Temsirolimus is a promising immunomodulatory agent for enhanced transplantation outcomes.

BACKGROUND: Identifying effective immunosuppressive strategies is critical for addressing immunological rejection following organ transplantation. This study explores the potential immunosuppressive effects and mechanisms of temsirolimus, a rapamycin derivative, in organ transplantation. METHODS: A mouse cardiac allograft model was established using a cervical cannula technique with BALB/c donors and C57BL/6 recipients. Mice were administered temsirolimus intragastrically and graft survival was evaluated. Histological staining was used to assess pathological changes. The BrdU assay was used to measure splenic T cell proliferation. Flow cytometry was used to quantify regulatory T cells (Tregs), CD4+ T cells, and CD8+ T cells. ELISA and qPCR assays were used to determine Foxp3, IL-4, IFN-γ, and TGF-ß expression. RESULTS: Temsirolimus displayed potent immunosuppressive effects at 20 mg/kg/day, significantly inhibiting T cell proliferation (84.6%, P < 0.0001) and prolonging graft survival (median 49 days vs. 8.5 days in controls, P < 0.0001). However, median survival decreased to 34.5 days upon withdrawal. Temsirolimus also reduced splenic CD4+ and CD8+ T cells (2.85% and 2.92%, P < 0.001) and antibody levels (IgM, IgG1, IgG2) by 11.85-29.09% (P < 0.0001) and increased Tregs, Foxp3, IL-4 (P < 0.01), and TGF-ß (P < 0.05), while decreasing IFN-γ (P < 0.001). CONCLUSIONS: Temsirolimus exhibited potent immunosuppressive effects, emerging as a strong candidate to mitigate organ transplant rejection.

Long-term exposure to azoxystrobin induces immunodeficiency in fish that are vulnerable to subsequent rhabdovirus infection.

Azoxystrobin (AZ) is one of the most widely used strobilurin fungicides in the world, and its residue has seriously endangered aquatic ecological security. Our previous data showed that AZ exposure may reduce the resistance of fish to rhabdovirus infection in aquatic environment. Here, we further reported a potential long-term adverse effect of AZ exposure on the antiviral and immunosuppressive recovery in fish, and observed that mitochondrial dynamic balance was disturbed by AZ in which excessive mitochondrial fission occurred in response to decreased ATP levels. When a recovery operation was performed in AZ-exposed cells and fish, infectivity of our model virus, spring viraemia of carp virus (SVCV), was significantly decreased in vitro (using the epithelioma papulosum cyprini [EPC] fish cell line) and in vivo (using zebrafish) in a time-dependent manner. Also, the expression of eight innate antiviral immune genes (IFNs, ISG15, MX1, RIG-I, IRF3, Nrf2 and HO-1) showed a similar change to SVCV replication between the longer exposure period and the expression recovery. Additionally, AZ facilitated horizontal transmission of SVCV in a static cohabitation challenge model, predicting the increase of the potential for the viral outbreak. Therefore, our data suggest that long-term effect of AZ on irreparable impairment in fish made AZ residue potentially greater for ecological risks.

Sis-25, a meroditerpenoid derivative with a cyclobutane scaffold, inhibits activated T cell proliferation by targeting GSK3ß in vitro and in vivo.

A series of novel scopariusicide derivatives were designed and synthesized starting from the main diterpenoid from the aerial parts of Isodon scoparius. Sis-25 was the most effective compound among them. The potential mechanism(s) of its immunosuppressive activity in vitro, as well as its effects on delayed type hypersensitivity (DTH) reaction and imiquimod-induced dermatitis in vivo were investigated in this study. Sis-25 inhibited anti-CD3/anti-CD28 mAbs, PHA or alloantigen-induced T cell proliferation without obvious cytotoxicity. Sis-25 was a highly selective inhibitor of GSK3-ß and inhibited the mTOR/p70S6K pathway but not the PI3K/Akt, p38 MAPK/ERK 1/2 and JAK3/STAT5 pathways. Furthermore, Sis-25 significantly inhibited IFN-γ, IL-6 and IL-17 expression but not IL-10 expression in activated T cells. Finally, Sis-25 treatment mitigated the DNFB-induced DTH reaction and ameliorated imiquimod-induced dermatitis. In summary, Sis-25 exerted significant immunosuppressive activity by targeting GSK3ß in vitro and in vivo. Sis-25 may guide the design of new drugs for more effective and safer treatments of autoimmune diseases and provide new insight into developing utilizations of Isodon scoparius.

Preparation of the sphingolipid fraction from mycelia of Cordyceps sinensis and its immunosuppressive activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Cordyceps sinensis (CS) is an herbal tonic in traditional Chinese medicine and is used to treat a wide range of disorders, including immune, kidney, respiratory, lung and cardiovascular diseases, in China. Most studies are focused mainly on nucleotides and polysaccharides from CS and consider them to be the main active ingredients, while other ingredients are often disregarded. Hundreds of sphingolipids have been identified from CS and showed inhibitory effects on mouse splenic lymphocytes. AIM OF THE STUDY: This study aimed to establish a method for preparing a fraction of sphingolipids from the mycelial powder of CS and evaluate its immunosuppressive activity. MATERIALS AND METHODS: Fraction of sphingolipids (Fr-SPLs) were prepared by silica gel chromatography and reversed-phase chromatography. Its components were identified and quantified by Quadrupole-Orbitrap UHPLC-MS/MS. PBMCs were prepared from human blood, and splenic lymphocytes, B cells, and T cells were prepared from mouse spleens. The inhibitory effect of Fr-SPLs on cell viability was evaluated by CCK-8 assay. PBMC apoptosis and the ratio of CD4+ T cells and CD8+ T cells were quantified by flow cytometry analysis. The expression of IL-2, IL-10, and TNF-α in PBMCs was detected by ELISA kits. RESULTS: A fraction containing 84.83% of sphingolipids (SPLs) was prepared from the mycelia of CS and named Fr-SPLs. 15 SPLs were identified from the Fr-SPLs. Fr-SPLs significantly inhibited the viability of human peripheral blood mononuclear cells (PBMCs) with an IC50 value of 9.82 µg/mL and promoted PBMC apoptosis in a dose-dependent manner. Moreover, Fr-SPLs inhibited the viability of mouse splenocytes, as well as that of B cells and T cells derived from splenocytes. Furthermore, Fr-SPLs reduced the production of IL-2, IL-10, and TNF-α in PBMCs. CONCLUSIONS: Fr-SPLs show immunosuppressive activity, and this study will be useful for preparing immunosuppressive components from CS and its mycelia for hyperimmune disease.

Antibacterial and Immunosuppressive Effects of OPS-2071, a Candidate Therapy for Inflammatory Bowel Disease.

BACKGROUND: Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract. Although many types of drug are used, clinical outcomes are still unsatisfactory. Previous studies have suggested that intestinal bacteria are involved in the pathogenesis of IBD. Accordingly, in an IBD model we evaluated the therapeutic effects of OPS-2071, a low-absorption quinolone antibacterial agent indicated for intestinal infection, and investigated its mechanism of action. METHODS: The therapeutic effects of OPS-2071 and comparison therapies were evaluated using naive CD4 + T cell-transfer IBD model mice. In vitro inhibition of LPS-induced TNF-α production and inhibitory effects on T cell responses stimulated using anti-CD3/CD28 antibody-loaded beads were evaluated using mouse splenocytes and human peripheral blood mononuclear cells. In addition, in vitro activities against bacteria implicated in IBD pathogenesis were tested. RESULTS: OPS-2071 dose-dependently decreased both colonic weight/length ratio and the colitis histological score as compared with the vehicle group. The therapeutic effect of OPS-2071 was equivalent to that of anti-IL-12/23 (p40) antibody. In vitro, OPS-2071 suppressed TNF-α production induced by LPS stimulation and T cell responses in a dose-dependent manner. At high concentrations, these effects were comparable to those of existing immunosuppressive agents, such as prednisolone, in both mouse and human cells. OPS-2071 also showed antibacterial activity against IBD-related bacteria. CONCLUSIONS: Our results suggest that OPS-2071 had both immunosuppressive and antibacterial effects. This dual effect makes OPS-2071 a unique and promising candidate for IBD.

The treatment effect on peripheral B cell markers in antibody positive myasthenia gravis patients.

B cells play a major role in the pathophysiology of myasthenia gravis (MG) with their ability to produce disease specific, pathogenic antibodies. However, their status during disease development and follow-up stages of the disease in the peripheral blood may need further studies to determine useful markers. In this study, we aimed to detect B cell associated factors concerning immunosuppressive treatment in generalized non-thymomatous MG patients. Although CD19+ B cell distribution did not vary among disease subgroups, expressions of both CD38 and BAFFR were altered on B cells in MG patients under immunosuppressive therapy. Serum levels of BAFF were elevated in untreated MG patients as compared to treated MG patients and healthy controls. B cell activation factors may show profound alterations due to immunosuppression.

Anti-thymocyte globulin in haematology: Recent developments.

Anti-thymocyte globulin (ATG) is a polyclonal antiserum introduced into clinical medicine more than 30 years ago. It induces a broad non-specific immunosuppression. In haematology, standard indications are severe aplastic anaemia and prophylaxis and treatment of graft-versus-host disease (GVHD) (after allogeneic transplantation). For aplastic anaemia, ATG from horses has been found to be superior to ATG from rabbits. In the situation of allogeneic transplantation, ATG lessens the risk of chronic GVHD but may not improve survival. There is current controversy regarding which patients benefit most from ATG and what the ideal dosage is. It is likely that in the coming years a more specific immunosuppressive will be developed that will minimize GVHD while maintaining the graft-versus-malignancy effect.

Immunosuppressive Effect of Exosomes from Mesenchymal Stromal Cells in Defined Medium on Experimental Colitis.

BACKGROUND AND OBJECTIVES: The exosomes released by mesenchymal stromal cells (MSCs) in classical FBS-containing media have been demonstrated as an alternative, cell-free therapy in various diseases including inflammatory bowel disease (IBD). It has been found that the function of exosomes is affected by culture condition. We previously developed a serum-free, xeno-free and chemically defined medium, and umbilical cord-derived MSCs in this medium retained the immunosuppressive capability. METHODS: In this study, we evaluated the immunosuppressive function of exosomes from MSCs (MSC-Exo) in defined medium and their therapeutic effect on treating colitis. RESULTS AND CONCLUSIONS: In vitro studies indicated that MSC-Exo reduced the concentration of pro-inflammatory cytokines IFN-γ, TNF-α and IL-1ß, and increased the secretion of anti-inflammatory cytokines TGF-ß1 and IL-10, but no significant change of inhibitory effect on peripheral blood mononuclear cells proliferation was shown. In vivo experimental colitis showed that administration of MSC-Exo was able to significantly ameliorate the disease activity index score, weight loss, colon shortening, and the histological colitis score through up-regulation anti-inflammatory responses and down-regulation of inflammatory responses. Moreover, the use of MSC-Exo (200 µg) led to an improved therapeutic efficacy when compared with MSCs at a dose of 1×106 cells. Our findings indicate that the exosomes from MSCs in defined medium possess a certain degree of immunosuppressive effect in vitro and exhibit a therapeutic capability in a mouse model of DSS-induced colitis through suppressing inflammation mechanism.

Toll-like receptor 4 activation on human amniotic epithelial cells is a risk factor for pregnancy loss.

BACKGROUND: Maternal-fetal tolerance plays a fundamental role in the maintenance of pregnancy. However, this immunological tolerance can be influenced by intrauterine infections. Human amniotic epithelial cells (hAECs) have immunomodulatory effects and respond to invading pathogens through expressing various toll-like receptors (TLRs). We hypothesize that bacteria or bacterial products affect the immunosuppressive effects of hAECs through TLR stimulation. Here, we investigated how a successful pregnancy can be threatened by TLR4 activation on hAECs on lipopolysaccharide (LPS) engagement. MATERIALS AND METHODS: hAECs were isolated from the amniotic membrane received from six healthy pregnant women. The immunophenotyping of hAECs was studied by flow cytometry. The isolated hAECs (4 × 105 cells/ml) were cultured in 24-well plates in the presence or absence of LPS (5 µg/ml). After 24, 48, and 72 h of incubation, the culture supernatants of hAECs were collected, and the levels of interleukin-5 (IL-5), IL-6, IL-1ß, tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta 1 (TGF-ß1), and prostaglandin E2 (PGE2) were measured by enzyme-linked immunosorbent assay. RESULTS: TLR4 activation showed a stimulatory effect on TGF-ß1 production of hAECs (P < 0.001-0.05). PGE2 production of LPS-stimulated hAECs was significantly increased (P < 0.01-0.05). Moreover, TLR4 could induce TNF-α and IL-1ß production of hAECs (P < 0.0001-0.01), while this effect was not observed on IL-6 production of hAECs. The IL-5 was produced at a very low level in two culture supernatants of hAECs, in which its production was independent of LPS effect. CONCLUSION: TLR4 activation by bacterial components on hAECs may be a potential risk factor for pregnancy complications.

Plant-derived alkaloid sinomenine potentiates glucocorticoid pharmacodynamics in mitogen-activated human peripheral blood mononuclear cells by regulating the translocation of glucocorticoid receptor.

Sinomenine has been used as an antirheumatic drug in China. Glucocorticoid combined with sinomenine could be an alternative therapeutic approach. In this study, we evaluated the sinomenine potential effect on glucocorticoid pharmacodynamics in vitro using a human peripheral blood mononuclear cell (PBMC) culture system. We also disclosed the possible action mechanism of sinomenine with a focus on P-glycoprotein function and glucocorticoid receptor (GR) translocation into nucleus. The median (range) of methylprednisolone IC50 values against the PBMC proliferation was 3.18 (0.45-6.81) ng/mL, whereas the median (range) IC50 values of methylprednisolone combined with 0.03, 0.3, 3, and 30 µM sinomenine were 1.85 (0.05-5.15), 0.83 (0.10-3.90), 0.56(0.09-1.62), and 0.59(0.05-1.30) ng/mL, respectively. Sinomenine significantly decreased the IC50 values of methylprednisolone and enhanced the immunosuppressive effect of methylprednisolone (p < 0.05). Sinomenine alone regulated the GR translocation in both Jurkat T cells and normal human PBMCs, and the combination of sinomenine and methylprednisolone showed stronger GR-modulatory activity than methylprednisolone alone. Thus, the additive effect of sinomenine to promote the methylprednisolone immunosuppressive efficacy was suggested to be related to nuclear GR-translocation. However, sinomenine did not significantly inhibit the P-glycoprotein function in the activated PBMCs, suggesting that sinomenine's additive effect seemed to be unrelated with the P-glycoprotein inhibition.

Immunosuppressive Effect of Exosomes from Mesenchymal Stromal Cells in Defined Medium on Experimental Colitis

BACKGROUND AND OBJECTIVES: The exosomes released by mesenchymal stromal cells (MSCs) in classical FBS-containing media have been demonstrated as an alternative, cell-free therapy in various diseases including inflammatory bowel disease (IBD). It has been found that the function of exosomes is affected by culture condition. We previously developed a serum-free, xeno-free and chemically defined medium, and umbilical cord-derived MSCs in this medium retained the immunosuppressive capability.METHODS: In this study, we evaluated the immunosuppressive function of exosomes from MSCs (MSC-Exo) in defined medium and their therapeutic effect on treating colitis.RESULTS AND CONCLUSIONS: In vitro studies indicated that MSC-Exo reduced the concentration of pro-inflammatory cytokines IFN-γ, TNF-α and IL-1β, and increased the secretion of anti-inflammatory cytokines TGF-β1 and IL-10, but no significant change of inhibitory effect on peripheral blood mononuclear cells proliferation was shown. In vivo experimental colitis showed that administration of MSC-Exo was able to significantly ameliorate the disease activity index score, weight loss, colon shortening, and the histological colitis score through up-regulation anti-inflammatory responses and down-regulation of inflammatory responses. Moreover, the use of MSC-Exo (200 μg) led to an improved therapeutic efficacy when compared with MSCs at a dose of 1×10⁶ cells. Our findings indicate that the exosomes from MSCs in defined medium possess a certain degree of immunosuppressive effect in vitro and exhibit a therapeutic capability in a mouse model of DSS-induced colitis through suppressing inflammation mechanism.

Immunosuppressive effects of hydroxychloroquine and artemisinin combination therapy via the nuclear factor-κB signaling pathway in lupus nephritis mice.

Lupus nephritis (LN) is one of the most common and severe manifestations of systemic lupus erythematosus, leading to permanent renal damage and chronic kidney disease. Hydroxychloroquine (HCQ) serves a protective role against lupus-associated clinical manifestations and medical complications; however, it results in numerous adverse reactions, limiting its long-term use. The aim of the present study was to investigate the combined effect of HCQ and artemisinin (ART) on LN, and to elucidate the underlying mechanisms. An in vivo LN mouse model was prepared, and the animals were administered prednisone (PDS; serving as a positive control), high-dose HCQ (H-HCQ) or low-dose HCQ combined with ART (L-HCQ + ART) once daily for 8 weeks. The body weight, serum biochemical parameters, immune and inflammatory indicators, renal and spleen histological alterations, and mRNA expression levels of Kruppel-like factor 15 (KLF15) and nuclear factor-κB (NF-κB) were analyzed. It was observed that L-HCQ + ART and H-HCQ ameliorated the LN-induced body weight decrease, and significantly decreased the levels of anti-double stranded DNA, antinuclear antibodies, immunoglobulin G, interferon γ, tumor necrosis factor-α and transforming growth factor-ß1, as well as improved the kidney and spleen pathology, when compared with the model group. In addition, L-HCQ + ART and H-HCQ treatments induced KLF15 upregulation and NF-κB downregulation. These results indicated that treatment with L-HCQ + ART exerted renoprotective effects by regulating the expression levels of cytokines, KLF15 and NF-κB. This combination treatment may have a similar immunosuppressive effect as PDS and H-HCQ, and may be a promising alternative for LN treatment.

Immunosuppressive Effects of Bryoria sp. (Lichen-Forming Fungus) Extracts via Inhibition of CD8+ T-Cell Proliferation and IL-2 Production in CD4+ T Cells.

Lichen-forming fungi are known to have various biological activities, such as antioxidant, antimicrobial, antitumor, antiviral, anti-inflammation, and anti proliferative effects. However, the immunosuppressive effects of Bryoria sp. extract (BSE) have not previously been investigated. In this study, the inhibitory activity of BSE on the proliferation of CD8+ T cells and the mixed lymphocytes reaction (MLR) was evaluated in vitro. BSE was non-toxic in spleen cells and suppressed the growth of splenocytes induced by anti-CD3. The suppressed cell population in spleen cells consisted of CD8+ T cells and their proliferation was inhibited by the treatment with BSE. This extract significantly suppressed the IL-2 associated with T cell growth and IFN-γ as the CD8+ T cell marker. Furthermore, BSE reduced the expression of the IL-2 receptor alpha chain (IL-2Rα) on CD8+ T cells and CD86 on dendritic cells by acting as antigen-presenting cells. Finally, the MLR produced by the co-culture of C57BL/6 and MMC-treated BALB/c was suppressed by BSE. IL-2, IFN-γ, and CD69 on CD8+ T cells in MLR condition were inhibited by BSE. These results indicate that BSE inhibits the MLR via the suppression of IL-2Rα expression in CD8+ T cells. BSE has the potential to be developed as an anti-immunosuppression agent for organ transplants.

Immunosuppressive effect of curcumin on skin transplantation in mice / 药物评价研究

Objective To investigate the immunosuppressive effect ofcurcumin on xenogenic skin transplantation in mice.Methods The skin transplantation model was established by an operation,tergal skin flaps from BALB/c donor mice transplanted to the back of C57BL/6 recipient mice.Then recipient mice were divided into five groups at random,namely sham,model,curcumin (50 mg/kg),Cyclosporin A (10 mg/kg) and curcumin + Cyclosporin A (50 mg/kg + 5 mg/kg).All mice were ip administered once daily for 10 d.The survival days of skin graft were recorded in all groups.The interleukin-2 (IL-2) levels in plasma of all mice were determined by ELISA 4 and 8 d after the operation,respectively.Results The mean survival time of skin graft in curcumin,Cyclosporin A and curcumin + Cyclosporin A groups were 14.77,16.81 and 19.96 d,respectively,which showed significant.differences comparing with 12.10 d of model group (P ＜ 0.01).Combination of curcumin and Cyclosporin A administration showed a longer mean survival days than curcumin or Cyclosporin A group (P ＜ 0.05 or 0.01).The IL-2 levels in plasma of mice in curcumin,Cyclosporin A and curcumin + Cyclosporin A groups on postoperative day 4 were 3.68,2.05 and 2.70 ng/mL,respectively,which were significantly reduced than 4.76 ng/mL of model group (P ＜ 0.05 or 0.01).The IL-2 levels in plasma of mice in curcumin,Cyclosporin A and combination of curcumin and Cyclosporin A groups on postoperative day 8 were 4.06,2.11 and 2.95 ng/mL,respectively,which were significantly reduced than 5.85 ng/mL of model group (P ＜ 0.01).Conclusion Curcumin may have a good immunosuppressive effect on mice with xenogenic skin transplantation.

Immunosuppressive effect of curcumin on skin transplantation in mice / 药物评价研究

Objective To investigate the immunosuppressive effect ofcurcumin on xenogenic skin transplantation in mice.Methods The skin transplantation model was established by an operation,tergal skin flaps from BALB/c donor mice transplanted to the back of C57BL/6 recipient mice.Then recipient mice were divided into five groups at random,namely sham,model,curcumin (50 mg/kg),Cyclosporin A (10 mg/kg) and curcumin + Cyclosporin A (50 mg/kg + 5 mg/kg).All mice were ip administered once daily for 10 d.The survival days of skin graft were recorded in all groups.The interleukin-2 (IL-2) levels in plasma of all mice were determined by ELISA 4 and 8 d after the operation,respectively.Results The mean survival time of skin graft in curcumin,Cyclosporin A and curcumin + Cyclosporin A groups were 14.77,16.81 and 19.96 d,respectively,which showed significant.differences comparing with 12.10 d of model group (P ＜ 0.01).Combination of curcumin and Cyclosporin A administration showed a longer mean survival days than curcumin or Cyclosporin A group (P ＜ 0.05 or 0.01).The IL-2 levels in plasma of mice in curcumin,Cyclosporin A and curcumin + Cyclosporin A groups on postoperative day 4 were 3.68,2.05 and 2.70 ng/mL,respectively,which were significantly reduced than 4.76 ng/mL of model group (P ＜ 0.05 or 0.01).The IL-2 levels in plasma of mice in curcumin,Cyclosporin A and combination of curcumin and Cyclosporin A groups on postoperative day 8 were 4.06,2.11 and 2.95 ng/mL,respectively,which were significantly reduced than 5.85 ng/mL of model group (P ＜ 0.01).Conclusion Curcumin may have a good immunosuppressive effect on mice with xenogenic skin transplantation.

Experimental Research of Immunosuppressive Effect of Sinomenine on Skin Transplantation / 中国药师

Objective: To investigate the immunosuppressive effect of sinomenine (SIN) on xenogenic skin transplantation in mice.Methods: BALB/c→C57BL/6 tergal skin transplantation model was established by an operation, and then the mice were divided into 5 groups at random, namely sham group, model group, SIN group (30 mg·kg-1), ciclosporin A group (CsA) (10 mg·kg-1) and combination of SIN and CsA group (SIN 30 mg·kg-1 and CsA 5 mg·kg-1) (n=10).All the mice were intraperitoneally administered once a day for ten days.The survival days of skin graft were recorded, and the IL-2 levels in plasma were determined by ELISA respectively on the 4th and 8th day after the operation.Results: The mean survival days of skin graft in the groups treated with different drugs were significantly prolonged when compared with that in the model group (P<0.01).The combination of SIN and CsA administration showed longer mean survival days than SIN or CsA (P<0.05 or P<0.01).The IL-2 levels in plasma in the groups treated with different drugs were significantly reduced than those in the model group on the 4th and 8th day after the operation (P<0.01).Conclusion: SIN may have a good immunosuppressive effect in the mice with xenogenic skin transplantation, and the combination of SIN and CsA shows a synergistic effect.

Eutypenoids A-C: Novel Pimarane Diterpenoids from the Arctic Fungus Eutypella sp. D-1.

Eutypenoids A-C (1-3), pimarane diterpenoid alkaloid and two ring A rearranged pimarane diterpenoids, were isolated from the culture of Eutypella sp. D-1 obtained from high-latitude soil of the Arctic. Their structures, including absolute configurations, were authenticated on the basis of the mass spectroscopy (MS), nuclear magnetic resonance (NMR), X-ray crystallography, and electronic circular dichroism (ECD) analysis. The immunosuppressive effects of eutypenoids A-C (1-3) were studied using a ConA-induced splenocyte proliferation model, which suggested that 2 exhibited potent immunosuppressive activities.

Comparison of Immune Responses Induced by Deferoxamine and Deferasirox

BACKGROUND: The iron chelating agents (ICA) have various biological effects besides iron chelation. We investigated the immunomodulatory effects of Deferasirox (DFS) compared to Deferoxamine (DFO). METHODS: Spleen cells (SP) were obtained from 5 week-old C57/BL6 (H-2(b)). The cytotoxicity of ICAs was examined using the CCK8 method. For the cell proliferation assay, SP were cultured with irradiated in addition to 10, 50, 100micrometer of DFS or DFO and 200ng/mL of cyclosporin A (CSA). Cytokines and nitrite levels were evaluated from supernatants by ELISA. RESULTS: The viability of ICA was reported to be over 100%. Both DFS and DFO inhibited cell proliferation in a manner comparable to CSA. Cell proliferation without iron was reduced at the concentration of 100micrometer of DFO. With iron treatment, the reduction of the stimulation index was dependent on DFO concentrations. DFS decreased the proliferation without reference to the concentrations. After stimulation of phytohemagglutinin, the nitrite concentrations increased with iron. With lipopolysaccharides, the nitrite levels were higher in DFO with iron than control, but similar in DFS regardless of iron treatment. The levels of interleukin-2 were not different. Interleukin-10 was more abundantly produced in 50micrometer of DFO compared to DFS. Transforming growth factor-beta was higher in DFS than DFO at the low concentration, but opposite at the high concentration. CONCLUSION: These data suggested that both iron chelating agents possessed immune suppressive effects comparable to CSA. The immunosuppressive effect of DFS may be distinct from DFO. More experiments are required to determine the exact mechanism of the immunosuppressive effect of DFS.

Effects of Serial Bathing in a Cold Spring on Immunological Parameters of Patients with Rheumatoid Arthritis. Supplements of the study on Kan no Jigoku Spa / 日本温泉気候物理医学会雑誌

Previously one of our colleagues reported that the serial bathing (twice a day for 3 weeks) in a cold spring, Kan-no-Jigoku (simple hydrogen sulfide spring of 14°C) resulted in clinical improvements for patients with rheumatoid arthritis (RA). In that study, the effect on immune functions was also investigated, since RA is characterized by immune abnormalities. The following results were obtained.<br>1. No change was observed in serum gamma globulin levels and hemolytic complement activities.<br>2. Rheumatoid factor titers after the latex fixation test were improved in 2 out of 8 cases, by 1-2 steps after 2 weeks of bathing.<br>3. Circulating immune complex levels, which were significantly higher initially, fell gradually during 3 weeks of bathing, but insignificantly.<br>4. OKT4T cells decreased significantly after 3 weeks of bathing, while OKT3 and OKT8T cells decreased insignificantly. The OKT4/OKT8 ratio was elevated slightly after serial bathing of 3 weeks.<br>5. Plasma prostaglandin E levels were elevated significantly after 2 weeks, but returned to the initial levels after 3 weeks of bathing, although all the levels were within normal range. No such changes of them were observed by a hot spring bathing.<br>6. Plasma cyclic AMP levels, which were a little higher than the normal range in 3 out of 9 cases initially, were also elevated significantly after 1 week of bathing and returned to the initial levels thereafter gradually, while no significant changes of them were observed by a hot spring bathing.<br>7. Urinary hydroxyproline excretion was not changed by the serial bathing.<br>From the above results it was suggested that a cold spring bathing may give an immunosuppressive effect to a living body, resulting in benefit for RA patients.