Abel et al. in Germany discovered a new dioxin-responsive gene, which has later been identified as rpt-1 (regulatory protein T-lymphocyte 1). While it is speculated that rpt-1 may play a role in signal transduction and carcinogenesis, its roles and functions remain unknown. The present study attempted to analyze functions of rpt-1 in human epithelial cells following the xenobiotic exposures. While German counterpart analyzed expressionn of rpt-1 in spleen and thymus cells from mouse and rat and characterizes molecular properties of the gene, our work mainly focused on analyzing function of rpt-1 in human skin cells. Expression of rpt-1 in human cells were analyzed by western and northern blot RT-PCR analysis. Expression of rpt-1 as well as Staf-50 in human cells with or without exposure to environmental pollutants were also analyzed by northern blot analysis, since Staf-50 is homologous with rpt-1 and found in human cells. To help study roles of rpt-1 in human cell system, retroviral vector system carrying rpt-1 gene under the CMV promoter were constructed and transfected. Cells overexpressing the gene after the transfection showed an increase of cell density and soft agar colony formations, as compared to the control cells, suggesting that rpt-1 may play a certain role in the transformation processes of human cells. While the expression of rpt-1 in spleen and thymus is known to be strong in the laboratory animals, both the basal and TCDD-induced expression of rpt-1 in the current cellular system remained insignificant. It is speculated that the expression pattern of rpt-1 may be tissue- and species-specific. The present study demonstrated a strong expression of rpt-1 protein in the brain of SD rat model. Since there is no previous report on the expression of rpt-1 in the brain tissue, the result may play a significant role in understanding dioxin-induced neurotoxicities in the future. The present study provides an opportunity to understand a role of rpt-1 in human cell system and suggest a possible lead and basis for the future study of dioxin-induced neurotoxicities.
Kim, Sung-Ho;Oh, Heon;Lee, Song-Eun;Kim, Soon-Tae;Jo, Sung-Kee;Hyun, Byung-Hwa;Ryu, Si-Yun;Daynes, Raymond A.
Korean Journal of Veterinary Pathology
/
v.1
no.1
/
pp.7-12
/
1997
Lethally irradited C3H/HeN mice were transplanted with syngeneic bone marrow. The B cell regeneration levels of spontaneous serum Ig, fecal igA and specific ig to diphtheria toxoid were determined at various time points. The number of B220+ cells reached normal range at 4 weeks after bone marrow transplantation(BMT) in spleen and lymph node. The B cell number of spleen returned to normal relatively soon than in the lymph node. Within 5 to 7 weeks after BMT, the transplanted mice contained nearly normal levels of spontaneous serum IgA, IgG2b and fecal IgA, but 2 fold lower levels of serum IgG2a, IgM and IgG3. Especially IgG3 levels were within low-normal range throughout the study. One to two weeks after immunization the predominant anti-diphtheria toxoid subtype was IgM. The levels of specific serum Ig were very low and after booster immunization at week 6, the short-lasting increase of Ig production was notd.
This study was performed to produce monoclonal antibodies (mAb) specifically reacting with chicken leukocyte surface antigens. Popliteal lymph node cells of BALB/c mice previously immunized through foot-pad with peripheral blood mononuclear cells (PBMC) of chickens separated by Ficoll-Histopaque method. They were fused with P3X63Ag14 mouse myeloma cells. A total of 34 hybridomas secreted antibodies specifically binding to the PBMC. According to the reactivity patterns with PBMC, the mAbs were divided into 4 groups. Group 1 mAbs (IIB3, IIB10, IIE10) specifically reacted with non-adherent lymphocytes but not with adherent cells which were mainly composed of thrombocytes and monocytes in PBMC culture. These mAbs were reactive with 25-59% of thymus cells and 42-64% of spleen cells of chickens. They did not show any significant reactivity with cells in the bursa of Fabricius, T-cell (MDCC-MSB1) and B-cell (LSCC-1104B1) lines. These results indicate that Group I mAbs specifically reacted with T-lymphocyte subpopulation. Monoclonal antibodies in Group II (IC6, IG2-2 and IID9) showed specific reactivity with monocytes but not with thrombocytes or non-adherent cells in PBMC culture. These mAbs, though not reacted with the chicken macrophage cell line, HD11, also bound to macrophages of the spleen and lung in immunohistochemical staining. Five mAbs in Group III showed characteristics of binding to lymphocytes and monocytes, but not to thrombocytes. Twenty-three mAbs in Group IV showed specific reactivity to lymphocytes, monocytes, and thrombocytes. Two mAbs (IC3 and IE9) in Group IV reacted with most of PBMC.
In fish both humoral and cell mediated immune responses have been reported whereas antibodies recognizing specific cellular populations have not yet been developed except for ones recognizing surface Ig molecules on B lymphocytes. Our aim was to develop and characterize monoclonal antibodies (Mabs) specific for the immune-related cells. Mabs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously sensitized against Israeli carp (I. carp) kidney mononuclear cells. We obtained 44 Mabs positively reacting with I. carp kidney mononuclear cells and partially characterized 7 Mabs in the morphological and mitogen-based proliferative aspects. Fluorescence-activated cell sorter (FACS) analysis against I. carp kidney cells by using 7 different Mabs showed 80.3% for ICK 17-4, 65.1% for ICK 2-3, 64.1% for ICK 25-1, 67.5% for lCK 22-1, 70.8% for ICK 16-2, 76.8% for ICK 13-2, 79.7% for ICK II-I. Panning method was used for the isolation of Mabs specific mononuclear carp spleen cells followed by Wright's stain. The stained cell populations were identified as monocytes (ICK 17-4, ICK 2-3, ICK 25-1, ICK 22-1 and ICK 16-2), lymphocytes (ICK 11-1), and a mixed cell population of monocytes and lymphocytes (ICK 13-2). In cell proliferation assay, monocytes purified by ICK 17-4, 2-3 and 22-1 efficiently responded to Con A and PHA, while ones separated by ICK 25-1 did not react with any mitogens. Lymphocytes isolated by ICK 11-1, though it is not known whether they are T or B cells, were more responsive to Con A than PHA or LPS, suggesting that fish immune cells are somewhat different from mammalian cells in responding to mammalian T or B cell mitogens.
Ha, Jong-Cheon;Kim, Young-Hyun;Woo, Won-Hong;Nam, Sang-Yun
Korean Journal of Pharmacognosy
/
v.32
no.3
s.126
/
pp.226-232
/
2001
To explore the possible cancer therapeutic application of "Bo-yang-hwan-oh-tang" (BH), a herbal medicinal recipe used for improvement of blood stasis, we have examined its direct cytotoxicity against tumor cell, and induction of cytotoxic activity of lymphocytes. Water extract of BH alone did not exhibit direct cytotoxicity to Yac-1 target cells even with high concentrations (10 mg/ml). By exposure for 3 days, BH did not induce any nonspecific cytotoxic activity of mouse spleen cells, either, when assessed in a 4 hr $^{51}Cr-release$ assay. However, when BH was added during CD3 stimulation of non-adherent spleen cells, non-specific CTL activity was markedly promoted in a dose dependent manner. In contrast, BH did not alter activated NK cell activity following IL-2 stimulation. These data suggest that BH does not induce but upregulates non-specific CTL effecter function and that activated NK cell does not respond to BH. For elucidation of the mechanism underlying this function of BH, time kinetic study for IL-2 production using ELISA was undertaken. IL-2 production following CD3 stimulation was significantly augmented and higher level of IL-2 is sustained over 3 days in the culture medium by BH treatment. Moreover, addition of exogenous IL-2 during CD3 stimulation resulted in a similar level of cytotoxicity between control and BH-treated culture. These data indicate that the BH-mediated upregulation of non-specific CTL activity is contributed by augmentation of IL-2 production. Our data imply the possible application of BH for combination therapy of cancer with non-specific activator.
It is noted that Dioscorea quinqueloba is a medicinal herb that is widely used to treat cardiovascular disease and is assessed as useful to treat other various medical conditions. The immunopotentiating effects of the protein extract (DQP-1) from Dioscorea quinqueloba were thus formally investigated in vivo under incident of cold stress. In this case study, the spleen and thymus weight in mice was shown to have decreased after a measured exposure to cold stress, while the adrenal gland weight in the mice was shown to have increased. The systematic oral administration of DQP-1 significantly recovered the weight loss of the spleen and suppressed the adrenal gland hypertrophy during the association with cold stress. Additionally, the DQP-1 also restored the ascorbic acid level in the adrenal gland reduced after cold stress. The cold stress exposure lowered the percentage of $CD4^+$ and $CD8^+$ cells in the mouse thymus as determined by the flow cytometric analysis, as well as the levels of some serum immunological cytokines(interleukin-2, interleukin-12, and interferon-${\gamma}$) in the studied mice. The resulting identified weakened immunity caused by cold stress was also recovered by a treatment with DQP-1. The DQP-1 significantly suppressed the formation of serum enzymes of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase, which were systematically elevated during the cold stress episode. These results indicate that DQP-1 can improve immunity in mice that are characteristically weakened under stress.
PARK JONG-HWAN;LEE YEONHEE;MOON ENPYO;SEOK SEUNG-HYEOK;BAEK MIN-WON;LEE HUI-YOUNG;KIM DONG-JAE;KIM CHANG-HWAN;PARK JAE-HAK
Journal of Microbiology and Biotechnology
/
v.15
no.3
/
pp.603-608
/
2005
We recently isolated a novel probiotic strain, Lactobacillus fermentum PL9005 (KCCM-10250), from infant feces and showed that it had a potential immunoenhancing effect. In the present study, a safety assessment of the bacteria was performed using a BALB/c mouse model. Mice were administered with L. fermentum PL9005 daily for 28 days. There were no detectable changes in body weight, feed intake, or clinical signs, and no significant difference in hematological parameters or blood biochemistry between the L. fermentum PL9005-fed and control groups. Bacterial translocation was detected in the mesenteric lymph nodes, liver, and spleen of some mice with and without L. fermentum PL9005 feeding, however, the organisms were not related to ingestion of L. fermentum PL9005; this was confirmed by PCR using a species-specific primer. No gross lesions were detected in the liver, spleen, or intestine of L. fermentum PL9005-fed or control mice. Mucosal thickness in the ileum, cecum, and colon of L. fermentum PL9005-fed mice was not significantly different from that of corresponding organs in control mice. No inflammation or epithelial cell degeneration in the intestines was observed in any mice. These results indicate that ingestion of L. fermentum PL9005 is safe in mice and can be applied in the functional food market.
Kim, Sung-ho;Oh, Heon;Kim, Se-ra;Jo, Sung-kee;Byun, Myung-woo;Shin, Dong-ho
Korean Journal of Veterinary Research
/
v.40
no.2
/
pp.221-228
/
2000
We performed this study to determine the effect of Bu-Zhong-Yi-Qi-Tang, as a prescription of traditional Oriental medicine, and its major ingredients on jejunal crypt survival, endogenous spleen colony formation, apopotosis in jejunal crypt cells, lethality and hematological change of mice irradiated with high and low dose of Y-radiation. Bu-Zhong-Yi-Qi-Tang administration before irradiation protected the jejunal crypts (p<0.0001), increased the formation of endogenous spleen colony (p<0.05) and reduced the frequency of radiation-induced apoptosis (p<0.05). The survival rate and mean survival time of the groups treated with Bu-Zhong-Yi-Qi-Tang within 30 days after the treatment were far better than the irradiation control group. In the experiment on the effect of ingredients of Bu-Zhong-Yi-Qi-Tang, the result indicated that the extract of Rensan (Panax ginseng), Danggui (Angelica sinensis), Shengma (Cimicifuga heracleifolia) and Chaihu (Bupleurum falcatnosa) might have a major radioprotective effect. Although the mechanisms of this inhibitory effect remain to be elucidated, these results indicated that BU-Zhong-Yi-Qi-Tang might be a useful radioprotector, especially since it is a relatively nontoxic natural product. Further studies are needed to characterize better the protective nature of Bu-Zhong-Yi-Qi-Tang extract and its ingredients.
Kim, Gi-Yeol;Kim, Jong-Dae;Jeong, Ji-Cheon;Nam, Kyung-Soo
The Journal of Korean Medicine
/
v.19
no.2
/
pp.313-325
/
1998
The effect of Holotrichia on natural killer cell activity in normal mouse were studied. 1. The oral administration of Holotrichia increased spleen weight about 21.1% and also cell numbers of spleen compared to control mice group. 2. The cytotoxicity of effector cell was most effectively induced in a ratio of 50 : 1(effector/target cell). 3. Cytotoxicity of effector cells was. increased about 24% as compared with control group in in vivo test. 4. On the other hand, the administration of Holotrichia original solution showed significant increase the cytotoxicity. The cytotoxicity was increased concentration dependently. 5. The cytotoxicity by $^{3}H-thymidine$ incorporation assay showed similar effect with LDH enzyme method. 6. In the purified NK cells, the cytotoxicity was increased about 31% as compared with control group in in vivo system and the ratio of cytotoxicity was generally more increased than that of partially purified NK cell. 7. In vitro experimet of the purified NK cells, the cytotoxicity was increased 11.8% as compared with control group and the ratio of cytotoxicity was also more increased than that of partially purified NK cell. These results suggest that Holotrichia is administrated to mice with malignant tumors, the increase of NK cell activity may occur and affect tumor cells.
Heo, Moon-Young;Kwon, Chang-Ho;Sohn, Dong-Hun;Lee, Su-Jun;Kim, Sung-Wan;Kim, Jung-Han;William W. Au
Archives of Pharmacal Research
/
v.16
no.3
/
pp.196-204
/
1993
Flavonol derivatives were tested for their anticlastogenic effect against induction of micronuclei by n-methyl-n'-nitor-n-nitorsoguanidine(MNNG), and against induction of chromosome aberration by bleomycin or MNNG.belomycin. For micronudeus assay, each flavonol derivative (0, 0.001, 0.01, 0.1, 1, 10 and 100 mg/kg) was administered orally twice with 24 h interval, together with intraperitioneally administered MNNG(150 mg/kg). The result showed that msot flavonol derivatives tested were effective in suppresing the frequencies of micronude induced by MNNG. For chromosome aberration assy, each flavonol derivative (0, 0.1, 1, 10m and 100 mg/kg)was administered to mice orally in vivo, and then mice were sacrificed and spleen lymphocyte cultures were made. Bleomycin $(3\;\mu$g/ml) was treated to the mouse spleen hymphocyte cultures at 24 h after con A initiation. There wre nomarked decrease tendencies in chromosome aberration unless all doses of galangin and some doses of several flavonol derivatives tested. In the another experiment, we have evaluated the effect of flavonol derivatives on the enhancement of bleomycin-induced chromsome aberration by MNNG. Most of flavonol derivtives reduced the incidence of chromosome aberration induced by in vitro treatment of bleomycin followed by in vivo treatment of MNNG. Galangin particulary showed a dose-dependent decrease tendency. Other flavonol derivative showed slightly suggest that most of flavonol derivatives may be capable of protecting the inhibition of suggest that most of flavonol derivatives may be capable of protecting the inhibition of DNA-repair by MNNG. Our data indicate clearly that flavonol derivatives can suppress MNNG-induced genotoxicity such as an induction of MNPCEs. Therfore, our results could suggest that flavonol derivtives may be useful as a chemopreventive agent of MNNG.
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