Research on principle of benzoxazole derivative PO-291 inhibiting T cell proliferation / 中国药理学通报

Objective To investigate the immunosup-pressive activity of benzoxazole derivative PO-291 in inhibiting human activated T cell proliferation and function. Methods Human T cells were isolated and purified by the immunomagnetic microbeads and acti-vated by anti-CD3/anti-CD28 mAbs or alloantigen. Cell proliferation, the expression of CD25 and CD69, cell cycle and apoptosis were measured by flow cytome-try. Secretion levels, including IL-2, IL-4, IL-6, IL-10, IL-17 and IFN-γ were determined by ELISA. The expression and phosphorylation of STAT5 and p70S6K of activated T cells were detected by Western blot. Re-sults PO-291 significantly inhibited human T cell proliferation with anti-CD3/anti-CD28 mAbs or alloan-tigen stimulation without obvious cytotoxicity. PO-291 did not affect CD25, CD69 and IL-2 expression, but induced T cell cycle arrest in G0/G1 phase. PO-291 significantly inhibited IL-17, IFN-γ and IL-6 expres-sion, but not IL-2, IL-4 and IL-10. PO-291 did not affect STAT5 and p70S6K expression, but inhibited STAT5 phosphorylation and enhanced p70S6K phos-phorylation. Conclusions PO-291 inhibits human ac-tivated T cell proliferation by affecting the JAK3/STAT5 pathway. PO-291 represents a potential lead compound for the design and development of new im-munosuppressive drugs for the treatment of organ trans-plantation and autoimmune diseases.

Effects of Periplaneta americana extract Ento-A on immune function in immunosuppressed mice / 中国药理学通报

Aim To investigate the effects of Periplane-ta americana extract Ento-A on the immune function in immunosuppressed mice . Methods Immunosup-pressed mouse model was induced by intraperitoneal injection of cyclophosphamide in KM mice .To evalu-ate the effects of Ento-A on the immune function in im-munosuppressed mice , neutral red method and MTT assay were used respectively to detect the effects of En-to-A on the phagocytosis of peritoneal macrophages and T cell proliferation rate in mice; with sheep red blood cell as immunogen , the effects of Ento-A on the pro-duction of serum hemolysin were evaluated;peripheral blood was tested and immune organ index calculated . Results Compared with model control group , the high, medium and low doses of Ento-A could improve the expression of serum hemolysin in immunosup-pressed mice ( P<0.01 ) , and increase the spleen in-dex(P<0.01) and thymus index (P>0.05), signifi-cantly increased the content of WBC ( P<0.01 ) , PLT ( P<0.01 ) , HGB ( P<0.01 ) , while the contents of RBC was on the rise , with no significant difference ( P>0.05 ) in peripheral blood , significantly enhanced phagocytic function and T lymphocyte proliferative abil-ity in a dose-dependent manner ( P<0.01 ) .Conclu-sion Ento-A can enhance the immune function of im-munosuppressed mice .

Effects of lung cancer cell-associated B7-H1 on T-cell proliferation in vitro and in vivo

B7 homolog 1 (B7-H1) is the most potent immunoinhibitory molecule in the B7 family. In this study, we examined the effects of tumor-associated B7-H1 on T-cell proliferation in lung cancer. The expression of B7-H1 in human adenocarcinoma A549 and mouse Lewis lung carcinoma (LLC) cells were examined by flow cytometry. To assess the in vitro effect of tumor-associated B7-H1 on T-cell proliferation, we isolated T cells from peripheral blood mononuclear cells (PBMCs) of healthy individuals, labeled them with carboxyfluorescein succinimidyl ester, and co-cultured them with A549 cells in the absence or presence of anti-B7-H1 antibody. For in vivo analysis, LLC cells were subcutaneously injected into mice treated or not with anti-B7-H1 antibody. T-cell proliferation in both in vitro and in vivo assays was analyzed by flow cytometry. In vitro, co-culturing T cells with A549 cells significantly inhibited the proliferation of the former compared with the proliferation of T cells alone (P<0.01), and the addition of B7-H1 blocking antibody dramatically reversed the inhibition of T-cell proliferation by A549 cells. Similarly, in mice bearing LLC-derived xenograft tumors, in vivo administration of anti-B7-H1 antibody significantly increased the total number of spleen and tumor T cells compared to levels in control mice that did not receive anti-B7-H1 antibody. Functionally, in vivo administration of anti-B7-H1 antibody markedly reduced tumor growth. Tumor-associated B7-H1 may facilitate immune evasion by inhibiting T-cell proliferation. Targeting of this mechanism offers a promising therapy for cancer immunotherapy.

Inhibitory effect of small molecule compound BD691 on activated T cell proliferation and its mechanism / 医学研究生学报

Obej ctive Abnormal proliferation of T cells plays an important role in the development of autoimmune diseases. The article aimed to study the inhibitory effect of small molecule compound BD691 on T cell proliferation and its mechanism. Methods Human peripheral blood T-lymphocytes were isolated and purified by the immunomagnetic microbeads,then T cells were ac-tivated with anti-CD3/CD28 mAbs or alloantigen.The inhibitory effect of BD691 on activated T cell proliferation, the cytotoxic effect BD891 on resting T cells and the expression of activated T cells marker CD25 were measured by flow cytometry.Furthermore, ELISA was used to detect the secretion of cytokines associated with T cell differentiation. Results BD691 significantly inhibited the prolif-eration of T cells being stimulated by anti-CD3/CD28 mAb or alloantigen in a dose-dependent manner, and IC50 values are (8.5 ± 1.5)μmol/L and (7.2 ±1.3)μmol/L, respectively.However, BD691 had no obvious cytotoxic effects on resting T cells and periph-eral blood mononuclear cells, even at a high concentration ( up to 100μmol/L) .In T cells which were not activated by anti-CD3/CD28 mAb, the percentage of CD25+T cells is only 1.6%of the total cells, while the number increased to 68% after activating treatment.Mean-while, in T cells which were activated by 0, 3.3, 10, 30μmol/L BD691, no obvious change of CD25 expression were observed, while immunosuppressant FK506 (0.1μmol/L) significantly decreased the expression of CD25 +T cells (14.9%).In unactivated T cells, 95.6%cells were at G0/G1 phase, while after activation, the percentage of cells at G0/G1 phase reduced to 57.7%.In addition, BD691 inhibited the secretion of IFN-γ, IL-6 and IL-17 in activated T cells, but had no effects on the secretion of IL-2, IL-4 and IL-10. Co nclusion BD691 exerts no effects on T cell activation, but it inhibits T cell proliferation by inducing T cell cycling arrest at G0/G1 phase.Moreover, BD691 inhibits the secretion of key cytokines (such as IFN-γ, IL-6, IL-17) closely related to the differ-entiation of Th1 and Th17 cells.The results suggest that BD 691 is a potential lead compound to develop a new immunosuppressant for the inhibition of abnormal proliferation and differentiation of T cells.

Suppressive effect of a novel benzothiazole derivative BD759 on T cell proliferation / 中国药学杂志

OBJECTIVE To determine the immunosuppressive activity of a novel benzothiazole derivative BD759 on T cell proliferation and its potential mode of action. METHODS: T cell proliferation, CD25 expression and cell cycle distribution were measured by flow cytometer. Cytokine levels, including IL-2, IL-4, IL-6, IL-10, IL-17A and IFN-γ, were determined by ELISA. RESULTS: BD759 significantly inhibited human T cell proliferation, stimulated either by anti-CD3/anti-CD28 monoclonal antibodies or by an al-loantigen, in a dose-dependent manner with IC50 values of (3.5 ± 0.7) and (3.3 ± 0.9) μmol · L-1, respectively. No obvious cytotoxic effects of BD759 were observed on human resting naive T cells and peripheral blood mononuclear cells in our experimental conditions. Furthermore, BD759 did not inhibit CD25 expression or IL-2, IL-4 and IL-10 secretion, but inhibited IL-6, IL-17A and IFN-γ production and induced cell cycle arrest at the G0/G1 phase in activated T cells. CONCLUSION: These data indicate that BD759 has no effect on T cell activation, but induces T cell cycling arrest at G0/G1 phase. BD759 also inhibits the secretion of inflammatory cytokines, such as IL-6, IL-17A and IFN-gamma. Thus, BD759 has the potential to be used as a lead compound for the design and development of new immunosuppressants for treating autoimmune diseases and preventing graft rejection.

Establishment of an anti-CD45 monoclonal antibody that inhibits T-cell proliferation / 中国实验动物学报

Objective To identify molecules that modulate T-cell functions and serve the studies on T-cell media-ted autoimmune diseases.Methods Bone marrow-derived dentritic cells were collected from BALB/c mice to immunize Wistar rats, and to establish many hybridoma cell lines.Many hybridoma cell lines which could modulate T-cell functions were obtained.One of the cell lines, most actively inhibiting T-cell proliferation, was further studied.Results The anti-CD45 mAb recognized CD45 and significantly suppressed T-cell proliferation in proliferation assays.Conclusions Our re-sults indicate that the anti-CD45 mAb can effectively suppress T-cell proliferation, and is promising to be used in the pre-vention and treatment of T-cell mediated autoimmune diseases in the future.

5-fluorouracil inhibits the myeloid-derived suppressor cells and reduces their immunosuppressive ac-tivities in mice with liver cancer / 中华微生物学和免疫学杂志

Objective To investigate the effects of 5-fluorouracil (5-FU) on the distribution, pro-portion and immunosuppressive activities of myeloid-derived suppressor cells (MDSCs) in a mouse model of liver cancer.Methods Orthotopic transplantation of H22 cell line was performed to establish the mouse model of orthotopic liver cancer.Forty mice were randomly divided into four groups (n=10) on the 7th day after transplantation.Three different doses of 5-FU (10 mg/kg, 30 mg/kg and 50mg /kg) and 0.4 ml of PBS were respectively given to the mice in each group for 10 consecutive days .A control group without canc-er cell transplantation was set up correspondingly .The distribution of MDSCs in mice liver tissues was detec-ted by immunohistochemistry .The proportions of MDSCs in spleen tissues were tested by flow cytometry . The levels of IFN-γand Arginase-I ( Arg-I) in serum samples were analyzed by enzyme-linked immunosor-bent assay.The activities of nitric oxide synthetase (NOS) in serum samples were tested by chemical colori-metry.The proliferation of T lymphocytes was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltet-razolium bromide (MTT) test.Results 5-FU reduced the tumor size and alleviated the adhesion between liver tissues and abdominal cavity in mice with liver cancer in a dose-dependent manner .The proportions of MDSCs in liver and spleen tissues from mice in four model groups were significantly higher than those in con - trol group (P0.05),but that were decreased in model groups than those in control group after 48 hours of culture (P<0.05).T cells from mice in PBS treated group showed the lowest level of proliferation .T cell proliferation was enhanced upon the treatment of 5-FU in a dose-dependent manner .Except for comparing between PBS treated group and 10 mg/kg of 5-FU treated group, significant differences with T cell proliferation were observed among other groups (P<0.05).The proportions of MDSCs in liver and spleen tissues were positively correlated with the levels of Arg -I and the activities of NOS, but negatively correlated with IFN-γlevel and T cell proliferation at the time point of 48 h.Conclusion To a certain extent , 5-FU could reduce the numbers of MDSCs in liver and spleen tis-sues from mice with liver cancer , inhibit immunosuppressive activities of MDSCs and restrain the growth of the tumor in a dose-dependent manner .

Stimulation of Allogenic lymphocytes by Dendritic Cells Derived from Human Umbilical Cord Blood Fused with Breast Cancer Stem Cells.

Aims: Cancer stem cells (CSCs) are cancer cells that possess characteristics associated with normal stem cells. CSCs represent a minor subset of cells in the tumor and are thought to be the reason for the initiation of disease, resistance to cancer treatment, and the occurrence of metastasis. Therefore, breast cancer stem cells (BCSCs) targeting therapies are considered as the promising therapy for breast cancer treatment. This research aims to evaluate the tumor associated antigens presentation of dendritic cells fused with breast cancer stem cells in dendritic cells based therapies. Methodology: Human breast cancer stem cells are isolated from malignant breast tumors by enrich culture and fluorescent activated cell sorting. Human dendritic cells are isolated from umbilical cord blood by culturing CD14 monocytes in induced medium. Electrocution is used to fuse breast cancer stem cells and dendritic cells. Fusion cells are used to evaluate functions of dendritic cells (DCs) and also to stimulate T cells. Results: These findings indicate that fusion cells have the ability to present the antigens of breast cancer stem cell to T-cells, and regarding functionality. Conclusion: They appear to be very good candidates for antitumor vaccine in breast cancer.

Dynamics of immunosuppression in hamsters with experimental visceral leishmaniasis

Immunosuppression has been reported to occur during active visceral leishmaniasis and some factors such as the cytokine profile may be involved in this process. In the mouse model of cutaneous leishmaniasis using Leishmania (Leishmania) major, the Th1 response is related to protection while the Th2 response is related to disease progression. However, in hamsters, which are considered to be an excellent model for the study of visceral leishmaniasis, this dichotomy is not observed. Using outbred 45- to 60-day-old (140 to 150 g) male hamsters infected intraperitoneally with 2 x 10(7) L. (L.) chagasi amastigotes, we evaluated the immune response of spleen cells and the production of cytokines. We used 3 to 7 hamsters per group evaluated. We detected a preserved response to concanavalin A measured by index of proliferation during all periods of infection studied, while a proliferative response to Leishmania antigen was detected only at 48 and 72 h post-infection. Messenger RNA from cytokines type 1 (IL-2, TNF-α, IFN-γ) and type 2 (IL-4, IL-10 and TGF-β) detected by reverse transcriptase polymerase chain reaction and produced by spleen cells showed no qualitative difference between control non-infected hamsters and infected hamsters during any period of infection evaluated. Cytokines were measured by the DNA band intensity on agarose gel using the Image Lab 1D L340 software with no differences observed. In conclusion, the present results showed an antigen-dependent immunosuppression in hamsters with active visceral leishmaniasis that was not related to the cytokine profile.

Fulminant Epstein-Barr Virus-associated T-cell Lymphoproliferative Disorder in an Immunocompetent Middle-aged Man Presenting with Chronic Diarrhea and Gastrointestinal Bleeding

The World Health Organization (WHO) recently defined systemic Epstein-Barr virus (EBV)-positive T-cell lymphoproliferative disorders (LPD) of childhood as a life-threatening illness. However, this rare disease has not been extensively studied. Here we report a case of systemic EBV-positive T-cell LPD in a previously healthy middle-aged man with a chief complaint of chronic diarrhea. The initial colon biopsy showed focal infiltration of EBV-positive small lymphocytes without any atypia. However, the disease rapidly progressed and the patient required a total colectomy due to severe gastrointestinal bleeding. Three and half months after admission, the patient died from a complication of disseminated intravascular coagulation. The resected colon showed diffuse infiltration of EBV-positive atypical lymphocytes with ischemic change. Most atypical lymphocytes were CD3+ or CD5+. The monoclonality of EBV was demonstrated by sequence variation analysis of the latent membrane protein 1 (LMP1) gene in the colectomy specimen as well as in the initial biopsy.

Effects of mitogen-activated protein kinase p38 on CD3/CD28-stimulated T-cell proliferation and IL-2 expression enhauced by shockwaves / 吉林大学学报(医学版)

Objective To investigate the effects of mitogen-activated protein kinase p38(p38 MAPK)on CD3/CD28-stimulated T-cell proliferation and IL-2 expression which were enhanced by shockwaves.Methods Jurkat T cells or peripheral blood mononuclear cells(PBMC)were pretreated with the p38 MAPK-inhibitor(SB203580)(The Jurkat T cells or PBMC of the control groups were not pretreated with SB203580),then treated with shockwaves,and stimulated respectively with a suboptimal dose of anti-CD3 and anti-CD28 antibodies or PHA.Finally the IL-2 productions of Jurkat T cells or the proliferation of PBMC were respectively measured.The protein tyrosine phosphorylation of p38 MAPKin Jurkat T cells treated either with shockwaves or not was measured by Western blotting with anti-phosphotyrine antibodies(Thr180/Tyr182).The expressoins of p38 MAPK in Jurkat T cells treated either with shockwaves or not were determined by Western blotting with anti-p38 MAPK antibodies.Results Compared with negative control group without shockwave treatment,the 3H-TdR incorporation of the phytohemagglutinin-stimulated PBMC,which were treated with low dose shockwaves(LDSWs)of 100,150,200,250,300,330 impulses at(0.180?0.015)mJ?mm2,increased significantly(P

Newly dentified members of the TNF recept or superfamily (mTNFRH1 and mTNFRH2) inhibit T-cell proliferation

By searching an EST database, we identified two TNF receptor superfamily members (named mTNFRH1 and mTNFRH2). Amino acid sequences are highly conserved between the two receptors (78% identity). The chromosomal loci of mTnfrh1 and mTnfrh2 genes are found in distal chromosome 7 in the mouse. mTNFRH1 and mTNFRH2 do not contain the cytoplasmic domain, indicating that they might function as decoy receptors. Furthermore, an alternatively spliced form of mTNFRH1 was found which contains neither the transmembrane domain nor the cytoplasmic domain, thus presumably existing as a soluble form. Northern blot analysis showed that mTnfrh1 mRNA was negligibly expressed in tissues, while mTnfrh2 mRNA was strongly expressed in spleen, lung, liver, kidney, and testis. When the extracellular domains of mTNFRH1 and mTNFRH2 were expressed in bacteria, their molecular weight of extracellular region was approximately 15 kDa. Both of the soluble forms were effective in inhibiting T-cell proliferation stimulated by anti-CD3 monoclonal antibody. Our data suggest that mTNFRH1 and mTNFRH2 may be implicated in exerting a modulatory role in the immune response.

Effect of Acupuncture on Leukocyte and Lymphocyte Subpopulation in Human Peripheral Blood. Quantitative discussion / 日本温泉気候物理医学会雑誌

To evaluate the effects of acupuncture on the immune system, the leukocyte, monocytes, lymphocyte and lymphocyte surface markers, CD2, CD4, CD8, CD11b, CD16, CD19 and CD56 in the peripheral blood of seventeen healthy volunteers were counted. The leukocyte above CD⁺ cell counts significantly increased after acupuncture. The results indicate that acupuncture may regulate the immune system and can increase the activity of cellular and humoral immunity and NK cell.<br>According to the percentage of lymphocytes or granulocytes, volunteers were divided into two types, those with more than 70% of granulocyte were recognized as G type and those with more than 40% of lymphocyte were divided into L type. Interestingly, before and after the treatment of acupuncture, the number of granulocytes and lymphocytes had a negative relationships. Namely we found an increase in the lymphocytes as well as a decrease in the granulocytes in the G type. On the other hand in the L type, we found an increase in the granulocytes and a decrease in the lymphocytes. Therefore we suggest that acupuncture can enhance the activity that maintains the balance of the immune function.

Properties of Israeli carp ( Cyprinus carpio L . ) sIgM + and sIgM - Cells in a Cell - mediated Immunity / 대한면역학회지

No abstract available.

Effect of ?-aminobutyric acid on proliferation of cytokine-induced killer cells / 中国免疫学杂志

Objective:To study the effect of GABA and A receptor agonist THIP on proliferation and apoptosis of cytokine-induced killer (CIK) cells.Methods:CIK cells were cultured by routine method,and then treated with different concentrations of GABA and THIP.The proliferation of CIK cells were investigated by MTT assay.The apoptosis,cell cycle and immunophenotype were investigated by flow cytometry.Results:GABA could inhibit the proliferation of CIK cells in a dose-dependent manner and affect the distribution of cell cycle of CIK cells(P

Astragalus polysaccharides(APS) induces the cord blood monocytes into mature dendritic cells(DCs) directly and its impact on T cells proliferation / 中国免疫学杂志

Objective:To observe the effect of Astragalus polysaccharides(APS) on inducing the cord blood monocytes into mature dendritic cells(DCs) in vitro and to investigate their morphous,cellar immunological characteristics,and contribution to T cell proliferation.Methods:①The cord blood monocytes were isolated by lymphocyte isolating solution under axenic condition,and three groups were divided.②Cells cultured with APS in concentration of 100 mg/L as the experiment group, that with the cytokines of GM-CSF/IL-4/TNF-? as the positive control, and another without either GM-CSF/IL-4/TNF-? or APS as the negative control, respectively. The morphology of DCs was identified by inverted optical microscope or scanning electron microscope. The phenotype of 12 days cultures of DCs(CD1a, CD80, CD86 and CD83) were identified by flowcytometry. The DCs preparations from the experiment group were treated with mitomycin for 45 min to remove their proliferative activity as incentive cells in the mixed cultures with allogenic peripheral blood mononuclear cells from healthy volunteers as responders. T cell proliferation induced with the DCs preparations was detected by MTT chromatometry.Results:After cultured for 72 hours, the cell of both the experiment group and the positive control grew dusteringly and began to change from round to irregularin shapes. The longer the cells were cultured, the more obvious the dendritic structure is. The cells of experiment group and the positive control group when cultured for 12 days had the most typical dendritic structure. The negative control group cells had no dendritic structure and became macrophages when cultured for 12 days. The experiment group cells cultured for 10 days showed typical dendritic morphology by SEM. The experiment group cells and the positive group cells cultured for 12 days significantly expressed high level of the phenotypes of DCs(CD1a, CD80, CD86 and CD83) by flowcytometry.And the difference exhibitied statistical significance when compared with the negative control group(P0.05).The mixed lymphocyte reaction showed that the DCs induced by APS trigerred proliferation of allogenic T cells obviously.Conclusion:Both APS and cytokine could induce the cord blood monocytes to differentiate into functional DCs committedly. DCs reduced by APS stimualate proliferation of the allogenic T cells obviously.