• IMMUNE NETWORK
• /
• v.5 no.2
• /
• pp.68-77
• /
• 2005

Background: Costimulation is a critical process in Ag-specific immune responses. Both B7.1 and CD28 molecules have been reported to stimulate T cell responses during antigen presentation. Therefore, we tested whether Ag-specific immune responses as well as protective immunity are influenced by coinjecting with B7.1 and CD28 cDNAs in a mouse HSV-2 challenge model system. Methods: ELISA was used to detect levels of antibodies, cytokines and chemokines while thymidine incorporation assay was used to evaluate T cell proliferation levels. Results: Ag-specific antibody responses were enhanced by CD28 coinjection but not by B7.1 coinjection. Furthermore, CD28 coinjection increased IgG1 production to a significant level, as compared to pgD+pcDNA3, suggesting that CD28 drives Th2 type responses. In contrast, B7.1 coinjection showed the opposite, suggesting a Th1 bias. B7.1 coinjection also enhanced Ag-specific Th cell proliferative responses as well as production of Th1 type cytokines and chemokines significantly higher than pgD+pcDNA3. However, CD28 coinjection decreased Ag-specific Th cell proliferative responses as well as production of Th1 types of cytokines and chemokine significantly lower than pgD+pcDNA3. Only MCP-1 production was enhanced by CD28. B7.1 coimmunized animals exhibited an enhanced survival rate as well as decreased herpetic lesion formation, as compared to pgD+pcDNA3. In contrast, CD28 vaccinated animals exhibited decreased survival from lethal challenge. Conclusion: This study shows that B7.1 enhances protective Th1 type cellular immunity against HSV-2 challenge while CD28 drives a more detrimental Th2 type immunity against HSV-2 challenge, supporting an opposite role of B7.1 and CD28 in Ag-specific immune responses to a Th1 vs Th2 type.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.9
• /
• pp.4415-4420
• /
• 2016

We aimed to investigate any association between hepatitis C virus (HCV) infection and non-Hodgkin's lymphoma (NHL) in the view of cytokines that control inflammation/angiogenesis and their correlation with certain CD markers. NHL patients with or without HCV infection were studied. CD5, CD30, CD3, CD20 and CD45 were immunohistochemically evaluated. Plasma levels of vascular endothelial and platelet derived growth factors (VEGF, and PDGF), tumor necrosis factor (TNF-${\alpha}$), transforming growth factor (TGF-${\beta}$), interleukin-6 (IL-6), IL-8, IL-4, IL-12 and interferon gamma (IFN-${\gamma}$) were detected by enzyme-linked immunosorbent assay (ELISA). HCV+ve NHL patients showed a significant reduction in VEGF, PDGF, IFN-${\gamma}$, CD5 and CD45 and a significant increase in IL-12 and IL-8. In conclusion, there was a significant change in cytokine secretion and expression of CD markers in HCV+ve NHL patients. Based on our results, HCV infection in NHL patients requires more in-depth investigations to explore any role in lymphoma progression.

• Journal of Dairy Science and Biotechnology
• /
• v.14 no.2
• /
• pp.229-245
• /
• 1996

1. CM-Sephadex C-50-120 column을 사용하여 HLF를 효과적으로 정제하였다. CM-Sepha-dex 50-120 column chromatography를 수행한 결과, HLF는 NaCl 350 mM-500 mM 사이에서 용출되었으며, 용출된 분획을 SDS-PAGE를 수행해서 단일 band를 확인하였다. Anti-HLF antibody를 이용한 Western blotting 결과 nitrocellulose paper 상에 단일 band 가 나타나므로 HLF 가 효과적으로 정제되었다는 것을 알 수 있었다. 2. Con A, PWM, PWA LPS 등의 자극원으로 단핵구와 대식세포를 자극한 다음 BLF를처리한 배 양액을 IL-1 bioassy한 결과는 Con A 33%, PMA 33%, PWM 15%, LPS 35% 이고, HLF로 처리하여 IL-1 bioassay를 한 결과는 Con A 15%, PMA 22%, PWM 10%, LPS 5%로 감소된 것으로 나타났다. 3. K-562 세포를 이용한 colony forming assay에서 BLF가 10 g/ml일 때는 30%, 30 g/ml일 때는 35%, HLF는 10 g/ml일 때는 5%,30 g/ml일 때는 30%의 저해를 나타냈다. 4. Lactoferrin의 면역증강효과를 알아보기 위하여 hapten인 VCR-BSA를 투여 한 후, 생성되는 항체생성능력을 ELISA로 비교하였다. 그 결과 HLF 및 BLF 투여군에서 대조군에 비하여 adjuvant 효과를 관찰할 수 있었다. 또한 이때 macrophage 수는 대조군에 비하여 HLF와 BLF를 투여한 군이 증가됨을 알 수 있었다. 이러한 결과로 미루어 볼 때 LF는 macrophage를 활성화 시켜서 항체 생성 능력을 증가시키는 효과가 있음을 관찰할 수 있었다. 5. Balb/c mouse의 thymus로부터 분리한 CD4- CD8- 세포를 BLF로 처리하여 24 시간 배양한 후 CB4$^-$ CD8$^-$ 세포의 분화를 측정한 결과, CD4$^-$를 CD4$^+$ 로 분화하였다. 그리고 HLF로 처리하여 24 시간 배양 후 CD4$^-$ CD8$^-$ 세포의 분화를 측정한 결과, CD4$^-$ CD8$^-$를 CD4$^+$ CD8$^+$ 로 분화하였다. 6. Lactoferrin이 T cell의 IL-2 production에 미치는 영향은 PMA 처리군, PMA+OKT3처리군, LF 단독 처리군 보다 PMA+OKT3+LF를 처리한 군이 IL-2 생성에 영향을 미쳤다. 7. Lfcin B의 단일크론항체에 의해 인식되어지는 Lfcin B의 항원결정기는 ‘QWR’로 밝혀졌다.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.21 no.1
• /
• pp.145-152
• /
• 2007

Following CNS injury, inhibitory influences at the site of axonal damage occur. Glial cells become reactive and form a glial scar, gliosis. Astrocyte-rich gliosis relates with up-regulation of GFAP and CD81, and eventually becomes physical and mechanical barrier to axonal regeneration. It is postulated that the astrocytic reaction is absent, regeneration of axons can occur. And it was reported that treatment with anti CD81 antibodies enhanced functional recovery in the rat with spinal cord injury. So in this current study, the author investigated the effect of the water extract of Persicae Semen on the regulation of GFAP and CD81 that increase when gliosis occurs. Persicae Semen decreased the expression of GFAP and CD81 in astrocyte cell by ELISA method. Persicae Semen decreased the RNA expression of CD81 and GFAP. The proteins that separate in whole cell were analaysed by western blot, and the expression of GFAP and CD81 was decreased. In vivo, rats brains were peformed cortical stab wound, the water extracts of Persicae Semen were injected for 7 days, 30 days. As a result, GFAP and CD81 expression were decreased in immunohistochemistry. These findings demonstrate that Persicae Semen decreases GFAP and CD81 expression. Accordingly, Persicae Semen could be a candidate for promotion of axon regeneration after CNS injury.

• Journal of Korean Neurosurgical Society
• /
• v.58 no.2
• /
• pp.101-106
• /
• 2015

Objective : The aim of this study was to explore the immunity in rats transplanted with adipose-derived mesenchymal stem cells (ADSCs) and acellular nerve (ACN) for repairing sciatic nerve defects. Methods : ADSCs were isolated from the adipose tissues of Wistar rats. Sprague-Dawley rats were used to establish a sciatic nerve defect model and then divided into four groups, according to the following methods : Group A, allogenic nerve graft; Group B, allograft with ACN; Group C, allograft ADSCs+ACN, and Group D, nerve autograft. Results : At the day before transplantation and 3, 7, 14, and 28 days after transplantation, orbital venous blood of the Sprague-Dawley rats in each group was collected to detect the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets using flow cytometry and to determine the serum concentration of interleukin-2 (IL-2), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and $interferon-{\gamma}$ ($IFN-{\gamma}$) using enzyme-linked immunosorbent assay (ELISA). At each postoperative time point, the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets and the serum concentration of IL-2, $TNF-{\alpha}$, and $IFN-{\gamma}$ in group C were all near to those in group B and group D, in which no statistically significant difference was observed. As compared with group A, the proportion of $CD3^+$, $CD4^+$, and $CD8^+$ subsets and the serum concentration of IL-2, $TNF-{\alpha}$, and $IFN-{\gamma}$ were significantly reduced in group C (p<0.05). Conclusion : The artificial nerve established with ADSCs and ACN has no obvious allograft rejection for repairing rat nerve defects.

• The Journal of Internal Korean Medicine
• /
• v.30 no.1
• /
• pp.24-35
• /
• 2009

Object : In conditions of brain infarction, irreversible axon damage occurs in the central nerve system (CNS), because gliosis makes physical and mechanical barriers. If gliosis formation could be suppressed, irreversible axon damage would be reduced. This could mean that an injured CNS could be regenerated. CD81 and GFAP have close relationships to gliosis. The increase in glial cells at CNS injury gives rise to the expression of CD81 and GFAP. CD81 was postulated to play a central role in the process of CNS scar formation. Method : In this study, the author investigated the effect of the water extract of the Moutan Radicis Cortex on regulation of CD81 and GFAP expression in injured CNS cells. MTT assay was used to examine cell viability, while RT-PCR and ELISA methods were carried out to measure the expression of CD81 and GFAP in the astrocyte. Results : We observed that water extract of the Moutan Radicis Cortex increased cell viability under hypoxia induced by $CoCl_2$ and suppressed the expression of CD81 and GFAP up-regulated by hypoxia. Conclusion : These results suggest that the Moutan Redicis Cortex could promote neural regeneration as a consequence of protecting CNS cells from hypoxia and suppressing the reactive gliosis following CNS injury.

• IMMUNE NETWORK
• /
• v.1 no.1
• /
• pp.87-94
• /
• 2001

Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.

• The Journal of Internal Korean Medicine
• /
• v.31 no.2
• /
• pp.189-200
• /
• 2010

Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

• The Korea Journal of Herbology
• /
• v.23 no.2
• /
• pp.111-121
• /
• 2008

Objectives : The present study was carried out to investigate the effects of Astragali radix on the asthma treatment. EAR-I is an extract of astragali radix cultured for one year and EAR-III is an extract of astragali radix cultured for three years. Methods : Two asthma-induced groups mice group was treated respectively with EAR- I and EAR- III. The other group was not. Each group was analyzed in vivo and in vitro experiments. Results : In asthma-induced mice by OVA treatment, the weight of lung, total cell number of leukocyte and eosinophil in BALF, both EAR- I and EAR- III treated groups were significantly decreased compared to control group. IL-4, IL-5, IL-13, histamine, IgE in BLAF of group treated with both EAR-I and EM-III were significantly decreased compared to control group. In FACS analyzing, granulocytes, $CD3e^-$/$CCR3^+$, $CD3^+$/$CD19^$, $CD3e^+$/$CD69^+$, $CD4^+$, $CD8^+$ and $GR-1^+$/$CD11b^+$ were significantly increased in asthma induced mice group by OVA treatment compared to control group, and decreased in group treated with both EAR- I and EAR-III. Conclusions : The present data proves that extract of astragali radix has an effect on the inhibiting asthma. Moreover, astragali radix cultured for three years was proved to be superior to the one cultured for one year.

• IMMUNE NETWORK
• /
• v.5 no.3
• /
• pp.137-143
• /
• 2005

Background: Millions of people in the world are suffering from atopic dermatitis (AD), which is a chronic inflammatory skin disease triggered by Th2 immune responses. The NC/Nga mouse is the most extensively studied animal model of AD. Like human AD, NC/Nga mice demonstrate increased levels of IgE, a hallmark of Th2 immune responses. Adaptive immunity cannot be generated without help of innate immunity. Especially natural killer T (NKT) cells and marginal zone B (MZB) cells have been known to play important roles in linking innate immunity to adaptive immunity. Methods: Through flow cytometric analysis and ELISA assay, we investigated whether these lymphocytes might be altered in number in NC/Nga mice. Results: Our data demonstrated that the number of NKT cells was reduced in NC/Nga mice and IFN${\gamma}$ production by NKT cells upon ${\alpha}-GalCer$ stimulation decreased to the levels of CD1d KO mice lacking in NKT cells. However, reduction of NKT cells in NC/Nga mice was not due to CD1d expression, which was normal in the thymus. Interestingly, there was a significant increase of $CD1d^{high}B220^+$ cells in the spleen of NC/Nga mice. Further, we confirmed that $CD1d^{high}B220^+$ cells are B cells, not dendritic cells. These $CD1d^{high}B220^+$ B cells show $IgM^{high}CD21^{high}CD23^{low}$, a characteristic phenotype of MZB cells. Conclusion: We provide the evidence that there are decreased activities of NKT cells and increased number of MZB cells in the NC/Nga mice. Our findings may thus explain why NC/Nga mice are susceptible to AD.