Sang-Yun Nam

The nitric oxide-producing activities of Scutellaria baicalensis.

Scutellaria baicalensis (SB) has antibacterial and antiviral activities. Nitric oxide (NO) as a potent macrophage-derived effector molecule against a variety of bacteria, viruses and tumors has received increasing attention. The objective of this study was to determine the effect of SB on the production of NO. Stimulation of mouse peritoneal macrophages with SB after the treatment of recombinant interferon-gamma (rIFN-gamma) resulted in the increased NO production. SB had no effect on NO production by itself. When SB was used in combination with rIFN-gamma, there was a marked cooperative induction of NO production in a dose-dependent manner. The optimal effect of SB on NO production was shown 6 h after treatment with rIFN-gamma. NO production by SB was inhibited by N(G)-monomethyl-L-arginine. The increased production of NO from rIFN-(gamma) plus SB-stimulated cells was decreased by the treatment of protein kinase C inhibitor such as staurosporin. In addition, synergy between rIFN-gamma and SB was mainly dependent on SB-induced tumor necrosis factor-alpha (TNF-alpha) secretion. All the preparations of SB were endotoxin free. These results suggest that the capacity of SB to increase NO production from rIFN-gamma-primed mouse peritoneal macrophages is the result of SB-induced TNF-gamma secretion.

Antiasthmatic Activity and Selective Inhibition of Type 2 Helper T cell Response by Aqueous Extract of Semen Armeniacae Amarum

Semen armeniacae amarum (SAA) has long been used to control asthma in Korean traditional medicine. However, its antiasthmatic action still remains poorly understood. In the current study, effective mechanism of SAA was investigated in a mouse model of allergic asthma induced by repeated sensitization and intranasal challenge with OVA. Airway hyperreactivity (AHR) measured by β-methacoline-induced airflow obstruction and airway recruitment of leukocytes including eosinophils were significantly reduced by oral treatment of SAA water extract. Level of interleukin (IL)-4, but not Interferon gamma (IFN-γ), in the bronchoalveolar lavage fluid (BALF) also appeared considerably lower in SAA-treated mice than in controls. Collectively, these data show that SAA suppresses type 2 helper T cell (Th2), but not type 1 helper T cell (Th1), response. This hypothesis was supported further by the data of ex vivo cytokine production of peribronchial lymph node cells. Thus, oral administration of SAA attenuates asthmatic manifestations including AHR and airway inflammation, which possibly result from selective inhibition of Th2 response to allergen. Our data strongly suggest that SAA may be effectively applied to control other Th2-related diseases as well as allergic asthma.

Amphiphilic triblock copolymer poly(p-dioxanone-co-L-lactide)-block-poly(ethylene glycol), enhancement of gene expression and inhibition of lung metastasis by aerosol delivery.

We describe the development of an aerosol system for topical gene delivery to the lungs of C57BL/6 mice. This system is based on the combination of the commercial cationic lipid Lipofectin with a novel amphiphilic triblock copolymer, poly(p-dioxanone-co-L-lactide)-block-poly(ethylene glycol) (PPDO/PLLA-b-PEG, and abbreviated in the text as polymeric micelles). After optimizing conditions for DNA delivery to the lungs of mice using the combination of polymeric micelles with Lipofectin and LacZ DNA, we used the Lipofectin/polymeric micelle system to deliver the tumor suppressor gene PTEN to the lungs of C57BL/6 mice bearing the B16-F10 melanoma. Lipofectin/PTEN/polymeric micelles significantly improved gene expression of PTEN in the lungs of mice with no evidence of cell toxicity or acute inflammation. Importantly, lung metastasis, as measured by lung weight, was significantly reduced (P<0.001), as were total tumor foci in the lungs (P<0.001) and size of individual tumor nodules in animals treated with Lipofectin/PTEN/polymeric micelles compared with control animals. Survival time was also extended. These results suggest that the Lipofectin/polymeric micelle system is appropriate for enhancing gene delivery in vivo and that it can be applied as a non-invasive gene therapy for lung cancer.

c-myb stimulates cell growth by regulation of insulin-like growth factor (IGF) and IGF-binding protein-3 in K562 leukemia cells

c-myb plays an important role in the regulation of cell growth and differentiation, and is highly expressed in immature hematopoietic cells. The human chronic myelogenous leukemia cell K562, highly expresses IGF-I, IGF-II, IGF-IR, and IGF-induced cellular proliferation is mediated by IGF-IR. To characterize the impact of c-myb on the IGF-IGFBP-3 axis in leukemia cells, we overexpressed c-myb using an adenovirus gene transfer system in K562 cells. The overexpression of c-myb induced cell proliferation, compared to control, and c-myb induced cell growth was inhibited by anti-IGF-IR antibodies. c-myb overexpression resulted in a significant increase in the expression of IGF-I, IGF-II, and IGF-IR, and a decrease in IGFBP-3 expression. By contrast, disruption of c-myb function by DN-myb overexpression resulted in significant reduction of IGF-I, IGF-II, IGF-IR, and elevation of IGFBP-3 expression. In addition, exogenous IGFBP-3 inhibited the proliferation of K562 cells, and c-myb induced cell growth was blocked by IGFBP-3 overexpression in a dose-dependent manner. The growth-promoting effects of c-myb were mediated through two major intracellular signaling pathways, Akt and Erk. Activation of Akt and Erk by c-myb was completely blocked by IGF-IR and IGFBP-3 antibodies. These findings suggest that c-myb stimulates cell growth, in part, by regulating expression of the components of IGF-IGFBP axis in K562 cells. In addition, disruption of c-myb function by DN-myb may provide a useful strategy for treatment of leukemia.

Regulation of lymphocyte clustering by CD30-mediated ICAM-1 up-regulation.

CD30 is expressed transiently on activated B and T lymphocytes and constitutively on several B- and T cell lymphomas. CD30 functions include participation in negative selection of thymocytes, costimulation of activated T cells, isotype switching of B cells, and regulation of the effector activity of cytotoxic lymphocytes. Although CD30 is not a marker for T helper 2 (TH2) cells, it may participate in the polarization of TH1 and TH2 cells. The pleiotropic functions of CD30 are initiated by interaction of CD30-expressing cells with other immune competent cells expressing CD30-L and providing the signals for modulation of effector cell activity. Here, we report that CD30 signals generated by anti-CD30 on activated, normal murine T cells strongly up-regulate the expression of intercellular adhesion molecule 1 (ICAM-1, CD54), and to a lesser extent, ICAM-2 (CD102). CD30 signals moreover delay the subsequent decline of ICAM expression. CD30 cross-linking did not alter the expression of CD11a/CD18 (LFA-1), the counter receptor for ICAM abundant on T cells. CD30-mediated ICAM-1 up-regulation is independent of cytokine secretion and appears to be transmitted directly through NF-kappaB activation. CD30-mediated up-regulation of ICAM-1 expression led to a significant increase in cluster formation of lymph node cells. Increased lymphocyte self-aggregation mediated by CD30 may set the stage for fraternal signaling to modulate lymphocyte function.

In vitro immunomodulatory activity of Bo-Yang-Hwan-O-Tang

Bo‐yang‐hwan‐o‐tang (BHT), an herbal decoction has been mainly used for improvement of blood flow in oriental medicine. Its in vivo immunomodulation was recently demonstrated but the effective mechanisms have not been described. This study was carried out to evaluate in vitro immunomodulatory activity of BHT. Water extract of BHT significantly promoted in vitro proliferative responses of mouse spleen cells (SPC) and also further enhanced the proliferation of SPC stimulated with anti‐CD3 antibody. Unexpectedly, addition of BHT extract did not affect proliferation of both resting and CD3‐activated T cells, whereas it showed a strong mitogenic activity on B cells. Flow cytometric analysis of CFSE‐stained SPC showed that BHT‐mediated enhancement of CD3‐activated SPC proliferation is due to T cell, but not B cell, division. Mixed culture experiment combining T and mitomycin C‐treated B cells demonstrated that BHT‐mediated enhancement of CD3‐activated T cell proliferation was dependent on the presence of B cells. However, B cell‐derived factors were not involved in BHT effect on T cell proliferation. In the presence of B cells, BHT treatment resulted in a great enhancement in IL‐2 production of CD3‐activated T cells, and BHT effect on T cell proliferation was completely abrogated by addition of exogenous IL‐2, indicating that IL‐2 plays a critical role in BHT‐mediated enhancement of CD3‐activated T cell proliferation. Taken together, our data revealed that BHT possesses a potent B cell mitogenic activity and also can enhance activated T cell response through B cell regulation.

Expression of CD30 in testis and Epididymis of adult mice

CD30 is a member of tumor necrosis factor receptor (TNFR) superfamily and has pleiotropic functions including cell activation, proliferation, differentiation, and death, depending on cell types and stage of differentiation. Although CD30 expression has been described mainly in hematopoietic tissues, several types of nonhematopoietic tumors including embryonic carcinoma and germ‐cell tumors express CD30. We examined CD30 distribution in the testis and epididymis from wild type and CD30‐deficient mice. In the testis, spermatogonia, spermatocytes and Sertoli cells expressed CD30, but not in spermatids. Spermatogonia and spermatocytes near the basement membrane strongly reacted to anti‐CD30. In the epididymis, CD30 expression was exclusively observed in luminal epithelia and some interstitial cells. Taken together, these results show a spatio‐temporal regulation of CD30 expression in mouse testis and epididymis and suggest a possible role of CD30 in spermatogonia and spermatocytes.