Objectives & Methods : The purpose of this study is to observe the effects of Phellodendri Cortex Herbal-acupuncture solution (PC-HAS) at Joksamni (ST36) on collagen II induced arthritis in DBA-1J mice. The author performed several experimental items to analyze arthritis evaluation, change of weight, spleen size and adhesion rate, change of cytokine level, IgG, IgM and anti-collagen II, chang of immunocyte count, histological change of CIA mouse joint. Results : 1. In the PC-HA group, arthritis index, the incidence of arthritis and joint edema were significantly decreased. 2. In the PC-HA group, the change of spleen size, spleen adhesion rate and the knee joint were significantly decreased. 3. The levels of $IL-1{\beta}$, IL-6 and INF- in serum of the CIA mouse were significantly decreased by PC-HA. 4. The levels of IgG, IgM and anti-collagen II in serum of the CIA mouse were significantly decreased by PC-HA. 5. In the CIA mouse spleen cell culture, the levels of IFN- , IFN- / IL-4, IL-10 were significantly decreased by PC-HA, but the level of IL-4 was significantly increased by PC-HA. 6. In the PC-HA group, the ratios of $CD3e^+$ to $CD45R^+$ cell, $CD4^+$ to $CD8^+$ cell and $CD4^+/CD25^+$ cell were similarly maintained as normal group in the CIA mouse spleen cell. 7. In the PC-HA group, $CD4^+CD25^+$ and $CD45R^+/CD69^+$ cell were significantly decreased in the lymph nodes. 8. In the PC-HA group, $CD3^+/CD69^+$ and $CD11b^+/Gr-1^+$ cell were significantly decreased in knee joint. 9. In histology, the cartilage destruction and synovial cell proliferation in the PC-HA group were similar with that of the normal group and the collagen fiber expressions in the PC-HA group were similar with that of the normal group. Conclusions : Form the result above, the results suggest that the PC-HA at ST36 has significant effect on collagen-induced arthritis, and can be put to practical use in the future rheumatoid arthritis clinic.
A polysaccharide fraction, AIP1, purified from Artemisia iwayomogi was shown to have immunomodulating and anti-tumor activities in mice. In order to determine how the AIP1 fraction exhibits the immunomodulating activity, the effect of the fraction on the apoptosis of mouse spleen cells was investigated. Treatment of the mouse spleen cells with the AIP1 fraction resulted ,in the suppression of apoptotic death and an extension of cell survival in culture, indicating that the fraction might modulate the death of spleen cells. Treatment of the mice with the AIP1 fraction in vivo also resulted in less apoptosis of the spleen cells, which indicates the physiological relevance of the anti-apoptosis effect of the fraction in vitro. A mouse gene array was used to determine the profile of the gene expression change showing a pattern of up- and down-regulated genes by the AIP1 treatment. This study provides preliminary information regarding the immunomodulatory mechanism of the AIP1 fraction.
TGF-$\beta$3 is among five TGF-$\beta$ isolorms and shows 80% sequence identity to TGF-$\beta$I, a prototype of TGF--$\beta$. It has been reported that TGF-$\beta$I, particularly in the presence of IL-2 or L-5, increases the pmduction of IgA and IgG2b isoiypes by LPS-actwated murine B cells. We examined the effect of TGF-P3 on Ig synlhesis by B cells from different lymphoid origins. IgA induction by TGP-$\beta$3 was mardnal in LPS-activated spleen B cell culture, while 1gA production was markedly enhanced in the culture shulated with TGF-$\beta$P3 and L-5. In addition, number of IgA secreting cells was increased by TGF-$\beta$P3. Under the same conditions, TGP-$\beta$3 alone was enough to increase IgG2b production but IgM and 1gGl. Sirmlar patiem of IgA and IgGZb enbancement by TGF-$\beta$3 and L-5 was observed in the cullures of mesenteric lymph node B cells. Thus, overall effect of TGF-$\beta$3 on Ig synthesis was quite similar to that of TGF-$\beta$I. Nonetheless, it remains to be underslood whether TGF-$\beta$3 is an important modulator in B cell differentiation since regulation of TGF-$\beta$3 expression is considered to differ from that of TGF-$\beta$I
1. Objectives: The purpose of this study is to investigate the effect of Suhwagije-tang(SGT) distillate on the immune activity of spleen cells of aged SD rats. 2. Methods: We used 10, 50, 72 weeks old SD rats in this study. Spleen cells from SD rats were stimulated with ConA and treated with 1% Vitamin C(Vit.C) or Suhwagijetang distillate(SGT). After 24 hours, the concentrations of IL-2, IL-4, IL-10, IFN-${\gamma}$ in the cell culture supernatant were measured by ELISA. 3. Results and Conclusions 1) At all concentration of SGT distillate, survival rates of liver cells were higher than the control group. In addition, 50% SGT distillate group's cell survival rates were significantly higher than other groups. 2) In 10 weeks SD rats(SGT group), the concentration of IL-2 significantly decreased in comparison with ConA group, Vit.C group. In 52 weeks SD rats(SGT group), the concentration of IL-2 significantly decreased in comparison with ConA group. 3) In 10, 52 weeks SD rats(SGT group), the concentration of IL-4 significantly decreased in comparison with ConA group. 4) In 10 weeks SD rats(SGT group), the concentration of IL-10 significantly decreased in comparison with ConA group. And in 72 weeks SD rats(SGT group), the concentration of IL-10 significantly increased in comparison with Vit.C group. 5) In 52, 72 weeks SD rats(SGT group), the concentration of IFN-${\gamma}$ significantly decreased in comparison with 10 weeks SD rats(SGT group). These results suggest that Suhwagije-tang(SGT) distillate has the effect of increasing the immune activity of spleen cells of aged SD rats.
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.
To investigate the influence of age and dietary fat level on the immune function and the growing potential of the fibroblast cells, male rats of 2 month, 6 month and 30 month of age were fed either 6% or 30% fat diet for 16 weeks. The weight of thymus decreased linearly with increasing age. And this age-dependent degeneration of thymus was delayed in rats fed low fat diets. The blastogenesis of spleen lymphocytes to PHA, ConA, and PWM was decreased with increasing age, however, no effect of dietary fat level was observed. The age-related decline in ratios of PHA/ConA response may suggest that T suppressor cell activity increases with age. In cell culture system, lung fibroblast cells from 30M rats showed lower plating efficiency. longer doubling time. and shorter cumulative doubling potential than those from 2M or 6M animals. Also. the morphology of fibroblasts from 30M rats was tended to be rouned rather than flattened and more variable in size and being generally larger. wherease those from 2M and 6M rats were uniform in size and adhered tightly to the culture vessel in ordered arrays. Therefore fibroblast cell culture system tried in this study reflects the changes of cellular aging.
Objectives : The aim of the present study was to observe the effect of Astragali Radix Herbal-Acupuncture Solution(AR-HAS) at $ST_{36}$(jok-samni, $Z\acute{u}s\bar{a}n$ Li) on collagen- II -induced arthritis(CIA) in mice. Methods : DBA/1J mice were immunized with bovine type II collagen(CII) on days 0 and 21 to induce arthritis. The mice were divided into 5 groups : normal group(no CIA), control group(CIA+no treatment), needle prick group(CIA+single prick with an injection needle), saline group(CIA+saline injection) and ARHA group(CIA+ R-HA treatment). The needle prick, saline injection, and AR-HA groups were injected on the right $ST_{36}$(jok-samni) of mice for 9 weeks, 3 times a week, beginning 4 weeks after the booster immunization. Results : 1. The incidence of arthritis, AI(arthritis index), and joint edema decreased in the AR-HA group. 2. Weight gain, hypertrophy of the spleen, adhesion of the tissues, and transformation of the joint were restrained in the AR-HA group. 3. The concentrations of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, $IFN-{\gamma}$ in CIA mouse serum and $IFN-{\gamma}$, IL-10 in the CIA mouse spleen cell culture decreased significantly at $ST_{36}$ in the AR-HA group. 4. Total cell counts increased significantly, and the ratio of $CD3e^+$ to $CD45R^+$, $CD4^+$ to $CD8^+$, and $CD4^+$ to $CD25^+$ cells decreased significantly at $ST_{36}$ in the CIA mouse spleen cell culture of the AR-HA group. 5. Total cell counts decreased significantly, and $CD4^+/CD25^+$ and $CD45R^+/CD69^+$ decreased significantly at $ST_{36}$ in the CIA mouse lymph nodes of the AR-HA group. 6. $CD3e^+/CD69^+$ and $CD11b^+/Gr-1^+$ cells decreased significantly at $ST_{36}$ in the CIA mouse joints of the AR-HA group. 7. The histological examination showed that cartilage destruction and synoviocyte proliferation decreased in the CIA mouse joints of the AR-HA group, and collagen fiber was expressed similar to that seen in the normal group. Conclusions : Our experiments show that at $ST_{36}$, an anti-inflammatory mechanism of AR-HA controls synovial cell proliferation and protects against cartilage destruction in rheumatoid arthritis.
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.
The case reports for clostridium type A enterotoxemia in Formosan deer have rarely been reported. This paper describes a natural case of type A enterotoxemia in farmed Formosan deer in Cheongwon-gun. A dead, male 10-month-old Formosan deer was submitted to Chungbuk Livestock and Veterinary Research Institute, March 24, 2001 and examined. That deer was fed with assorted grain feed, oak leaves, acorn and bean curd. Grossly there was no visible external change. Despite of the carcass being examined within 12 hours of death, there was a quite degree of posonortem decomposition. There was severe hemorrhage in the serosa of abomasum and small intestine. Much blood tinged and watery contents were contained in those organs. Also there were severe swelling of spleen, some red foci in hepatic parenchyma. Microscopically there were severe congestion and hemorrhage in mucosa submucosa, muscular layer, and serosa of abomasum and small intestine. Also spleen and pancreas showed severe Congestion and hemorrhage. There were multifocal hemorrhage with hepatic necrosis in periportal area and focal mononuclear cell deposition in sinusoid. In bacterial culture for small intestine, Cl perfringens was isolated. By toxin typing for the strain, that had $\alpha$ -toxin belonged to type A. In electronmicroscopy for feces, no vims particle was detected. Considering clinical signs, gross lesions, microscopic lesions, bacterial culture, and toxin typing of the isolate, this case was diagnosed as enterotoxemia by Cl perfringens type A.
This study was carried out to determine effectively the sex of rabbit embryos using H-Y antiserum. H-Y antiserum was obtained from inbred SD strain female rat which was immunized by injection of testis cell of inbred SD strain male rate into its spleen. The titer of antiserum was identified by sperm cytotoxicity test and culture of rabbit embryos with antiserum. The developed or undeveloped embryos were separated by exposure the embryos to the antiserum with H-Y antibody. Developed embryo were transferred to the recipients and sex of offspring were examined. 1. In the sperm cytotoxicity test, the rate of dead sperm showed no difference between two antisera from spleen and testis cell as antigens. It is confirmed that H-Y antibody in antiserum was absorbed by H-Y antigen in male rat spleen cells. 2. When rabbit morulae were exposed to antiserum and complement, the rate of embryos developed or arrested was 51 and 49% respectively and the rate was closely same as natural sex ratio of 50:50%. 3. When rabbit morulae were cultured for 12h in the medium containing antiserum produced by antigen of testis cell, the rate of embryos developed or arrested was 48 and 52% respectively and the rate was closely same as natural sex ratio of 50:50%. 4. Eighty rabbit embryos which were not affected by H-Y antiserum were transferred to four recipients. Two recipients were pregnant and born 13 pups among which 2 (14%) were male and 11 (86%) were female. In conclusion, existence of H-Y antibody in the serum from female rat immunized by injecting testis cell from newborn male rat to the spleen of the female rat was confirmed. When rabbitmorulae were exposed to H-Y antiserum and complement, about a half of embryos were developed to blastocysts. When the rabbit embryos not affected by H-Y antiserum were transferred, the rate of female offspring was 86%. Therefore, it was identified that most of embryos which were not affected by H-Y antiserum were female.
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