The level of regulatory T lymphocyte detected by flow cytometry in chimeric antigen receptor T cell immunotherapy and its clinical significance / 中华检验医学杂志

Objective:To investigate the characteristics of changes in peripheral blood regulatory T lymphocyte (Treg) levels in patients with B-cell lymphoma who received chimeric antigen receptor (CAR)-T cell immunotherapy, and the relationship between Treg levels and optimal efficacy and treatment response.Methods:The data of 23 patients with relapsed/refractory B-cell malignancies who received CD19/CD22 CAR-T cell immunotherapy in Wuhan Tongji Hospital from 2019 to 2021 were retrospectively studied. The enrolled patients were divided into complete remission (CR) group (8 cases), partial remission (PR) group (7 cases) and no response(NR) group (8 cases) according to Lugano′s revised lymphoma efficacy evaluation criteria. A total of 16 patients with B-cell lymphoma who did not receive CAR-T cell immunotherapy during the same period in Wuhan Tongji Hospital were collected as the control group.In different periods during CAR-T cell immunotherapy, multicolor flow cytometry(MFC) was used to dynamically detect peripheral blood the proportion of Treg in CD4 +T cells (Treg/CD4 +T), the proportion of lymphocytes (Treg/Lym), the proportion of Treg in white blood cells (Treg/WBC), and the absolute number of Treg (Treg#). The trend of Treg levels over time, as well as the differences in Treg levels in patients with different prognosis groups in different periods were analyzed.According to the proportion of Treg and the median level of absolute number within 1 to 15 days after CAR-T cell infusion, the patients were divided into a low-level group with 11 cases and a high-level group with 12 cases. The statistical differences in the peak value of CAR-T copy, iron protein, and IL-6 were compared between various groups. Independent samples t test, Mann-Whitney U test, Cox-Stuart trend existence test and one-way analysis of variance was used in statistical analysis. Results:In the 23 patients who received CAR-T cell immunotherapy, the mean values of Treg/CD4 +T and Treg/Lym before CAR-T cell infusion were (20.42±7.96)% and (13.61±7.13)%, respectively, which were significantly higher than that of the control group [(7.33±3.61)%, t=5.893, P<0.001; (1.91±0.90)%, t=6.53, P<0.001]. The number of Treg in the meantime was significantly lower [(1.81±1.52)/μl<(13.66±9.89)/μl, t=4.261, P<0.001]. After infusion, Treg/CD4 +T and Treg/Lym all remarkably decreased ( P<0.001),Treg/WBC increased significantly( P=0.01). The mean values of Treg/CD4 +T (12.87±1.93)%, Treg/Lym (6.35±2.84)%, and Treg/WBC (0.05±0.05)% in the patients with CR as the best response group were lower than those in the PR group [(29.68±5.49)%( P<0.01), (21.85±2.1)%( P<0.01), 0.50±0.69( P<0.05)] before CAR-T cell immunotherapy. Patients with lower mean Treg/CD4 +T within 1 to 15 days after reinfusion of CAR-T cells had higher peak CAR-T copy number ( P<0.05). Conclusion:Treg/CD4 +T and Treg/Lym were increased and then decreased during CAR-T treatment in B cell malignancies. The patients with lower proportions of Treg before infusion have favorable treatment efficacy. Besides, patients with lower Treg/CD4 +T after infusion have better CAR-T cell expansion. In the process of CAR-T cell immunotherapy, the use of MFC to dynamically monitor the proportion of Treg has certain clinical significance for the prediction of the optimal efficacy of immunotherapy and the prediction of treatment response.

Role of Treg in the immune pathogenesis of subacute thyroiditis / 基础医学与临床

Objective To explore the role of regulatory T-lymphocytes(Treg) in the immune pathogenesis of suba-cute thyroiditis (SAT). Methods The proportion of Treg in CD4+T cells in peripheral blood of 46 SAT patients and15 controls was detected using flow cytometry. And the concentration of interleukin-10(IL-10), transforming growth factor-beta1(TGF-β1) and prostaglandin E2(PGE2) in serum of 46 SAT patients and 15 controls was measured with ELISA. In addition, the Forkhead box protein 3 (Foxp3) positive cells in thyroid tissue of 29 SAT patients and20 controls was detected by immunohistochemistry. Results The proportion of Treg in peripheral blood of SAT pa-tients was significantly lower than that of controls (P＜0.05). And the concentration of TGF-β1 in serum of SAT patients was apparently higher than that of controls(P＜0.05). Additionally, the positive rate of Foxp3 in thyroid tissue of SAT patients was markedly higher than that of controls(P＜0.05).Conclusions The decrease of Treg may play an important role in the immune pathogenesis of SAT.

Mecanismos da imunoterapia alérgeno-específica / Mechanisms of allergen-specific immunotherapy

A imunoterapia alérgeno-específica tem sido usada há mais de 100 anos como um tratamento de dessensibilização para doenças alérgicas, representando um método potencialmente curativo e específico. O presente estudo tem como objetivo revisar os mecanismos da imunoterapia alérgeno-específica, através de revisão bibliográfica com base em artigos publicados entre 1998 e 2016, disponíveis no banco de dados PubMed. Os mecanismos de ação da imunoterapia incluem modulação de linfócitos T e B, produção de IgG4 alérgeno-específica e redução de IgE alérgenoespecífica, migração de eosinófilos, basófilos e mastócitos nos tecidos, bem como a liberação de seus mediadores. As células T reguladoras (Treg) suprimem as células dendríticas responsáveis pela geração de células T efetoras, inibindo TH1, TH2 e TH17. As células Treg foram identificadas como peças-chave no processo de indução de tolerância periférica aos alérgenos.

Allergen-specific immunotherapy has been used for more than 100 years as a desensitizing therapy for allergic diseases, representing a potentially curative and specific method. The aim of the present study was to review the mechanisms of allergenspecific immunotherapy based on papers published between 1998 and 2016 and available in the PubMed database. The mechanisms of action of immunotherapy include modulation of T-and B-cell responses, induction of allergen-specific IgG4 and suppression of allergen-specific IgE production, migration of eosinophils, basophils, and mast cells to tissues, as well as release of their mediators. Regulatory T cells (Treg) suppress dendritic cells that support the generation of effector T cells, inhibiting TH1, TH2, and TH17 cells. Treg cells have been identified as key regulators of the induction process of peripheral allergen tolerance.

Targeting the epitope spreader Pep19 by naïve human CD45RA⁺ regulatory T cells dictates a distinct suppressive T cell fate in a novel form of immunotherapy

PURPOSE: Beyond the limited scope of non-specific polyclonal regulatory T cell (Treg)-based immunotherapy, which depends largely on serendipity, the present study explored a target Treg subset appropriate for the delivery of a novel epitope spreader Pep19 antigen as part of a sophisticated form of immunotherapy with defined antigen specificity that induces immune tolerance. METHODS: Human polyclonal CD4⁺CD25⁺CD127(lo−) Tregs (127-Tregs) and naïve CD4⁺CD25⁺CD45RA⁺ Tregs (45RA-Tregs) were isolated and were stimulated with target peptide 19 (Pep19)-pulsed dendritic cells in a tolerogenic milieu followed by ex vivo expansion. Low-dose interleukin-2 (IL-2) and rapamycin were added to selectively exclude the outgrowth of contaminating effector T cells (Teffs). The following parameters were investigated in the expanded antigen-specific Tregs: the distinct expression of the immunosuppressive Treg marker Foxp3, epigenetic stability (demethylation in the Treg-specific demethylated region), the suppression of Teffs, expression of the homing receptors CD62L/CCR7, and CD95L-mediated apoptosis. The expanded Tregs were adoptively transferred into an NOD/scid/IL-2Rγ(−/−) mouse model of collagen-induced arthritis. RESULTS: Epitope-spreader Pep19 targeting by 45RA-Tregs led to an outstanding in vitro suppressive T cell fate characterized by robust ex vivo expansion, the salient expression of Foxp3, high epigenetic stability, enhanced T cell suppression, modest expression of CD62L/CCR7, and higher resistance to CD95L-mediated apoptosis. After adoptive transfer, the distinct fate of these T cells demonstrated a potent in vivo immunotherapeutic capability, as indicated by the complete elimination of footpad swelling, prolonged survival, minimal histopathological changes, and preferential localization of CD4⁺CD25⁺ Tregs at the articular joints in a mechanistic and orchestrated way. CONCLUSIONS: We propose human naïve CD4⁺CD25⁺CD45RA⁺ Tregs and the epitope spreader Pep19 as cellular and molecular targets for a novel antigen-specific Treg-based vaccination against collagen-induced arthritis.

Effects of Shen-Fu Injection () on apoptosis of regulatory T lymphocytes in spleen during post-resuscitation immune dysfunction in a porcine model of cardiac arrest / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate whether Shen-Fu Injection (, SFI) reduces post-resuscitation immune dysfunction in a porcine model of cardiac arrest by modulating apoptosis of regulatory T lymphocytes (Treg) in the spleen.</p><p><b>METHODS</b>After 8-min untreated ventricular fibrillation and 2-min basic life support, 24 pigs were divided into 3 groups with a random number table, i.e. SFI group, epinephrine (EP) group, and saline (SA) group (8 in each group), which received central venous injection of SFI (1.0 mL/kg), EP (0.02 mg/kg) and SA, respectively. The same procedure without CA initiation was achieved in the sham-operated (sham) group (n=6). After successful return of spontaneous circulation (ROSC), apoptosis rate of splenic Treg was detected by flow cytometry; and the mRNA expression of forkhead/winged helix transcription factor (Foxp3) of splenic Treg was detected by real time-polymerase chain reaction; and the levels of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in porcine splenic Treg were detected by using enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Compared with the sham group, the apoptosis rate of Treg was significantly decreased, and the levels of Foxp3 mRNA expression, IFN-γ, IL-4 and IFN-γ/IL-4 were increased in the SA group (P<0.05 or P<0.01). Compared with the EP and SA groups, SFI treatment increased the apoptosis rate of Treg and reduced the levels of Foxp3 mRNA expression, IFN-γ and IFN-γ/IL-4 (P<0.05).</p><p><b>CONCLUSIONS</b>SFI has signifificant effects in attenuating post-resuscitation immune dysfunction by modulating apoptosis of Treg in the spleen.</p>

Canine adipose tissue-derived mesenchymal stem cells ameliorate severe acute pancreatitis by regulating T cells in rats

Severe acute pancreatitis (SAP) is associated with systemic complications and high mortality rate in dogs. Mesenchymal stem cells (MSCs) have been investigated for their therapeutic potential in several inflammation models. In the present study, the effects of canine adipose tissue-derived (cAT)-MSCs in a rat model of SAP induced by retrograde injection of 3% sodium taurocholate solution into the pancreatic duct were investigated. cAT-MSCs labeled with dioctadecyl-3,3,3′-tetramethylindo-carbocyanine perchlorate (1 × 10⁷ cells/kg) were systemically administered to rats and pancreatic tissue was collected three days later for histopathological, quantitative real-time polymerase chain reaction, and immunocytochemical analyses. Greater numbers of infused cAT-MSCs were detected in the pancreas of SAP relative to sham-operated rats. cAT-MSC infusion reduced pancreatic edema, inflammatory cell infiltration, and acinar cell necrosis, and decreased pancreatic expression of the pro-inflammatory cytokines tumor necrosis factor-α, interleukin (IL)-1β, -6, -12, -17, and -23 and interferon-γ, while stimulating expression of the anti-inflammatory cytokines IL-4 and IL-10 in SAP rats. Moreover, cAT-MSCs decreased the number of clusters of differentiation 3-positive T cells and increased that of forkhead box P3-positive T cells in the injured pancreas. These results indicate that cAT-MSCs can be effective as a cell-based therapeutic strategy for treatment of SAP in dogs.

Stem cell therapy in animal models of allergic airway diseases

Allergic airway diseases are characterized by T-helper type 2 (Th2)-skewed eosinophilic inflammation, mucus hypersecretion, and airway hyperresponsiveness. The excessive activation of Th2 cells due to insufficient suppression of regulatory T cells (Tregs) is thought to play a major role in the initiation and development of allergic airway disease. Several studies have shown that stem cells provide a significant reduction in allergic airway inflammation and improve lung function in animal models. The immunomodulatory effects of stem cells in allergic airway disease may be mediated by the up-regulation of Tregs and increases in several soluble factors, such as prostaglandin E2, transforming growth factor-β, interleukin-10, and indoleamine 2, 3-dioxygenase. This review examines the current understanding of the immunomodulatory properties of stem cells and its therapeutic implication in allergic airway disease. Furthermore, we will discuss mechanisms by which stem cells inhibit allergic airway inflammation via immunomodulation from a Th2- to a Th1-biased response.

Detection and clinical significance of regulatory T cells in peripheral blood of elderly patients with type 2 diabetic peripheral neuropathy / 第二军医大学学报

Objective To investigate the changes of CD4+CD25+Foxp3+ regulatory T cell (Treg) in the peripheral blood of elderly patients with type 2 diabetic peripheral neuropathy (DPN) and its clinical implication. Methods Treg cells were detected by flow cytometry in the peripheral blood of elderly patients with type 2 DPN (DNP group, 20 cases), type 2 diabetes mellitus (T2DM) with no complications (T2DM group, 20 cases) and age-matched healthy volunteers (CON group, 20 cases). The results were statistically analyzed and compared between different groups. Results The numbers of CD4+CD25+Foxp3+ Treg in DPN group and T2DM group were significantly less than that in the healthy volunteers (P<0.05), and the number of Treg cells in DPN group was significantly less than that in T2DM group (P<0.05). The proportion of CD4+CD25+Foxp3+ Treg was negatively associated with Michigan neuropathy screening instrument (MNSI) scores in DPN group, but with no statistical significance; however, the proportion had a significantly negative correlation with Michigan diabetic neuropathy score (r=-0.339, P<0.05). Conclusion The number of CD4+CD25+Foxp3+ Treg is decreased in the peripheral blood of elderly patients with DPN, which may play a role in the development and progression of DPN in elderly patients.

Mesenchymal stem cells for treatment of experimental autoimmune encephalomyelitis and the immunoregulation mechanism / 第二军医大学学报

Objective To observe the efficacy of mesenchymal stem cells (MSC) for treatment of experimental autoimmune encephalomyelitis (EAE) and the related immunoregulation mechanism. Methods EAE models of C57BL/6 mice were established with MOG35-55 /CFA emulsion and the EAE mice were randomly divided into MSC treatment group and EAE model group. MSCs were purified and cultured from the bone marrow of GFP-C57BL/6 mice. On the 15th day of EAE model establishment, MSCs (1 X 106/400 µL) were injected via the tail vein for the MSC treated group, and 400 µL PBS were injected for the EAE model mice. Then we recorded the clinical scores and analyzed the pathological changes of spinal cord so as to evaluate the state of EAE in each group. The peripheral blood tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-4 (IL-4) and transforming growth factor-β (TGF-β) cytokines were evaluated by ELISA. The percent of CD4+Foxp3+ Treg cells and GFP+ cells were analyzed by Flow cytometry. Results Compared with EAE model group, the MSC treated group had significantly decreased clinical score (P < 0. 05) and less T-cell infiltration in the spinal cord. MSC treated group had increased peripheral blood IL-4, TGF-β and decreased IFN-γ, TNF-α levels. Moreover, the MSC treated group also had greatly increased ratio of CD4+ Foxp3+ T cells (Treg). The MSC cells almost disappeared 10 days after transplantation. Conclusion Mesenchymal stem cell transplantation can effectively treat experimental autoimmune encephalomyelitis in mice. MSC may exert immunoregulation effect through increasing blood anti-inflammatory cytokines (IL-4, TGF-β) and decreasing proinflammatory cytokines (IFN-γ, TNF-α), which can prompt the naive cell differentiation into Treg cells. The effect of MSC remains even after disappearing for a certain time period.

Correlation between Treg/Th17 and Traditonal Chinese Medicine syndrome differentiation classification in patients with immune thrombocytopenia / 第二军医大学学报

Objective To explore the role of Treg/Th17 cell ratio imbalance in the pathogenesis of idiopathic thrombocytopenic purpura (ITP) patients with different Traditional Chinese Medicine (TCM) syndrome differentiation classifications: bleeding due to blood-heat, Yin deficiency with fire hyperactivity and Qi deficiency-caused bleeding. Methods A total of 92 patients were divided into the bleeding due to blood-heat group (n=30), Yin deficiency with fire hyperactivity group (n=31) and Qi deficiency-caused bleeding group (n=31) according to the TCM syndrome differentiation classification. The peripheral blood samples were obtained from the patients and 30 volunteers served as healthy controls. The percentages of Treg cells and Th17 cells in the blood samples were analyzed by flow cytometry, and the mRNA levels of Foxp3 and ROR-γt were analyzed by RT-PCR. Results The percentages of Treg cells in the peripheral blood of 3 different TCM syndrome ITP groups were significantly lower than those of control group (P<0.05), that of the bleeding due to blood-heat group was significantly lower than that of Yin deficiency with fire hyperactivity group and Qi deficiency-caused bleeding group (P<0.05), and that of Qi deficiency-caused bleeding group was signficantly lower than that of Yin deficiency with fire hyperactivity group (P<0.05). The percentages of Th17 cells in peripheral blood of 3 different TCM syndrome ITP groups were significantly higher than that of control group(P<0.05), and that of Yin deficiency with fire hyperactivity group was signficanlty higher than that of Qi deficiency-caused bleeding group (P<0.05). The ratios of Treg/Th17 of the 3 different TCM syndrome ITP groups were significantly lower than that of control group (P<0.05), that of the bleeding due to blood-heat group was significantly lower than that of Qi deficiency-caused bleeding group and Yin deficiency with fire hyperactivity group (P<0.05), and that of Qi deficiency-caused bleeding group was significantly lower than that of Yin deficiency with fire hyperactivity group (P<0.05). The Foxp3 mRNA levels of 3 different TCM syndrome ITP groups were significantly lower than that of control group (P<0.05), and there were significant differences between each two groups by the pairwise comparison (P<0.05). While ROR-γ mRNA levels of the 3 different TCM syndrome ITP groups were significantly higher than that of the control group (P<0.01). Conclusion The decreased Treg cells in patients with ITP contributes to the development and progression of ITP. The imbalance of Treg/Th17 ratio may play a critical role in the pathogenesis of ITP. The distribution of the percentages of Treg cells, the ratio of Treg/Th17 and the mRNA level of Foxp3 in a increasing order is: bleeding due to blood-heat group < Qi deficiency-caused bleeding group < Yin deficiency with fire hyperactivity group.

Avaliação de células T regulatórias e pefil de ativação de linfócitos T na ceratoconjuntivite seca associada ao HTLV-1 / Regulatory T cells evaluation and T lymphocyte activation profile in keratoconjunctivitis sicca associated with HTLV-1

O HTLV-1 é o agente etiológico da leucemia /linfoma de células T do adulto (ATLL), da paraparesia espástica tropical/ mielopatia associada ao HTLV-1 (HAM/TSP) e da uveíte. Além destas, a ceratoconjutivite seca (CCS), doença multifatorial da lágrima e da superfície ocular, tem sido descrita com maior frequência em indivíduos infectados pelo HTLV-1. Assim como em outras doenças associadas, a CCS tem sido relacionada a uma elevada carga proviral. As células T regulatórias (Treg) são importantes na manutenção da homeostase do sistema imunológico e um comprometimento da imunorregulação exercido por elas pode contribuir para o ambiente inflamatório observado na CCS. Este estudo objetivou avaliar os linfócitos Treg de pacientes com CCS associada à infecção pelo HTLV-1. Foram realizados ensaios de imunofenotipagem por citometria de fluxo para avaliar a frequência de linfócitos T ativados (HLA-DR+) e de células T CD4+ e CD8+ regulatórios (FOXP3+), bem como a produção de IL-10 e TGF-β por estas células. Foram avaliados 37 pacientes infectados pelo HTLV-1 e assintomáticos para HAM/TSP, sendo 27 com diagnóstico positivo para a manifestação ocular (CCS), 10 com diagnóstico negativo (ASS), além de 17 voluntários não infectados pelo vírus (NI). As frequências de linfócitos T CD4+FOXP3+, CD8+FOXP3+, CD4+HLA-DR+ e CD8+HLA-DR+ foram significativamente maiores nos grupos CCS e ASS, quando comparados aos indivíduos não infectados. Quanto à produção das citocinas imunossupressoras, foi observada uma maior frequência de linfócitos T CD4+FOXP3+ duplo produtores de IL-10 e TGF-β no grupo CCS quando comparado ao grupo ASS. Com relação aos linfócitos CD8+FOXP3+, o grupo CCS apresentou uma maior frequência de células mono produtoras de IL-10 quando comparado ao ASS. Nossos resultados sugerem que a menor frequência de células Treg CD8+ produtoras de TGF-β em indivíduos infectados pelo HTLV-1 com CCS, pode contribuir para a intensificação da ativação celular e fisiopatologia da doença.

HTLV-1 is the causative agent of leukemia/lymphoma adult T-cell (ATLL), tropical spastic paraparesis / myelopathy associated with HTLV-1 (HAM / TSP) and uveitis. In addition, keratoconjunctivitis sicca (KCS), a multifactorial disease of the tear and of the ocular surface, has been more frequently reported in patients infected with HTLV-1. As for other HTLV-1-associated diseases, KCS has been related to a high proviral load. Regulatory T (Treg) cells are important in maintaining the homeostasis of the immune system. An impairment in the immunoregulation function of Treg may contribute to the inflammatory environment observed in the KCS. This study aimed to evaluate the Treg cells of patients with KCS associated with HTLV-1. Frequency of activated T cells (HLA-DR+) and CD4+ and CD8+ Treg cells (FOXP3+), as well as IL-10 and TGF-β production by Treg were quantified using flow cytometry. Thirty-seven HTLV-1 individuals were included (27 asymptomatic for HAM/TSP with positive diagnosis of ocular manifestation (KCS), 10 with negative diagnosis (ASS - asymptomatic). Seventeen non-infected individuals were included as controls (NI). The frequencies of CD4+ FOXP3+ T cells, CD8+FOXP3+, CD4+HLA-DR+ and CD8+HLA-DR+ were significantly higher in KCS and ASS groups when compared to non-infected individuals. As the production of immunosuppressive cytokines, a higher frequency of CD4+ FOXP3+ double producers of IL-10 and TGF-β in the KCS group was observed when compared to group ASS. Regarding the CD8+FOXP3+ lymphocytes, the KCS group had a higher frequency of mono cells producing IL-10 when compared to the ASS. Our results suggest that the lower frequency of Treg cells CD8+ TGF-β-producing in individuals infected with HTLV-1 with KCS, may contribute to the intensification of cellular activation and pathophysiology of the disease.

Role of IL-2 1 in the Pathogenesis of Primary Biliary Cirrhosis / 现代检验医学杂志

Objective To investigate the pathogenesis of interleukin-21 (IL-21)role in primary biliary cirrhosis (PBC).Meth-ods 38 cases of PBC were selected for the study.35 caeses of healthy subjects were selected as the control group in January 2012 to January 2014 from Shenzhen Fuyong People’s Hospital of Baoan District and the Third People’s Hospital of Shenz-hen.The levels of IL-21,IL-17 of two groups were measured with Enzyme-linked immunosorbent assay (ELISA).The levels of T helper lymphocytes 17 (Th17),regulatory T cells (Treg)cells ratios of two groups were measured with flow cytome-try.The relationship of IL-2 1 and liver function were analyzed with pearson correlation.Another analysis of pyruvate through the establishment of in vitro deaminase complex E2 subunit (PDC-E2)peptides relationship with Th17 and Treg cell.Results The levles of IL-17 (18.23±4.12)ng/L,IL-21 (512.25±18.25)ng/L,Th17 (3.98±0.98)%,Th17/Treg (39.25±3.25)% of PBC group were significantly higher than the control group (7.89±3.26)ng/L,(212.98±21.52)ng/L,(0.92±0.21)% and (12.98±3.12)%,while the PBC group Treg (3.11±0.85)% were lower than the control group (8.36±1.12)%.IL-21 were positive correlation with IL-17,Th17 (t=8.886,7.021,4.448,10.112,5.125,P<0.05).In vitro experiments showed that the addition of IL-21 in the culture system,the levels of Th17 (2.41±0.82)%,Th17/Treg cells(32.98±3.12)% were significantly higher than the control group(0.98±0.21)%,(12.25±3.25)%,while Treg (4.75 ±0.68)% were lower than the control group(11.85±0.96)%,the difference was statistically significant (8.521,6.489, 8.236,P<0.05).Conclusion IL-21 levels increased with the pathogenesis of PBC has a close relationship,which can pro-mote the reaction of PDC-E2 Th1 7/Treg immune imbalance caused thereby occurs mediated by PBC.

Change and significance of the regulatory T lymphocytes expression in rats model of chronic abacterial prostatitis / 中华泌尿外科杂志

Objective To investigate the change and significance of regulatory T lymphocytes in peripheral blood of rats model of chronic abacterial prostatitis.Methods Twelve Wistar rats with weight of approximate 390 g were randomly divided into two groups,model group and control group.Rats in the model group was injected subcutaneously 17β-estradiol(0.25 mg/day,for 30 days) after castration to establish rat model of chronic abacterial prostatitis.Flow cytometry was applied to detect the frequency of CD+4 CD+25 cells and CD+8CD-28 cells in peripheral blood of rats after model establishment.Results Compared with control group (11.63±1.36)％,the proportion of CD+4 CD+25T lymphocytes in model group (7.90±1.74)％ significanlty decreased (P＜0.01).Compared with control group (24.64±4.76)％,the proportion of CD+8CD-28T lymphocytes in model group (17.18±2.83) ％ also significantly decreased (P＜0.01).Conclusions The changes of the ratio of CD+4 CD+25T lymphocytes and CD+8CD-28T lymphocytes in peripheral blood of rats model of chronic abacterial prostatitis provided evidences for pathogenic mechanism of regulatory T lymphocytes participating in the development of chronic abacterial prostatitis.

Effects of type-3-polysaccharide on airway inflammation and Treg/Thl7 cells in bronchial asthmatic mice / 第二军医大学学报

Objective To investigate the effects of type-3-polysaccharide (T3P) on airway inflammation and Treg/Th17 cells in bronchial asthmatic mice. so as to explore the immunoregulation mechanism of T3P in asthmatic mice. Methods BALB/c mice were randomly divided into 3 groups, with 8 mice in each group. The control group received PBS treatment, the asthma group were sensitized and challenged with ovalbumin(OVA),and the T3P group received a pretreatment with T3P by subcutaneous injection before sensitization with OVA. The pulmonary histological changes were observed and differential cell counts in bronchoalveolar lavage fluid (BALF) were performed. Serum OVA-IgE and IFN-γ IL-4, IL-17 and IL-10 levels in the BALF were detected by enzyme-linked immunosorbent assay (ELISA). The levels of Foxp3 and RORγt mRNA expression were measured by real-time fluorescence-based quantitative PCR. The proportions of Treg and Th17 cells in CD4+cells were assessed by flow cytometric analysis. Results The inflammatory degree of pulmonary tissue, serum OV-IgE, and total cell number, proportion of eosinophils, and IL-4, IL-17 levels in BALF were significantly lower in T3P group than in the asthma group (P<0. 05). TheBALF levels of IFN-y and IL-10, Foxp3 mRNA expression and proportion of Treg cells among CD4+ cells were significantly higher in the T3P group than in the asthma group (P<0. 05), while RORut mRNA expression and the proportion of Th17 cells among CD4+ cells were significantly lower than in the asthma group (P<0. 05). The count of BALFeosinophils was negatively correlated with the ratios of Foxp3-mRNA/RORut~mRNA and Treg/Th17 in asthma and T3P groups (P<0. 05). Conclusion T3P may inhibit airway inflammation by inhibiting OVA-IgEand regulating Th1/Th2 and Treg/Th17 cell balance in asthmatic mouse model.

Linfócitos T CD4+CD25+ e a regulação do sistema imunológico: perspectivas para o entendimento fisiopatológico da sepse / CD4+CD25+ T lymphocytes and regulation of the immune system: perspectives for a pathophysiological understanding of sepsis

A resposta inflamatória sistêmica representa o evento patogênico central da sepse, subjazendo às manifestações clínicas e aos achados laboratoriais presentes nos enfermos. Inúmeras pesquisas têm demonstrado que os linfócitos T CD4+CD25+ - também conhecidos como células T reguladoras (Treg) - participam dos processos de desenvolvimento da sepse, em virtude de sua capacidade de suprimir a resposta imune. Com base nessas ideias, propôs-se, no presente artigo, a discussão do papel dos linfócitos Treg na sepse, com base na revisão da literatura com estratégia de busca definida (LILACS, PubMed e SciELO), tendo em vista duas abordagens principais: a participação dessas células nos processos de inflamação e imunidade, e as perspectivas de investigação fisiopatológica computacional da condição mórbida.

The systemic inflammatory response represents the core pathogenic event of sepsis, underlying clinical manifestations and laboratory findings in patients. Numerous studies have shown that CD4+CD25+ T lymphocytes, also known as regulatory T lymphocytes (Treg), participate in the development of sepsis due to their ability to suppress the immune response. The present article discusses the role of Treg lymphocytes in sepsis based on a specific search strategy (Latin American and Caribbean Health Sciences / Literatura Latino-americana e do Caribe em Ciências da Saúde - LILACS, PubMed, and Scientific Electronic Library Online - SciELO) focusing on two main topics: the participation of Treg cells in inflammation and immunity as well as perspectives in the computational physiological investigation of sepsis.

Inmunopatogenia de las enfermedades autoinmunes / Immunophat ogenesis of aut oimmune diseases

Las enfermedades autoinmunes son patologías de gran complejidad clínica, difícil diagnóstico y complejo tratamiento cuya etiología permanece aún desconocida pese a los múltiples avances realizados en los últimos años. En la génesis de estas enfermedades participan múltiples factores que conflyen entre sí para dar origen a cada una de las patologías autoinmunes conocidas, sean estas órgano-específicas o sistémicas. Entre estos elementos se incluyen la pérdida de los mecanismos de tolerancia, factores de susceptibilidad genética (polimorfismos HLA, genes no HLA y mecanismos epigenéticos), factores ambientales (agentes vivos de enfermedad, agentes inorgánicos, hormonas y otros) y factores inmunológicos (linfocitos reguladores, citoquinas y moléculas coestimulatorias, entre otros). La identificación de estos factores permitirá mejorar el conocimiento de los variados mecanismos que median estas complejas enfermedades, facilitando no sólo el entendimiento de su etiología sino también perfeccionar las herramientas terapéuticas para enfrentarlas.

Autoimmune diseases are pathologies of great clinical complexity, difficult diagnosis and treatment complex which etiology still remains unknowns despite the many advances made in recent years. In the genesis of these diseases involves multiple factors that converge together to give rise to each of the autoimmune diseases knows, whether organ specific or systemic. These elements include loss of tolerance mechanisms, genetic susceptibility factors (HLA polymorphisms, genes non-HLA and epigenetic mechanisms), environmental factors (living agents of disease, inorganic agents, hormones, etc.) and immunologic factors (regulators lymphocyte, cytokines, costimulatory molecules and others). Identifying these factors will improve the knowledge of the various mechanisms that mediate these complex diseases facilitating not only the understanding of the etiology but also improve the therapeutic tools to address them.

Role of PLC/PIP2 signaling pathway in radiation-induced NRP1 + Treg cells / 中华放射医学与防护杂志

Objective To explore the role of PLC/PIP2 signal pathway in the changes of mouse thymus CD4 + CD25 + NRP1 + Treg and TGF-β1 after different doses of X-ray irradiation Methods 36 ICR mice were randomly divided into 6 groups according to the irradiation doses of 0,0.5,1.0,2.0,4.0 and 6.0 Gy,respectively.Flow cytometry was used to detect the expression of NRP1 + Treg,and ELISA was used to detect the expression of TGF-β1 in mouse thymocytes at 16 h post-irradiation.The EL-4 cells were irradiated by X-rays at the dose of 4.0 Gy after co-cultured with the PMA and TMB-8 for 2 hours.Flow cytometry was used to detect the expression of NRP1 + Treg,and ELISA was used to detect the changes of TGF-β1 at 48 h post-irradiation.Results The NRP1 + Treg appeared a transient decrease at both 0.5 and 1.0 Gy irradiation and reached its valley value at 1.0 Gy (t =6.96,P ＜ 0.01),then showed a dosedependent increase and reached its peak at 6.0 Gy (t =6.70,P ＜ 0.01).The TGF-β1 level decreased after 0.5 Gy X-rays (t =12.53,P ＜0.01),then increased at a dose-dependent manner and reached its peak at 4.0 Gy (t =10.40-19.56,P ＜ 0.01).Compared with the sham-irradiation,NRP1 + Treg was decreased significantly after PMA treatment (t =3.06,P ＜ 0.01),while it was up-regulated significantly after irradiation in the presence of PMA (t =8.27,P ＜ 0.01).TGF-β1 was reduced in the presence of PMA with or without irradiation (t =10.46-39.69,P ＜ 0.01).NRP1 + Treg and TGF-β1 were increased significantly after TMB-8 treatment (t =5.53-44.26,P ＜ 0.01).Conclusions NRP1 + Treg cells and TGF-β1 were up-regulated after a high dose radiation,and the PLC/PIP2 signal pathway may participate in the regulation.

Foxp3 expression is associated with aggressiveness in differentiated thyroid carcinomas

OBJECTIVES: Forkhead box P3 (FoxP3) expression has been observed in human cancer cells but has not yet been reported in thyroid cells. We investigated the prognostic significance of both FoxP3 expression and intratumoral FoxP3+ lymphocyte infiltration in differentiated thyroid carcinoma cells. METHODS: We constructed a tissue microarray with 385 thyroid tissues, including 266 malignant tissues (from 253 papillary thyroid carcinomas and 13 follicular carcinomas), 114 benign lesions, and 5 normal thyroid tissues. RESULTS: We determined the expression of FoxP3 in both tumor cells and tumor-infiltrating lymphocytes using immunohistochemical techniques. Cellular expression of FoxP3 was evident in 71% of benign and 91.9% of malignant tissues. The nuclear and cytoplasmic expression patterns were quantified separately. A multivariate logistic regression analysis indicated that cytoplasmic FoxP3 expression is an independent risk factor for thyroid malignancy. Cytoplasmic FoxP3 staining was inversely correlated with patient age. Nuclear FoxP3 staining was more intense in younger patients and in tumors presenting with metastasis at diagnosis. FoxP3+ lymphocytes were more frequent in tumors smaller than 2 cm, those without extrathyroidal invasion, and in patients with concurrent chronic lymphocytic thyroiditis. CONCLUSIONS: We demonstrated FoxP3 expression in differentiated thyroid carcinoma cells and found evidence that this expression may exert an important influence on several features of tumor aggressiveness.

Mechanism of experimental autoimmune encephalomyelitis in Lewis rats: recent insights from macrophages / 대한해부학회지

Experimental autoimmune encephalomyelitis (EAE) in Lewis rats is an acute monophasic paralytic central nervous system disease, in which most rats spontaneously recover from paralysis. EAE in Lewis rats is induced by encephalitogenic antigens, including myelin basic protein. EAE is mediated by CD4+ Th1 cells, which secrete pro-inflammatory mediators, and spontaneous recovery is mediated by regulatory T cells. Recently, it was established that classically activated macrophages (M1 phenotype) play an important role in the initiation of EAE, while alternatively activated macrophages (M2 phenotype) contribute to spontaneous recovery from rat EAE. This review will summarize the neuroimmunological aspects of active monophasic EAE, which manifests as neuroinflammation followed by neuroimmunomodulation and/or neuroprotection, with a focus on the role of alternatively activated macrophages.

Application of Regulatory T Cells in Transplantation Field / 대한이식학회지

The development of immunosuppressant treatments has enabled remarkable progress in the tissue and organ transplantation field by helping to prevent acute graft rejection. However, complications related to transplantation, such as infection by bacteria and viruses, and the occurrence of cancers resulting from prolonged immune suppression are major obstacles to overcome. Therefore, transplantation immunology research efforts should focus on the induction of donor-specific immune tolerance which preserves patient immune competence which promotes infection and cancer surveillance. Additionally, lifelong administration of immunosuppressants should be forgone in preference to short term therapies. In the 1990s, Dr. Shimon Sakaguchi identified the CD4+CD25+ regulatory T cells which develop in the thymus, and demonstrated that these cells play crucial roles in the maintenance of immune self tolerance. Studies which followed proved that these regulatory T cells are important to the control of autoimmune disease and prevention of graft rejection. Regulatory T cells have also been found to induce immune tolerance in rodent models. In this review, we discuss several considerations for the use of regulatory T cell therapy in the clinical transplantation field.