All-transRetinoic Acid Regulates Th1/Th2 Balance in CD4+T cells When GATA-3 is Deficient

The essential effect of vitamin A on immune function occurs through various mechanisms including direct effect on ThloTh2 balance modulation. However, it is unclear whether or not vitamin A can regulate Thl-Th2 balance under a strong Thl-polarizing condition. Therefore, the purpose of our study was to examine the effect of vitamin A metabolite allotrans retinoic acid （ATRA） on ThloTh2 differentiation in CD4~ T cells under GATA-3 deficiency, which can induce Thl-polarizing condition. In the present study, GATA-3 deficiency T cells were induced by siRNA and checked by real-time quantitative PCR and western blot. GATA-3 deficiency CD4＋ T cells and normal CD4＋ T were treated for 48 h with or without ATRA.

Myricetin alleviates ovalbumin-induced allergic rhinitis in mice by regulating Th1/Th2 balance

Objective:To evaluate the effect of myricetin on ovalbumin(OVA)-induced allergic rhinitis in mice.Methods:Mice were sensitized and challenged using OVA(5%,500 mL)intraperitoneally and intranasally,respectively,on an alternative day for 14 days,followed by administration of myricetin(50,100,and 200 mg/kg)till day 21.Nasal symptoms,biochemical parameters,protein expressions,and histopathology were observed.Results:OVA-induced increased nasal symptoms including rubbing,sneezing,and discharge were significantly reduced by myricetin(100and 200 mg/kg)(P<0.05).Myricetin also protected against histamine challenge and attenuated elevated serum immunoglobulin E(IgE;total and OVA-specific),total IgG1,andβ-hexosaminidase levels,as well as leukotriene C4 and interleukins levels in nasal lavage fluid(P<0.05).Western blot analysis showed that myricetin significantly upregulated the protein expression of T-box expressed in T cells,while downregulating the protein expression of GATA binding protein 3,NF-κB,and IκB-α(P<0.05).Additionally,OVA-induced histopathological abberations in the nasal mucosa was markedly ameliorated by myricetin treatment(P<0.05).Conclusions:Myricetin exerts anti-allergic effects against OVAinduced allergic rhinitis via regulating Th1/Th2 balance.

Effects of seasonal changes on T-helper 1/T-helper 2 immune balance and eczema onset in rats

Objective: To explore the impact of seasonal changes on the T-helper 1(Th1)/T-helper 2(Th2) immune balance of the body in order to elucidate the internal immunological mechanisms underlying seasonal eczema.Methods: Eighty 4-week-old male Sprague-Dawley rats were divided into 5 season groups kept in corresponding season simulation environments, and subdivided into the model group and the control group. The Eczema Area and Severity Index(EASI) and scratching frequency were evaluated. The serum levels of immunoglobulin E(IgE), interleukin-2(IL-2), interleukin-12(IL-12), and interferon-gamma(IFN-γ), interleukin-4(IL-4), interleukin-25(IL-25), and interleukin-31(IL-31), and melatonin(MT), as well as the MT receptor(MTR) levels in the spleen, were detected by enzyme-linked immunosorbent assay analysis. The mRNA expression levels of T-bet and GATA3 were detected by quantitative real-time polymerase chain reaction.Results: EASI scores and the scratching frequency of the model group were higher in the long summer than in the other 4 seasons. Meanwhile, the serum levels of IgE and the Th2 cytokines IL-4, IL-25, and IL-31, as well as GATA3 mRNA expression levels, were high during the long summer in both groups. However, the serum levels of the Th1 cytokines IL-2, IL-12, and IFN-γ, as well as MT, MTR, and T-bet mRNA levels, were lower during the long summer. In all 5 seasonal groups(spring, summer, long summer, autumn, and winter), the levels of all immune factors, especially IL-4 and IL-31, were higher in the model group than those in the control group,while the concentrations of MT and MTR were lower.Conclusion: Under long light, hot, and humid conditions in the long summer, the body is more likely to suffer from Th2-dominated immune imbalance. This is the internal mechanism behind the high incidence and severity of eczema during the long summer. MT and MTR play a key role in the seasonal onset of eczema.

Analysis of Specific Th1/Th2 Helper Cell Responses and IgG Subtype Antibodies in Anti-CD4 Monoclonal Antibody Treated Mice with Autoimmune Cardiomyopathy

The cytokine repertoire of ADP/ATP carrier-specific humoral immune responses and the cytokine-dependent anti-ADP/ATP carrier antibody IgG subclasses were examined in a cohort of ADP/ATP carrier-immunized BALB/c mice treated with anti-CD4 monoclonal antibody. Eighteen male BALB/c mice （6–8 weeks old） were randomized into 3 groups： dilated cardiomyopathy （DCM） group, DCM-tolerance （Tol） group and control group. The mice in DCM group were immunized with the peptides derived from human ADP/ATP carrier protein for 6 months and mice in the control group were sham-immunized, while the mice in DCM-Tol group were immunized with ADP/ATP carrier protein and anti-CD4 McAb simultaneously. Serum autoantibody against ADP/ATP carrier and IgG subclasses were measured by ELISA, intracellular cytokines IFN-γ and IL-4 of Th cells were moni- tored with flow cytometry, and splenic T cell cytokines IFN-γ, IL-2, IL-4 and IL-6 were detected by using real-time fluorescent quantitative PCR. The results showed that the autoantibody against ADP/ATP carrier was found in all mice in DCM group, and the antibody level, serum IgG1 and IgG2a subclasses, cytokines in T cells and Th cells were all elevated in DCM group, as compared with those in control group （P〈0.01）. On the other hand, in DCM-Tol group, the autoantibody level and contents of all the cytokines were significantly different from those in DCM group （P〈0.01）, and were close to those in control group. And the levels of IgG1, IgG2a, IgG2b and IgG3 were influenced, to varying degrees, by anti-CD4 McAb as compared with those in DCM group. All these four types of IgG subclasses were substantially decreased in DCM-Tol group as compared with DCM group. It is concluded that the treatment with anti-CD4 McAb could prevent the activation of T cells, reverse the abnormal secretion of cytokines and the imbalance between Th1/Th2 cell subsets and abnormal production of autoantibody against ADP/ATP carrier, and eventually avoid myocardial injuries.

Effect of Th1/Th2 balance on serum prostaglandin,COX-2,AVP and OT in primary dysmenorrhea rats with cold coagulation and blood stasis syndrome

Objective:Explore the role of Th1/Th2 balance in the pathogenesis of primary dysmenorrhea through study the effect of Th1/Th2 balance drift on serum prostaglandin F_(2α),prostaglandin,6-keto-prostaglandin F_(1α),TXB2,COX-2,AVP and OT in Primary Dysmenorrhea Rats with cold coagulation and blood stasis syndrome.Method:The rat model of primary dysmenorrhea with cold coagulation and blood stasis syndrome was established.On the basis of this model,Th1 shift model and Th2 shift model were established.The rats were divided into blank group,model group,Th1 shifting group and Th2 shifting group.The writhing reaction of rats in each group was compared,and the contents of PGF_(2α),PGE_(2),6-keto-PGF_(1α),TXB2,COX-2,AVP and OT in serum were detected.Results:Writhing latency:There was no significant difference between Th1 and Th2 shifting groups(P>0.05),Th2 shifting group was shorter than Th1 shifting group(P<0.05).Writhing times in 30 minutes:Compared with the model group,the writhing times of Th1 shifting group decreased significantly(P<0.01),while Th2 shifting group increased significantly(P<0.01).Compared with the blank group,the contents of PGF_(2α),TXB2,COX-2,AVP and OT were increased(P<0.05,P<0.01),and the contents of 6-keto-PGF_(1α)and PGE_(2)were decreased(P<0.01).Compared with the model group,the contents of PGF_(2α),TXB2,COX-2 and AVP in Th1 shifting group were decreased(P<0.05,P<0.01),and the contents of 6-keto-PGF_(1α)and PGE_(2)were increased(P<0.01),while the contents of PGF_(2α),TXB2,COX-2,AVP and OT in Th2 shifting group were increased(P<0.01),and the contents of 6-keto-PGF_(1α)and PGE_(2)were decreased(P<0.01).Conclusion:The intervention of Th1/Th2 cell balance on the expression of PGF_(2α),PGE_(2),6-keto-PGF_(1α),TXB2,COX-2,AVP and OT may be one of the pathogenesis of PD with cold coagulation and blood stasis syndrome.

Effect of glycyrrhetinic acid on Th1/Th2 balance in cough variant asthma mice

Objective:To evaluate the therapeutic effect of Glycyrrhetinic Acid on cough variant asthma(CVA)mice and to investigate the possible mechanism in reducing lung inflammation.Methods:48 young female Balb/c mice were divided into Control,CVA,Prednisone Acetate,Glycyrrhetinic Acid high-dose,Glycyrrhetinic Acid middle-dose and Glycyrrhetinic Acid lowdose groups randomly,with 8 mice in each group.The CVA mice model was established by ovalbumin(OVA)sensitization and OVA challenge,the animal asthma behavior was observed after drug administration,and the index of the lung of mice were recorded.The level of OVAsIgE in the bronchoalveolar lavage fluid(BALF)was tested by ELISA.The pathological changes of the lung tissue were observed by Hematoxylin and Eosin(H&E)staining.The protein expressions of T-bet,IFN-γ,Gata3,IL-4 and IL-13 in the lung tissue were determined by Western blot.Results:Compared with the CVA group,the index of lung of mice,the OVA-sIgE level in BALF and expression levels of Th2-related factor in the lung tissue of mice in Prednisone Acetate and Glycyrrhetinic Acid groups were significantly decreased(P<0.05 or P<0.01),the infiltration of inflammatory cells in the lung tissue was reduced,while expressions of Th1-related factor in the lung tissue was significantly increased(P<0.05 or P<0.01).Conclusion:Glycyrrhetinic acid has therapeutic effect on CVA mice,the underlying mechanism of Glycyrrhetinic acid alleviating lung impairment and airway inflammation may be associated with mediating the Th1/Th2 imbalance in the lung tissue.

Correlation of serum TGF-β1 content with liver fibrosis and Th1/Th2 immune levels in patients with chronic hepatitis b

Objective:To investigate the correlation of serum TGF-β1 content with liver fibrosis and Th1/Th2 immune levels in patients with chronic hepatitis b.Methods: A total of 178 patients who were diagnosed with chronic hepatitis b in our hospital between February 2015 and August 2017 were selected as chronic hepatitis b group, and 100 healthy volunteers who received physical examination in our hospital during the same period were selected as normal control group. The differences in serum levels of TGF-β1, liver fibrosis indexes and Th1/Th2 cytokines were compared between the two groups, and the Pearson test was adopted to evaluate the correlation between serum TGF-β1 content and disease severity in patients with chronic hepatitis b.Results: Serum TGF-β1 level of chronic hepatitis b group was higher than that of normal control group;serum liver fibrosis indexes CⅣ, PC-Ⅲ, HA and LN levels were higher than those of normal control group;serum Th1 cytokines IFN-γ and IL-12 levels were lower than those of normal control group whereas Th2 cytokines IL-4 and IL-13 levels were higher than those of normal control group. Pearson test showed that the serum TGF-β1 level in patients with chronic hepatitis b was positively correlated with the degree of liver fibrosis and Th1/Th2 immune imbalance.Conclusions: Serum TGF-β1 expression is abnormally high in patients with chronic hepatitis b and directly correlated with the degree of liver fibrosis and immune imbalance.

Effect of dexmedetomidine on Th1/Th2 cytokine and immune function in patients undergoing radical mastectomy

Objective: To investigate the effect of dexmedetomidine on Th1/Th2 cytokines and immune function in patients with breast cancer after radical mastectomy. Methods: In our hospital from July 2016 to July 2017 undergoing radical mastectomy for breast cancer were studied in 79 patients, were randomly divided into observation group and control group. Two groups of patients with routine preoperative preparation, monitoring blood pressure, electrocardiogram, heart rate, pulse, oxygen saturation, establish vein channel, using propofol, remifentanil, vecuronium induced anesthesia, observation group before induction of anesthesia, dexmedetomidine 1 μg/kg, 10 min after infusion, followed by 0.5 μg/kg/h continuous infusion to the end of the operation, the control group with normal saline continuous infusion till the end of the operation. Two groups of patients before induction of anesthesia (T0), at the end of operation (T1), 6 h after operation (T2), 24 h after operation (T3), 72 h after operation (T4) from peripheral venous blood determination of interleukin-2 by ELISA method (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon gamma (IFN-γ), calculated IFN-γ/IL-4 in T0, T2, T3, T4from peripheral blood. CD3+, CD4+, CD8+, NK cells were determined by flow cytometry and CD4+/CD8+ were calculated. Results: Two groups of IL-2 and IFN- in T1, T2, T3gamma, T4is higher than T0, IL-10 less than T0, and the observation group IFN-γ/IL-4 is higher than T0, the control group was lower than that of T0when compared with T0significant difference, 2 in group IL-4 had no obvious changes were observed in group IL-2;IFN-γ, IFN-γ/IL-4 in T1, T2, T3, T4higher than the control group, IL-10 was lower than the control group, significant difference between the 2 groups. CD3+, CD4+, CD4+/CD8+, NK cells in T2group was lower than that of T0, T3, and CD8+ had no obvious change, compared with T0significant difference;the observation group CD3+, CD4+, CD4+/CD8+, T2, T3in NK cells was higher than the control group, significant difference between the 2 groups. Conclusion: Dexmedetomidine can inhibit the stress response during the perioperative period of radical mastectomy, correct the balance disorder of Th1/Th2, improve the level of T lymphocyte subsets, and exert better immune protection function.

Immune response to inactivated bacterial vector carrying the recombinant K39 antigen of Leishmania infantum in mice

Objective:To evaluate the immunological response elicited by an inactivated bacterial vector carrying the K39 antigen of Leishmania infantum,and a purified antigen.Methods:Mice were subjected to the following treatments:(1)Purified recombinant K39(rK39)protein at a 20μg dose with complete Freund’s adjuvant;(2)Inactivated Escherichia coli(BL21 DE3)carrying the K39 protein at an equivalent total protein content of 200μg;(3)Inactivated bacteria lacking the K39 protein;(4)Non-immunized control animals.Serological monitoring was performed.All groups were challenged by intraperitoneal injection of 10^(7) Leishmania infantum promastigotes.After euthanasia,the liver and spleen were collected to analyze the levels of TNF,IFN-γ,IL-12,IL-4,and IL-10.Results:Mice immunized with purified rK39 or the inactivated bacterial vector carrying the K39 antigen of Leishmania infantum showed a long-lasting immune response with high levels of polyclonal antibodies specifically recognizing the recombinant proteins.The IgG1 subclass was the predominant immunoglobulin;however,the induction of IgG2a and the profile of cytokines produced were indicative of the induction of a mixed-type response.Conclusions:The inactivated bacterial vector carrying the K39 antigen,as well as the purified antigen can induce a long-lasting immune response in immunized mice,predominantly favouring a Th2 profile response.

Potential immunomodulatory role of sesamin in combating immune dysregulation associated with COVID-19

The ongoing outbreak of novel coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),has caused an unprecedented global health crisis.Development of a cure for this devastating disease is currently at full speed,with several vaccines against COVID-19 already authorized and administered.Currently,demand for these vaccines far exceeds supply.As such,boosting immunity represents a viable route to halt the rapid spread of SARS-CoV-2 and limit fatalities until vaccines become more readily and widely available.The use of phytochemicals appears to be a promising panacea.Sesamin,a lignan isolated from Sesamum indicum seeds,is known for its potent pharmacological properties,and is therefore hypothesized as a potential candidate in the therapeutic regimen against COVID-19.Herein,we highlight the confirmed therapeutic anti-inflammatory and immune-modulatory potential of sesamin against myriads of respiratory disorders,and tentatively suggest that sesamin may exert similar potent effects against COVID-19.Precisely,we speculate that sesamin may help alleviate COVID-19 via restoring Th1/Th2 balance and preventing inflammation and cytokine storm development.Additionally,we further support the promising role of sesamin against COVID-19 by underscoring the direct evidence,which suggests that sesamin may demonstrate promising inhibitory potential against three important SARS-CoV-2 targets,namely main protease,spike protein,and angiotensin-converting enzyme 2 receptor.Although preliminary,there is ample evidence to propose sesamin as a potential phytotherapeutic and prophylactic candidate against COVID-19.Further in vitro,in vivo,and preclinical studies are required to further substantiate the role of sesamin in the prevention and/or treatment of COVID-19.

Correlation of serum YKL-40 content with eosinophil function as well as Th1/Th2 and Treg/Th17 immunity in patients with allergic rhinitis

Objective: To discuss the correlation of serum YKL-40 content with eosinophil function as well as Th1/Th2 and Treg/Th17 immunity in patients with allergic rhinitis. Methods: A total of 310 patients with allergic rhinitis who were treated in the hospital between January 2015 and May 2017 were selected as allergic rhinitis group and 100 healthy volunteers who received physical examination in the hospital during the same period were selected as normal control group. The differences in serum YKL-40 content, eosinophil function as well as Th1/Th2 and Treg/Th17 cytokine contents were compared between the two groups. Pearson test was used to assess the correlation between serum YKL-40 content and disease severity in patients with allergic rhinitis. Results: Serum YKL-40 content of allergic rhinitis group was higher than that of normal control group;serum eosinophil function IgE and Eotaxin-2 contents of allergic rhinitis group were higher than those of normal control group;serum Th1 cytokines IFN-γand IL-12 contents were lower than those of normal control group while Th2 cytokines IL-4 and IL-13 contents were higher than those of normal control group;serum Treg cytokine TGF-β content was lower than that of normal control group while Th17 cytokine IL-17 content was higher than that of normal control group. Pearson test showed that serum YKL-40 content in patients with allergic rhinitis was directly correlated with the eosinophil function as well as Th1/Th2 and Treg/Th17 immunity. Conclusion: Serum YKL-40 content is abnormally high in patients with allergic rhinitis, and the specific expression is positively correlated with the disease severity.

Effects of intravenous inhalational anesthesia on the hemodynamic homeostasis as well as postoperative brain function and Th1/Th2 immunity in elderly patients with femoral neck fracture

Objective: To explore the effects of intravenous inhalational anesthesia on the hemodynamic homeostasis as well as postoperative brain function and Th1/Th2 immunity in elderly patients with femoral neck fracture. Methods: A total of 176 elderly patients with femoral neck fracture who received hip replacement in our hospital between July 2016 and June 2017 were divided into the total intravenous anesthesia group (n=86) and the intravenous inhalational anesthesia group (n=90) according to the anesthesia solution. The differences in introperative hemodynamic parameter levels as well as postoperative brain function index and Th1/Th2 cytokine contents were compared between the two groups. Results: During operation, hemodynamic parameters MAP and HR levels in intravenous inhalational anesthesia group were lower than those in total intravenous anesthesia group. 24 h after operation, serum brain function indexes MBP, S100B and NSE contents in intravenous inhalational anesthesia group were lower than those in total intravenous anesthesia group;serum Th1 cytokines IFN-γand IL-2 contents were higher than those in total intravenous anaesthesia group whereas Th2 cytokines IL-4 and IL-13 contents were lower than those in total intravenous anesthesia group. Conclusion: The intravenous inhalational anesthesia can effectively stabilize the intraoperative hemodynamics and reduce the postoperative brain function and Th1/Th2 immune function injury in elderly patients with femoral neck fracture.

Effect of Early Intensive Insulin Therapy on Immune Function of Aged Patients with Severe Trauma

This study examined the effect of intensive insulin therapy on immune function and inflammatory factors at the early phase after severe trauma. At day 1, 3, 5, 7 after admission, subsets of CD4＋ helper T lymphocytes （Th1/Th2） and human leukocyte antigen （HLA）-DR expression on CD14＋ monocytes were flow cytometrically measured. Levels of cytokines, including tumor necrosis factor-alpha （TNF-α）, interleukin-6 （IL-6）, interleukin-10 （IL-10） and other immunity markers, such as IgA, IgG, IgM, C3, C4 and C reaction protein （CRP） were examined in two groups. The results showed that TNF-α, IL-6 and CRP levels in the intensive insulin therapy group were significantly lower than those in the conventional therapy group, whereas IL-10 levels were substantially increased after intensive insulin therapy. C3 level at day 3, 5, 7 and C4 levels at day 5, 7 were lower in the intensive therapy group than in the conventional therapy group. Th1/Th2 ratios decreased gradually over time in both groups, and were much lower at day 3, 5, 7 in intensive therapy group. There were significant differences among day 3 to day 7 after admission in HLA-DR expression in CD14＋ monocytes. It was concluded that the intensive insulin therapy could decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in the elderly suffering from severe trauma, at the same time, with complement recovery being delayed. Moreover, intensive insulin therapy promoted immune suppression and, therefore, measures need be taken to address the issue.

Scutellarin alleviates complete freund’s adjuvant-induced rheumatoid arthritis in mice by regulating the Keap1/Nrf2/HO-1 pathway

Scutellarin(SCU)is a herbal flavonoid glucuronide with multiple pharmacological activities,including antioxidant,anti-inflammation,vascular relaxation,anti-platelet,and myocardial protection.However,the effect of SCU on complete Freund’s adjuvant(CFA)-induced rheumatoid arthritis(RA)had not been studied.In this study,we investigated the beneficial effects of SCU in the CFA-induced RA mice model and the anti-arthritic activity was evaluated by paw edema.Enzyme-linked immunosorbent assay(ELISA)was carried out to evaluate the plasma levels of immunoglobulin(Ig)G,IgE,tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,receptor activator of nuclear factor-κB ligand(RANKL),and osteoprotegerin(OPG).Histological slides were prepared from the harvested paws of mice to determine the pathological changes in the joints.The proportions of T helper type 1(Th1)and T helper type 2(Th2)cells of CD4+T lymphocyte subsets were analyzed by flow cytometry.The expression of Kelch-like ECHassociated protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)was analyzed using real-time quantitative PCR(RT-qPCR)and western blotting assays.The present study demonstrated that SCU prevented CFA-induced RA,and inhibited the expression of inflammation factors,IgG,IgE,TNF-α,IL-1β,and IL-6.While SCU also reduced the RANKL level,it increased OPG expression in RA mice.The Th1/Th2 ratio was significantly lower in mice treated with SCU.Additionally,HO-1 expression was reduced while the expression of Keap1 and Nrf2 was elevated following SCU treatment.Results provide preliminary evidence to employ SCU in arthritis treatment which might be related to the regulation of Th1/Th2 balance and the Keap1/Nrf2/HO-1 pathway.

Research progress of ICOSL/ICOS pathway in maternal-fetal immune tolerance

Co-signaling molecules are molecules whose ligands on the surface of cells interact with receptors on the surface of T cells to convey stimulatory or inhibitory signals to regulate immune responses.Co-signaling molecules play an important role in tumor and autoimmune diseases.Lately,studies have shown that co-signaling molecules are also involved in the regulation of maternal-fetal immune tolerance,and abnormalities of co-signaling molecules may lead to the imbalance of maternal-fetal immune tolerance,resulting in recurrent abortion,eclampsia and other pregnancy complications.ICOSL/ICOS is a ligand and receptor of costimulatory signals,which regulates maternal and fetal immune tolerance by participating in T cell differentiation and Th1 and Th2 cytokine secretion.Therefore,this article reviews the structure of ICOSL/ICOS,the distribution of ICOSL/ICOS at the maternal-fetal interface and its immune regulation during pregnancy,in order to provide new ideas for the future study of immunotherapy of pregnancy complications caused by abnormal co-signaling molecules.

Effects of combined immune therapy on survival and Th1/Th2 cytokine balance in rat orthotopic liver transplantation

Background The induction of immune tolerance and suppression of allograft rejection has become the focus in the study of liver transplantation. The effect of immune therapy with anti-CD40L mAb alone or in combination with cyclosporine A （CsA） on the recipient survival and Th1/Th2 cytokine profile was studied to elucidate its immunological mechanism and role in rat orthotopic liver transplantation.Methods The model of rat orthotopic liver transplantation was established by modified Kamada＇s technique. Recipients were divided into group A （control group）： SD→SD; group B （group of rejection）： SD→Wistar without any treatment; group C： SD→Wistar with CsA monotherapy from day 1 to day 5; and group D： SD→Wistar with CsA from day 1 to day 5 and anti-CD40L mAb on day 0 and day 2. The survival of the recipients in all groups was observed and ELISA technique was used to detect the level of cytokines in peripheral blood on post-transplant day 7.Results The survival period of recipients in groups A （〉60 days） and D （〉60 days） was significantly longer than that in group B （13.8±2.4 days）. The serum levels of interleukin 2 （IL-2） and interferon y in group B were significantly higher than those in other groups; the level of tumor necrosis factor a was higher but not statistically significant. In contrast, the serum levels of IL-4 and IL-10 in group D were elevated more significantly than those in group B （P〈0.05）.Conclusions Combined immune therapy can prolong the survival of al log rafts. Increased expression of Th2 cytokines, which is closely related to the induction of tolerance and suppression of rejection, is beneficial to the long-term survival of recipients and allografts.

High mobility group box 1 protein(HMGB1)as an immune-modulating factor for polarization of human T lymphocytes

Objective This study was performed to investigate the effect of high mobility group box-1 protein(HMGB1)on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells(PBMCs)were isolated,then rhHMGB1 was added to PBMCs.Four-color flow cytometric(FCM)analysis was used for the measurement of intracellular cytokine including interleukin IL-4 and interferon IFN-?ELISA kits were employed for the determination of IL-2 and sIL-2R protein levels in cell culture supernatants.Results(1)Different stimulating time and dosage of rhHMGB1 did not alter the number of IFN-αpositive cells(Th1).rhHMGB1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th1 to Th2.(2)Compared with the untreated cells,when the cells were coincubated with rhHMGB1(10-100ng/ml)for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGB1.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.

Hydrolyzed seawater pearl tablet modulates the immunity via attenuating Th1/Th2 imbalance in an immunosuppressed mouse model

OBJECTIVE: To investigate whether Hydrolyzed Seawater Pearl tablet(HSPT) could modulate the Th1/Th2 imbalance in an immunosuppressed mouse model with Th1 to Th2 shift induced by Cyclosporine A(Cs A) which can be used in the clinical treatment of Th2 to Th1 shift diseases, and explore the possible mechanism for the adjuvant therapeutic efficacy of HSPT on recurrent respiratory infections(RRI) and acquired immune deficiency syndrome(AIDS).METHODS: The mice were randomly divided into six groups of five animals each, namely normal group, model group, lentinan polysaccharide tablet(LPT) group and three HPST treated groups. HPST treated groups were administered with HPST(0.51,1.02, 2.04 g/kg) via intragastric gavage(i.g) for 30 consecutive days. LPT used as reference drug for positive control, LPT group was administered with LPT(8.2 mg/kg) for 30 consecutive days. Normal group and model group were received distilled water. The animals in model group, LPT group and HPST treated groups were injected intraperitoneally with Cs A(50 mg/kg) to establish the immunosuppressed mice model with Th1 to Th2 shift on the20 th, 22 nd and 24 th day, one hour after the administration of the respective treatment. Animals were sacrificed one hour after the last administration to collect blood and splenic tissue. The proportion of T cells including CD8+ and CD4+ T cells, Th1 and Th2 in peripheral blood of experimental mice were measured by flow cytometric. The protein level in serum and m RNA level in splenic tissue of experimental mice for interleukin(IL)-2, IL-12, interferon-γ(IFN-γ), IL-4, IL-6, IL-10 and IL-13 were measured by enzyme linked immunosorbent assay and fluorescence quantitative polymerase chain reaction respectively.RESULTS: HSPT elevated the proportion of T cells including both CD8+ and CD4+ T cells, in which the proportion of Th1 and Th2 cells increased, while the ratio of Th1/Th2 cells decreased in peripheral blood of the immunosuppressed mouse model with Th1 to Th2 shift induced by Cs A. Furthermore,HSPT elevated both protein and m RNA level of Th1-type cytokines IL-2 and IFN-γ, while had no significant effect on protein and m RNA level of Th1-type cytokine IL-12 and Th2-type cytokines IL-4, IL-6, IL-10, IL-13 in mouse model.CONCLUSION: Our findings suggest that HSPT can increase proportion of T cells including both CD8+and CD4+T cells and induce Th2 to Th1 shift in both cells and cytokines, which probably was the mechanism to account for the adjuvant therapeutic efficacy of HSPT on RRI and AIDS.

Study on prevention effect of Zishen Yutai pill combined with progesterone for threatened abortion in rats

Objective:To observe preventive and therapeutic effects and the mechanism of actions on Zishen Yutai pill combined with progesterone on threatened abortion in rats.Methods:After pregnancy,50 SPF female SD rats were selected and divided into control group,model group,progesterone group,Zishen Yutai pill group,and progesterone plus Zishen Yutai pill group(combination group),with 10 rats in each group.The rats of control group and abortion model group were lavaged with 2 mL/kg normal saline on pregnancy day 1 for continuous 10 days.Rats in Zishen Yutai pill group were given 1.575 g/kg/d of Zishen Yutai pill for intragastric administration for continuous 10 days.Rats in progesterone group were given intramuscular injection treatment of 0.1 mL/d progestin,continuous for 10 days.Rats in combination group were given injection therapy of aqueous solution of Zishen Yutai pill for continuous 10 days,and other treatments were the same as previous two groups.Abortion model were established then and live births,numbers of abortion and average rate of abortion were compared between the five groups.Peripheral blood was collected to detect the estradiol(E2) and progestational hormone(P),and obtain ratio of Th1/Th2 cytokines(IL-2,INF-γ,IL-4,IL-10).Results:Significant more live births of rats were found in the control group compared with other four groups(P<0.05).The numbers of live births of the rats in abortion model group were significantly less than that of progesterone group,Zishen Yutai pill group and joint group(P<0.05).The numbers of live births of rats in joint group were significant more than that of progesterone group and Zishen Yutai pill group.The serum E2 level of P of rats in the control group,progesterone group,Zishen Yutai pill group and joint group were significant higher than that of abortion model group(P<0.05).Serum levels of E2 and P of rats in the control group,progesterone group,Zishen Yutai pill group and joint group were not significant different(P>0.05) but these levels in the control group and Zishen Yutai pill group were significant lower than that of progesterone group and joint group(P<0.05).The maternal-fetal interface IL-4/IL-2,IL-10 / IL-2 of model group were balanced deviating to Th1,while the IL-4/IL-2,IL-10/IL-2 of progesterone group,Zishen Yutai pill group and joint group were balanced deviating to Th2,and joint group' deviation was better than progesterone group and Zishen Yutai pill group(P<0.05).Conclusions:Zishen Yutai pill combined with progesterone has a significant control effect for threatened abortion,which can obviously increase contents of maternal serums E2 and P,and regulate the Thl/Th2 balance with a remarkable effect.

Wang-Bi tablet, a patented Chinese medicine, maintains the balance of Th1/Th2 in mice with collagen-induced arthritis

OBJECTIVE: To investigate the pharmacological mechanism of Wang-Bi tablets(WBTs), a Chinese patented medicine, in rheumatoid arthritis(RA) using mice with collagen-induced arthritis(CIA).METHODS: A mouse model of CIA was induced using bovine type Ⅱ collagen. WBT treatment was administered and efficacy was evaluated. The levels of interferon-γ(IFN-γ), interleukin-2(IL-2), and interleukin-4(IL-4) were examined using an enzyme-linked immunosorbent assay, and the proportions of Th1 and Th2 were detected using flow cytometry. T-bet and GATA-binding protein 3(GATA3)expression were demonstrated using Western blot analysis.RESULTS: Paw swelling and the arthritis index decreased significantly following WBT treatment. Histopathological analysis revealed markedly alleviated damage to synovium tissue in the WBT and methotrexate treatment groups. WBT regulated the production of IFN-γ, IL-2, and IL-4 and modulated Th1 and Th2 cell populations, which might have been induced by the attenuation of Th1 and Th2 cell differentiation through a decrease in the expression of T-bet and an increase in the expression of GATA3 in the synovial tissue in CIA mice.CONCLUSION: These results indicate that WBT may produce a therapeutic effect on CIA through maintaining the balance of Th1/Th2 cells, which could result in a decrease in the autoinflammatory disorder observed in RA.