• Korean Journal of Microbiology
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• v.54 no.3
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• pp.222-227
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• 2018

Mycobacterium tuberculosis (MTB)-originated lipid antigen is presented on the antigen-presenting cell surface with CD1b. When monocyte-derived dendritic cells phagocytosed MTB H37Rv (Multiplicity of infection 10, infectivity: 46.89%), the CD1b expression level decreased slowly. Since this was just a live MTB-mediated phenomenon, it was not detected from heat-killed MTB or mycolic acid, which is a unique antigen of MTB. We confirmed that the phosphorylation of CD1b molecules using 2D electrophoresis with staining could phosphorylate and induce the presentation of the lipid antigen using the phagocytosis assay.

• Biomedical Science Letters
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• v.2 no.2
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• pp.275-282
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• 1996

The authors inquired into what reactions comprise the response of mice(as a model) CD3, CD4 and CD8 monoclonal antibodies in spleen tissue when injected intraperitoneally by antigens of Clonorchis sinensis. The author is objective was focused on investigating the property of cellular immunity for liver fluke. In particular, the results of having examined the phenotype of the tissue of spleen were revealed as follows: a certain length of time after having been intraperitoneally injected with antigens of Clonorchis sinensis and Freund's adjuvant, the tissue of spleen was embedded and immunohistochemically stained by the avidin-biotin complex method. A strong reaction in response to CD3, while a feeble reaction resulted from CD4 and CD8. The tissue region showed a positive reaction to all antibodies, especially from capsules, vascular areas, white pulps and membrane of blood cells.

• Journal of the Korean Society of Visualization
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• v.13 no.1
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• pp.35-42
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• 2015

CD4+ T-cell count determines the effectiveness for antiretroviral therapy (ART) in patients with human immunodeficiency virus (HIV). Although ART slows the progression of HIV to AIDS, rapid counting of CD4+ T lymphocytes with a drop of patient's blood sample is urgently needed to ensure timely ART treatment in rural areas. Recently point-of-care CD4 testing devices have been developed by using non-flow based imaging cytometer incorporated with a sample cartridge where CD4+ T cells are reacted with fluorescently tagged specific antibodies. Here we conducted an experimental study using a conventional fluorescence microscope-based imaging system to quantitate the interaction of CD4 antibodies with CD4+ T cells at different reaction conditions. We demonstrated that a fast and affordable point-of-care CD4 test is feasible with a far less amount of antibodies and a shorter incubation time compared with a conventional sample preparation protocol for flow cytometry. We also proposed a general method to evaluate and compare the detection limit across different CD4 counting platforms by using fluorescently labelled microbeads for intensity calibration.

• Journal of Dairy Science and Biotechnology
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• v.14 no.2
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• pp.229-245
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• 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 Ginseng Research
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• v.31 no.4
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• pp.175-180
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• 2007

To evaluate whether there is a relation between Korean red ginseng (KRG)-intake and the suppression of immune hyperactivation in HIV-1-infected patients, we measured serum soluble CD8 (sCD8) over 31-48 months in 168 patients. They were divided into four groups; HIV-1-infected control (n = 49), zidovudine (ZDV) group (n = 22), KRG group (n = 48), and combination of KRG and ZDV group (n = 49). In control, sCD8 and the ratio of sCD8/CD8+ T cells significantly increased by 33% (paired t-test, P < 0.05) and 54% over $21\;{\pm}\;13$ months (P < 0.001), respectively. In ZDV group, sCD8 decreased within first 6 months and then showed steady increase and the ratio also increased over $19\;{\pm}\;10$ months. In KRG group, sCD8 and the ratio of sCD/CD8+ T cells continuously decreased by 45% (P < 0.01) and 19% over $19\;{\pm}\;11$ months (P < 0.05), respectively. In combination group, sCD8 gradually decreased by 29% (P < 0.01). There was a clear difference in the changes in serum sCD8 over time among 4 groups. There was no rebound phenomenon in KRG group as shown in ZDV group. These results suggest that KRG-intake suppresses immune hyperactivation state by HIV antigen itself in the HIV-infected patients.

• Journal of Life Science
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• v.16 no.6
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• pp.904-910
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• 2006

Dendritic cells (DCs) are the only antigen presenting cells (APCs) capable of initiating immune responses, which is crucial for priming the specific cytotoxic T lymphocyte (CTL) response and tumor immunity. Upon activation by DCs, CD4+ helper T cells can cross-prime CD8+ CTLs via IL-12. However, recently activated DCs were described to prime in vitro strong T helper cell type 1 $(Th_1)$ responses, whereas at later time points, they preferentially prime $Th_2$ cells. Therfore, we examined in this study the optimum kinetic state of DCs activation impacted on in vivo priming of tumor-specific CTLs by using ovalbumin (OVA) tumor antigen model. Bone-marrow-derived DCs showed an appropriate expression of surface MHC and costimulatory molecules after 6 or 7-day differentiation. The 6-day differentiated DCs pulsed with OVA antigen for 8 h (8-h DC) and followed by restimulation with LPS for 24 h maintained high interleukin (IL)-12 production potential, accompanying the decreased level in their secretion by delayed re-exposure time to LPS. Furthermore, immunization with 8-h DC induced higher intracellular $interferon(IFN)-{\gamma}+/CD8+T$ cells and elicited more powerful cytotoxicity of splenocytes to EG7 cells, a clone of EL4 cells transfected with an OVA cDNA, than immunization with 24-h DC. In the animal study for the evaluation of therapeutic or protective antitumor immunity, immunization with 8-h DC induced an effective antitumor immunity against tumor of EG7 cells and completely protected mice from tumor formation and prolonged survival, respectively. The most commonly used and clinically applied DC-based vaccine is based on in vitro antigen loading for 24 h. However, our data indicated that antigen stimulation over 8 h decreased antitumor immunity with functional exhaustion of DCs, and that the 8-h DC would be an optimum activation state impacted on in vivo priming of tumor-specific CTLs and subsequently lead to induction of strong antitumor immunity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.10
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• pp.1322-1328
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• 2006

In our previous study, a novel herb mixture (HIM-I) of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect= the intestinal and immune systems and to promote their recovery from radiation damage. A new herbal composition (HemoHIM) with the high immune modulating activity was developed from HIM-I. In the present study, we examined the effects of HemoHIM on the maturation process of murine bone marrow (BM)-derived dendritic cells (DC). BM cells were cultured in the presence of iL-4 and GM-CSF and the generated immature DC were stimulated with HemoHIM for 24 hours. HemoHIM significantly enhanced the expression of co-stimulatory molecules, CD80 and CD86, especially. The activation capacity of HemoHIM-treated DC was significantly higher than that of immature DC, as analyzed by IL-2 and $IFN-\gamma$ production and proliferation of the responding T cells in the co-culture with allogeneic T cells. The antigen-presenting capacity of HemoHIN-treated DC was also increased by the co-culture with OVA-specific T cells (HS-1), as analyzed by IL-2 and $IFN-\gamma$ production and the proliferation. These results indicate that HemoHIM causes the maturation and ;Activation of DC, which may be a part of mechanisms of immunomodulation by HemoHIM.

• Korean Journal of Poultry Science
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• v.28 no.3
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• pp.231-237
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• 2001

To examine the effects of feed additives on the expression of perpheral blood cell surface molecules, phagocytosis and antigen specific antibody formation, broilers were randomly assigned to $T_{1}$ , $T_{2}$ , $T_{3}$ , and $T_{4}$ groups. $T_{1}$ group was fed diet without any additives for 13 weeks, $T_{2}$ was fed diet with full fat flax, $T_{3}$ was fed diet with full fat flax containing $\alpha$-tocopherol, and $T_{4}$ was fed diet with full- fat flax containing $\alpha$-tocopherol and selenium. Since 5 weeks feeding the data were examined by flow cytometry using a panel of monoclonal antibodies. The expression of monocyte in all treated groups was significantly increased, in which the ratio of expression in $T_{3}$ group was especially evident. B cell expression of all treated groups was increased more than 2 fold. The expression of CD4+(helper T cell) cell and CD8+(cytotox$ic^pressor T cell) cell of all treated groups also was increased.ed.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.342-347
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• 2011

T cells play a key role in viral infection. However, in patients with chronic hepatitis C virus (HCV) infection, HCV-specific T cells are dysfunctional and impaired in the liver, which is the primary site for HCV replication. There are multiple potential mechanisms for HCV-specific T cell dysfunction including induction of immune inhibitory pathways (program death-1; PD-1, cytotoxic t lymphocyte associated antigen-4; CTLA-4) and immune tolerance induced specific for the liver. However, the interaction between hepatocytes and HCV-specific CD8 T cells has not clearly established. In this study, we confirmed huh (human hepatoma) 7.5 cells expressing HLA (human leukocyte antigen) A2 presented antigen to activate HCV-specific CD8 T cells in HLA A2-restricted manner and expression of PD-L (program death ligand) 1 on huh7.5 cells reduced HCV-specific CD8 T cell activation, suggesting an immune modulatory activity. Loss of HCV-specific tetramer responses following antigenic stimulation correlated with increased caspase-3 activity. In addition, PD-L1 on huh7.5 cells rescued HCV-specific CD8 T cells from apoptosis. Our results suggest that the interaction between PD-L1 and PD-1 can recover the function of HCV-specific CD8 T cells in the liver, which could be applied in therapy of HCV chronic infection.

• Biomedical Science Letters
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• v.2 no.1
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• pp.13-20
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• 1996

A Recent discovery of surface antigens in cells has led to the success of quantitative measurement of T-cell subpopulations, and this has especially opened the way for an epoch-making development in the understanding and classification of cellular immune mechanisms. It is known that phenotypes of T-cell subpopulations exist in many forms according to the variation of species or animal experimental models. In Korea, Clonorchis sinensis still gives rise to public concern as it infects more than eighty million people and threatens the public by causing cirrhosis of the liver, or liver cancer when liver infection becomes prolonged and chronic. Up until now there has been much progress in research and improvement in the classification system of Clonorchis sinensis in the area of humoral immunity, but as for research in the area of cellular immune mechanisms, there is almost none. Knowing all these circumstances, the authors delved for the characterization of Iymphocyte subpopulations with mice as Clonorchis sinensis in the area of cellular immunity, and obtained the following results. That is, we injected Clonorchis sinensis antigens mixed in Freund's ajuvant solution intraperitoneally in mice and measured the T-cell subpopulation characterization of spleen lymphocytes with flow cytometry. The results of these measurements showed that CD2, CD5 and CD8 decreased early following injections but then in-creased again seven weeks after the injections. CD4, however, showed a slight increase shortly after the injection but then a fair increase seven weeks after the injection.