OBJECTIVE: To analyze the Epstein-Barr virus (EBV) load in different lymphocyte subsets, as well as clinical characteristics and outcomes in patients with hematologic malignancies experiencing EBV reactivation. METHODS: Peripheral blood samples from patients were collected. B, T, and NK cells were isolated sorting with magnetic beads by flow cytometry. The EBV load in each subset was quantitated by real-time quantitative polymerase chain reaction (RT-qPCR). Clinical data were colleted from electronic medical records. Survival status was followed up through outpatient visits and telephone calls. Statistical analyses were performed using SPSS 25.0. RESULTS: A total of 39 patients with hematologic malignancies were included, among whom 35 patients had undergone allogeneic hematopoietic stem cell transplantation (allo-HSCT). The median time to EBV reactivation was 4.8 months (range: 1.7-57.1 months) after allo-HSCT. EBV was detected in B, T, and NK cells in 20 patients, in B and T cells in 11 patients, and only in B cells in 4 patients. In the 35 patients, the median EBV load in B cells was 2.19×10⁴ copies/ml, significantly higher than that in T cells (4.00×10³ copies/ml, P <0.01) and NK cells (2.85×10² copies/ml, P <0.01). Rituximab (RTX) was administered for 32 patients, resulting in EBV negativity in 32 patients with a median time of 8 days (range: 2-39 days). Post-treatment analysis of 13 patients showed EBV were all negative in B, T, and NK cells. In the four non-transplant patients, the median time to EBV reactivation was 35 days (range: 1-328 days) after diagnosis of the primary disease. EBV was detected in one or two subsets of B, T, or NK cells, but not simultaneously in all three subsets. These patients received a combination chemotherapy targeting at the primary disease, with 3 patients achieving EBV negativity, and the median time to be negative was 40 days (range: 13-75 days). CONCLUSION In hematologic malignancy patients after allo-HSCT, EBV reactivation commonly involves B, T, and NK cells, with a significantly higher viral load in B cells compared to T and NK cells. Rituximab is effective for EBV clearance. In non-transplant patients, EBV reactivation is restricted to one or two lymphocyte subsets, and clearance is slower, highlighting the need for prompt anti-tumor therapy.
Humans ; Hematologic Neoplasms/virology* ; Herpesvirus 4, Human/physiology* ; Epstein-Barr Virus Infections ; Hematopoietic Stem Cell Transplantation ; Virus Activation ; Lymphocyte Subsets/virology* ; Flow Cytometry ; Killer Cells, Natural/virology* ; Male ; Female ; B-Lymphocytes/virology* ; Viral Load ; Adult ; T-Lymphocytes/virology* ; Middle Aged

Hepatitis B is a global public health concern. Inducing hepatitis B surface antibody (HBsAb) through vaccination is a crucial preventive strategy. However, individuals show varying immune responses to the hepatitis B vaccine. Based on HBsAb levels, individuals can be categorized as high responders, low responders, or non-responders. T cells and their subsets play critical roles in modulating this response, and the composition of the T cell receptor (TCR) repertoire also influences immune responsiveness. Investigating the characteristics of T cells, their subsets, and TCR repertoires in individuals with differential responses post-vaccination may provide theoretical guidance for optimizing vaccine design and immunization strategies.
Humans ; Hepatitis B Vaccines/immunology* ; Hepatitis B/immunology* ; Vaccination ; Hepatitis B Antibodies/blood* ; T-Lymphocytes/immunology* ; Receptors, Antigen, T-Cell/immunology* ; Hepatitis B Surface Antigens/immunology* ; T-Lymphocyte Subsets/immunology*

OBJECTIVE: To investigate the distribution of bone marrow lymphocyte subsets in patients with myelodysplastic syndrome(MDS),the proportion of activated T cells with immunophenotype CD3⁺HLA-DR⁺ in the lymphocytes and its clinical significance, and to understand the effects of different types of MDS, different immunophenotypes, and different expression levels of WT1 on the proportion of lymphocyte subsets and activated T cells. METHODS: The immunophenotypes of 96 MDS patients, the subsets of bone marrow lymphocytes and activated T cells were detected by flow cytometry. The relative expression of WT1 was detected by real-time fluorescent quantitative PCR, and the first induced remission rate (CR1) was calculated, the differences of lymphocyte subsets and activated T cells in MDS patients with different immunophenotype, different WT1 expression, and different course of disease were analyzed. RESULTS: The percentage of CD4⁺T lymphocyte in MDS-EB-2, IPSS high-risk, CD34⁺ cells >10%, and patients with CD34⁺CD7⁺ cell population and WT1 gene overexpression at intial diagnosis decreased significantly (P<0.05), and the percentage of NK cells and activated T cells increased significantly (P<0.05), but there was no significant difference in the ratio of B lymphocytes. Compared with the normal control group, the percentage of NK cells and activated T cells in IPSS-intermediate-2 group was significantly higher(P<0.05), but there was no significant difference in the percentage of CD3⁺T, CD4⁺T lymphocytes. The percentage of CD4⁺T cells in patients with complete remission after the first chemotherapy was significantly higher than in patients with incomplete remission(P<0.05), and the percentage of NK cells and activated T cells was significantly lower than that in patients with incomplete remission (P<0.05). CONCLUSION In MDS patients, the proportion of CD3⁺T and CD4⁺T lymphocytes decreased, and the proportion of activated T cells increased, indicating that the differentiation type of MDS is more primitive and the prognosis is worse.
Humans ; Lymphocyte Subsets ; Myelodysplastic Syndromes/diagnosis* ; Bone Marrow ; B-Lymphocytes ; Killer Cells, Natural ; Flow Cytometry ; T-Lymphocyte Subsets

OBJECTIVES: To investigate the change in the distribution of memory B cell subsets in children with frequently relapsing nephrotic syndrome (FRNS) during the course of the disease. METHODS: A total of 35 children with primary nephrotic syndrome (PNS) who attended the Department of Pediatrics of the Affiliated Hospital of Xuzhou Medical University from October 2020 to October 2021 were enrolled as subjects in this prospective study. According to the response to glucocorticoid (GC) therapy and frequency of recurrence, the children were divided into two groups: FRNS (n=20) and non-FRNS (NFRNS; n=15). Fifteen children who underwent physical examination were enrolled as the control group. The change in memory B cells after GC therapy was compared between groups, and its correlation with clinical indicators was analyzed. RESULTS: Before treatment, the FRNS and NFRNS groups had significantly increased percentages of total B cells, total memory B cells, IgD⁺ memory B cells, and IgE⁺ memory B cells compared with the control group, and the FRNS group had significantly greater increases than the NFRNS group (P<0.05); the FRNS group had a significantly lower percentage of class-switched memory B cells than the NFRNS and control groups (P<0.05). After treatment, the FRNS and NFRNS groups had significant reductions in the percentages of total B cells, total memory B cells, IgM⁺IgD⁺ memory B cells, IgM⁺ memory B cells, IgE⁺ memory B cells, IgD⁺ memory B cells, and IgG⁺ memory B cells (P<0.05) and a significant increase in the percentage of class-switched memory B cells (P<0.05). The FRNS group had a significantly higher urinary protein quantification than the NFRNS and control groups (P<0.05) and a significantly lower level of albumin than the control group (P<0.05). In the FRNS group, urinary protein quantification was negatively correlated with the percentage of class-switched memory B cells and was positively correlated with the percentage of IgE⁺ memory B cells (P<0.05). CONCLUSIONS Abnormal distribution of memory B cell subsets may be observed in children with FRNS, and the percentages of IgE⁺ memory B cells and class-switched memory B cells can be used as positive and negative correlation factors for predicting recurrence after GC therapy in these children.
Child ; Humans ; B-Lymphocyte Subsets/metabolism* ; Immunoglobulin E ; Immunoglobulin M ; Nephrotic Syndrome/immunology* ; Prospective Studies ; Glucocorticoids/therapeutic use*

Objective:b> T lymphocyte exhaustion is an important component of immune dysfunction. Therefore, exploring peripheral blood-exhausted T lymphocyte features in patients with hepatitis B virus-related acute-on-chronic liver failure may provide potential therapeutic target molecules for ACLF immune dysfunction. Methods:b> Six cases with HBV-ACLF and three healthy controls were selected for T-cell heterogeneity detection using the single-cell RNA sequencing method. In addition, exhausted T lymphocyte subpopulations were screened to analyze their gene expression features, and their developmental trajectories quasi-timing. An independent sample t-test was used to compare the samples between the two groups. Results:b> Peripheral blood T lymphocytes in HBV-ACLF patients had different differentiation trajectories with different features distinct into eight subpopulations. Among them, the CD4(+)TIGIT(+) subsets (P = 0.007) and CD8(+)LAG3(+) (P = 0.010) subsets with highly exhausted genes were significantly higher than those in healthy controls. Quasi-time analysis showed that CD4(+)TIGIT(+) and CD8(+)LAG3(+) subsets appeared in the late stage of T lymphocyte differentiation, suggesting the transition of T lymphocyte from naïve-effector-exhausted during ACLF pathogenesis. Conclusion:b> There is heterogeneity in peripheral blood T lymphocyte differentiation in patients with HBV-ACLF, and the number of exhausted T cells featured by CD4(+)TIGIT(+)T cell and CD8(+)LAG3(+) T cell subsets increases significantly, suggesting that T lymphocyte immune exhaustion is involved in the immune dysfunction of HBV-ACLF, thereby identifying potential effective target molecules for improving ACLF patients' immune function.
Humans ; Hepatitis B virus ; Acute-On-Chronic Liver Failure/pathology* ; Hepatitis B, Chronic ; T-Lymphocyte Subsets/pathology* ; Receptors, Immunologic

OBJECTIVE: To explore the expression of CD40/CD40L in multiple myeloma(MM) patients and its influence on prognosis. METHODS: Thirty patients with MM treated in Cangzhou People's Hospital from May 2016 to June 2017 were selected and divided into MM group, then 30 healthy people with a physical examination in our hospital at the same time were selected as the normal group. The serum CD40/CD40L levels of the patients in the two groups was detected by flow cytometry, and its correlation with the lymphocyte population, pathological grade and prognostic significance of MM patients was anaysis. RESULTS: The expression of CD40 in serum of the patients in MM group was significantly higher than those in normal group (P<0.05). The expression of CD40L in serum of the patients in MM group showed no significant difference as compared with those in normal group (P>0.05). The levels of CD40 and CD40L in the patients before and after chemotherapy showed no difference(P>0.05). The levels of Ts and NK cells in the patients of MM group were lower than those in normal group (P<0.05). The proportion of total B lymphocytes, Th and Th/Ts cells between the two groups showed no significant difference (P>0.05). The CD40 level was correlated with the serum total B lymphocyte level of the patients in MM group (r=0.877, P=0.005). There was a correlation with CD40L and Th cells in the serum of MM patients (r=-0.783, P=0.035). The expression of serum CD40 in the patients at phase III-IV was higher than those of the patients at phase I-II, the levels of serum CD40L in MM patients at different periods showed no significant difference(P>0.05). The survival rate of MM patients with high CD40 expression was lower than that of MM patients with low CD40 expression (χ CONCLUSION The increasing of CD40 level in MM patients is related to the pathological grade of the patients. Chemotherapy can reduce the level of CD40. The increasing of CD40 is an important factor for the poor prognosis of MM patients. CD40L level is not meaningful for MM treatment and prognosis.
B-Lymphocytes ; CD40 Antigens ; CD40 Ligand ; Humans ; Lymphocyte Subsets ; Prognosis

OBJECTIVE: To explore the significance of lymphocytes in systemic sclerosis (SSc), by detecting the levels of T lymphocytes, B lymphocytes and natural killer (NK) cells, and analyzing the correlation between the lymphocytes and clinical laboratory indexes. METHODS: The numbers and proportion of T, CD4⁺T, CD8⁺T, B, and NK cells were detected by flow cytometry in peripheral blood of 32 SSc patients who had taken immunosuppressive drugs and 30 healthy controls (HC). The comparison of the lymphocyte subsets in SSc with them in the HC groups, and the correlation between the lymphocytes and other clinical and laboratory indicators were analyzed by the relevant statistical analysis. RESULTS: Compared with the HC group, the numbers of T, CD4⁺T, CD8⁺T, and NK cells in peripheral blood of SSc group, who had taken immunosuppressive drugs, were significantly decreased (P < 0.05). More-over, the proportion of NK cells in peripheral blood of the SSc group was also significantly lower than that in the HC group (P=0.004). In addition, all the lymphocyte subsets were decreased in peripheral blood of more than 65% of the SSc patients who had taken immunosuppressive drugs. Compared with CD4⁺T normal group, the positivity of Raynaud's phenomenon, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) was significantly increased in CD4⁺T reduction group, respectively (P＝0.024, P < 0.001, P=0.018). ESR was higher in CD8⁺T reduction group than CD8⁺T normal group (P＝0.022). The prevalence of fingertip ulcer was significantly increased in B cell decrease group (P=0.019). Compared with NK cell normal group, the prevalence of fingertip ulcer was significantly increased in NK cell lower group (P=0.033), IgM was remarkablely decreased yet (P=0.049). The correlation analysis showed that ESR was negatively correlated with the counts of T lymphocytes (r＝－0.455, P＝0.009), CD4⁺T lymphocytes (r＝－0.416, P＝0.018), CD8⁺T lymphocytes (r＝－0.430, P=0.014), B cells (r＝－0.366, P＝0.039). CONCLUSION The number of CD4⁺T, CD8⁺T, B, and NK cells significantly decreased in peripheral blood of SSc patients who had used immunosuppressive drugs, some lymphocyte subsets might be related with Raynaud's phenomenon and fingertip ulcer, and reflected the disease activity by negatively correlated with ESR and CRP; the numbers of lymphocyte subsets in peripheral blood should be detected regularly in SSc patients who had taken immunosuppressive drugs.
B-Lymphocytes ; Flow Cytometry ; Humans ; Killer Cells, Natural ; Lymphocyte Subsets ; Scleroderma, Systemic ; T-Lymphocyte Subsets ; T-Lymphocytes

lymphocyte ratio (NLR), and lymphocyte subsets have been reported to be associated with the clinical outcomes in colorectal cancer (CRC). However, studies on the clinical impact of each parameter have produced controversial results. Moreover, there is a paucity of comprehensive studies regarding these parameters in Korean CRC patients.METHODS: Sixty-eight CRC patients who underwent surgical resection were recruited for this study. NLR was measured using an automated blood cell counter. Flow cytometric analysis was performed to determine lymphocyte subsets and identify MDSCs during the diagnostic stage. Clinical and laboratory data were analyzed according to each blood parameter.RESULTS: The distribution of lymphocytes, MDSCs, and NLR were not associated with TNM stages. Large tumor sizes (P=0.042) and greater perineural invasion (P=0.031) were significantly associated with high CD19+ B-cell populations. Elevated granulocytic MDSCs (P=0.234), total MDSCs (P=0.234), and NLR (P=0.062) were associated with the poorly differentiated type of CRC, albeit without statistical significance. Additionally, patients in the high CD19+ B-cell group (P=0.012) revealed a moderately inferior relapse-free survival.CONCLUSIONS: Our findings indicate that preoperative evaluation of CD19+ B-cell proportion is recommended to predict the clinical outcomes of patients with stage II-III CRC.]]>
B-Lymphocytes ; Blood Cell Count ; Colorectal Neoplasms ; Humans ; Lymphocyte Subsets ; Lymphocytes

OBJECTIVE: To study the distribution of peripheral blood lymphocyte subsets in healthy children aged 0-6 years. METHODS: A total of 826 healthy Han children aged 0-6 years were recruited. According to their age, the children were divided into four groups: newborn, infant, toddler and preschool. Their peripheral blood samples were collected to measure the percentages of lymphocyte subsets by flow cytometry. RESULTS: There were significant differences in the percentages of CD3 T cells, CD3CD4 T cells and CD3CD19 B cells and the CD4/CD8 ratio between boys and girls (P<0.05). The girls had a lower percentage of CD3CD19 B cells, higher percentages of CD3 T cells and CD3CD4 T cells and a higher CD4/CD8 ratio than the boys. The newborn group had the highest percentages of CD3 T cells and CD3CD4 T cells and the highest CD4/CD8 ratio (P<0.05). The percentage of CD3CD4 T cells and the CD4/CD8 ratio gradually decreased with age and the preschool group had the lowest values (P<0.05). The newborn group had the lowest percentages of CD3CD19 B cells and CD3CD16CD56 NK cells (P<0.05). The percentage of CD3CD16CD56 NK cells gradually increased with age and the preschool group had the highest percentage (P<0.05). The percentage of CD3CD19 B cells reached the peak in the toddler period and then decreased with age (P<0.05). The preschool group had the highest percentage of CD3CD8 T cells (P<0.05). The variation trend of distribution of lymphocyte subsets in boys from different age groups was consistent with that in children from different age groups. For girls, the newborn group had the highest percentage of CD3CD4 T cells and CD4/CD8 ratio (P<0.05). CONCLUSIONS The distribution of peripheral blood lymphocyte subsets in healthy children is significantly different across ages and sexes. Therefore, the reference values should be established according to age and sex.
Antigens, CD19 ; B-Lymphocytes ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Infant, Newborn ; Killer Cells, Natural ; Lymphocyte Count ; Lymphocyte Subsets ; Male

OBJECTIVE: To investigate the changes of T lymphocyte subsets, B lymphocyte and NK cells in peripheral blood of patients with acute leukemia at different periods and their significance. METHODS: The peripheral T lymphocyte subsets and B lymphocyte of 95 patients with acute leukemia [(43 cases of acute lymphoblastic leukemia (ALL), 52 cases of acute myeloid leukemia (AML)] and 50 normal people were detected by flow cytometry respectively. RESULTS: The positive rate of CD3, CD3CD4, CD3CD8, NK cells and CD4/CD8 in the patients with newly diagnosed acute leukemia were significantly lower than those in normal controls (P<0.05) , but increased obviously after complete remission. The positive rate of Treg cells in the patients with newly diagnosed leukemia group was significantly higher than that in normal controls (P<0.01) , but decreased obviously after complete remission. Positive rate of CD3CD19 cells in the patients with newly diagnosed acute myeloid leukemia was significantly lower , but higher in the patients with newly diagnosed acute lymphoblastic leukemia than that in normal controls with statistical significant difference (P<0.05) , and increased obviously in the patients with acute myeloid leukemia (P<0.05) after complete remission , but decreased in the patients with acute lymphoblastic leukemia (P<0.01) after complete remission or no-remission. CONCLUSION Changes of T lymphocyte subsets, B lymphocytes and NK cells in the patients with newly diagnosed acute leukemia are significant, thus the detection of T lymphocyte cell subsets, B lymphocytes and NK cells can provide some evidences. for evaluation of the disease severity, curative efficiency and prognosis of patients with acute leukemia.
B-Lymphocytes ; Flow Cytometry ; Humans ; Killer Cells, Natural ; Leukemia, Myeloid, Acute ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; T-Lymphocyte Subsets