Th17/Treg balance and macrophage polarization ratio in lower extremity arteriosclerosis obliterans

@#Objective: To explore the balance of peripheral blood T helper 17 cells/regulatory T cell (Th17/Treg) ratio and the polarization ratio of M1 and M2 macrophages in lower extremity arteriosclerosis obliterans (ASO). Methods: A rat model of lower extremity ASO was established, and blood samples from patients with lower extremity ASO before and after surgery were obtained. ELISA was used to detect interleukin 6 (IL-6), IL-10, and IL-17. Real-time RCR and Western blot analyses were used to detect Foxp3, IL-6, IL-10, and IL-17 expression. Moreover, flow cytometry was applied to detect the Th17/Treg ratio and M1/M2 ratio. Results: Compared with the control group, the iliac artery wall of ASO rats showed significant hyperplasia, and the concentrations of cholesterol and triglyceride were significantly increased (P<0.01), indicating the successful establishment of ASO. Moreover, the levels of IL-6 and IL-17 in ASO rats were pronouncedly increased (P<0.05), while the IL-10 level was significantly decreased (P<0.05). In addition to increased IL-6 and IL-17 levels, the mRNA and protein levels of Foxp3 and IL-10 in ASO rats were significantly decreased compared with the control group. The Th17/Treg and M1/M2 ratios in the ASO group were markedly increased (P<0.05). These alternations were also observed in ASO patients. After endovascular surgery (such as percutaneous transluminal angioplasty and arterial stenting), all these changes were significantly improved (P<0.05). Conclusions: The Th17/Treg and M1/M2 ratios were significantly increased in ASO, and surgery can effectively improve the balance of Th17/Treg, and reduce the ratio of M1/M2, and the expression of inflammatory factors.

Progress in the mechanism of regulatory B cells and its role in human diseases / 中华微生物学和免疫学杂志

Regulatory B cells (Bregs) are a group of cells with immunoregulatory function. They can regulate the progression of diseases by inhibiting excessive inflammatory responses and serve as critical protective cells in immune disorders and other conditions. This article reviewed the source, function and mechanism of Bregs and its role in autoimmune diseases, infection and tumor.

The therapeutic effect of budesonide on childhood asthma and its influence on growth and development 1 to 2 years after the treatment / 药学实践杂志

Objective To explore the effect of budesonide suspension for inhalation in the treatment of childhood asthma and its influence on growth and development in 1-2 years. Methods The 68 children with asthma admitted to our hospital from October 2016 to January 2017 were selected. Every patient had acute attacks and received continued medication. 34 patients treated with salbutamol sulfate inhaled aerosol were used as the control group. 34 patients treated with budesonide suspension combined with salbutamol sulfate aerosol were classified as the observation group. The interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), high-sensitivity-C-reactive protein (hs-CRP), maximum respiratory flow, IS maximum expiratory volume, regulatory B cell ratio, wheezing disappearance time, shortness of breath relief time, wet rales disappearing time, cough disappearing time, and the two year follow-up indicators of growth and development were compared. Results After medication, IL-6, TNF-α, hs-CRP, regulatory B cell ratio, wheezing disappearance time, shortness of breath relief time, moist rales disappearance time, and cough disappearance time were lower in the observation group(P<0.05). The maximum respiratory flow and IS maximum expiratory volume in the observation group were higher than those in the control group (P<0.05). The GH level, height, and weight obtained from two year follow up in the observation group were lower than those in the control group (P<0.05). Conclusion Budesonide suspension combined with salbutamol sulfate aerosol inhalation therapy can alleviate the inflammatory reaction, improve the lung function and immune function of children, and accelerate the disappearance of clinical symptoms, but it will affect the growth and development of children to a certain extent.

Expression of regulatory B cells in rheumatoid arthritis and its correlation with disease severity / 临床检验杂志

Objective@#To investigate the IL-10+CD19+ regulatory B cells (Breg) in peripheral blood of patients with rheumatoid arthritis (RA), and analyze the correlation between the expression of IL-10+CD19+ Breg cells and disease severity. @*Methods@#A total of 40 patients with active RA and 30 healthy individuals (healthy controls, HC) were involved in this study. The peripheral blood mononuclear cells (PBMC) were isolated. The isolated PBMC were co-cultured with CpG oligodeoxynucleotide 2006 (CpG ODN 2006) and phorbol myristate acetate (PMA) in vitro. Flow cytometry was employed to analyze the expression of Breg before and after stimulation. The correlations of Breg expression with ESR, CRP and DAS28 were analyzed. @*Results@#Before stimulation in vitro, the expression of Breg in PBMC of RA group and HC group were 1.92%(1.58%, 2.56%) and 2.04%(1.73%, 2.93%) respectively. There was no significant difference between the two groups (P=0.258). After induction in vitro, the expression of Breg of RA group [13.00%(9.75%, 14.85%)] was significantly higher than that of HC group [9.12%(6.83%, 10.22%)], P＜0.001. Spearman correlation analysis showed that Breg expression was negatively correlated with DAS28 (P=0.002), but not with ESR and CRP (P＞0.05). @*Conclusion@#The expression of Breg was significantly increased after stimulation in vitro and negatively correlated with DAS28, which indicated Breg may play important regulatory roles in pathogenesis and development of RA.

Low frequency of regulatory B-cells and increased CD4+ and CD8+ interferon-γ-producing cells in patients with tropical spastic paraparesis associated with human T-cell lymphotropic virus type

Abstract INTRODUCTION: Tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM) is a disease caused by human T-cell lymphotropic virus type 1 (HTLV-I) that mainly infects CD4 T cells-for example, those of the CD4+CD25hiFOXP3+ [Treg] phenotype-where it inhibits forkhead box protein P3 (FOXP3) expression and promotes interferon-γ (IFN-γ) expression. However, the role it exerts on regulatory B cells (CD19+CD24hiCD38hi; Breg) is unknown. METHODS: The frequencies of Treg and Breg cells was evaluated and the Th1 profiles were assessed in TSP/HAM patients and healthy control subjects. RESULTS: Low percentages of Breg cells and high production of IFN-γ were observed in patients compared to those in healthy control subjects. CONCLUSIONS: The low percentage of Breg cells in patients and the increase in the frequency of Th1 cells suggest an imbalance in the control of the inflammatory response that contributes to the immunopathogenesis of TSP/HAM.

Impact and Modulations of Peripheral and Edaphic B Cell Subpopulations in Chronic Rhinosinusitis With Nasal Polyposis

OBJECTIVES: The pathophysiological mechanisms of chronic rhinosinusitis with nasal polyposis (CRSwNP) still are discussed controversially. Regulatory B cells (Breg) are responsible for the suppression of T cell activity: deficiencies for Breg have been demonstrated to contribute to autoimmune disorders, e.g., systemic lupus erythematosus. In order to evaluate the influence of B cell subpopulations, especially Breg, on the etiology of this disease, the aim of this study was to characterize subpopulations of peripheral and edaphic B cells in CRSwNP. METHODS: Polypoid tissue and blood samples were collected from 10 patients undergoing paranasal sinus surgery and lymphocytes were analyzed by multicolor flow cytometry. RESULTS: There was a significantly lower frequency of B cells in nasal polyps compared to peripheral blood mononuclear cells (PBMC) in patients with CRSwNP. Mature resting B cells were the main population within B cells in PBMC, and memory B cells in nasal polyps. Remarkably, Breg and mature B cells significantly decreased in nasal polyps compared to PBMC. Memory B cells significantly increased and represented the main subpopulation in nasal polyps in patients with CRSwNP. CONCLUSION: In this study a detailed contemporary characterization of B cell subpopulations in patients with CRSwNP is presented. The influence of edaphic B cells could play a key role in the maintenance of this chronic infectious disease.

Analysis of Peripheral B Cell Subsets in Patients With Allergic Rhinitis

PURPOSE: Recent evidence suggests that B cells can both promote and inhibit the development and progression of allergic disease. However, the characteristics of B cell subsets in patients with allergic rhinitis (AR) have not been well documented. This study aimed to analyze the characteristics of B cell subsets in the peripheral blood of AR patients. METHODS: Forty-seven AR patients and 54 healthy controls were enrolled in this study, and the B cell subsets in peripheral blood of all subjects were analyzed by flow cytometry. Moreover, the serum total immunoglobulin E (IgE) and IgE concentrations secreted into the cultured peripheral blood mononuclear cells (PBMCs) were measured by using enzyme-linked immunosorbent assay. RESULTS: We found the peripheral blood of AR patients contained higher percentages of memory B cells, plasma cells, and CD19+CD24hiCD27+ regulatory B cells (Bregs) than those of age-matched healthy controls (P < 0.05), while the percentages of naïve B cells and CD19+CD24hiCD38hi Bregs were significantly lower in AR patients than in healthy individuals (P < 0.05). In addition, the serum total IgE and IgE concentrations secreted into the cultured PBMCs were elevated in AR patients than in the healthy controls (P < 0.05). CONCLUSIONS: Our findings indicate that AR patients were characterized by increase in terminally differentiated memory B cells or plasma cells and decreases in CD19+CD24hiCD38hi Breg cells in the peripheral blood.

Effect of estrogen on IL-10 production in mouse splenic B cells in vitro / 国际药学研究杂志

Objective To ascertain whether estrogen could induce B cell to produce interleukin(IL)-10. Methods C57BL/6 splenic B cells were purified by magnetic activated cell sorting(MACS)method,followed by estrogen treatment for 3 days. The secretion of IL-10 from cultured cell supernatant was tested by ELISA technique. The abundance of mRNA for IL-10、PD-L1 and RBM47 in B cells with estrogen treatment was tested by real-time PCR method. The intracellular IL-10 expression and the surface PD-L1 expression of treated B cells were tested by fluorescence activated cell sorting(FACS)method. And the expression of RBM47 in B cells by estrogen treatment was tested using Western blotting method. Results Estrogen could induce B cell to produce IL-10 in a dose-dependent manner. Estrogen treatment could increase the percentage of IL-10+B cells,the abundance of mRNA for IL-10,PD-L1 and RBM47 in B cells,as well as the expression of PD-L1 on B cell surface. Furthermore,our experimental result indicated the upreg?ulation of RBM47 expression in B cells by estrogen treatment. Conclusion Estrogen treatment in vitro can induce the upregulation of IL-10+regulatory B cells(Breg). Upregulation of RBM47 in the treated B cells might participate in this process by stabilizing IL-10 mRNA.

The changes of regulatory B cell subset in children with type 1 diabetes mellilus / 中华内分泌代谢杂志

[Summary] The proportions of CD19+CD24 hi CD38 hi regulatory B cells ( Breg ) and CD4+CD25+Treg in peripheral blood of children with type 1 diabetes mellitus ( T1DM) were detected by flow cytometric analysis. The concentration of interleukin 10 (IL-10) protein was measured by ELISA. The mRNA expression levels of Foxp3, cytotoxic T lymphocyte-associated antigen-4 ( CTLA-4 ) , and glucocorticoid-induced tumor necrosis factor receptor ( GITR) in CD4+T cells and IL-10 in CD19+ B cells were evaluated using real-time PCR. The results showed that the proportions of CD19+CD24hiCD38hiBreg in peripheral blood, the mRNA and protein levels of IL-10 in B cells from patients with T1DM were lower than those from healthy controls (P<0. 05). The mRNA expression levels of Treg associated factors such as Foxp3, CTLA-4, and GITR were lower in CD4+ T cells from children with T1DM compared with the controls(P<0. 05). These results suggest that Breg cell deficiency and dysfunction might be one of the important factors causing cellular immune dysfunction in patients with T1DM.

Correlation of regulatory B cells and regulatory T cells in patients with chronic hepatitis C / 中国基层医药

Objective To investigate expression and correlation of IL -10 +CD +19 regulatory B cells (Breg)and CD +4CD25high regulatory T cells (Treg)in patients with chronic hepatitis C.Methods 45 patients with chronic hepatitis C and 25 healthy individuals were involved in this study.Flow cytometry was used to analyze the expression of Breg and Treg.ELISA was employed to determine serum IL -10 and TGF -β.Results The expression of Breg in group of HCV was higher than that of the health controls[3.55%(2.40% -4.14%)vs.1.66%(1.24% -2.02%),t =6.78,P <0.001],and Treg in patients with chronic hepatitis C was also higher than that of the health controls[4.55%(3.35% -5.88%)vs.2.05%(1.39% -2.61%),t =7.17,P <0.001].Breg was correlated with Treg positively in group of chronic hepatitis C (r =0.374,P =0.019)and no correlation was found in the health con-trols.Serum IL -10 in patients infected with HCV was significantly higher than that in the health controls[(78.65 ± 17.21)pg/mL vs.(53.21 ±11.47)pg/mL,t =7.51,P <0.001].Conclusion Breg and Treg may play regulatory roles by IL -10 in patients with chronic hepatitis C promote persistence of HCV infection and exhaustion of effector T cells.

Change of the peripheral blood immune pattern and its correlation with prognosis in patients with liver cancer treated by sorafenib

OBJECTIVE@#To study the change of the peripheral blood immune pattern and its correlation with prognosis in patients with liver cancer after treated by sorafenib.@*METHODS@#Patients with advanced liver cancer admitted in our hospital were enrolled and treated with sorafenib. After two months of the treatment, their peripheral blood was collected. The immune cell subset and cytokines level were determined by flow cytometry and luminex technology. According to the reaction expressed by patients towards sorafenib, patients were divided into the response group and the no response group. The changes of the peripheral blood immune pattern and its correlation with prognosis of patients in the two groups were compared.@*RESULTS@#Before and after treatment of sorafenib, there was no significant difference in the ratios of T cells, NK cells and their subtypes in peripheral blood of patients between the two groups; while after treatment the ratio of B cells and regulatory B cells (Breg) of patients in the response group was significant higher than that of the no response group (P < 0.05), and the prognosis conditions of patients with decreased ratio of Breg cells were better than other patients after undergoing chemotherapy. The levels of plasma cytokines IL-6, IL-10, IL-12, IL17, FIL-3L, IFN-γ, TNF-α, MCP-1 and VEGF showed no significant differences.@*CONCLUSIONS@#After treatment of sorafenib, the prognosis conditions of patients of advanced liver cancer with a reduced Breg ratio are better than patients with an unaltered or increased Breg ratio. The ratio of Breg in peripheral blood may be considered as early biological indicator for the prediction of the curative effects of sorafenib.

Effect of estrogen on IL-10 production in mouse splenic B cells in vitro / 国际药学研究杂志

Objective To ascertain whether estrogen could induce B cell to produce interleukin (IL) -10. Methods C57BL/ 6 splenic B cells were purified by magnetic activated cell sorting (MACS) method, followed by estrogen treatment for 3 days. The secretion of IL- 10 from cultured cell supernatant was tested by ELISA technique. The abundance of mRNA for IL- 10, PD- L1 and RBM47 in B cells with estrogen treatment was tested by real-time PCR method. The intracellular IL-10 expression and the surface PD-L1 expression of treated B cells were tested by fluorescence activated cell sorting (FACS) method. And the expression of RBM47 in B cells by estrogen treatment was tested using Western blotting method. Results Estrogen could induce B cell to produce IL- 10 in a dose-dependent manner. Estrogen treatment could increase the percentage of IL-10+ B cells, the abundance of mRNA for IL-10, PD-L1 and RBM47 in B cells, as well as the expression of PD-L1 on B cell surface. Furthermore, our experimental result indicated the upregulation of RBM47 expression in B cells by estrogen treatment. Conclusion Estrogen treatment in vitro can induce the upregulation of IL- 10+ regulatory B cells (Breg). Upregulation of RBM47 in the treated B cells might participate in this process by stabilizing IL- 10 mRNA.

Protective effect of estrogen on experimental autoimmune encephalomyelitis / 国际药学研究杂志

Objective To explore the effect and mechanism of estrogen in mouse model with experimental autoimmune encephalomyelitis (EAE). Methods A mouse model with EAE was induced in female C57BL/6J mice by immunizing with MOG35-55 and CFA. The immunized mice were randomly divided into two groups. In treatment group, estrogen at 50 μmol/L was injected into mice by s.c. in a consecutive fashion from the day of myelin oligodendroglia glycoprotein (MOG) immunization until the day of experiment completion. Meanwhile, in control group, solvent was injected into mice by the same means. The disease score for EAE was recorded everyday. The secretion of TNF-α and IL-17A from cultured mouse splenic cell supernatant was tested by ELISA technique. The abundance of mRNA for TNF-α, IL-17A, PD-1 and PD-L1 in spinal cord was tested by real-time PCR method. The intracellular expression for TNF-α and IL-17A and the surface expression for CD4/PD-1, CD19/PD-L1, CD4/CD25/Foxp3, CD19/CD21/CD23 as well as CD19/ CD5/CD1dhigh of mouse splenocytes were tested by fluorescence activated cell sorting (FACS) method. Results The prophylactic treatment of estrogen could delay the progress in mouse EAE. Histopathological evaluation of mouse spinal cord specimen showed reduced demyelination and inflammatory cell invasion by estrogen treatment. Furthermore, both TNF-α and IL-17A production in estrogen treated mice was significantly downregulated compared to those in control group mice. However, the transcription and expression level for PD-1 in CD4+T cells and that for PD-L1 in CD19+B cells were observed to be upregulated in estrogen treated mice. Also, the percentages of CD19+CD21highCD23low and CD19+CD5+CD1dhigh Breg cells in splenocytes were augmented by estrogen treatment. In contrast, no changes were observed for the proportion of the splenic CD4+CD25+Foxp3+Tregs in mice composed to estrogen treatment with comparison to those in mice with vehicle treatment. Conclusion The prophylactic treatment of estrogen can delay the disease progress in EAE mice, likely through the upregulation of PD-1/PD-L1 pathway and Breg cells.

Change of the peripheral blood immune pattern and its correlation with prognosis in patients with liver cancer treated by sorafenib

Objective To study the change of the peripheral blood immune pattern and its correlation with prognosis in patients with liver cancer after treated by sorafenib. Methods Patients with advanced liver cancer admitted in our hospital were enrolled and treated with sorafenib. After two months of the treatment, their peripheral blood was collected. The immune cell subset and cytokines level were determined by flow cytometry and luminex technology. According to the reaction expressed by patients towards sorafenib, patients were divided into the response group and the no response group. The changes of the peripheral blood immune pattern and its correlation with prognosis of patients in the two groups were compared. Results Before and after treatment of sorafenib, there was no significant difference in the ratios of T cells, NK cells and their subtypes in peripheral blood of patients between the two groups; while after treatment the ratio of B cells and regulatory B cells (Breg) of patients in the response group was significant higher than that of the no response group (P < 0.05), and the prognosis conditions of patients with decreased ratio of Breg cells were better than other patients after undergoing chemotherapy. The levels of plasma cytokines IL-6, IL-10, IL-12, IL17, FIL-3L, IFN-γ, TNF-α, MCP-1 and VEGF showed no significant differences. Conclusions After treatment of sorafenib, the prognosis conditions of patients of advanced liver cancer with a reduced Breg ratio are better than patients with an unaltered or increased Breg ratio. The ratio of Breg in peripheral blood may be considered as early biological indicator for the prediction of the curative effects of sorafenib.

Regulatory B cells: Research advances / 国际药学研究杂志

B cells notionly can regulate the immune responses by producing antibody, presenting antigens to target immune cells, but also have the function of immune suppression. A small quantity of regulatory B (Breg) cells in vivo is capable of performing regulatory functions by producing of inhibitory cytokines,such as interleukin -10, or by direct interaction with other immune cells. In vivo, Breg can maintain immune tolerance and inhibit excessive inflammatory responses efficiently. Clinical studies indicate that Breg are associated with some autoimmune diseases, cancers, infections, and transplantation tolerance. The review focuses on molecules associated with the differentiation and function of the Breg. And also, we discuss the mechanisms and the relations of Breg to clinical diseases.

Toxoplasma gondii Infection Suppresses House Dust Mite Extract-Induced Atopic Dermatitis in NC/Nga Mice

PURPOSE: Toxoplasma gondii (T. gondii) is an obligate intracellular protozoan parasite that infects humans and animals via congenital or postnatal routes, and it is found worldwide. Modulation of the immune system by parasite infection is proposed to suppress allergic inflammation. Growing evidences have shown that interleukin (IL)-10-producing regulatory B cells (B(regs)) and CD4+CD25+FoxP3+ regulatory T cells (T(regs)) induced by parasite infection play a critical role in allergic or autoimmune diseases because these cells regulate negatively cellular immune responses and inflammation. Currently, the role of IL-10-producing regulatory B cells in host immune response during T. gondii infection is unknown. In this study, we investigate whether T. gondii infection can suppress the development of unrelated atopic dermatitis (AD)-like lesions. METHODS: AD is a chronically relapsing inflammatory skin disease accompanied by severe itching; for this, we used NC/Nga mice, a well-known experimental model of systemic AD. Repeated exposure to Dermatophagoides farinae crude extract (DfE), known as a major environmental allergen, evokes AD-like skin lesions in NC/Nga mice under specific pathogen-free conditions. NC/Nga mice were intraperitoneally infected with 10 cysts of T. gondii. RESULTS: T. gondii infection significantly ameliorated AD-like skin lesions in NC/Nga mice. The subpopulation of B(regs) and T(regs) in the AD mice was expanded in the course of T. gondii infection. In addition, T. gondii infection inhibited Th2 and enhanced Th1 immune response in the DfE-treated AD mice. CONCLUSIONS: We have experimentally demonstrated for the first time that T. gondii infection ameliorated AD-like skin lesions in a mouse model of AD. Our study could in part explain the mechanisms of how parasite infection prevents the development of allergic disorder. Therefore, these immunemechanisms induced by T. gondii infection may be beneficial for the host in terms of reduced risk of allergic immune reactions.

The effect of umbilical cord mesenchymal stem cells transplantation for the GD mice on regulatory B cells / 重庆医学

Objective To investigate the therapeutic mechanism of umbilical cord mesenchymal stem cell(UC-MSCs)trans-plantation for the graves disease(GD)mice .Methods Thirty two mice were divided into 3 groups as following :normal control group (G0) ,GD control group (G1) ,UC-MSCs group(G2) .Enzyme linked immunosorbent assay(ELISA)was used to measure the level of TSAb in blood serum and the expression of FT4 was measured by chemiluminescence .Thyroid sections were stained with hema-toxylin and eosin(HE)for histological examination .Splenocytes were stained with multicolor immunofluorescence and detected by flow cytometry to analyze the percentages of CD1dhiCD5+CD19+ regulatory B cells(Bregs) .Expressions of IL-10 and TGF-βmR-NA in spleen organization were measured by Real-time PCR .Results At 26 weeks ,the level of TSAb in blood serum in G2 was more significantly decreased than in G1(P<0 .05) ,and the level of CD19+ B in spleen in G2 was also more significantly decreased than in G1(P<0 .05) ,however ,the percentage of CD1dhiCD5+CD19+ Bregs splenocytes and the levels of IL-10 and TGF-βmRNA in spleen organization were more significantly increased than in G1(P<0 .05) .The concentration differences of TSAb in serum was negatively correlated with the percentage differences of CD1dhi CD5+ CD19+ Bregs ,however ,positively correlated with the expres-sion differences of IL-10 and TGF-βmRNA in spleen before and after transplantation .Conclusion Activation of Bregs may be one of the mechanisms of UC-MSCs therapeutic effect on GD mice .

Regulatory B cells:research advances / 国际药学研究杂志

B cells not only can regulate the immune responses by producing antibody, presenting antigens to target immune cells, but also have the function of immune suppression. A small quantity of regulatory B ( Breg) cells in vivo is capable of performing regulatory functions by producing of inhibitory cytokines,such as interleukin -10, or by direct interaction with other immune cells. In vivo, Breg can maintain immune tolerance and inhibit excessive inflammatory responses efficiently. Clinical studies indicate that Breg are associated with some autoimmune diseases, cancers, infections, and transplantation tolerance. The review focuses on molecules associated with the differentiation and function of the Breg. And also, we discuss the mechanisms and the relations of Breg to clinical diseases.

Effect of helminth infection on allergic and autoimmune diseases / 中国人兽共患病学报

ABSTRACT:Many studies show that helminth‐derived molecules can induce immunoregulatory cells to form immune net‐work‐mediated immune ,thereby inhibiting allergic and autoimmune diseases .Helminth parasites can induce immune cell activa‐tion and produce cytokine .And parasites play an inhibitory effect to affect other immune related diseases .However ,the associ‐ation between helminths infections and immune related diseases does not always have an unequivocal outcome .While some hel‐minths infections protect against allergic diseases ,other helminth can exacerbate this immunopathology .

Study on the levels of CD19+CD5+CD1dhigh B10 cells in NOD mice / 中华微生物学和免疫学杂志

Objective To study the alterations of CD19+CD5+CD1dhigh B, Th1 and Th17 cells in non-obese diabetic ( NOD) mice and the correlation between B10 cells and type 1 diabetes in NOD mice. Methods Flow cytometry ( FCM) was used to measure the levels of CD19+CD5+CD1dhigh B, CD19+IL-10+B, CD4+IFN-γ+Th1, CD4+IL-17+Th17 and CD4+CD25+Foxp3+T cells in NOD mice ( 4 weeks old NOD mice:group A, n=10;8 weeks old NOD mice:group B, n=10; NOD mice with diabetes: group C, n=10) and age-matched C57BL/6 mice ( control group, n=20 ) .Hematoxylin-eosin staining of pancreatic tissues was performed for histopathological assessment of the development of insulitis in NOD mice.Results (1) Histopathological analysis showed that mice from A, B and C groups respectively showed no insulitis, insulitis and obvious insulitis with no intact islets.(2) The highest levels of B10 cells in NOD mice were ob-served in group B, followed by those in group C and group A (P0.05).More B10 cells were detected in pancreatic lymph nodes than in other tissues of 8 weeks old NOD mice, the levels of which were also higher than those in pancreatic lymph nodes of mice form group C ( P0.05).Conclusion The percentages and distribution of B10 cells in NOD mice changed with age and the development of insulitis.The decrease of B10 cells might participate in the development of type 1 diabe-tes in NOD mice.