A pathogenic free-living amoeba, Naegleria fowleri, is a causative protozoan parasite of primary amebic meningoencephalitis in human and experimental animals. It is known that humoral and cellular immunity contribute as the defence mechanism of host against this organism. Recently splenectomy has been argued on its effect on host defence mechanisms. The present study was aimed to observe the enact of immunization in splenectomized mice. For immunization, $5~10{\times}10^5$ trophozoites of Naegleria fewleri o 359 were intraperitoneally inoculated once a week for two weeks to BALB/c mice, and $5~10{\times}10^4$ of ameba trophozoites were intranasally inoculated for infection after splenectomy and/or immunization. ELISA technique was applied for the detection of seum IgG antibody levels. Experimental animals were divided into 4 groups; I. splenectomized and immuniEed; ll. splenectomized only; III. immunized only; IV. not splenectomized nor immunized. The results obtained were as follows: 1. Mortality rates of splenectomized and immunized mice in group I (38.1%) and immunized only in group III (25.0%) were lower than those of not immunized mice in group II (50%) and control group, IV (46.4%). 2. Survival times of mice in group I, II, III and IV were $20.1{\pm}3.6$, $17.3{\pm}4.5$, $20.4{\pm}7.0$ and $19.6{\pm}7.6$ days respectively, and there were no significant differences between them. 3. ELISA values (absorbance at 492nm) of group I (1, $10{\pm}0.29$) and group III ($1.31{\pm}0.28$) were significantly higher than that of group IV($0.24{\pm}0.37$) at day 31 of infection (p<0.05). Conclusively, it is presumed that humoral immunity against N. fowleri may operate as ever, after immunization, even though the mouse was splenectomized.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.14
no.1
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pp.129-153
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2001
Background: SMG (升麻葛根湯加味方) is an herbal medicine which has been used in oriental medicine as a traditional therapeutic agent of pruritus and skin disease. Objective: This study was performed to investigate the effect of SMG on the anti-hypersensitivity and immune response in the murine of type I hypersensitivity induced by the experiment. Materials and Methods: Laboratory rats were primary sensitized with OA (ovalbumin); on day 1, rats of a Control group and Sample group (SMG group) were systemically immunized by subcutaneous injection of 1mg OA and 300mg of Al(OH)3 in a total volume of 2ml saline. The rats of the sample group were orally administered with an SMG water extract for 14 days after primary immunization. On day 14 after the systemic immunization, rats received local immunization by inhaling $0.9\%$ saline aerosol containing $2\%$(wt/vol) OA. A day after local immunization, BAL fluid and peripheral blood were collected from the rats. Total cell, lymphocyte, $CD4^+\;T\;cell,\;CD8^+\;T\;cell,\;CD4^+/CD8^+$ ratio in the BALF, and IgE, $CD4^+\;T\;cell,\;CD8^+$ T cell in the peripheral blood were measured and evaluated. Results: SMG showed a suppressive effect on the immune response in the rats. 1. Total Cells in the BALF decreased in the SMG treated group in comparison group, but statistic differences were not observed. 2. Total lymphocytes in the BALF were statistically decreased in SMG treated group in comparison to the control group. 3. CD4+ T cells in the BALF were statistically decreased in SMG treated group in comparison to the control group. 4. CD8+ T cells in the BALF were decreased in SMG treated group in comparison to the control group, but statistic differences were not observed. 5. The ratio of CD4+/CD8+ in the BALF was statistically decreased in SMG treated group in comparison to the control group. 6. The IgE level in serum was statistically decreased in SMG treated group in comparison to the control group. 7. The ratio of CD4+ and CD8+ in peripheral blood showed undetectable differences between each group of rats. From the experiment cited above, this study shows that SMG has both anti-hypersensitivity effects and immunoregulatory effects when administered to rats. Based on this experiment, it is suggested that SMG could be a useful immunomodulator and anti-allergy agent.
Background: SBP(桑白皮)is an herbal medicine which has been used in oriental medicine as a traditional therapeutic agent of bronchial asthma. Objective: This study was performed to investigate the effect of SBP on the anti-hypersensitivity and immune response in the murine of type I hypersensitivity induced by the experiment. Materials and Methods: Laboratory rats were primary sensitized with OA(ovalbumin); on day 1, rats of a Control group and Sample group (SBP group) were systemically immunized by subcutaneous injection of 1 mg OA and 300mg of Al(OH)3 in a total volume of 2ml saline. The rats of the sample group were orally administered with an SBP water extract for 14 days after primary immunization. On day 14 after the systemic immunization, rats received local immunization by inhaling 0.9% saline aerosol containing 2%(wt/vol) OA. A day after local immunization, BAL fluid and serum were collected from the rats. Total cell, lymphocyte, CD4+ T cell, CD8+ T cell, CD4+/CD8+ ratio in the BALF, and IgE level in serum were measured and evaluated. Results: SBP showed a suppressive effect on the immune response in the rats. 1. Total cells in the BALF decreased in the SBP treated group in comparision to the control group, but statistic differences were not observed. 2. Total lymphocytes in the BALF were statistically decreased in SBP treated group in comparision to the control group. 3. CD4+ T cells in the BALF were statistically decreased in SBP treated group in comparision to the control group. 4. CD8+ T cells in the BALF were not statistically different in SBP treated group and the control group. 5. The ratio of CD4+/CD8+ in the BALF was statistically decreased in SBP treated group in comparision to the control group. 6. The IgE level in serum decreased in the SBP treated group in comparision to the control group, but statistic differences were not observed.
Objectives: Maekmoondong-tang and Jeongcheonhwadamgangki-tang are herbal decoctions which have been used as traditional therapeutic agents for asthma. This study was performed to investigate the effects of Maekmoondong-tang and Jeongcheonhwadamgangki-tang on immune Cell & serum OA-specific 19B in broncho-alveolar lavage fluid (BALF) in rat asthma model. Methods: Rats were sensitized with ovalbumin (OA); at day I the sensitized group and Maekmoondong-tang and Jeongcheonhwadamgangki-tang groups were systemically immunized by subcutaneous injection of 1mg OA and 300mg of $Al(OH)_3$ in a total volume of 2ml. At the same time, 1ml of 0.9% saline containing $6{\times}10^9$ B. pertussis bacilli was injected by i.p. for 14 days. After systemic immunization, the rats received local immunization by inhaling 0.9% saline aerosol containing 2% (wt/vol) OA. A day after local immunization, BAL fluid was collected from the rats. A day after local immunization, rats were orally administered with each of Maekmoondong-tang and Jeongcheonhwadamgangki-tang extract for 14 days. Lymphocyte, CD4+T-cell CD8+ T-cell counts, CD4+/CD8+ ratio in BALF, change of serum OA-specific IgE level, and CD4+T-cell CD8+T-cell percentage in the peripheral blood were measured and evaluated. Results: Maekmoondong-tang and Jeongcheonhwadamgangki-tang showed a suppressive effect on the rat asthma model. Maekmoondong-tang decreased total cell, lymphocyte, CD4+T-cell, CD8+T-cell in BALF, and serum OA specific 19B level as compared with the control group, whereas Maekmoondong-tang decreased CD4+/CD8+ ratio in BALF with statistical nonsignificance as compared with the control group. 1 decreased total cell, CD4+T-cell, CD4+/CD8+ ratio in BALF, and serum OA specific IgE level, whereas Jeongcheonhwadamgangki-tang decreased ymphocyte, and CD8+T-cell in BALF with statistical nonsignificance as compared with the control group. CD4+T-cell and CD8+ T-cell percentage in peripheral blood were not changed significantly in all the experimental groups. Conclusions: This study shows that Maekmoondong-tang and Jeongcheonhwadamgangki-tang have a suppressive effect on asthma. Maekmooruiong-tang and Jeongcheonhwadamgangki-tang would be useful asthma treatment agents.
Seo, Byung-Kwan;Baek, Yong-Hyun;Choi, Do-Young;Park, Dong-Suk
Journal of Acupuncture Research
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v.22
no.2
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pp.19-32
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2005
Objectives : to evaluate the analgesic effect of bee venom acupuncture on Choksamni (ST36) in the collagen-induced arthritis rats and investigate the role played by serotonergic receptor subtypes (5-HT1a, 5-HT2a) in the antinociceptive effect of bee venom acupuncture in a thermal hyperalgesia test Methods : Experiments were performed on 5 week-aged 60 male Sprague-Dawley rats according to National Institute of Health guidelines and the ethical guidelines of the International Association for the Study of Pain (IASP). Arthritis was induced with arthrogenic collagen emulsion (Bovine type II collagen ${\mu}g$ with incomplete Freund's adjuvant $100\;{\mu}g$). The onset of arthritis was considered to be present when erythema and swelling were detected in at least one joint. The thermal hyperalgesia was evaluated weekly with tail flick test in the rats of severity grade 3 without any injury at tail and foot (including inflammation, ulceration, snap). In the fourth week after first immunization, the analgesic effect of bee venom acupuncture (Choksamni, ST36) was measured with consecutive tail flick latency after intraperitoneal injection of spiroxatrine (1mg/kg) and spiperone (1mg/kg). Results : Chronic inflammatory pain was induced as time elapsed after the immunization of arthrogenic collagen and the maximum value was reached from third to fifth week. Chronic inflammatory pain induced by CIA was inhibited by bee venom acupuncture on the left ST36. The analgesic effect of bee venom acupuncture was inhibited by intraperitoneal injection of 5-HT1a antagonist spiroxatrine and 5-HT2a antagonist spiperone. Conclusions : Therefore, a conclusion. that the analgesic effect of bee venom acupuncture in the chronic inflammatory pain is partially mediated by 5-HT1a and 5-HT2a receptors can be made.
To detect the effect of ginseng saponin on the immune response, mice were immunized with a protein antigen (gamma-globulin of chick). Blood was then drawn from them twice, after 10 days of the first immunization and after 10 days of the second immunization respectively, and measurements were made by ELISA method of the antibody titer in antiserum. In addition, mice that has been immunized with the same antigen were treated with immunosuppressor to suppress the immune system of the mice. After the immune system was suppressed, the effect of ginseng saponin on the recovery of immune response was measured by the same method. The experimental groups those were given ginseng saponin (10 mg/kg/day) showed a little variance between-individuals, however showed much higher antibody titer than the control groups those were given the saline solution. Moreover, there was a little recovery from the immune suppression. Although the mechanism of the effect of ginseng saponin on immune response was not well loom, it is believed that ginseng saponin has the effect of increasing the synthesis of serum protein together with its action as one of the immunostimulators.
Objective: To investigate the analgesic effect and its serotonergic mechanism, especially related with 5-HTI and 5-HT2 receptor, of electroacupuncture(EA) in the rat model of collagen-induced arthritis(CIA). Methods : Immunization of male Sprague-Dawley rats with bovine type II (C II) collagen emulsified in Freund's incomplete adjuvant, followed by a booster injection 14 days later, leads to development of arthritis in more than 70% of rats by 21 days postinjection. After three weeks of first immunization, EA stimulation(2 Hz, 0.07 mA, 0.3 ms) was delivered into Jogsamni($ST_{36}$) for 30 minutes. Analgesic effect was evaluated by tail flick latency(TFL). We compared the analgesic effect of EA with TFLs between pretreatment of normal saline and pretreatment of spiroxatrine (5-HT1 receptor antagonist, 1mg/kg, intraperitoneal) and spiperone (5-HT2 receptor antagonist, 1 mg/kg, intraperitoneal) in CIA. Results : 1. TFLs were gradually decreased in CIA as increasing severity of arthritis. 2. Jogsamni($ST_{36}$) EA stimulation in CIA increased TFLs and the effect lasted for 60 minutes. 3. Increased TFLs with Jogsamni($ST_{36}$) EA stimulation were inhibited by pretreatment of spiroxatrine and spiperone in CIA. Conclusions : Jogsamni($ST_{36}$) EA showed analgesic effects in CIA The analgesic effects of Jogsamni($ST_{36}$) EA were inhibited by spiroxatrine and spiperone pretreatment. These observations suggest that 5-HT1 and 5-HT2 serotonergic receptor, which involve the release of serotonin neurotransmitter, play an important roles in analgesic mechanism of EA stimulation.
The adjuvant effect of a muramyl dipeptide (MDP) derivative, MDP-Lys(L18), on enhancing of antitumor immunity induced by X-irradiated tumor cells against highly metastatic B16-BL6 melanoma cells was examined in mice. Mice immunized intradermally (i.d.) with a mixture of X-irradiated B16-BL6 cells and MDP-Lys (L18) [Vac+MDP-Lys (L18)] followed by an intravenous (i.v.)inoculation of $10^4$ viable tumor cells 7 days after immunization, showed a significant inhibition of experimental lung metastasis of B16-BL6 melanoma cells. The most effective immunization for the prophylactic inhibition of tumor metastasis was obtained from the mixture of $100{\;}\mu\textrm{g}$ of MDP-Lys (L18) and $10^4$ X-irradiatied tumor vaccine. Furthermore, immunization of mice with Vac+MDP-Lys(L18), 3 days after tumor challenge, resulted in a significant inhibition of lung metastasis of B16-BL6 melanoma cells in an experimental lung metastasis model. Similarly, the administration of Vac+MDP-Lys(L18), 1 or 7 days after tumor removal, markedly inhibited tumor metastasis of B16-BL6 in a spontaneous lung metastasis model. When Vac+MDP-Lys (L18) was i.d. administered 3 days after subcutaneous (s.c.) inoculation of tumor cells ($5{\times}10^5/site$) on the back, mice treated with Vac+MDP-Lys(L18) showed inhibition of significantly tumor growth on day 20. These results suggest that MDP-Lys (L18) is able to enhance antitumor activity induced by X-irradiated tumor vaccine to reduce lung metastasis of tumor cells, and is a potent immunomodulating agent which may be applied prophylactically as well as therapeutically to treatment of cancer metastasis.
Adrenal glucocorticoids, secreted by the stimulus of adrenocorticotropic hormone (ACTH), are catabolic hormones in the pig. The present study was conducted to find whether active immunization against ACTH would suppress cortisol secretion accompanied by an increased growth rate in growing-finishing barrows. ACTH was conjugated to keyhole limpet hemocyanin or human histone using glutaraldehyde or 3-maleimidobenzoic acid N-hydroxysuccinimide, under a 2 (ACTH vs no hapten)${\times}$2 (carrier)${\times}$2 (crosslinker) factorial arrangement of treatments. Cross-bred barrows weighing approximately 25 kg were injected with an ACTHcarrier or carrier only conjugate every 4th wk and slaughtered at approximately 110 kg body weight. Antibodies against ACTH were detected in serum, as determined by $[^{125}I]$ACTH-binding activity, in most animals immunized against the ACTH conjugate, but not in carrier only-injected animals, except for the animals which had received the hapten conjugated to histone via glutaraldehyde. The $[^{125}I]$ACTH-binding activity of serum increased after the second booster injection, but overall ACTH antibody titer was very low. Main effect was not detected not only for the carrier and crosslinker but for the hapten in serum cortisol concentration, ADG, loin muscle area, backfat thickness and longissimus muscle composition including fat and protein. In addition, bound $[^{125}I]$ACTH percentage had no relation to cortisol concentration or to any of the above growth-related variables. Results suggest that ACTH or its conjugates used in the present study were not immunogenically potent enough to affect the glucocorticoid secretion and thus the growth of the immunized pigs.
This study was performed to determine the effect of maternal protein intake on 1) the concentration of immune substances in milk 2) degree of passive immunity to pups via lactation, and 3) specific antibody production to a specific antigen, $\beta$-lactoglobulin(BLG). 4) the effect of passive immunity that pups received from mother during lactation on the production of antibodies when the pups were challenged to the same antigen. Part of the female rats were immunized with BLG before and during pregnancy. The pregnant rats were placed into either 25% or 10% isolated soy protein diet throughout gestation and lactation. After weaning, pups from each group continued to be fed the same diet. At 18 weeks of age, all the pups were challenged with BLG. Total IgA and IgG, lysozyme, BLG-specific IgA and IgG were measured in dam's serum, dam's milk, and pup's serum. Total IgG, and lysozyme in dam's serum and milk were higher in high protein group. Total IgA and IgG in pup's serum remained higher in high protein group from 5 to 18 weeks of age. BLG-specific antibodies were found in the milk and serum of immunized dams, and in serum of pups born to immunized dams but not in the non-immunized group. BLG-specific IgA and IgG were again higher in high protein group and declined with time. The concentration decreased faster in the low proetein group than in the high protein groups. After immunization the pups with LBG, serum BLG-specific antibodies were not differ between rats born to immunized dams and those born to non-immunized dams. Therefore passive immunity rats received via milk as a pup had no effect on the BLG-specific antibody production later in life. This study shows the importance of protein status of mother and strongly support to the endorsement of breast feeding.
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