• IMMUNE NETWORK
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• 제5권2호
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• pp.89-98
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• 2005

Background: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. Methods: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, $gB_{498-505}$ DNA) and recombinant vaccinia virus $(vvgB_{498-505})$ expressing epitope $gB_{498-505}$ (SSIEF ARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with $gB_{498-505}$ epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. Results: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and $vvgB_{498-505}$, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with $vvgB_{498-505}$ as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly $gB_{498-505}$ DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and $vvgB_{498-505}$. Of particular interest CD8+ T cell-mediated immunity was optimally induced when $vvgB_{498-505}$ was used to prime and gB DNA was used as alternative boost. Especially CD7+ T cell responses induced by such protocol was longer lasted than other protocols. Conclusion: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.

• 생명과학회지
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• 제14권4호
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• pp.670-675
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• 2004

본 연구에서 인간의 백혈병세포주인 Jurkat T 세포에서 인터페론 감마(INF-${\gamma}$ 유전자의 methylation에 대한 S-nitroso-N-acetylpenicillamine (SNAP), 프로스타글란딘 $E_2$ (PG $E_2$) 그리고 dibutric cyclic AMP (dbcAMP)의 효과를 조사하였다. 인터페론 감마 유전자의 프로모터기능에 아주 중요한 디뉴클레오티드인 CpG는 SNAP, PG $E_2$, 그리고 dbcAMP를 각각 처리하였을 때 methylation되었다. PG $E_2$에 의해서 유도된 그 methylation은 아데닐산 사이클라제의 저해제의 하나인 2',5'-dideoxyadenosine (DDA)에 의해서 억제되었지만, SNAP에 의해서 유도된 methylation은 DDA에 의해서 억제되지 않았다. PG $E_2$나 dbcAMP를 처리한 세포에서 일산화질소(NO)의 생성의 증가는 나타나지 않았으며, PG $E_2$나 dbcAMP에 의해 유도된 인터페론 감마유전자의 methylation도 일산화질소합 성효소의 저해제인 $N^{G}$ -methyl-L-arginine (L-NMMA)에 의해서 억제되지 않았다. 따라서 인간의 Jurkat T 세포에서 PG $E_2$에 의한 인터페론 감마 유전자의 발현 억제는 세포내의 cAMP생성경로를 통한 인터페론 감마 유전자의 methylation과 연관되어있으나 일산화질소의 생성경로와는 무관한 것으로 보인다.화질소의 생성경로와는 무관한 것으로 보인다.

• Food Science and Biotechnology
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• 제17권5호
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• pp.947-952
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• 2008

The unripe fruit of Momordica charantia (MC) has been shown to possess antidiabetic activity. However, the mechanism of its antidiabetic action has not been fully understood. In this study, the effects of the aqueous ethanolic extract of MC (AEE-MC) were evaluated on the apoptosis in pancreatic $\beta$-cells treated with a combination of the cytokines, interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-$\alpha$, and interferon (IFN)-$\gamma$. In MIN6N8 cells, the inhibitory effect of AEE-MC was significantly observed at 2 to 50 ${\mu}g/mL$: a 26.2 to 55.6% decrease of cytoplasmic DNA fragments quantified by an immunoassay. The molecular mechanisms, by which AEE-MC inhibited $\beta$-cell apoptosis, appeared to involve the inhibition on the expression of p21, Bax, and Bad, the up-regulation of Bcl-2 and Bcl-$X_L$, and the inhibition on the cleavage of caspase-9, -7, and -3 and poly (ADP-ribose) polymerase. This study suggests that MC may inhibit cytokine-induced apoptosis in $\beta$-cells and, thus, may contribute via this action to the antidiabetic influence in diabetes.

• 생약학회지
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• 제36권4호통권143호
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• pp.332-337
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• 2005

We here demonstrate the evidence of increased anti-tumor and immunostimulating activities of crude extracts (GAL) from Galium. aparine L. In experimental lung metastasis of colon26-M3.1 carcinoma or B16-BL6 melanoma cells, prophylactically intravenous (i.v) administration of GAL significantly inhibited lung metastasis in a dose-dependant manner. In an in vitro cytotoxicity analysis, GAL at the concentration up to $500\;{\mu}g/ml$ did not affect the growth of B16-BL6 melanoma cells. In contrast, GAL showed the enhancement of splenocyte proliferating activity in a dose-dependent manner. Peritoneal macrophages stimulated with GAL produced various cytokines such as $1L-1{\beta},\;TNF-{\alpha},\;IFN-{\gamma}$ and IL-12. These data suggest that GAL has an antitumor activity to inhibit tumor metastasis, and its antitumor effects is associated with activation of nonspecific immnune related cells.

• 한국환경성돌연변이발암원학회지
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• 제22권2호
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• pp.112-124
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• 2002

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been reported to exert detrimental toxicities on various organ systems including reproductive, cardiovascular, nervous, or dermal system. Immunomodulatory effects of TCDD is thymic atrophy, downregulation of cytotoxic T or B lymphocyte differentiation and activation, which were demonstrated using experimental animals, whereas immunotoxicity in human has not been investigated well. This study was proceeded to evaluate general immunologic spectrum of the Korean Vietnam War veterans exposed to TCDD during their operation, and compare with that of the non-exposed control subjects with similar age. Regarding composition and quantity, immune cells in peripheral blood collected from the TCDD-exposed was not much different from those of the control except decreased red blood cell, hemoglobin and hematocrit level. Furthermore, plasma IgG2, G3, and G4 isotype distribution was similar between two groups, but IgG1 level was significantly lowered in the TCDD-exposed, indicating a TCDD-mediated functional alteration of B cells. Significantly enhanced level of IgE in plasma, a hallmark of dermal or respiratory allergic response, was also observed in the TCDD-exposed compared with that of the control. Elevated generation of IL-4 and IL-10 was resulted from in vitro stimulation of T cells with PMA plus ionomycin or PHA, respectively, from the TCDD-exposed in comparison to those of the control, suggesting a skewed type-2 response. In addition, the level of IFN${\gamma}$, a multifunctional cytokine for T cell-mediated immunity, was lowered in the TCDD-exposed with upregulation of tumor necrosis factor $\alpha$. The present study suggests that TCDD exposure disturbs immunohomeostasis in humans observed as an aberrant plasma IgE and IgG1 levels and dysregulation of T cell activities.

• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7965-7970
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• 2014

One of the goals of tumor immunotherapy is to generate immune cells with potent anti-tumor activity through in vitro techniques using peripheral blood collected from patients. However, cancer patients generally have poor immunological function. Thus using patient T cells, which have reduced in vitro proliferative capabilities and less tumor cell killing activity to generate lymphokine-activated killer (LAK) cells, fails to achieve optimal clinical efficacy. Interleukin-2 (IL-2) is a potent activating cytokine for both T cells and natural killer cells. Thus, this study aimed to identify optimal donors for allogeneic LAK cell immunotherapy based on single nucleotide polymorphisms (SNP) in the IL-2 and IL-2R genes. IL-2 and IL-2R SNPs were analyzed using HRM-PCR. LAK cells were derived from peripheral blood mononuclear cells by culturing with IL-2. The frequency and tumor-killing activity of LAK cells in each group were analyzed by flow cytometry and tumor cell killing assays, respectively. Regarding polymorphisms at IL-2-330 (rs2069762) T/G, LAK cells from GG donors had significantly greater proliferation, tumor-killing activity, and IFN-${\gamma}$ production than LAK cells from TT donors (P<0.05). Regarding polymorphisms at IL-2R rs2104286 A/G, LAK cell proliferation and tumor cell killing were significantly greater in LAK cells from AA donors than GG donors (P<0.05). These data suggest that either IL-2-330(rs2069762)T/G GG donors or IL-2R rs2104286 A/G AA donors are excellent candidates for allogeneic LAK cell immunotherapy.

• Clinical and Experimental Pediatrics
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• 제45권7호
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• pp.875-883
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• 2002

목 적 : 기관지 천식은 Th2 반응 우세로 인한 기도의 호산구증과 IgE 증가를 특징으로 하는 질환이다. 박테리아 DNA 혹은 합성 ODNs인 unmethylated CpG motifs(CpG)은 Th1 사이토카인인 $INF-{\gamma}$와 IL-12 유도와 Th2 사이토카인을 하강 조절하여 기도 내의 호산구증 및 전신적 IgE 생성을 억제하며, 메타콜린 흡입에 의한 기관지 과민성을 방지한다. 저자들은 기도 염증을 유발한 생쥐의 비강내에 합성 CpG ODNs의 점적 투여로 기도의 호산구증과 그 외 면역학적 기전에 대한 영향을 연구하고자 한다. 방 법 : BALB/c 생쥐에 주당 2번씩 면역보강제 없이 OVA를 $10{\mu}g$을 피하 주사하여 감작 시켰다. 10주째에 $50{\mu}g$ OVA를 백서의 비강내로 주입하여 기도 염증을 유도한 후, 4주 뒤인 14주째 다시 생쥐의 비강내에 OVA＋CpG ODNs과 OVA를 점적 흡입 주입하였다. 마지막 항원 유발(14주) 후 4일째에 기관지 폐포 세척액을 추출하여 세포를 염색한 후 호산구를 측정하였고 동시에 비장 세포를 배양하여 사이토카인 측정을 하였다. 비장 세포에서 OVA에 의해 생성된 사이토카인 즉 Th1($INF-{\gamma}$, IL-12), Th2(IL-4, IL-5, IL-13) 사이토카인을 ELISA에 의해 측정하였다. 혈청 항원 특이 IgE, IgG2a 또한 ELISA에 의해 측정하였다. 결 과 : 1) 기관지폐포 세척액의 호중구수 : CpG ODNs을 투여한 군이 7.6[5.8-9.4]%, CpG ODNs을 투여하지 않은 군이 42.0[38.2-44.6]%로 기도 염증 세포내의 호산구증을 현저히 억제시켰다(P<0.01). 2) 혈청 OVA-specific IgE와 IgG2a : CpG ODNs으로 투여한 군과 투여하지 않은 군에서 측정한 혈청 OVA-specific IgE와 IgG2a는 음성 대조군에 비해 현저히 증가하였고, CpG ODNs을 비강내 투여 후 4일째 측정한 혈청 OVA-specific IgE와 IgG2a 수치는 CpG ODNs 투여군과 비투여군 사이에 유의한 차이가 없었다. 3) 사이토카인 분석 : CpG ODNs으로 투여한 군은 투여하지 않은 군에 비해 IL-4, IL-5, IL-13의 생성이 감소하였고 IL-12 생성은 증가하였으나, $INF-{\gamma}$의 상향조절은 보여주지 못하였다. 결 론: CpG ODNs의 백신 접종은 면역반응을 조절하여 기도 염증이 형성된 생쥐에서 기도 호산구증을 감소시키는데 매우 효과적이며 천식 치료에 유용할 수 있을 것으로 사료된다.

• 동의생리병리학회지
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• 제21권5호
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• pp.1233-1242
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• 2007

Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which might be mediated by the altered activation of Immune system, ultimately leading to the destruction of cartilage and bone. To examine effects of GHS on rheumatoid arthritis DBA/1J mice were immunized with bovine type II collagen to induced arthritis and then treated with GHS once a day for 7 weeks. Oral administration of GHS (200 mg/Kg) significantly suppressed the progression of CIA, which extend is comparable to that of methotrexate (MTX, 0.3 mg/Kg), a positive control. The severity of arthritis within the knee joints, which was evaluated by histological assessment of cartilage destruction and pannus formation, was also lowered by GHS. The production of TNF-and IL-6 in serum was significantly suppressed. The levels of IFN-g in the culture supernatant of splenocytes stimulated with CD3/CD28 or collagen were dramatically decreased, while those of IL-4 was increased. The levels of IgG and IgM RA factor were also decreased in the serum. FACS analysis indicated that B cells (in DLN), CD3+ T cells (in spleen, and paw joint), CD11b+Gr-1+ cells (in paw joint), CD3+CD49b(DX5) (in PBMC) were decreased and there was increased proportion of CD3+, CD4+, CD8+, CD4+CD25+ T cells in DLN. In conclusion, our results demonstrates that GHS significantly suppressed the progression of CIA and this action was characterized by the decreased production of TNF-a, IL-6, and rheumatoid factors, and modulations of immune cell populations.

• 생약학회지
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• 제42권4호
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• pp.354-360
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• 2011

In order to screen new allogeneic mixed lymphocyte culture (allo-MLR) inhibitor which is expected to be immunomodulating drug lead, we have investigated allo-MLR inhibitory activity on the marine-derived symbiotic microorganisms (1,895 strains) from the marine algae. The potent inhibitory activities (over 45% inhibition of proliferation at 10 and 2 ${\mu}g/ml$) without cytotoxicity were observed in the extracts of 46 strains. While, the significant stimulating activities (over 100% proliferation at 10 and 2 ${\mu}g/ml$) without cytotoxicity were observed in the extracts of 5 strains. In the second assay using 46 bioactive strains, 14 strains exhibited again significant allo-MLR inhibitory activity. Finally, 11 strains among the 14 strains inhibited proliferation and IFN-${\gamma}$ production of CD4+ T cells during the stimulation with specific antigen in the third assay. On the basis of above results, the marine algae is nice source for isolation of immunomodulating microorganism, and the marine algae-associated microorganism is also nice target for development of the new immunomodulating drug lead.

• 생약학회지
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• 제41권4호
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• pp.275-281
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• 2010

To evaluate the immunomodulatory activity of a water extract (KMF-WE) of Korean mistletoe (Viscum album var. coloratum) fruit, we examined its ability to induce humoral and cellular immune response against keyhole limpet hemocyanine (KLH). Immunized mice with KLH admixed with KMF-WE (KLH/KMF-WE) showed significant induction of KLH-specific antibodies compared to mice immunized with KLH alone. The assay for determining isotypes of antibodies revealed that KMFWE augmented KLH-specific-IgG1 and -IgG2a production. In vitro T lymphocyte proliferation analysis against KLH revealed that the splenocytes of mice immunized with KLH/KMF-WE showed a significantly higher proliferative ability than those from mice immunized with KLH alone. The culture supernatants of splenocytes, which were harvested from mice immunized with KLH/KMF-WE, showed higher levels of both Th-1 type (IL-2, IFN-${\gamma}$) and Th-2 type (IL-4) cytokines in response to KLH stimulation compared to those from mice immunized with KLH alone. Also, in delayed-type hypersensitivity (DTH) assay, mice immunized with KLH/KMF-WE showed a significantly higher reaction to KLH than mice treated with KLH alone. These results suggest that KMF-WE possess immunoadjuvant activity to enhance both antigen-specific humoral and cellular immune responses against protein antigens (KLH).