Objective: To investigate the effect of epirubicin on soluble CD25 (sCD25) secretion by CD4+CD25+ regulatory T (Treg) cells isolated from diffuse large B-cell lymphoma (DLBCL) patients. Methods: Treg cells were isolated from the peripheral blood mononuclear cells isolated from the newly diagnosed DBLCL patients. The concentration of sCD25 in the supernatant was determined with a commercial sCD25 (IL-2R) enzyme-linked immunosorbent assay (ELISA) kit. The fluorescence intensity of CD25 was detected by flow cytometry. Results: Cell survival rate was significantly decreased along with the increase of epirubicin concentration after treatment for 24 h. There was also a significant difference in the concentration of sCD25 between the epirubicin group and the control group (P<0.01). A positive correlation between the Treg cells survival rate and the concentration of sCD25 was detected (r=0.993, P<0.01). When equal numbers of CD4+CD25+ Treg cells of the epirubicin group and the control group were cultured for another 24 h without epirubicin the CD25 fluorescence intensity on the surface of Treg cells was obviously higher in the epirubicin group than that in the control group (P<0.01), while the sCD25 concentration in the supernatant in the epirubicin group was significantly lower than that in the control group (P<0.05). Conclusion: Epirubicin may improve the body's immune functions by inhibiting the sCD25 secretion by Treg cells in DLBCL patients.
We previously reported that 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG) accelerates hematopoiesis and has an improving effect on animal disease models such as sepsis and asthma. The effects of PLAG supplementation on immune modulation were assessed in healthy men and women. The objective was to evaluate the effects of PLAG supplementation on immune regulatory functions such as activities of immune cells and cytokine production. A randomized double blind placebo-controlled trial was conducted. Seventy-five participants were assigned to one of two groups; all participants had an appropriate number of white blood cells on the testing day. The PLAG group (n=27) received oral PLAG supplements and the control group (n=22) received oral soybean oil supplements. IL-4 and IL-6 production by peripheral blood mononuclear cells (PBMC) were lower (p<0.001 and p<0.001, respectively) with PLAG than with soybean oil. However, the production of IL-2 and IFN-$\gamma$ by PBMC was unaltered with PLAG supplementation. The B cell proliferation decreased significantly in the PLAG group compared to the soybean oil control (p<0.05). The intake of PLAG in healthy adults for 4 weeks was deemed safe. These data suggest that PLAG has an immunomodulatory function that inhibits the excessive immune activity of immunological disorders such as atopic and autoimmune diseases. PLAG could improve the condition of these diseases safely as a health food supplement.
Yan, Hai-Dun;Kim, Charn;Kim, Ji-Mok;Lim, Won-Il;Kim, Sang-Jeong;Kim, Jun
The Korean Journal of Physiology
/
제30권1호
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pp.105-116
/
1996
Rendering the brain ischemic would evoke the cerebral ischemic reflex which is characterized by an arterial pressor response, apnea and bradycardia. Since the rostral ventrolateral medulla (RVLM) is known to play a key role in the maintenance of normal cardiopulmonary activity, during the cerebral ischemic reflex some cardiac related cells should be excited and respiration related cells inhibited. In this context, the responses of RVLM neurons to systemie and focal hypoxia were analyzed in the present study. Twenty-five adult cats of either sex were anesthetized with ${\alpha}-chloralose$ and the single neuronal activities were identified from RVLM area. For the induction of focal hypoxia in the recording site, sodium cyanide was applied iontophoretically and for systemic hypoxia the animal was ventilated with nitrogen gas for a twenty-second period. Cellular activities were analyzed in terms of their discharge pattern and responses to the hypoxia by using post-stimulus time and single-pass time histograms. Of eighteen cardiac related cells recorded from the RVLM area, twelve cells were excited by iontophoresed sodium cyanide and of twenty-five respiration related cells, fourteen cells were excited by iontophoresed sodium cyanide. Remaining cells were either inhibited or unaffected. Eight of fifteen cells tested with iontophoresed sodium lactate were excited and remaining seven cells were inhibited. Systemic hypoxia induced by nitrogen gas inhalation elevated the arterial blood pressure, but excited, inhibited or unaffected the single neuronal activities. Some cells showed initial excitation followed by inhibition during the systemic hypoxia. Bilateral vagotomy resulted in a decrease of arterial pressor response to the systemic hypoxia, and a slight decrease in the rhythmicity related to cardiac and/or respiratory rhythms. The single neuronal responses to either systemic or focal hypoxia were not affected qualitatively by vagotomy. From the above results, it was concluded that the majority of the cardiac- and respiration- related neurons in the rostral ventrolateral medulla be excited by hypoxia, not through the mediation of peripheral chemoreceptors, and along with the remaining inhibited cells, all these cells be involved in the mediation of cerebral ischemic reflex.
Type II collagen (CII), major component of hyaline cartilage, has been considered as an auto-antigen in rheumatoid arthritis (RA). However, the clinical and biological significances with regard to the CII autoimmunity need to be clarified in human RA. The presence of antibodies to CII has been identified in sera, synovial fluid, and cartilage of patients with RA. In our study, the increased titer of IgG anti-CII in sera was well correlated with C-reactive protein, suggesting that this antibody may reflect the inflammatory status of RA. The titer of anti-CII antibodies (anti-CII Abs) tended to be higher in early stages of diseases. In our extending study, among 997 patients with RA, 269 (27.0%) were positive for circulatory IgG antibody to CII, those levels were fluctuated over time. It is hard to assess the significant amount of T cell responses to CII and CII (255~274) in RA. By using a sensitive method of antigen specific mixed lymphocyte culture, we can detect the presence of CII-reactive T cells in peripheral blood mononuclear cells of RA patients. Sixty seven (46.9%) of 143 patients showed positive CII reactive T cell responses to CII or CII (255~274). The frequencies of CII reactive T cells were more prominent in inflamed synovial fluid (SF) than in peripheral blood. These T cells could be clonally expanded after consecutive stimulation of CII with feeding of autologous irradiated antigen presenting cells (APC). Moreover, the production of Th1-related cytokine, such as IFN-${\gamma}$, was strongly up-regulated by CII reactive T cells. These data suggest that T cells responding to CII, which are probably presenting the IFN-${\gamma}$ producing cells, may play an important role in the perpetuation of inflammatory process in RA. To evaluate the effector function of CII reactive T cells, we investigated the effect of CII reactive T cells and fibroblasts-like synoviocytes (FLS) interaction on the production of pro-inflammatory cytokines. When the CII reactive T cells were co-cultured with FLS, the production of IL-15 and TNF-${\alpha}$ from FLS were significantly increased (2 to 3 fold increase) and this increase was clearly presented in accord to the expansion of CII reactive T cells. In addition, the production of IFN-${\gamma}$ and IL-17, T cell derived cytokines, were also increased by the co-incubation of CII reactive T cells with FLS. We also examined the impact of CII reactive T cells on chemokines production. When FLS were co-cultured with CII stimulated T cells, the production of IL-8, MCP-1, and MIP-1${\alpha}$ were significantly enhanced. The increased production of these chemokines was strongly correlated with increase the frequency of CII reactive T cells. Conclusively, immune response to CII was frequently found in RA. Activated T cells in response to CII contributed to increase the production of proinflammatory cytokines and chemokines, which were critical for inflammatory responses in RA. The interaction of CII-reactive T cells with FLS further augmented this phenomenon. Taken together, our recent studies have suggested that autoimmunity to CII could play a crucial role not only in the initiation but amplification/perpetuation of inflammatory process in human RA.
Objective : We aimed to identify the effects of Bogimakseong-bang(BGMSB) treatment on cBSA-induced MN in a mouse model. Methods : We divided 20 mice into 4 groups. The normal group (NR) had no treatment. We used cBSA to induced MN to the other 3 groups. One group (CT) was treated with cBSA (7mg/kg i.p) only. The second (BG-250) was treated with cBSA (7mg/kg i.p) and BGMSB extract (250mg/kg, p.o). The third group (BG-500) was treated with cBSA(7mg/kg i.p) and BGMSB extract (500mg/kg, p.o). After cBSA and BGMSB extract treatment for 4 weeks, proteinuria, serum albumin, total cholesterol, serum creatinine, BUN, total nucleated spleen cell number and total infiltrated kidney cell number of all groups were measured. CD3e+/CD19+ and CD4+/CD8 cells ratio of peripheral blood, kidney and spleen of all groups were analyzed. $IL-1{\beta}$ and $TNF-{\alpha}$, IL-6, IgG, IgM, and $IFN-{\gamma}$ levels of all groups were gauged. Histological analysis of kidney tissue and immunohistochemical staining (CD4 CD8) of kidneys were observed. Results : Proteinuria significantly decreased and serum albumin increased in groups treated with cBSA and BGMSB extract compared with the control. Total cholesterol decreased but not significantly. CD3e+/CD19cells ratio of peripheral blood decreased. CD3e+/CD19+ and CD4+/CD8 cells percentage of kidney and spleen showed no significant change. Level of $IL-1{\beta}$, $TNF-{\alpha}$ and IL-6 significantly decreased. and $IFN-{\gamma}$ increased but has not significantly. Concentration of IgG and IgM significantly decreased compared with control. Thickness of GBM decreased on histological analysis of kidney. Deposition of CD4 and CD8 decreased on immunohistochemical staining of kidney. Conclusions : According to the above result, BGMSB had a significant effect for treating MN which is cBSA-induced.
To determine if initial infection with Mycobacterium tuberculosis changes the balance of cytokines between T cells and macrophages, we evaluated interferon (IFN)-${\gamma}$), interleukin-12 (IL)-12, and tumor necrosis factor (TNF)-${\alpha}$ productions by peripheral blood mononuclear cells (PBMC) from 15 untreated active pulmonary tuberculosis (TB) patients and 12 healthy tuberculin reactors (HTR). Freshly isolated PBMC were stimulated with Triton X-100 solubilized protein (TSP), 30-kDa or purified protein derivatives (PPD) antigen for 6, 18 and 96 hours. IL-12 p40 production by antigen-stimulated PBMC from TB patients was significantly decreased compared with that in HTR. In addition, IFN-${\gamma}$ production was significantly depressed in TB patients than that in HTR at a 96-hr stimulation. However, TNF-${\alpha}$ production was significantly higher in antigen-stimulated PBMC from TB than that of HTR. A pronounced increase in IFN-${\gamma}$ protein followed neutralization of IL-10 in early TB patients. However, neutralization of TNF-${\alpha}$ did not significantly alter IFN-${\gamma}$ induction in PBMC from TB patients. There were no significantly differences in the cytokine productions among three proteins, TSP, 30-kDa or PPD antigen. These results indicate that development of TB may be strongly associated with dysregulated productions of IL-12, IFN-${\gamma}$ and TNF-${\alpha}$, during the initial immune responses to M. tuberculosis. Further understanding of operative cytokine networks during human immune cell responses to protein antigens of M. tuberculosis may improve strategies for vaccine development.
Peripheral blood mononuclear cells (PBMCs) discriminate microbial pathogens and induce T-cell responses of appropriate effector phenotype accordingly. Toll-like receptors (TLRs), in part, mediate this microbial recognition and differentiation while the development of T-cell effector functions critically depends on the release of Th1- or Th2- type cytokines. In the present study, buffalo PBMCs were stimulated under in vitro culture conditions by Bacillus subtilis cell wall petidoglycan, a TLR2 ligand, in a dose- and time- dependent manner. The expression of TLR2 as well as the subsequent differential induction of the Th1 and Th2 type cytokines was measured. Stimulation was analyzed across five doses of peptidoglycan ($10{\mu}g/ml$, $20{\mu}g/ml$, $30{\mu}g/ml$, $40{\mu}g/ml$ and $50{\mu}g/ml$) for 3 h, 12 h, 24 h and 36 h incubation periods. We observed the induction of TLR2 expression in a dose- and time-dependent manner and the peptidoglycan induced tolerance beyond $30{\mu}g/ml$ dose at all incubation periods. The correlation between peptidoglycan stimulation and TLR2 induction was found positive at all doses and for all incubation periods. Increased production of all the cytokines was observed at low doses for 3 h incubation, but the expression of IL-4 was relatively higher than IL-12 at the higher antigen doses, indicating tailoring towards Th2 response. At 12 h incubation, there was a pronounced decrease in IL-4 and IL-10 expression relative to IL-12 in a dose- dependent manner, indicating skewing to Th1 polarization. The expression of IL-12 was highest for all doses across all the incubation intervals at 24 h incubation, indicating Th1 polarization. The relative expression of TNF-${\alpha}$ and IFN-${\gamma}$ was also higher while that of IL-4 and IL-10 showed a decrease. For 36 h incubation, at low doses, relative increase in the expression of IL-4 and IL-10 was observed which decreased at higher doses, as did the expression of all other cytokines. The exhaustion of cytokine production at 36 h indicated that PBMCs became refractory to further stimulation. It can be concluded from this study that the cytokine response to sPGN initially was of Th2 type which skews, more pronouncedly, to Th1 type with time till the cells become refractory to further stimulation.
본 연구에서는 영장류에서 감염성 면역결핍 증상을 일으키는 type D simian retrovirus (SRV)를 검출하기 위해 SRV env 유전자의 특정 지역을 선정하여 증폭, 검색하는 중합효소 연쇄반응 (PCR)법을 개발하였다. 증폭반응의 대상부위인 SRV env 유전자의 3' 후반부는 서로 다른 3 종류의 SRV subtype 1, 2, 그리고 subtype 3에 걸쳐 높은 보존율을 보이고 있다. 반응 결과, 1차 PCR 만으로 3 종류의 SRV subtypes를 동시에 검출, 증폭하였으며 SRV와 더불어 영장류에서 감염성 면역결핍을 유도하는 주요 바이러스 simian immunodeficiency virus 또는 simian T-Iymphotropic virus type 1에 감염된 영장류의 peripheral blood mononuclear cells (PBMCs)을 비교, 완전한 증폭 특이성이 확인되었고 교차반응으로 인한 위양성반응은 발견되지 않았다. 한편, 위 증폭반응의 검출 민감도 측정을 위해 준정량적 적정 PCR을 수행하였으며 SRV 게놈과 증폭 대상 부위의 분자량을 기준으로 한 본 PCR법의 검출 한계는 단 한번의 증폭과 간단한 ethidium bromide 염색만으로 적어도 $5-7{\times}10^4$ 개의 PBMCs 중 한 개의 감염세포를 검출할 수 있는 것으로 확인되었다. 본 연구에서 확인된 신속성과 반응 특이성, 그리고 높은 민감도의 본 PCR 법은 현재 유일한 AIDS 연구모델인 영장류의 SIV 감염 연구 전후에 반드시 필요한 효과적인 SRV 감염검색에서 ELISA법의 위양성에 의한 약점을 보완하고 보다 높은 검출 민감도를 확보함으로써 연구모델 실험의 오류를 최소화하는 데 중요한 역할을 하게 될 것이다.
Background: Bronchoalveolar lavage (BAL) is a useful technique to recover lower airway fluid and cells involved in many respiratory diseases. Miliary tuberculosis is potentially lethal, but the clinical manifestations are nonspecific and typical radiologic findings may not be seen until late in the course of disease. In addition, invasive procedures are often needed to confirm disease diagnosis. This study analyzed the cells and the T-lymphocyte subset in BAL fluid from patients with miliary tuberculosis to determine specific characteristics of BAL fluid that may help in the diagnosis of miliary tuberculosis, using a less invasive procedure. Methods: On a retrospective basis, we enrolled 20 miliary tuberculosis patients; 12 patients were male and the mean patient age was $40.5{\pm}16.2$ years. We analyzed differential cell counts of BAL fluid and the T-lymphocyte subset of BAL fluid. Results: Total cells and lymphocytes were increased in number in the BAL fluid. The percentage of CD4+ Tlymphocytes and the CD4/CD8 ratio in BAL fluid were significantly decreased and the percentage of CD8+ T-lymphocytes was relatively higher. These findings were more prominent in patients infected with the human immunodeficiency virus (HIV). In the HIV-infected patients, the proportion of lymphocytes was significantly higher in BAL fluid than in peripheral blood. There were no significant differences between the BAL fluid and the peripheral blood T-lymphocytes subpopulation. Conclusion: BAL fluid in patients with miliary tuberculosis demonstrated lymphocytosis, a lower percentage of CD4+ T-lymphocytes, a higher percentage of CD8+ T-lymphocytes, and a decreased CD4/CD8 ratio. These findings were more significant in HIV-infected subjects.
Background: Ginseng (G) and Ligustrum lucidum Ait (LLA) are core traditional Chinese medicines in treating myelosuppression formula. The present study was designed to profile effect of G and LLA herb pair (G-LLA) on myelosuppressed mice. Methods: The mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (Cy). Hematopoietic function of bone marrow was measured by hemopoietic progenitor cell culture and peripheral blood count, and serum hemopoietic factors were tested by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. HPLC was used to measure 20 potential chemical components related to myelosuppression, including ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd, Rk3, Rh4, 20 (S)-Rg3, 20 (R)-Rg3, Rk1, Rg5, salidroside, and so on. Results: G, LLA, and G-LLA improved the amount of peripheral blood cells and bone marrow cells of myelosuppressed mice (P < 0.01). They significantly increased the colony quantity of colony-forming unit-granulocyte macrophage, burst-forming unit-erythroid, colony-forming unit-erythroid, and colony-forming unit-megakaryocyte and amount of G2/M and S phase cells (P < 0.01). They also significantly decreased the amount of hematopoiesis-related cytokines (P < 0.01). The content of chemical components in G-LLA changed, and the change of rare saponin was the most obvious. Conclusion: These results show that G-LLA herb pair might produce synergistic or complementary compatibility effects on bone marrow suppression after chemotherapy. It suggests that the substance basis of G-LLA for treating bone marrow suppression may be effective chemical components.
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