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Background Thyroid autoantibody is a marker for the diagnosis of Hashimoto's thyroiditis (HT), and B cells are essential in the pathogenesis of HT. Enhancer of Zeste homolog 2 (EZH2), which is an important epigenetic regulator, plays an important role in the regulation of lymphocytes development and function. Objective To investigate EZH2 expression in plasmoblasts and plasma cells in HT, and further explore the therapeutic effect of EZH2 inhibitors in experimental autoimmune thyroiditis (EAT) model. Methods The thyroid tissues from 6 patients who underwent thyroidectomy (3 HT patients with PTC, 3 patients with PTC alone) in Peking University First Hospital between 2010 and 2020 were obtained from the contralateral lobe with thyroid cancer, and screened for the expression of B-lymphocyte-related genes by RNA-seq; thyroid tissues from 16 HT patients and 8 normal thyroid tissues were collected and verified for the the expression of EZH2 in B cells in HT thyroid tissues by immunohistochemistry or immunofluorescence, respectively. Fine-needle aspiration (FNA) samples from patients with HT (n=25), and peripheral blood from patients with HT (n=19) or healthy donors (n=12) were analyzed by flow cytometry to define altered EZH2 expression in plasmablasts and plasma cells. Fifteen seven-week-old NOD.H-2h4 mice were randomly divided into the control (n=5), EAT without injection group (n=5), and EZH2 inhibitor+ GSK126 injection group (10 mg/kg, intraperitoneal injection 3 times /week, n=5). The degree of thyroid inflammation and changes in TgAb levels were observed after 8 weeks. Results RNA sequencing analysis showed that EZH2 and genes associated with the B-cell phenotype such as CD19, CD27, CD38, CD52 were higher expressed in HT hyroid tissues compared with normal thyroid tissues. Immunohistochemical results showed that immunohistochemical staining for EZH2 in 16 HT thyroid tissue specimens was strongly positive with positive cells observed in the GC region, and no positive cells were observed in the staining of 8 normal thyroid tissues. EZH2 staining in HT thyroid tissue was highly expressed in the GC region, and EZH2 was specifically expressed in CD19+ B cells. The results of flow cytometry assay showed that the proportion of CD19+ B cells, plasmablasts and plasma cells in HT FNA samples was higher than that of HD peripheral blood and HT peripheral blood samples (P

Objective To investigate the clinical significance of peripheral B-cell subsets with CD27 and CD38 phenotypes in myasthenia gravis (MG) patients. Methods Peripheral blood samples of 43 MG patients and 38 health controls (HC) from January 2016 to Decamber 2018 were collected in this study. Peripheral CD27+CD38- , CD27+CD38+ , CD27-CD38+ and CD27-CD38- B-cell subsets were detected through flow cytometer and tricolor fluorescence labeling. Results Percentages of CD27+CD38- and CD27+CD38+ B lymphocytes were increased but CD27-CD38+B-cell percentage was decreased in MG patients compared with those in HC (P = 0.000; P = 0.000; P = 0.001). Elevated percentages of CD27+CD38-B cells were found in MG patients compared with MG patients (P = 0.031). However, patients with thymoma demonstrated had lower CD27-CD38+B-cell percentages than patients with normal thymus (P = 0.026). CD27+CD38- and CD27+CD38+ B cells were enriched in patients with myasthenic crisis compared with patients without crisis (P = 0.017; P = 0.011). In addition, after the treatment of methylprednisolone, the Quantitative Myasthenia Gravis Score (QMGS) was decreased (P = 0.012), the CD27+CD38+ B cells reduced (P = 0.034) and CD27+CD38- B celsl elevated (P = 0.006). Meanwhile, percentages of CD27+CD38- B cells were significantly correlated with disease severities in MG patients (rs = 0.394, P = 0.009). Conclusions Abnormal changes of B-cell subsets with CD27 and CD38 phenotypes were observed and closely associated with disease severities in MG patients. These findings suggested that aberrancy of B-cell subsets may be involved in the progression of MG. DOI：10.3969/j.issn.1672⁃6731.2019.11.013

CD93 is expressed in progenitor B (pro-B) cells, precursor B (pre-B) cells and various immature B cells. It can interact with moesin, MMRN2 and other molecules to participate in cell migration, adhesion and phagocytosis, so it plays an important role in inflammation and angiogenesis. Detection of CD93 + B cell subsets has a crucial role in the diagnosis, treatment and prognosis monitoring of inflammation and inflammation-related diseases, such as Helicobacter pylori-related gastritis, sepsis, non-obese diabetes and periodontitis.

Objective To investigate the clinical significance of peripheral B cell subsets with CD27 and CD38 phenotypes in myasthenia gravis (MG) patients. Methods Peripheral blood samples of 43 MG patients and 38 healthy controls from January 2016 to December 2018 were collected. Peripheral CD27+ CD38-, CD27+ CD38+, CD27-CD38+ and CD27-CD38-B cells were detected through flow cytometer and tricolor fluorescence labeling. Results Percentages of CD27+ CD38- and CD27+ CD38+ B cells were increased (P = 0.000, 0.000), while percentage of CD27-CD38+ B cells was decreased in MG patients compared with those in controls (P = 0.001). The percentage of CD27+ CD38+ B cells was higher in generalized GMG patients than that in OMG patients (P = 0.031). However, patients with thymoma had lower percentage of CD27-CD38+B cells than that in patients with normal thymus (P = 0.026). CD27+ CD38- and CD27+ CD38+ B cells were enriched in patients with myasthenic crisis compared with patients without crisis (P = 0.017, 0.011). In addition, after treatment of methylprednisolone, the Quantitative Myasthenia Gravis Score (QMGS) was decreased (P = 0.012), the CD27+ CD38+ B cells reduced (P = 0.034) and CD27+ CD38-B cells elevated (P = 0.006). Meanwhile, percentage of CD27+ CD38-B cells was postively correlated with disease severity in MG patients (rs = 0.394, P = 0.009). Conclusions Abnormal changes of B cell subsets with CD27 and CD38 phenotypes were observed and closely associated with disease severity in MG patients. All of these findings suggested that aberrancy of B cell subsets may be involved in the progression of MG. [ABSTRACT FROM AUTHOR]

Objective: To detect the levels of serum C-C chemokine ligand 19 (CCL19) in patients with systemic lupus erythematosus (SLE) and to evaluate the correlation between CCL19 expression and clinical features and laboratory parameters, trying to reveal the possible role of CCL19 in the pathogenesis of systemic lupus erythematosus., Methods: The levels of serum CCL19 were measured by enzyme linked immunosorbent assay (ELISA) in 90 patients with SLE and 30 healthy controls. These SLE patients included 75 patients who received treatment with glucocorticoids and disease-modifying anti-rheumatic drug (DMARD) and 15 patients without therapy. The frequencies of peripheral blood B cells and the B cell subsets were assessed in the patients with SLE by flow cytometry. The correlation between the clinical data, laboratory parameters, B cell subset frequencies and serum CCL19 levels were analyzed. Indepen-dent samples t test, paired t test, Pearson and Spearman correlation were used for statistical analyses., Results: The levels of CCL19 were markedly higher in the SLE patients without therapy and the patients with therapy than in the health controls[(596.25±409.19) ng/L and (422.90±395.84) ng/L vs. (157.79±125.23) ng/L, all P<0.001]. Serum CCL19 levels in the SLE patients without therapy were higher than the SLE patients who accepted glucocorticoids and DMARD treatment (P<0.05). The levels of serum CCL19 were positively correlated with anti-double stranded deoxyribonucleic acid (dsDNA), anti-nucleosome antibody (AnuA), IgA, IgG and IgM (r=0.38, P=0.007; r=0.332, P=0.029; r=0.519, P=0.007; r=0.461, P=0.018, respectively). Serum CCL19 levels in the SLE patients with photosensitivity, arthritis and secondary Sjögren's syndrome were higher than the SLE patients without photosensitivity, arthritis and secondary Sjögren's syndrome, respectively [(562.25±399.12) ng/L, (565.6±435.24) ng/L and (694.9±531.02) ng/L vs. (394.7±281.42) ng/L, (385.90±325.33) ng/L and (424.8±305.46) ng/L, all P<0.05]. The levels of serum CCL19 were positively correlated with the percentage of CD27-B cells and CD27-IgD-double-negative memory B cells (r=0.519, P=0.007; r=0.461, P=0.018, respectively). However, the levels of serum CCL19 were negatively correlated with the percentage of CD27 + memory B cells and CD27 + IgD - switched memory B cells (r=-0.433, P=0.027; r=-0.616, P=0.001, respectively)., Conclusion: The increased serum CCL19 levels in SLE patients were associated with the production of autoantibodies, and CCL19 might be involved in the pathogenesis of SLE by disturbing the homeostasis of B cell subsets.

Objective: To investigate the serum level of C-C chemokine ligand 19 (CCL19) and its clinical significance in rheumatoid arthritis., Methods: The serum CCL19 levels in both rheumatoid arthritis (RA) patients and health controls were detected by ELISA. The proportion of peripheral blood B cells and memory B cell subsets were also detected in some patients. Then the clinical and laboratory data of the patients were collected. The CCL19 levels in patients with different clinical features were analyzed. And the correlation between the clinical data, laboratory parameters, B cell subsets proportion and serum CCL19 levels were also analyzed. Independent samples t test, paired t test, Pearson and Spearman correlation were used for statistical analysis., Results: The levels of CCL19 was higher in the RA patients than the health controls (P<0.05). The serum CCL19 levels were decreased in the RA patients who accepted disease-modifying anti-rheumatic drugs (DMARDs) treatment for 6 months (P<0.001). Serum CCL19 levels were correlated with the titers of both rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) antibody (r=0.42, P=0.002; r=0.33, P=0.013), but not with erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and disease activity score in 28 joints (DAS28) (P>0.05). The levels of CCL19 were higher in the serum positive (RF and anti-CCP antibody) patients, but there were no differences between low and high disease activity RA, as well as early and non-early RA. There was no correlation between the serum CCL19 levels and the proportion of B cells as well as memory B subsets. All the proportion of peripheral blood CD27+ memory B cell subsets in RA was lower than the healthy controls, including CD27+IgD+, CD27+IgD- and CD27+ B cells., Conclusion: The increased serum CCL19 levels in RA patients are associated with the activity of B cells, so CCL19 might predict whether the RA type is a B cell mediated RA, and specify the treatment directions for the rheumatologist.

Objective: To explore the clinical significance of immunocyte subsets before and after immunosuppressive therapy in the peripheral blood of patients with immune thrombocytopenic purpura (ITP)., Methods: The percentages of immunocyte subsets in the peripheral blood of 35 patients with ITP and 20 healthy controls were detected by flow cytometry, including CD(3)(+), CD(4)(+), CD(8)(+), CD(+)(56), CD(19)(+) lymphocytes and CD(4)(+)/CD(8)(+)., Results: The percentages of CD(3)(+) T lymphocyte (61.58 ± 6.45)%, CD(4)(+) T lymphocyte (28.38 ± 4.89)% and the ratio of CD(4)(+)/CD(8)(+) 0.99 ± 0.22 in patients with ITP were lower than those in healthy controls [(67.85 ± 4.68)%, (38.00 ± 3.37)%, 1.54 ± 0.13, all P < 0.05]. After immunosuppressive therapy, the percentages of CD(3)(+)T lymphocyte (69.41 ± 5.03)%, CD(4)(+)T lymphocyte (38.17 ± 3.18)% and the ratio of CD(4)(+)/CD(8)(+) 1.60 ± 0.15 recovered to control levels. The percentages of CD(8)(+)T lymphocyte (29.20 ± 4.50)% and CD(19)(+)B lymphocyte (17.74 ± 4.14)% were higher than those in healthy controls [(24.82 ± 2.93)% and (12.09 ± 3.51)%, all P < 0.05]. After the immunosuppressive therapy, the percentages of CD(8)(+)T lymphocyte (24.06 ± 3.02)% and CD(19)(+)B lymphocyte (10.90 ± 3.55)% recovered to control levels. There were no significant difference of the percentage of CD(56)(+) lymphocyte among ITP patients (15.80 ± 2.85)%, ITP patients after immunosuppressive therapy (15.16 ± 2.77)% and healthy controls (16.36 ± 2.75)%., Conclusion: The aberrant immunocyte subsets are involved in the pathogenesis of ITP, and detection of immunocyte subsets might be helpful for the diagnosis and determination of therapeutic outcome of ITP.

Objective: To explore the clinical effect of Tripterygiitotorum (T II) combined with prednisone in treating patients with myasthenia gravis (MG) and the changes of immune function after treatment., Methods: Sixty-eight patients with MG were randomly divided into two groups, the 36 patients in the treated group were treated with T II plus prednisone and the 32 in the control group were treated with prednisone alone. The therapeutic effect, serum interleukin-6 (IL-6) and peripheral B lymphocyte levels were observed., Results: The therapeutic effect in the treated group was significantly higher than that in the control group (P < 0.05). There were significant decrease in serum IL-6 and peripheral B lymphocyte in both groups after treatment (P < 0.05), with the decrements more significant in the treated group (P < 0.05)., Conclusion: The therapy of T II plus prednisone is superior to that of prednisone alone in treating MG, it has a more potent effect of immunosuppression.