Research and application progress in adoptive re-transfusion of regulatory cells in organ transplantation / 器官移植

Rejection and adverse reactions caused by long-term use of immunosuppressants severely affect the survival rate and quality of life of organ transplant recipients. Immune tolerance induction plays a key role in improving the survival rate and quality of life of organ transplant recipients. In recent years, tremendous progress has been achieved in adoptive re-transfusion of regulatory cells. In this article, research progress in regulatory T cell (Treg), myeloid-derived suppressor cell (MDSC) and regulatory B cell (Breg) in animal experiment and clinical application was reviewed, and the main clinical problems of adoptive re-transfusion of regulatory cells, the application of chimeric antigen receptor Treg and the concept of cell therapy in immune evaluation were summarized, aiming to deepen the understanding of regulatory cell therapy, promote the application of regulatory cells in immune tolerance of organ transplantation, and improve clinical efficacy of organ transplantation and the quality of life of recipients.

Changes and significance of CDld^(hi)CD5⁺CD19⁺ regulatory B cells in peripheral blood of children with juvenile idiopathic arthritis-associated uveitis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To explore the changes and significance in JIA-U and to detect the levels of CDld^(hi)CD5⁺CD19⁺ regulatory B cells(Breg)in the peripheral blood of children with juvenile idiopathic arthritis-associated uveitis(JIA-U).<p>METHODS: From April 2018 to May 2020, 95 children with JIA-U were selected as JIA-U group; 70 children with juvenile idiopathic arthritis(JIA)were selected as JIA group, and all of them were diagnosed and treated in our hospital; another 75 healthy children in the same period were selected as the control group. The ratio of CDld^(hi)CD5⁺CD19⁺Breg in peripheral blood was detected by flow cytometry; the level of IL-10 in peripheral blood was detected by ELISA; the correlation between CDld^(hi)CD5⁺CD19⁺Breg ratio and the expression of IL-10, the severity of JIA-U patients was analyzed by Pearson correlation coefficient method; the influencing factors of JIA-U were analyzed by Logistic regression. <p>RESULTS: Compared with the control group, the proportion of CDld^(hi)CD5⁺CD19⁺Breg in peripheral blood of children in JIA group and JIA-U group was significantly lower(all <i>P</i><0.01), while the level of IL-10 was significantly higher(all <i>P</i><0.01); compared with JIA group, the proportion of CDld^(hi)CD5⁺CD19^+Breg in JIA-U group was significantly lower(<i>P</i><0.01), while the level of IL-10 was significantly higher(<i>P</i><0.01); compared with the stationary phase, the ratio of CD1d^(hi)CD5⁺CD19⁺Breg in the active phase of JIA and JIA-U patients was significantly reduced(all <i>P</i><0.01), and the IL-10 level was significantly increased(all <i>P</i><0.01); the ratio of CDld^(hi)CD5⁺CD19⁺Breg was negatively correlated with the expression of IL-10 and the severity of JIA-U; multivariate analysis showed that low proportion of CDld^(hi)CD5⁺CD19⁺Breg, less-joint JIA subtype, duration of arthritis < 4a, high levels of IL-10 were risk factors for JIA-U. <p>CONCLUSION: The proportion of CDld^(hi)CD5⁺CD19⁺Breg in peripheral blood of children with JIA-U is significantly decreased, which may be involved in the occurrence and development of JIA-U, and has the potential to be used as an index to judge the severity of JIA-U.

B cells and tumor immune escape / 中南大学学报(医学版)

B lymphocyte is an important component of the human immune system and it has a role in the process of the body's specific immunity. In recent years, the research on B cells and tumor immune escape has rapidly progressed. Studies have shown that different types of B cells play different roles in tumor microenvironment through a variety of mechanisms. B cells in the tertiary lymphatic structure promote anti-tumor immunity, while regulatory B cells promote tumor immune escape. Antibody drugs targeting B cells are a promising direction for tumor immunotherapy.

Effect and mechanism of human gingival mesenchymal stem cell on B cells / 器官移植

Objective To investigate the regulating function of human gingival mesenchymal stem cell (GMSC) on the proliferation and differentiation of B cells and its underlying molecular mechanism. Methods GMSC were isolated and B cells were isolated from peripheral blood. GMSC or fibroblasts were co-cultured with B cells in vitro and assigned into the GMSC group and fibroblast group. The proliferation of B cells was detected in two groups. The expression of IgG1 and IgM in the cell supernatants was measured between two groups. The secretion of interleukin (IL)-6, Perforin, interferon (IFN)-γ and tumor necrosis factor (TNF)-α was compared between two groups. The expression levels ofIL-10 and transforming growth factor (TGF)-β in B cells were detected between two groups. The expression of PC-1 in B cells was measured in two groups. The signaling pathway involved with the regulating effect of GMSC on B cell function was investigated. The regulating effect of GMSC on the role of B cells in activating T cell function was assessed. Results Compared with the fibroblast group, the proliferation of B cells was significantly weakened in the GMSC group (P < 0.05). Co-culture of GMSC and B cells significantly inhibited the secretion of IgG1 and IgM from B cells and the secretion ofIL-6, Perforin, IFN-γ and TNF-α (all P < 0.05). Compared with the fibroblast group, the secretion of IL-10 and TGF-βwas significantly higher in the GMSC group (both P < 0.05). The expression level of PC-1 in the GMSC group was significantly down-regulated (P < 0.05). After adding ALK5, an inhibitor of TGF-β receptor, the inhibitory effect of GMSC upon B cells was significantly weakened (P < 0.05). Compared with the fibroblast group, the ability of B cells to activate and proliferate T cells was significantly attenuated in the GMSC group (P < 0.05). Conclusions GMSC can inhibit B cells and their mediated immune responses. The activation of B cells and other related functions can be suppressed through the TGF-β signaling pathway.

The function of regulatory immunological cell in xenotransplantation immunity / 器官移植

Xenotransplantation is the most promising method to resolve the organ shortage problem in the future. In recent years, the advances in gene editing and immunological technique have driven the rapid development of xenotransplantation. However, there are still many insurmountable obstacles in the clinical application of xenotransplantation, among which the rejection is the most important cause of the xenotransplantation failure. Regulatory immunological cells are a group of immunological cells with the negative regulation function in the body, which can inhibit allotransplantation rejection and prolong the survival time of the graft. This paper summarized the research progress of regulatory immunological cells in the xenotransplantation application in recent years, providing reference for the prevention and treatment of xenotransplantation rejection.

Analysis of B cell subsets in rhumatoid arthritis patients and the effect of epigallocatechingallate on B cell subsets / 上海交通大学学报（医学版）

Objective:To explore the characteristics of B cell subsets in rheumatoid arthritis (RA) patients and the regulation of epigallocatechingallate (EGCG) on B cell subsets in RA patients. Methods:Twenty-nine age- and sex-matched RA patients and 29 healthy controls were selected, and the difference of B cell subsets in peripheral blood between the two groups was analyzed by paired t-test. According to the value of disease activity score in 28 joints (DAS28), RA patients were divided into active group (2.6 ≤ DAS28 0.05). There was no significant difference in the numbers and the proportions of total B cells and B cell subsets (except CD19+ IL-10+ Breg) between 10 RA patients of active group and 19 RA patients of highly active group (P>0.05). There was no significant difference in the number and the proportion of CD19+ IL-10+ Breg in lymphocytes between 6 RA patients of active group and 12 RA patients of highly active group (P>0.05). The proportion of total B cells was weakly positively correlated with IgG type rheumatoid factor (r=0.308). EGCG could significantly increase the proportion of CD19+ IL-10+ Breg (P0.05). Conclusion:B cells may play an auxiliary role in the development of RA. The number of CD19+ IL-10+ Breg in RA patients increases as a feedback. EGCG can promote Breg proliferation and suppress BAFF-R mRNA expression.

Role of regulatory B cells in children with newly diagnosed immune thrombocytopenia / 中华实用儿科临床杂志

Objective To investigate the role of regulatory B cells (Breg)in children with newly diagnosed immune thrombocytopenia (ITP).Methods A total of 35 newly diagnosed ITP children admitted to the Pediatric De-partment,the First Affiliated Hospital of Zhengzhou University from January to December 201 4 were recruited in this study,and another 20 gender -and age -matched healthy children from the Department of Medical Examination Center of the same Hospital were recruited as controls during the same period.Peripheral blood samples (3 mL from each chil-dren)were collected from all the newly diagnosed ITP children and the normal controls.Breg cells were tested by Flow Cytometry,and the expression levels of interleukin -1 0 (IL -1 0)and transforming growth factor -β1 (TGF -β1 ) mRNA were measured by real time fluorescence quantitative polymerase chain reaction.Meanwhile,the correlation be-tween Breg cells and the expression levels of IL -1 0,TGF -β1 mRNA were analyzed by Pearson correlation.Results The percentages of Breg cells in the peripheral blood of the newly diagnosed ITP children [(2.37 ±0.67)%]were sig-nificantly lower than those of the normal controls [(4.92 ±1 .32)%],and there was a significant difference (t =-7.47,P =0.000);the expression levels of IL -1 0 mRNA in the newly diagnosed ITP children(0.202 ±0.059) were significantly decreased compared with those of the normal controls(0.41 5 ±0.21 2),and there was a significant difference(t =-5.1 75,P =0.000);while the expression levels of TGF -β1 mRNA in the newly diagnosed ITP chil-dren(1 .587 ±0.823)were significantly increased than those in the normal controls(0.61 9 ±0.322),and there was a significant difference(t =4.081 ,P =0.001 ).There was a significant positive correlation between Breg cells and the ex-pression levels of IL -1 0 mRNA(rs =0.828,P <0.05),but no correlation between Breg cells and expression level of TGF -β1 mRNA was found (rs =0.527,P =0.1 1 7).Conclusions The decrease expressions of Breg cells can be found in the newly diagnosed ITP children,and the abnormal expression of Breg cells may play a key role in the immu-nological pathogenesis of the newly diagnosed ITP children.

The role of interleukin-35-producing regulatory B cells in immune pathogenesis of Kawasaki disease / 中华实用儿科临床杂志

Objective To investigate the role of IL-35-producing regulatory B cells(IL-35 + Breg)in immunological pathogenesis of Kawasaki disease (KD).Methods Thirty-two children with KD and 28 age-matched healthy children were allowed to participate in this study.Flow cytometry was performed to evaluate the proportions of IL-35 + Breg as well as requlatory T cells (Treg)and expression levels of associated molecules such as programmed death-ligand 1 (PD-LI),CD169,programmed death 1 (PD-1),CD43,IL-12p35,epstein-Barr virus induced 3 (IL-27 EBI3).IL-12 receptor beta 2 (IL-12 Rβ2),IL-27 receptor alpha (IL-27 Rα),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 3 (pSTAT3).Transcription levels of the Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),vav1 guanine nucleotide exchange factor(Vav) in CD19 + B cells were determined by using quantitative real-time PCR.Plasma concentrations of IL-35,tumor necrosis factor α(TNF-α) and IL-12 were measured by adopting enzyme-linked immunosorbent assay.Results (1) The proportions of IL-35 +Breg and its expressions of IL-12p35,IL-27EBI3 and IL-10 in patients with acute KD were lower than those of healthy controls [IL-35 + Breg:(5.79 ± 2.60) ％ vs (12.65 ± 5.34) ％;F =19.23,9.70,14.30.7.08;all P ＜ 0.05],but they were significantly increased after intravenous immune globulin (IVIG) treatment [IL-35 + Breg:(10.52 ± 4.95) ％;all P ＜ 0.05].(2) The proportions of Treg and its transcriptional levels of IL-12p35 and IL-27 EBI3 were down-regulated during acute KD [Treg:(4.12 ± 1.51) ％ vs (8.06 ± 3.32) ％;F =19.70,17.69,38.22;all P ＜ 0.05],but were increased after therapy [Treg:(7.39 ± 2.85) ％;P ＜ 0.05].A positive correlation was found between the proportions of Treg and IL-35 + Breg during acute KD (r =0.69,P ＜ 0.05).Meanwhile,plasma concentrations of IL-35 and expression levels of IL-12Rβ2,IL-27Rα,pSTAT1 and pSTAT3 in CD19 + B cells were significantly down-regulated in children with acute KD,but they were increased after treatment(F =8.09,7.54,7.69,5.89,12.59,all P ＜ 0.05).(3) Compared with healthy controls,expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated during acute KD (F =24.94,16.53,34.71,19.51;all P ＜ 0.05).Expression levels of PD-1,CD43 and its downstream molecules (SHP-2,PTEN,Vav) in CD19 + B cells were down-regulated during acute KD (F =6.43,5.57,19.52,10.37,11.37;all P ＜ 0.05),and restored remarkably after therapy (all P ＜ 0.05).Conclusion Insufficiency of IL-35 + Breg and its expression of IL-35 may be the important factors contributing to immunological dysfunction in KD.

The role of CD19+CD24hiCD27hi regulatory B cells in the severity assessment of acute pancreatitis / 中华微生物学和免疫学杂志

Objective To investigate the prognostic value of regulatory B cells (Bregs) in patients with acute pancreatitis .Methods Flow cytometry analysis was performed to detected the percentages of CD19+IL-10+and CD19+CD24hiCD27hi Bregs in peripheral blood samples collected from patients with acute pancreatitis (36 cases with mild acute pancreatitis and 15 cases with severe acute pancreatitis ) as well as the surface costimulatory molecules including CD80 and CD86 on CD19+CD24hiCD27hi Bregs.Their correlations with lymphocytes and C-reactive protein ( CRP) were further analyzed .Results The numbers of lympho-cytes, CD19+lymphocytes, CD19+IL-10+and CD19+CD24hi CD27hi Bregs in peripheral blood samples col-lected from patients with severe and mild acute pancreatitis as well as the mean fluorescence intensities ( MFI) of CD80 and CD86 were significantly lower than those from healthy subjects .Compared with patients with mild acute pancreatitis , the numbers of lymphocytes and CD 19+lymphocytes , the absolute numbers of CD19+IL-10+and CD19+CD24hiCD27hi Bregs as well as the mean fluorescence intensities (MFI) of CD80 and CD86 in patients with severe acute pancreatitis were significantly decreased .The percentages of CD19+IL-10+and CD19+CD24hiCD27hi Bregs in patients with mild acute pancreatitis were significantly increased af-ter an initial drop , but in patients with severe acute pancreatitis those values were continuously decreased along with the disease progression .The percentage of CD19+IL-10+Bregs was positively correlated with the percentage of CD19+CD24hiCD27hi Bregs and the absolute number of CD19+lymphocytes, but was negatively correlated with CRP .Conclusion The abnormal number and function of CD 19+IL-10+and CD19+CD24 hi CD27hi Bregs might be one of the important reasons causing immune dysfunction in patients with acute pan -creatitis.

Development and differentiation of B cells and autoimmune diseases / 中华实用儿科临床杂志

Of B cells derived from bone marrow hematopoictic stem cells,bone marrow (poultry,as the bursa of fabricius) is a place of B cells development and mature.The growth of B cells can experience from progenitor cell B,pre-B cells,immature B ceils and mature B cells stages in the bone marrow.The differentiation process of B cells can be divided into antigen independ stage and antigen depend stage.CD5 + B1 cells and B10 cells of B cell subsets probably associated with autoimmune disease,B1 cells may play roles of promotion in inflammatory reaction,and B10 cells may suppress autoimmune inflammation.

Dendritic cells phagocytized apoptotic lymphocytes induce the development of regulatory B cells with high secretion of interleukin-10 / 器官移植

Objective To investigate the feasibility of immature dendritic cells (imDC)phagocytized psoralen ultraviolet A (PUVA)-treated splenic lymphocytes (PUVA-SP DC)in mice inducing B lymphocytes to be regulatory B cells (Breg)with high secretion of interleukin (IL)-10 (IL-10 +Breg). Methods Bone marrow-derived DC of mice was cultured. Spleen lymphocytes of mice were isolated and treated by PUVA,and turned to be PUVA-SP. The bone marrow-derived imDC was co-cultured with PUVA-SP in vitro to obtain PUVA-SP DC. Splenic B lymphocytes of mice were separated by anti-CD19 magnetic beads and co-cultured with different kinds of DC for 48 hours. The levels of interferon (IFN )-γ,transforming growth factor (TGF)-β,IL-12p70,and IL-10 in the culture supernatant of B lymphocytes,imDC,imDC+B lymphocytes, PUVA-SP DC and PUVA-SP DC +B lymphocytes were measured by enzyme-linked immune absorbent assay (ELISA). The accounts of IL-10 +Breg in B lymphocytes,imDC+B lymphocytes,mDC+B lymphocytes and PUVA-SP DC+B lymphocytes were detected by flow cytometry. Results Compared with the other 4 groups, the level of IL-10 in cell culture supernatant of PUVA-SP DC+B lymphocytes was significantly higher (all in P<0.05). Compared with the other groups,the account of IL-10 +Breg in PUVA-SP DC+B lymphocytes was significantly higher. Conclusions PUVA-SP DC can induce splenic B lymphocytes to differentiate into IL-10 +Breg.

In Vitro Induction of Allergen-Specific Interleukin-10-Producing Regulatory B Cell Responses by Interferon-gamma in Non-Immunoglobulin E-Mediated Milk Allergy

PURPOSE: Specific oral immunotherapy (SOIT) using interferon-gamma (IFN-gamma) has been successful as a food allergy treatment. Interleukin-10 (IL-10)-producing regulatory B cells (Br1s) play a role in immune tolerance to food allergens. In addition, IFN-gamma shows tolerogenic effects on allergen-induced Br1 responses. METHODS: Eleven patients that were allergic to cow's milk and 12 milk-tolerant subjects were selected by double-blind placebo-controlled food challenge (DBPCFC) and clinical characteristics. The immunomodulatory effects of IFN-gamma on allergen-specific Br1 responses were evaluated in 6 milk allergy patients and 8 milk-tolerant subjects. Peripheral blood mononuclear cells (PBMCs) from subjects were stimulated with casein and/or IFN-gamma and analyzed by flow cytometry. RESULTS: IFN-gamma had no effect on total cell counts or the proportion of Br1 cells in PBMCs. IFN-gamma stimulation did not change total Br1 cell counts or the percentage of Br1s among CD5(+) B cells in the milk allergy or the milk-tolerant groups. In the milk allergy group, Br1 counts were not different between the control and the casein stimulation but significantly increased in the IFN-gamma + casein stimulated cells, and the Br1 fractions were decreased after casein stimulation and recovered in the addition of IFN-gamma for stimulation. In the milk-tolerant group, Br1 counts increased in the casein stimulated cells and in the IFN-gamma + casein stimulated cells, but the increase was significantly less when IFN-gamma was added, and the Br1 fractions were increased after casein stimulation and IFN-gamma + casein stimulation, that was not significant when IFN-gamma was added. CONCLUSIONS: IFN-gamma-induced allergen-specific Br1 responses in the PBMCs of milk allergy patients play a role in milk allergen-specific tolerance induction in vitro. Further investigations into the molecular immunological mechanisms underlying the induction of allergen-specific Br1 responses are needed.

IL-10 is Predominantly Produced by CD19(low)CD5(+) Regulatory B Cell Subpopulation: Characterisation of CD19 (high) and CD19(low) Subpopulations of CD5(+) B cells

IL-10 production by CD19(+)CD5(+) B cells was investigated, by determining the expression levels of CD19, a classical B cell marker. Peripheral mononuclear cells were stained with fluorescence-conjugated anti-CD5, anti-CD19, anti-IL-10, and Annexin V. Interestingly, IL-10-producing B cells were found to be localised within the CD19(low)CD5(+) B cell subset. Apoptotic changes were also observed mainly in CD19(low) cells among B cells. Thus, CD5(+) B cells should be classified as CD19(high) and CD19(low) cells, and the immunological significance of CD19 for the IL-10 production by CD5(+) B cells requires further studies.

Regulatory B Cells and Allergic Diseases

B cells are generally considered to positively regulate immune responses by producing antigen-specific antibodies. B cells are classified into classical CD5- conventional B cells and CD5+ B1 cells. The latter produce multi-specific autoantibodies and are thought to be involved in autoimmune diseases. However, evidence supporting a B cell negative regulatory function has accumulated over the past 30 years. Multiple reports have suggested that absence, or loss, of regulatory B cells exacerbates symptoms of both allergic (including contact hypersensitivity and anaphylaxis) and autoimmune (such as experimental autoimmune encephalomyelitis, chronic colitis, and collagen-induced arthritis) diseases, and in lupus-like models of autoimmunity. Regulatory B cells are characterized by production of the negative regulatory cytokines, IL-10 and TGF-beta. IL-10-producing B cells were the first regulatory B cells to be recognized and were termed 'B10' cells. IL-10-producing regulatory B cells are of the CD19(+)CD5(+)IgM(hi)IgD(lo)CD1d(hi) type. Recently, a TGF-beta-producing regulatory B cell subset, Br3, has been shown to be related to immune tolerance in food allergies. Moreover, forkhead box P3 (Foxp3)-expressing B cells have also been identified in humans and may act as regulatory B cells (Bregs). The functional image of regulatory B cells is similar to that of regulatory T cells. Because of the proliferative and apoptotic responses of Br1 and Br3 cells in immune tolerance in non-IgE-mediated food allergy, reciprocal roles and counter-regulatory mechanisms of Br1 and Br3 responses are also suspected. Additionally, different roles for regulatory B and T cells at different time points during initiation and progression of autoimmune disease are described.

Presence of Foxp3-expressing CD19(+)CD5(+) B Cells in Human Peripheral Blood Mononuclear Cells: Human CD19(+)CD5(+)Foxp3(+) Regulatory B Cell (Breg)

Foxp3 is a transcript factor for regulatory T cell development. Interestingly, Foxp3-expressing cells were identified in B cells, especially in CD19(+)CD5(+) B cells, while those were not examined in CD19(+)CD5(-) B cells. Foxp3-expressing CD5(+) B cells in this study were identified in human PBMCs and were found to consist of 8.5+/-3.5% of CD19(+)CD5(+) B cells. CD19(+)CD5(+)Foxp3(+) B cells showed spontaneous apoptosis. Rare CD19(+)CD5(+) Foxp3(+) regulatory B cell (Breg) population was unveiled in human peripheral blood mononuclear cells and suggested as possible regulatory B cells (Breg) as regulatory T cells (Treg). The immunologic and the clinical relevant of Breg needs to be further investigated.