circulating lymphocytes are readily available despite being valuable as benchmarks in a variety of transcriptomic studies. To begin to fill this gap, we purified 4 lymphocyte populations (CD4+ T, CD8+ T, NK, and Pan B cells) from the peripheral blood of a healthy male donor and obtained high-quality RNA (RIN &gt; 8) for isoform sequencing and parallel RNA-Seq analyses. Many novel polyadenylated transcript isoforms, supported by both isoform sequencing and RNA-Seq data, were identified within each sample. The datasets met several metrics of high quality and have been deposited to the Gene Expression Omnibus database (GSE202327, GSE202328, GSE202329) as both raw and processed files to serve as long-read reference transcriptomes for future studies of human circulating lymphocytes.

Effect of pembrolizumab on T lymphocyte subsets in patients with advanced oral cancer and its therapeutic effect.

Background:The aim of this study is to investigate changes of peripheral blood lymphocyte subsets before and after treatment with pembrolizumab for advanced oral cancer and its clinical sig-nificance.Methods:32 patients with advanced oral cancer who received pembrolizumab treatment were selected as observation group, 30 healthy people during the same period were selected as control group. Before treatment and in cycles 1, 2, 3 and 4 after treatment, fluid cytometry was used to detect changes in levels of lymphocyte subsets in peripheral blood of patients.Results:CD3+, CD4+, CD4+/CD8 + indexes of patients with advanced oral cancer before treatment were significantly lower than those in control group (P < .05), CD8 + level was significantly increased (P < .05); After 1 cycle of pembrolizumab treatment, there was no significant difference in changes of lymphocyte subsets compared with before immunotherapy; After 2 and 3 cycles of treatment, CD3+, CD4+, CD4+/CD8 + values were higher than before the treatment (P > .05), CD8 + index was slightly lower than before treatment (P < .05); After fourth cycle of treatment, CD3+, CD4+, CD4+/CD8 + values were significantly improved compared to before treatment (P < .05), CD8 + index was significantly lower than before treatment (P < .05); In treatment process of patients with stable disease (SD)/partial response (PR), the CD3+, CD4+, CD4+/CD8 + values of fourth cycles were higher than before treatment (P < .05), CD8 + index was lower than before treatment (P < .05); During treatment of progressive disease (PD) patients, changes of lymphocyte subsets in fourth cycles were not significantly different from those before treatment (P > .05). This article shows through analysis that expression of programmed cell death ligand 1 (PD-L1) and pathological types have no obvious influence on effect of immunotherapy. Multi-factor analysis shows that it is more meaningful to observe the changes of CD3+, CD4 + and CD8 + at the same time to predict effect of immunotherapy.Conclusion:Pembrolizumab can regulate changes of T lymphocyte subsets in patients with advanced oral cancer, improve immune status of patients, there is no obvious adverse reaction. Monitoring changes of lymphocyte subsets during treatment can predict effect of immunotherapy.

Adaptive immunity in different CT patterns of active tuberculosis and possible variability according to patients' geographic provenience.

BackgroundIt is still unclear if low lymphocyte levels are directly related to immunological modifications induced by the TB infection or if they depend on the general pre-existing health impairment of affected patients. Our aim was to detect eventual differences in the immunological status of patients with pulmonary TB compared to an age and sex-matched group of hospitalized patients with other bacterial community-acquired pneumonia (CAP). In addition, we tried to assess an association between alterations in the peripheral lymphocyte subsets and the development of different CT patterns of active TB and to discover differences in the immunological status and in the radiological patterns of TB presentation between patients of different geographic proveniences.MethodsThis observational study included 48 patients with TB and 48 sex- and age-matched patients affected by other bacterial CAP. The presence of HIV/AIDS, other immunocompromising conditions, and confounding chronic pulmonary comorbidities was excluded. Flow cytometry was performed on all the enrolled subjects at admission, before starting the appropriate antibiotic therapy. Patients with TB also underwent a computed tomography (CT) scan.ResultsPatients with TB showed a decrease in the absolute count of all the lymphocyte subsets compared to the CAP group. Only the reduction in the percentage of CD4+ T-lymphocytes was significant, while the percentage of CD8+ T-lymphocytes was significantly increased. Patients presenting exudative forms with atypical locations of TB showed a significant reduction in the absolute count and percentage of CD19+ B-lymphocytes compared to those affected by productive TB forms with the typical location. Despite being younger, our black Sub-Saharan Africans showed a significant reduction in the CD4+ T-lymphocytes compartment and a higher prevalence of atypical and exudative forms of TB compared with white Europeans.ConclusionTuberculosis itself may alter peripheral blood lymphocyte subsets compared to other CAP. An impaired CD19+ B-lymphocyte compartment may result in an abnormal exudative response in atypical locations and a suboptimal bacterial control. Other constitutive or environmental causes may influence immunological differences found in patients with TB, particularly in case of different geographic origins. Anyhow, flow cytometry may be of great value in evaluating the immune function of these patients.

Early alteration of peripheral blood lymphocyte subsets as a risk factor for delirium in critically ill patients after cardiac surgery: A prospective observational study.

ObjectiveThere is a high incidence of delirium among patients with organ dysfunction undergoing cardiac surgery who need critical care. This study aimed to explore the risk factors for delirium in critically ill patients undergoing cardiac surgery and the predictive value of related risk factors.MethodsWe conducted a prospective observational study on adult critically ill patients who underwent cardiac surgery between January 2019 and August 2021. Patients were consecutively assigned to delirium and non-delirium groups. Univariate analysis and multivariate logistic analysis were used to determine the risk factors for delirium. Receiver operating characteristic curves and a nomogram were used to identify the predictive value of related risk factors.ResultsDelirium developed in 242 of 379 (63.9%) participants. Acute Physiology and Chronic Health Evaluation II (APACHE II) and Sequential Organ Failure Assessment (SOFA) scores were 14.2 ± 5.6 and 18 ± 8.4, respectively. Patients with delirium had longer cardiopulmonary bypass time (149.6 ± 59.1 vs. 126.7 ± 48.5 min, p < 0.001) and aortic cross-clamp time (98.7 ± 51.5 vs. 86.1 ± 41.6 min, p = 0.010) compared with the non-delirium group. The area under the curve was 0.824 for CD4+ T cell count and 0.862 for CD4/CD8 ratio. Multivariate analysis demonstrated that age [odds ratio (OR) 1.030, p = 0.038], duration of physical restraint (OR 1.030, p < 0.001), interleukin-6 (OR 1.001, p = 0.025), CD19+ B cell count (OR 0.996, p = 0.016), CD4+ T cell count (OR 1.005, p < 0.001) and CD4/CD8 ratio (OR 5.314, p < 0.001) were independent risk factors for delirium. A nomogram revealed that age, cardiopulmonary bypass duration, CD4+ T cell count and CD4/CD8 ratio were independent predictors of delirium.ConclusionAge, duration of physical restraint, CD4+ T cell count and CD4/CD8 ratio were reliable factors for predicting delirium in critically ill patients after cardiac surgery. The receiver operating characteristic curves and nomogram suggested a potential role for CD4+ T cells in mediating potential neuroinflammation of delirium.

Evaluation of peripheral lymphocyte subsets in acne vulgaris patients before and after systemic isotretinoin treatment.

Isotretinoin (ISO) is a retinoic acid-derived molecule which is very efficient in the treatment of acne vulgaris (AV). Little is known regarding alterating affects of systemic ISO on immune system. Thirty-two patients with moderate and severe AV with indications for systemic ISO treatment were recruited. The evaluation of peripheral blood lymphocyte subsets was made on treatment initiation and at the end of the therapy by flow cytometry. CD16/56+ natural killer (NK) cell levels were higher in patients with severe acne compared to moderate acne (P = 0.009). CD45RA+ (naive) cell levels decreased significantly after systemic ISO treatment; the mean percentage was 66.9 ± 6.5 which decreased to 63.9 ± 7.5 after treatment (P = 0.013). CD19+ cell levels were also significantly increased; mean percentage (%) and absolute cell counts were 8.0 ± 3.1 and 161.3 ± 7.36 cells/μl which increased to 10 ± 2.7 and 227.7 ± 108.7 cells/μl, at the end of treatment, respectively (P = 0.007, P = 0.011). NK cell count may be a parameter related to acne severity. Systemic ISO may have an inhibitory effect on naïve T cells. Finally, systemic ISO seems to stimulate mature B-cell proliferation.

Prognostic Value of Pretreatment Neutrophil-to-Lymphocyte Ratio in HER2-Positive Metastatic Breast Cancer.

This study aimed to examine the prognostic value of the neutrophil-to-lymphocyte ratio (NLR) and other clinicopathological features in HER2+ MBC patients who received first-line anti-HER2 therapy. A total of 129 patients were assigned to NLR-low and NLR-high groups based on a cutoff value of 3.0 at baseline. Peripheral blood lymphocyte subsets and gene mutations in circulating tumor DNA were analyzed by flow cytometry and Next-generation sequencing, respectively. Survival was evaluated by the Kaplan–Meier method and Cox regression analysis. Of the 129 patients, 77 and 52 were assigned to the NLR-low (≤3) and NLR-high (>3) groups, respectively. Compared with NLR-high patients, the NLR-low patients had significantly longer median progression-free survival (PFS) (11.7 vs. 7.7 months) (p = 0.001, HR = 2.703 95% CI 1.543–4.736 and overall survival (OS) (37.4 vs. 28.7 months) (p = 0.044, HR = 2.254 95% CI 1.024–4.924). Furthermore, this association was independent of metastatic sites or estrogen receptor status. Peripheral blood CD3+ (p = 0.034) and CD4+ (p = 0.010) T cell numbers were significantly higher in the NLR-low group than the NLR-high group. The mutational profile of MBC was generally similar between the two groups. Baseline NLR was a prognostic factor of PFS and OS for patients with HER2+ MBC in the first-line setting. These results may facilitate the selection of patients who will benefit most from anti-HER2 treatment.

Establishing Reference Values for Peripheral Blood Lymphocyte Subsets of Healthy Children in China Using a Single Platform.

Lymphocyte subsets significantly change during childhood; thus, age-matched reference values derived from healthy children are crucial. We established reference values for lymphocyte subsets, including T cells (CD3+), CD4 T cells (CD3 + CD4+), CD8 T cells (CD3 + CD8+), double negative T (DNT) cells (CD3 + CD4-CD8-), B cells (CD3-CD19+), NK cells (CD3-CD56+), and NKT-like cells (CD3 + CD56+) in the peripheral blood of 813 healthy children. We used the method of the international standard document (Clinical Laboratory Standard Institute C28-A3) to establish reference intervals with a single platform. First, we used the Skewness and Kurtosis test to analyze the normality of the data. The nonnormally distributed data was transformed into approximately normal distribution by the Box-Cox transformation. Second, we used the Tukey's method to eliminate outliers. Further, all the subjects were grouped into subgroups according to sex (male and female) and age (0–1 month, 2–12 months, 1–3 years, 4–6 years, and 7–18 years). We used the standard normal deviation test (Z-test) to evaluate whether age and sex were possible grouping factors. The analyses indicated age to be an important factor associated with changes in lymphocyte subsets. The absolute number of lymphocyte subsets and total number of lymphocytes, T cells, CD4 T cells, CD8 T cells, and B cells gradually increase from birth to 12 months and then gradually decrease with age. Furthermore, CD4 T cells and the ratio of CD4+/CD8+ gradually decrease with age. In contrast, CD8 T and DNT cells gradually increase with age. The percentage and number of NK and NKT-like cells gradually increase with age and remain stable between 1 and 18 years of age. In conclusion, the age-related reference intervals established in healthy children in this study can aid in monitoring and assessing the changes in immune levels in diseased conditions.

Efficacy and safety of belimumab/low-dose cyclophosphamide therapy in moderate-to-severe systemic lupus erythematosus

ObjectivesWe have reported previously that Belimumab, a human monoclonal antibody that inhibits B-cell activating factor(BAFF) could be an effective and safe option to treat Neuropsychiatric manifestations of SLE (NPSLE). To avoid inadequate efficacy of Belimumab and significant adverse events of often-used dose of cyclophosphamide (CYC) for SLE, we evaluated the efficacy, safety, and possible immune mechanisms of Belimumab treatment in combination with intermittent low-dose intravenous CYC for moderate-to-severe SLE.MethodsIn this non blinded and parallel-group trial, we collected 82 cases of moderate-to-severe SLE patients, 40 received Belimumab treatment and 42 received conventional treatments as historical controls for 24 weeks. The demographic features, clinical manifestations, and laboratory indicators including peripheral blood lymphocyte subgroups or subsets were compared before and after the treatments.ResultsCompared with the baseline, 6 months post Belimumab group treatment, disease activity score SLEDAI (13.78 to 3.82, P<0.05) and BILAG scores (16.40 to 5.48, P<0.05) were reduced; C3 (0.19 to 1.14, P<0.05) and C4 (0.04 to 0.22, P<0.05) increased; the absolute numbers of B and T cells were the first decreased and then significantly increased, tended to balance. Moreover, Belimumab group treatment significantly reduced the serum levels of IL-6, the ratio of B and T cells, and the proportion of infections and menstrual disorders.ConclusionCompared with conventional treatment, Belimumab with low-dose intravenous CYC significantly reduced disease activity scores and maintained the B/T cell balance for SLE patients at 24 weeks. It was more efficacy and safe (adverse events such as infection were significantly lower). It should be the mechanism that Belimumab combined with low-dose intravenous CYC therapy restores the balance of T and B cells, which proposes a potential treatment strategyfor SLE.