Peripheral Lymphocyte Subsets Research Articles

#3767 THE IMPACT OF COVID-19 PANDEMICS ON THE OUTCOME OF 24 CHINESE PATIENTS DIAGNOSED OF MEMBRANOUS NEPHROPATHY RECEIVING RITUXIMAB THERAPY

Abstract Background and Aims Rituximab has been considered as the first-line immunosuppressive therapy in membranous nephropathy. Anti-CD20 depleting therapy is associated with prolonged B-cell depletion and has been reported to increase risk of severe COVID-19 infection in several rheumatic and neurologic diseases. However, few studies reported the impact of COVID-19 infection on membranous nephropathy in the status of B-cell depletion. Our study aimed to describe the impact of prior rituximab therapy on clinical outcomes of a group of Chinese membranous nephropathy within 6-month follow-up during COVID-19 pandemics. Method We prospectively conducted a cohort study of adult patients diagnosed of membranous nephropathy receiving rituximab therapy from Aug 1st to Nov 30th in 2022, in one tertiary hospital, in Beijing. Patients’ baseline characteristics (including demographic data, serum albumin, serum creatinine, 24-hour proteinuria, serum IgG level, peripheral lymphocyte subsets, medications, past medical history, prior COVID-19 vaccination history), clinical outcomes were recorded. We followed up all the patient regularly until Jan 31st, 2023. Results We included 24 patients with median age of 55 (27, 83) years old. 13 (54.2%) patients were on chronic immunosuppression with corticosteroids, calcineurin inhibitors, and/or mycophenolate mofetil. We followed up the patients within median 4.6 months since last rituximab infusion at the dose of 1 gram. During follow-up, 18 (75%) patients got COVID-19 infection including 16 non-severe cases and two severe cases requiring hospitalization and receiving intravenous dexamethasone. The overall COVID-19 vaccination rate was 33%. In subgroup analysis, the baseline characteristics were comparable between COVID-19 infection group and non-infection group. Vaccination rate in COVID-19 infection group was lower than that in non-infection group insignificantly (28% vs 50%, P=0.362), and the two cases of severe infection were unvaccinated with severe hypoimmunoglobulinemia. Circulating natural killer cell counts were significantly different between the two groups (365(63, 745) vs 194(23, 928)/μl, P=0.019), while neither CD4+T cells nor CD8+T cells counts showed statistically difference between the counterparts (CD4+T cells: 632 (227, 1231) vs 407 (230, 571)/μl, P=0.05; CD8+T cells: 496 (134, 1815) vs 306 (127, 728)/μl, P=0.179). No relapse of membranous nephropathy was observed after infection during follow-up. Conclusion COVID-19 infection was common in Chinese membranous nephropathy receiving recent administration of rituximab during COVID-19 pandemics. Low vaccination rate and prolonged B-cell depletion after rituximab infusion in this population may attenuate the protective effect of COVID-19 vaccine. Nature killer cells may play a key role in antiviral immunity. Patients should continue to maintain a high adherence to preventive measures against COVID-19.

Changes in peripheral blood TBNK lymphocyte subsets and their association with acute exacerbation of chronic obstructive pulmonary disease.

This study aimed to determine the changes in peripheral blood TBNK lymphocyte subsets in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and their relationship with the pathogenesis of AECOPD. A cross-sectional study of 1252 hospitalized patients in Zhejiang Hospital was conducted. There were 162 patients in the AECOPD group and 1090 in the non-chronic obstructive pulmonary disease (COPD) group. The proportions of peripheral blood T helper cells, cytotoxic T cells, total B cells, total natural killer (NK) cells, and total T cells in the two groups were determined, and the CD4/CD8 ratio was calculated. The proportions of men and total natural killer cells, and the mean age were significantly higher in the AECOPD group than in the non-COPD group. The T helper cell, total T cell, and CD4/CD8 ratios were significantly decreased in the AECOPD group. A multivariate logistic regression analysis showed that male sex, age, the total T cell ratio, and the CD4/CD8 ratio were significantly associated with the incidence of AECOPD. Cellular immune dysfunction in patients with AECOPD causes a decrease in total T lymphocytes and the CD4/CD8 ratio, which may be involved in the pathogenesis of AECOPD.

Initial results from a first in human dose escalation trial of a novel immune stimulating oncolytic adenovirus, AdAPT-001TGF-ß Trap.

2550 Background: As a class, oncolytic adenoviruses (OAV), modified agents of the common cold, specifically target and lyse tumor cells, and by expression of cytotoxic transgenes activate an immune response. AdAPT-001 is an OAV, which carries a transforming growth factor beta (TGF-ß) trap that neutralizes the immunosuppressive and profibrotic cytokine, TGF-ß. Preclinically, AdAPT-001TGF-ß Trap eradicates non-T-cell infiltrated tumors. The aim of this Phase 1 study was to assess the safety and tolerability and, secondarily, the efficacy of AdAPT-001 TGF-ß Trap after single intra-tumoral injection (IT) (Phase 1a) and multidose IT injection (Phase 1b) in patients with advanced refractory solid tumors. Methods: This was an open label, single-arm, multicenter dose escalation design trial. AdAPT-001 TGF-ß Trap was administered by intratumoral injection. 9 patients were in a single-dose group, where doses of 2.5 x 1011, 5.0 x 1011, 1.0 x 1012 viral particles (VPs), were tested, and 19 patients with a median age of 70, 5 median prior regimens, and a median ECOG score of 1 received AdAPT-001 every 2 weeks at 1.0 x 1012 VPs until progression. Results: AdAPT-001 TGF-ß Trap was well tolerated with local inflammation, fever, and fatigue as the main side effects. No dose limiting toxicities, no grade 4 toxicities, or treatment-related serious adverse events (SAEs) were seen. Tumor types were sarcoma (n = 8), melanoma (n = 2), head & neck (n = 5), gastric (n = 1), rectal (n = 1), breast (n = 1), fallopian (n = 1). Biological activity (virus replication, local reactions, and tumor shrinkage) was observed. The duration of local reactions and virus replication suggested that dosing every 2 weeks was appropriate. The overall responses to treatment were as follows: 5 patients had durable stable disease for 6 months or more, 1 patient had a partial response, and 2 patients demonstrated pseudoprogression with inflammation of uninjected as well as the injected tumor, 5 patients had stable disease for approximately 2 months and 15 patients had progressive disease. 2 abscopal responses have been seen to date. The data also suggest a dose-response relationship, with higher and more frequent doses associated with better outcomes. Peripheral lymphocyte subset analysis and serum cytokine analysis are ongoing. Conclusions: AdAPT-001 TGF-ß Trap is well tolerated and no dose limiting toxicities were observed with the single or multidosing protocol described. Evidence of an antitumor effect was seen. The recommended Phase 2 dose was 1.0 x 1012 VPs administered by intratumoral injection once every 2 weeks. Combination of AdAPT-001TGF-ß Trap with a checkpoint inhibitor is planned. Clinical trial information: NCT04673942 .

CD19 (+) B Cell Combined with Prognostic Nutritional Index Predicts the Clinical Outcomes of Patients with Gastric Cancer Who Underwent Surgery.

(1) Background: The aim of this study was to explore the predictive ability of lymphocyte subsets for the prognosis of gastric cancer patients who underwent surgery and the prognostic value of CD19 (+) B cell combined with the Prognostic Nutritional Index (PNI). (2) Methods: This study involved 291 patients with gastric cancer who underwent surgery at our institution between January 2016 and December 2017. All patients had complete clinical data and peripheral lymphocyte subsets. Differences in clinical and pathological characteristics were examined using the Chi-square test or independent sample t-tests. The difference in survival was evaluated using Kaplan-Meier survival curves and the Log-rank test. Cox's regression analysis was performed to identify independent prognostic indicators, and nomograms were used to predict survival probabilities. (3) Results: Patients were categorized into three groups based on their CD19 (+) B cell and PNI levels, with 56 cases in group one, 190 cases in group two, and 45 cases in group three. Patients in group one had a shorter progression-free survival (PFS) (HR = 0.444, p < 0.001) and overall survival (OS) (HR = 0.435, p < 0.001). CD19 (+) B cell-PNI had the highest area under the curve (AUC) compared with other indicators, and it was also identified as an independent prognostic factor. Moreover, CD3 (+) T cell, CD3 (+) CD8 (+) T cell, and CD3 (+) CD16 (+) CD56 (+) NK T cell were all negatively correlated with the prognosis, while CD19 (+) B cell was positively associated with the prognosis. The C-index and 95% confidence interval (CI) of nomograms for PFS and OS were 0.772 (0.752-0.833) and 0.773 (0.752-0.835), respectively. (4) Conclusions: Lymphocyte subsets including CD3 (+) T cell, CD3 (+) CD8 (+) T cell, CD3 (+) CD16 (+) CD56 (+) NK T cell, and CD19 (+) B cell were related to the clinical outcomes of patients with gastric cancer who underwent surgery. Additionally, PNI combined with CD19 (+) B cell had higher prognostic value and could be used to identify patients with a high risk of metastasis and recurrence after surgery.

The profile and prognostic value of circulating lymphocyte subsets in metastatic colon cancer.

Colon cancer (CC) are the most common malignant cancer in human digestive system, however, the profile and prognostic value of circulating lymphocyte subsets in CC patients has not been systemically clarified. In this study, 158 patients with metastatic CC were enrolled. Chi-square test was used to analyze the relationship between baseline peripheral blood lymphocyte subsets and clinicopathological parameters. Kaplan-Meier and Log-rank tests were used to analyze the relationship between clinicopathological parameters and baseline peripheral lymphocyte subsets and overall survival (OS) of patients with metastatic CC. Univariate/multivariate COX regression analysis was used to identify the independent factors in metastatic CC. The baseline peripheral blood CD3+T cells, CD4+T cells, NK cells and B cells of BRAF mutant patients were significantly lower than those in BRAF wild-type patients; The baseline CD8+T cells of KRAS mutation group was lower than that in KRAS wild type group. Peripheral blood CA19-9>27, left-sided colon cancer (LCC), KRAS and BRAF mutation were poor prognostic factors, and ALB>40, NK cells were protective prognostic factors for metastatic CC. In patients with liver metastases subgroup, higher NK cells also indicated a longer OS. Finally, LCC (HR=0.56), CA19-9 (HR=2.13), ALB (HR=0.46) and circulating NK cells (HR=0.55) were independent prognostic factors for metastatic CC. LCC, higher level of ALB and NK cells at baseline are protective factors, and higher CA19-9, KRAS/BRAF gene mutation are adverse prognostic factors. Sufficient circulating NK cells are independent prognostic factor for metastatic CC patients.

Predictive value of peripheral lymphocyte subsets for the disease progression in patients with sepsis.

To investigate the predictive value of peripheral lymphocyte subsets for sepsis progression. Patients with sepsis were divided into the improved group (n=46) and severe group (n=39) according to disease progression. Flow cytometric analysis was performed to enumerate absolute counts of peripheral lymphocyte subsets. Logistic regression analyses were conducted to identify clinical factors linked to sepsis progression. The absolute counts of peripheral lymphocyte subsets were markedly decreased in septic patients compared with healthy controls. After treatment, the absolute counts of lymphocytes, CD3+ T cells, and CD8+ T cells were restored in the improved group, and reduced in the severe group. Logistic regression analysis indicated that a low CD8+ T cells count was a risk factor for sepsis progression. Receiver operating characteristic curve analysis revealed that CD8+ T cells count had the greatest ability to predict sepsis progression. The absolute counts of CD3+ T cells, CD4+ T cells, CD8+ T cells, B cells, and natural killer cells were significantly higher in the improved group than the severe group. CD8+ T cells count was predictive of sepsis progression. Lymphopenia and CD8+ T cells depletion were associated with the clinical outcomes of sepsis, suggesting that CD8+ T cells have potential as a predictive biomarker and therapeutic target for patients with sepsis.

Short-term prognostic role of peripheral lymphocyte subsets in patients with gastric cancer.

Immune function is associated with clinical outcomes in patients with gastric cancer. This study aimed to explore the prognostic role of peripheral lymphocyte subsets in patients with gastric cancer after curative surgery. This retrospective study was conducted at a single tertiary referral hospital. We included patients diagnosed with gastric cancer who had undergone surgery and met the inclusion criteria. Clinicopathological characteristics and preoperative peripheral lymphocyte subset data were collected for the analysis. Recurrence-free survival (RFS) and overall survival were analyzed using the Kaplan-Meier curve and Cox hazard regression model. We used the Whitney test and Spearman test to analyze the correlation between lymphocyte subsets and clinicopathological characteristics. This study included 171 patients with gastric cancer who underwent curative surgery. Multivariate analysis revealed that carcinoembryonic antigen (p<0.01), carbohydrate antigen 19-9 (p<0.001), lymph node metastases (p<0.001), total T-cell count (p=0.02), B-cell count (p<0.01), and regulatory T-cell percentage (p<0.01) were independent predictive factors associated with RFS. Impaired immune function may lead to early recurrence following curative surgery. Our study showed that the characteristics of peripheral lymphocyte subsets (T, B, and Treg cells) were independent predictive factors for recurrence in patients with gastric cancer after surgery.

Kidney involvement of chronic active Epstein-Barr virus infection.

A 54-year-old Japanese woman presented with a fever of unknown origin and renal dysfunction. She had experienced intermittent fever for 10 years, but the frequency increased to a constant fever. Her renal function had gradually deteriorated over the past year, and serum creatinine and urinary α1-microglobulin levels were elevated to 2.23 mg/dL and 113 mg/L, respectively. Physical and laboratory examinations were unremarkable, except for mild anemia and elevated serum creatinine levels. Analysis of peripheral blood lymphocyte subsets showed 59% CD3, 35% CD4 and 31% CD8. Epstein-Barr virus (EBV) serology showed elevated levels of EBV capsid antigen IgG and EBV nuclear antigen (1:1280 and 1:40, respectively). Plasma EBV-DNA level was 5.63 log IU/mL, and an oligoclonal band was detected by Southern blot hybridization of EBV terminal repeat analysis. Quantification of EBV-DNA in peripheral lymphocyte subset using magnetic beads showed that EBV predominantly resided in CD4+ T-cells. (CD4, 258,768; CD8, 27,738; and CD56, 23,369; IU/µgDNA, respectively). T-cell receptor α gene rearrangement was undetected. Based on the above, she was diagnosed with chronic active EBV infection (CAEBV). 18F-fluorodeoxyglucose positron emission tomography/computerized tomography showed bilateral kidney enlargement and mildly enhanced uptake (maximum standardized uptake value 4.08 g/mL) (Figure 1 top left). Kidney biopsy specimen stained with hematoxylin and eosin showed diffuse tubulointerstitial infiltration of small to medium-sized lymphocytes in the renal parenchyma (Figure 1 top right). Massive infiltration of CD4+ T-cells was documented by immunohistochemical staining (Figure 1 bottom left). Furthermore, EBV-infected cells were detected by in situ hybridization with EBV-encoded RNA (EBER) (Figure 1 bottom right). Immunostaining of EBV nuclear antigen-2 and latent membrane protein 1 were negative. EBER-positive cells were distributed in the same region as CD4+ T-cells, consistent with the predominant presence of EBV-DNA in CD4+ T-cells. Her renal function had improved after initiating chemotherapy, and she is scheduled for hematopoietic stem cell transplantation. The clinical features of EBV infection vary according to the type of EBV-infected cells. The predominant EBV-infected cells in EBV-induced hemophagocytic lymphohistiocytosis are CD8+ T cells, whereas in CAEBV, CD4+ T cells are predominantly infected. Although rare, renal invasion of EBV should be considered in cases with renal impairment, and its clinical significance needs to be investigated. AF and SS wrote and edited the manuscript, provided patient care and reviewed the literature. KS and MT provided patient care. TS and KM wrote and edited the manuscript. All authors reviewed and approved the final version of the manuscript We thank Dr. Yuka Torii (Department of Pediatrics, Nagoya University, Japan) for quantification of the EBV viral load of lymphocyte subsets using magnetic beads. We also thank Dr. Hiroshi Kimura (Department of Virology, Nagoya University, Japan) for his helpful advice on patient care. We appreciate the staff of the Department of Hematology, Kameda Medical Center for providing patient care. We thank Editage (www.editage.jp) for English language editing. The authors declare they have no conflicts of interest. The authors received no specific funding for this work. This study was conducted according to the Declaration of Helsinki, and written consent was obtained from the patient for this report. The patient provided written informed consent for the publication of this case, with the removal of all identifying information to ensure anonymity and maintain privacy. The data that support the findings of this study are available from the corresponding author upon reasonable request. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

The alterations in peripheral lymphocyte subsets predict the efficacy and prognosis of immune checkpoint inhibitors in hepatocellular carcinoma.

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.

Effect of hyperglycemia on the immune function of COVID-19 patients with type 2 diabetes mellitus: a retrospective study.

To analyze the clinical characteristics and immune function parameters and to explore the effect of hyperglycemia on the immune function in patients with Corona Virus Disease 2019 (COVID-19) with type 2 diabetes mellitus (T2DM). This retrospective study included patients with COVID-19 with T2DM hospitalized in Renmin Hospital of Wuhan University between January 31, 2020, and February 10, 2020. The clinical data were collected and patients were divided into a well-controlled group (blood glucose 3.9-10.0 mmol/L) and a poorly-controlled group (blood glucose >10.0 mmol/L). The differences in routine blood tests, peripheral lymphocyte subsets, humoral immune components, C-reactive protein (CRP) level, and cytokines were compared, and the correlation between blood glucose and immune parameters as well as the severity of the disease was analyzed. A total of 65 patients with COVID-19 and T2DM were included in the final analysis. Compared with the well-controlled group, patients in the poorly-controlled group had decreased lymphocytes, CD16+ 56+ NK cells, CD3+ T cells, CD8+ T cells and increased neutrophil percentage, IL-6 levels, CRP levels and serum concentration of IgA. Blood glucose was inversely correlated with CD16+ 56+ NK cells, CD3+ T cells, CD4+ T cells, and CD8+ T cells and positively correlated with IL-6 and CRP levels. There was a positive correlation between blood glucose and the severity of the COVID-19. Hyperglycemia will aggravate the immune dysfunction of COVID-19 patients with T2DM and affect the severity of COVID-19.

Change of circulating lymphocyte subsets is related to disease activity and secondary infection in children with primary nephrotic syndrome-a retrospective study.

Primary nephrotic syndrome (PNS) is an immune-mediated glomerular disease that often reoccurs. However, the characteristics of circulating lymphocyte subsets in PNS children remain unclear. Immunosuppressive therapy can lead to temporary or persistent remissions, but also increases the risk of infection, and whether the circulating lymphocyte subsets can be used to predict the secondary infection also remains unclear. Here, we explored the distribution of lymphocyte subpopulations in the different stages of PNS, and its predictive value of secondary infection in pediatric patients. We included 89 children who were first PNS episodes or diagnosed with PNS admitted to Nanfang Hospital from September 2019 to April 2021, and 19 healthy children were recruited as controls (C). PNS patients were divided into three groups according to their serum biochemical tests: active group (A), partial remission (PR) group, and complete remission (CR) group. PNS patients with infection symptoms were divided into a co-infection group, others were divided into the non-infection group. The peripheral lymphocyte subsets were analyzed by flow cytometry. The relationship between the peripheral lymphocyte subsets and PNS activity or infection was analyzed. Compared to the healthy controls, the PNS patients' CD8+CD28+ T cell (TC) (C: 16.6%, 450.8/µL; A: 29.1%, P=0.000, 886.1/µL, P=0.012; PR: 25.7%, P=0.000, 817.3/µL, P=0.012; CR: 24.9%, P=0.001, 747.9/µL, P=0.020), and CD4+CD45RO+ ("memory" helper) T cells (C: 13.2%, 358.9/µL; A: 15.7%, P=0.036, 578.7/µL, P=0.001; PR: 17.6%, P=0.002, 610.0/µL, P=0.000; CR: 13.7%, P=0.676, 398.1/µL, P=0.525) were elevated. In addition, the regulatory T cells counts (non-infection: 117.9/µL; Co-infection: 73.3/µL, P=0.001) were significantly lower in patients with infection. We found that the predictive value measured by the area under the curve (AUC) showed that the AUC (t) Treg cell counts (61.5-84.5%) were almost always higher than the AUC for the (t) CD4+ T cell counts (55.1-77.1%). In this study, we found that T cell subpopulations had different characteristics in PNS during different disease phases. The CD8+CD28+ T cells, and CD4+CD45RO+ T cells increased at the disease quiescence of PNS. Moreover, CD4+ T cell subsets (regulatory T cell <82.5/µL) had higher predictive value than CD4+ T cell counts for PNS infection.

Absolute decrease in regulatory T cells and low-dose interleukin-2 therapy: restoring and expanding regulatory T cells to treat systemic sclerosis: a 24-week study.

Systemic sclerosis (SSc) is an autoimmune disease characterized by vascular lesions, immunological alterations and tissue fibrosis. There is some evidence of an imbalance between T-cell subsets in this disease. Interleukin (IL)-2 is a cytokine that can regulate the activity of immune cells and there is evidence that low-dose IL-2 therapy can be used to treat immune diseases. To investigate the changes of peripheral lymphocyte subsets, especially T helper (Th)17 and regulatory T (Treg) cells and the effects of low-dose IL-2 therapy in patients with SSc. In total, 66 patients with SSc and 49 sex- and age-matched healthy controls (HCs), were enrolled. The absolute numbers of peripheral lymphocyte subsets in these individuals were determined by flow cytometry. The 66 patients, were divided into 2 groups: 23 (the IL-2 group) were treated with low-dose (5.0 × 105 IU) IL-2 by subcutaneous injection daily for 5 days combined with conventional therapy, while the remaining 23 patients received conventional therapy only. Compared with HCs, the absolute numbers of peripheral T, CD4+ T, CD8+ T, natural killer and Treg cells were significantly lower in patients with SSc, with the most dramatic difference seen in both the absolute number and percentage of Treg cells in these patients, including new (previously untreated) cases, resulting in an imbalance (elevated ratio) between Th17 and Treg cells. At Week 24 after commencement of IL-2 treatment, Treg cells were markedly increased and tended to restore the balance of Th17 to Treg cells compared with baseline. Erythrocyte sedimentation rate, C-reactive protein, modified Rodnan Skin Score and visual analogue scale score were significantly decreased in both the IL-2 and non-IL-2 groups, indicating disease improvement. Notably, compared with those in the non-IL-2 group, patients treated with IL-2 had greater improvement. Our study showed that the absolute numbers of peripheral Treg cells together with total T, CD4+ T, CD8+ T and NK is significantly decreased, leading to an imbalance of Th17 to Treg cells in patients with SSc, and that low-dose IL-2 treatment could restore the balance of the two immune cells and reduce disease activity without obvious adverse effects.

Characteristics of lymphocyte subset alterations in COVID-19 patients with different levels of disease severity

BackgroundCoronavirus disease 2019 (COVID-19) is a respiratory disorder caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which had rapidly spread all over the world and caused public health emergencies in the past two years. Although the diagnosis and treatment for COVID-19 have been well defined, the immune cell characteristics and the key lymphocytes subset alterations in COVID-19 patients have not been thoroughly investigated.MethodsThe levels of immune cells including T cells, B cells, and natural killer (NK) cells in 548 hospitalized COVID-19 patients, and 30 types of lymphocyte subsets in 125 hospitalized COVID-19 patients admitted to Wuhan Huoshenshan Hospital of China were measured using flow cytometry. The relationship between lymphocytes subsets with the cytokine interleukin-6 (IL-6) and the characteristics of lymphocyte subsets in single-cell RNA sequencing (scRNA-seq) data obtained from peripheral blood mononuclear cells (PBMCs) were also analysed in COVID-19 patients.ResultsIn this study, we found that patients with critical COVID-19 infection exhibited an overall decline in lymphocytes including CD4+ T cells, CD8+ T cells, total T cells, B cells, and NK cells compared to mild and severe patients. However, the number of lymphocyte subsets, such as CD21low CD38low B cells, effector T4 cells, and PD1+ depleted T8 cells, was moderately increased in critical COVID-19 patients compared to mild cases. Notably, except for effector memory T4 cells, plasma blasts and Tregs, the number of all lymphocyte subsets was markedly decreased in COVID-19 patients with IL-6 levels over 30-fold higher than those in healthy cases. Moreover, scRNA-seq data showed obvious differences in the distribution and numbers of lymphocyte subsets between COVID-19 patients and healthy persons, and subsets-specific marker genes of lymphocyte subsets including CD4, CD19, CCR7, and IL7R, were markedly decreased in COVID-19 patients compared with those in healthy cases.ConclusionA comprehensive decrease in immune cell and lymphocyte subsets in critical COVID-19 patients, and peripheral lymphocyte subset alterations showed a clear association with clinical characteristics.

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.

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.

Alterations in bile acids as metabolic signatures in the patients with human adenovirus type 7 infection.

ObjectivesThe changes in metabolism by human adenovirus (HAdV) infection was unclear. The potential mechanism of HAdV-7 causing acute respiratory tract infection was explored.MethodsTotally 35 patients with HAdV-7 infection, 32 asymptomatic cases with HAdV-7 and 14 healthy controls were enrolled from an outbreak of HAdV-7 in the army. The serum samples were analyzed by untargeted and targeted metabolomics. The effects of differential metabolites were verified on HAdV-7 replication in an A549 cell line.ResultsThe untargeted metabolomics analysis revealed more significant changes in the classes of sphingolipids, polyketides, glycerolipids, fatty acyls, and carboxylic acids and their derivatives in the patients with HAdV-7 than in healthy controls. Two key metabolic pathways of secondary and primary bile acid biosynthesis were noted from pathway enrichment analysis. Targeted metabolomics analysis showed that the levels of unconjugated bile acids in the patients were significantly lower, while the levels of glyco- and tauro- conjugated bile acids in patients and asymptomatic cases were higher than those in the healthy controls. The profiles of cytokines and peripheral lymphocyte subsets obviously varied at different levels of bile acids, with significant differences after HAdV-7 infection. A cell verification test demonstrated that the replication of HAdV-7 significantly reduced when GCDCA and TCA were added.ConclusionBile acids inhibited HAdV-7 replication in vitro. Alterations in bile acids was metabolic signatures of HAdV-7 infected subjects, and our results suggested bile acids might play protective roles against HAdV-7 infection.

The Predictive Value of Changes in the Absolute Counts of Peripheral Lymphocyte Subsets for Progression and Prognosis in Breast Cancer Patients.

Background As the number and proportion of lymphocyte subsets are an important indicator of the immune function, an in depth understanding of the immune function of patients with malignant tumor has important clinical values for the treatment, prognosis, and evaluation of the disease. This retrospective study was to evaluate the clinical value of the absolute counts of lymphocyte subsets as potential blood biomarkers for progression and prognosis in breast cancer patients. Methods A total of 237 BC patients and 55 age-matched female normal healthy donors were included in this study. Flow cytometry was used to determine the absolute counts and the percentages of CD3+, CD4+, CD8+, B, and NK cells. The receiver operating characteristic curve (ROC) was used to evaluate the accuracy of absolute count of lymphocyte subsets in the curative efficacy assessment. The clinicopathological parameters influencing the disease progression were determined by Cox proportional hazards regression. Progression-free survival (PFS) was estimated using the Kaplan–Meier method with the log-rank test. Results: Compared with the healthy donors, the absolute counts of lymphocyte subsets in patients decreased significantly. ROC analysis showed that the area under the curve of the CD4+ absolute count was 90% (95% confidence interval 0.859–0.940), and the sensitivity and specificity were 80.9% and 85.3%, respectively. The analysis of Cox regression showed that the cutoff value of the CD4+ absolute count ≥451 cells/μL might be a favorable prognostic factor. Multivariate analysis of prognostic factors of PFS showed that the CD4+ and CD8+ absolute count were independent factors for predicting PFS. Conclusions The remarkably impaired absolute counts of the CD3+, CD4+, CD8+, B, and NK cells in patients with breast cancer can be used as potential susceptible biomarkers to evaluate the patient's immune status. The higher level of CD4+ and CD8+ absolute counts probably contributed to the longer PFS and favorable outcome of BC patients.

Evaluation of the immune feature of ACPA-negative rheumatoid arthritis and the clinical value of matrix metalloproteinase-3.

Anti-citrullinated protein antibodies (ACPAs) are highly specific for the diagnosis of rheumatoid arthritis (RA). However, about one-third of RA patients are negative for ACPAs, which presents a challenge to the early diagnosis of RA. The purpose of this study was to analyze differences in lymphocyte subsets and CD4+ T cell subsets between ACPA+ and ACPA- RA patients, and to evaluate the value of matrix metalloproteinase-3 (MMP-3) as a diagnostic and monitoring marker in ACA- RA patients. A total of 145 ACPA+ RA patients, 145 ACPA- RA patients, and 38 healthy controls (HCs) were included in this study. Peripheral lymphocyte subsets were detected using flow cytometry, and serum MMP-3 was detected using chemiluminescence. Information about joint symptoms, other organ involvement, and related inflammatory markers was also collected. The results showed that, compared to ACPA- RA patients, ACPA+ cases had greater imbalances between peripheral CD4+ T cell subsets, mainly manifested as an increase in T-helper 1 (Th1) cells (p < 0.001) and decrease in regulatory T (Treg) cells (p = 0.029). This makes these patients more prone to inflammatory reactions and joint erosion. MMP-3 levels in ACPA+ and ACPA- RA patients were significantly higher than in HCs (p < 0.001), and MMP-3 could effectively distinguish between ACPA- RA patients and HCs (area under the curve [AUC] = 0.930, sensitivity 84.14%, specificity 92.11%). MMP-3 was also a serum marker for distinguishing between RA patients with low and high disease activities. Further analysis showed that MMP-3 was positively correlated with the levels of inflammatory markers and disease activity, and negatively correlated with the levels of lymphocyte subsets. In addition, with improvements in the disease, MMP-3 levels decreased, and further increased as the patients started to deteriorate. In summary, our research showed that there was a mild imbalance between peripheral CD4+ T cell subsets in ACPA- RA patients. MMP-3 may be used as a potential marker for early diagnosis of ACPA- RA. MMP-3 was an important index for RA disease evaluation, disease activity stratification, and prognosis.

Immune cell subset profiling in multiple sclerosis after fingolimod initiation and continued treatment: The FLUENT study.

BackgroundFingolimod is a sphingosine 1-phosphate receptor modulator approved for relapsing MS. Long-term effects on the immunological profile are not fully understood.ObjectiveInvestigate fingolimod's temporal effects on immune cell subsets, and safety outcomes.MethodsIn FLUENT, a 12-month, prospective, non-randomized, open-label, phase IV study, adult participants received fingolimod 0.5 mg/day. Changes in immune cell subsets, anti-John Cunningham virus (JCV) antibody index, and serum neurofilament levels were assessed.Results165 fingolimod-naive and 217 participants treated for 2–12 years in routine clinical practice were enrolled. Levels of all monitored peripheral lymphocyte subsets were reduced from month 3 in fingolimod-naive participants. Greatest reductions occurred in naive and central memory CD4+ and CD8+ T cells, and in naive and memory B cells. Most lymphocyte subset levels remained stable in the continuous fingolimod group. Components of the innate immune system remained within reference ranges. No increase in JCV seropositivity was observed. No single cellular subset correlated with anti-JCV antibody index at any time point. Neurofilament levels remained within healthy adult reference limits throughout. No opportunistic infections were reported; no new or unexpected safety signals were observed.ConclusionFLUENT provides insights into the utility of immunological profiling to evaluate therapy response and potential infection risk.

Association of Peripheral Lymphocyte Subsets with Cognitive Decline and Dementia: The Cardiovascular Health Study

Association of Peripheral Lymphocyte Subsets with Cognitive Decline and Dementia: The Cardiovascular Health Study