Modulation of lung local immune responses by oral administration of a herbal medicine Sho-saiko-to

Sho-saiko-to (SST), a Chinese/Japanese herbal medicine (Kampo medicine) widely used to treat chronic hepatitis in Japan, is known to modulate immune responses, and thus its immunomodulating activity may be responsible for its bi-directional effects on the lungs as therapeutic efficacy in various lung diseases and involvement in development of interstitial pneumonia. We administered SST to BALB/c mice orally and examined the lung tissue levels of pro/anti-inflammatory cytokines, interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), and the effects of SST on acute lung injury induced by instillation of lipopolysaccharide (LPS) or IL-1. Although SST had no effect on lung TNF-alpha or IL-1beta level, it increased IL-6. Investigation of active fractions of SST suggested that multiple ingredients were supposed to be responsible for IL-6-inducing activity. Liquiritigenin, a metabolite of liquiritin which is one of the major ingredients in SST enhanced in vitro IL-6 production in anti-CD3 monoclonal antibody (anti-CD3 mAb)-stimulated lung mononuclear cells in a cell-type specific and dose-dependent manner. SST suppressed LPS-induced lung injury at the later phase when lung leak was evident while being ineffective on initial neutrophil sequestration to the lung in these models. These findings suggest that SST modulates lung inflammation by regulating local immune response.     

Modulation of antigen-specific immune responses by the oral administration of a traditional medicine,bo-yang-hwan-o-tang

Bo-yang-hwan-o-tang (BHT) has long been used to treat cancer in traditional Korean medicine and is believed to have immune-modulating activity. This study investigated the effect of BHT on the induction of antigen-specific immune responses using hen egg-white lysozyme (HEL) as a model antigen system. Oral administration of BHT enhanced both HEL-specific humoral and lymphocyte proliferative responses in HEL low-responder mice. Feeding BHT to the mice increased INF-gamma levels, but did not change IL-4 levels. Interestingly, however, the oral BHT feeding significantly increased HEL-specific antibodies of the IgG1, IgG2b, and IgG3 subtypes, which are associated with the direct stimulation of B cells. This indicates that BHT treatment enhances anti-HEL-specific humoral immune responses via the direct stimulation of B lymphocytes rather than by selective priming of specific subtypes of the helper T-cell population. This conclusion was supported by in vitro experiments, in which the presence of BHT significantly augmented B-cell mitogen-mediated proliferation of mouse splenocytes. This augmentation was closely associated with a glycoprotein with a molecular weight of around 100 kDa. The results suggest that BHT modulates antigen-specific immune responses, and might be used as a therapeutic agent for patients who need enhanced immune function.     

Modulation of systemic antigen-specific immune responses by oral antigen in humans

Oral antigen uptake can induce systemic immune responses ranging from tolerance to immunity. However, the underlying mechanisms are poorly understood, especially in humans. Here, keyhole limpet hemocyanin (KLH), a neoantigen which has been used in earlier studies of oral tolerance, was fed in a repeated low-dose and a single high-dose protocol to healthy volunteers. KLH-specific CD4(+) T-cell proliferation and cytokine production, as well as KLH-specific serum Ab and the effects of oral KLH on a subsequent parenterally induced systemic immune response, were analyzed. Repeated low-dose oral KLH alone induced antigen-specific CD4(+) T cells positive predominantly for the gut-homing receptor integrin β7 and the cytokines IL-2 and TNF-α; some CD4(+) T cells also produced IL-4. Oral feeding of KLH accelerated a subsequent parenterally induced systemic CD4(+) T-cell response. The cytokine pattern of KLH-specific CD4(+) T cells shifted toward more IL-4- and IL-10- and less IFN-γ-, IL-2- and TNF-α-producing cells. The parenterally induced systemic KLH-specific B-cell response was accelerated and amplified by oral KLH. The impact of single high-dose oral KLH on antigen-specific immune responses was less pronounced compared with repeated low-dose oral KLH. These findings suggest that oral antigen can effectively modulate subsequently induced systemic antigen-specific immune responses. Immunomodulation by oral antigen may offer new therapeutic strategies for Th type1-mediated inflammatory diseases and for the development of vaccination strategies.     

Modulation of cancer patients' immune responses by administration of anti-idiotypic antibodies

Thirty patients with Dukes stage D colon carcinoma who had undergone operative removal of the primary tumor and had growing hepatic metastases each received four intradermal injections of 0.5-4 mg of alum-precipitated goat anti-idiotypic antibodies (anti-Id). The anti-Id had been produced against murine monoclonal antibody (mAb) CO17-1A, which defines a human colon carcinoma associated antigen. All patients elaborated anti-anti-Id that shared idiotopes with mAb CO17-1A, bound to tumor cells and isolated tumor antigen, and competed with the mAb for binding to tumor cells. The clinical response was monitored by ultrasonography, CT, radionuclide scanning, and serum marker assays. Six patients had partial tumor responses; five of these had received additional booster anti-Id injections along with chemotherapy. Seven patients had stabilized tumor; six had received additional anti-Id, with chemotherapy also in four. Conclusions about the clinical role of such immunization await further study, but in demonstrating a specific response to anti-Id, our results support the use of this approach in human immunotherapy against tumors or pathogens.     

Modulation of immune responses in newborn and adult mice by interferon.

Interferon was found to have both suppressive and enhancing effects on the antibody response in newborn and adult mice. Evidence was obtained that these effects are primarily evoked during the initial steps controlling cell proliferation. Stimulation of thymus and spleen cells with a T-cell mitogen was enhanced by low doses and suppressed by high doses of interferon. Treatment of parental spleen cells with interferon before injecting them into immunized F1 hybrid mice resulted in an enhanced allogeneic effect. These results are compatible with the hypothesis that interferon affects T cells and has an immunoregulatory role, either by inhibiting the action of suppressor cells or by promoting immunological maturation.     

尼氟灭酸抑制哮喘小鼠气道黏液高分泌

目的观察小鼠钙激活Cl^-通道Ⅲ型（mCLCA3）阻断剂尼氟灭酸（NFA），对哮喘小鼠气道黏液高分泌的抑制作用。方法将BABL／c小鼠随机分为哮喘组、吸入NFA预防组和正常对照组。AB-PAS特染法检测各组小鼠小支气管杯状细胞的数量及黏液分泌状况。RT-PCR法检测各组肺组织中黏蛋白MUC5AC mRNA和mCLCA3 mRNA水平。结果正常对照组小支气管只有少量散在分布的杯状细胞，管腔干净；而哮喘组出现大量簇状分布的杯状细胞，胞质内富含着色颗粒，管腔具有着色黏液；NFA预防组的杯状细胞数量与正常对照组相近。哮喘组的MUC5AC及mCLCA3 mRNA水平均显著高于正常对照组、NFA预防组；NFA预防组的MUC5AC mRNA水平与正常对照组接近，而mCLCA3 mRNA水平稍高于正常对照组。结论NFA能有效抑制哮喘小鼠气道杯状细胞数量的增加及黏蛋白的合成，这一效果是通过阻断mCLCA3活性和降低其表达水平来实现的。     

Nonspecific and Candida-specific immune responses in mice suppressed by chronic administration of anti-mu

CBA/J mice were immunosuppressed by repeated administration of goat antibody specific for mu chain of immunoglobulin M (IgM) and tested for nonspecific and Candida albicans-specific immune responses. Immunosuppression was demonstrated by a dramatic reduction in the number of antibody-forming cells in the spleens of anti-mu-treated mice when immunized with sheep erythrocytes, by greatly reduced in vitro responsiveness of both spleen and lymph node lymphocytes from anti-mu-treated mice to lipopolysaccharide, and by a large reduction in the number of splenic IgM-positive cells. T cell function, on the other hand, appeared to be relatively unaltered in anti-mu-treated animals, in the cytotoxic T lymphocyte activity against an allogeneic target was similar in splenocyte cultures from anti-mu- and mock-treated animals, and splenic and lymph node lymphocytes proliferated in response to concanavalin A in a lymphocyte stimulation assay. Moreover, Candida-specific delayed hypersensitivity to two different Candida antigens, one cell wall-derived (GP) and the other cell membrane-derived (BEX), was of comparable intensity in immunosuppressed and normal animals. When anti-mu- and mock-treated mice were immunized by the cutaneous inoculation of viable C. albicans blastospores and then challenged intravenously to assess the development of protective immunity, only mock-treated animals, male and female, had significant (p less than or equal to 0.05) protective responses demonstrable by reduction in the number of colony-forming units cultured from their kidneys 28 days after intravenous challenge. If consideration was given to the number of animals which had cleared Candida completely from the kidney, however, there appeared to be protective responses operative in the female anti-mu-treated animals as well. Neither anti-mu-treated males nor females, when immunized and challenged with C. albicans, produced Candida-specific antibody detectable by counterimmunoelectrophoresis, whereas all immunized and challenged mock-treated animals produced antibody. The data are consistent with the hypothesis that anti-mu treatment has little effect on multiple cellular immune functions, including those specific for C. albicans, and the combination of antibody, cell-mediated immunity and innate defenses are responsible for solid systemic defense against the fungus.     

Modulation of innate immune responses by hantaviruses

Hantavirus infection can cause hantavirus cardiopulmonary syndrome or hemorrhagic fever with renal syndrome depending on the virus species involved. The determinants for the virus species specific virulence in humans are unclear. Successful infection is a conditio sine qua non for the virulence of a virus and it is well-known that the innate interferon (IFN) system generally plays a decisive role to prevent establishment of an infection. The importance of the IFN system is underscored by the fact that viruses have developed an amazing number of different escape mechanisms to enable replication in face of the antiviral host response. Interestingly, pathogenic hantaviruses escape induction of innate antiviral responses in the early phase of the infection, which are elicited in a pronounced manner by nonpathogenic hantaviruses in vitro. This differential response might be important for the pathogenicity of hantaviruses in humans. This review aims to summarize the current knowledge about the interaction between hantaviruses and the innate IFN system. Detailed characterization of the cellular sensors and pathways that lead to activation of the IFN system on one side and the viral escape mechanisms on the other might help to develop novel vaccination strategies and therapeutic approaches.     

Modulation of cellular immune responses in mice with disseminated histoplasmosis by recombinant interleukin-2.

Depression of the cellular immune responses in mice with disseminated histoplasmosis is associated with deficient production of interleukin-2 (IL-2) by splenocytes. Therefore, we examined whether a highly purified preparation of IL-2, recombinant human IL-2 (rIL-2), could modify the cellular immune responses in infected mice and whether this lymphokine could alter the severity of histoplasmosis in animals. Exogenous rIL-2, at concentrations of up to 1,000 U/ml, failed to augment the proliferative responses to concanavalin A by unfractionated splenocytes or splenic T cells from mice infected for 1 week. In addition, rIL-2 did not modulate the plaque-forming cell response to sheep erythrocytes by splenocytes from these same mice. However, at week 3, rIL-2 in concentrations ranging from 10 to 1,000 U/ml considerably augmented the proliferative response to concanavalin A and plaque-forming cell response to sheep erythrocytes by splenocytes from infected mice. Kinetics studies demonstrated that rIL-2 exerted maximal immunoregulatory activity when added on day 0 or 1 to cultures of splenocytes. In vivo administration of rIL-2, 200 to 20,000 U/day, for 10 days to normal and 3-week-infected mice did not alter the proliferative activity of splenocytes to concanavalin A; 200,000 U of rIL-2 per day actually depressed the proliferative responses of splenocytes from normal and infected mice. In vivo, rIL-2 did not modify delayed-type hypersensitivity responses to sheep erythrocytes or to histoplasmin by normal and infected mice. Moreover, treatment with rIL-2 in vivo did not reduce the number of Histoplasma CFU in spleens of mice. Thus, despite the immunoenhancing effect of rIL-2 in vitro, this lymphokine failed to exert similar effects in vivo.     

Modulation of immune responses by anabolic androgenic steroids

Anabolic androgenic steroids (AS) have recently been placed on the Food and Drug Administration's (FDA's) list of controlled substances, because of the adverse effects seen in athletes taking accelerated dosages in attempts to enhance performance. Reported deleterious effects on abusers include sterility, gynecomastia in males, acne, balding, psychological changes, and increased risks of heart disease and liver neoplasia. Considering the roles of the immune and neuroendocrine systems and their interactions in many of these pathologies, it is important to determine the effects of these derivitized androgens on this connection. Little is known in this respect. We therefore determined the effects of anabolic steroids on certain immune responses and their effects on the extrapituitary production of corticotropin by lymphocytes. We present evidence that (1) both 17-β and 17-α esterified AS, nandrolone decanoate and oxymethenelone, respectively, significantly inhibited production of antibody to sheep red blood cells in a murine abuse model; (2) the control androgens testosterone and dehydroepian-drosterone (DHEA) or sesame seed oil vehicle had no significant effects on antibody production; (3) nandrolone decanoate and oxymethenelone directly induced the production of the inflammatory cytokines IL-1β and TNF-α from human peripheral blood lymphocytes but had no effect on IL-2 or IL-10 production; (4) control androgens had no direct cytokine inducing effect; (5) nandrolone decanoate significantly inhibited IFN production in human WISH and murine L-929 cells; and (6) nandrolone decanoate significantly inhibited the production of corticotropin in human peripheral blood lymphocytes following vital infection. These data indicate that high doses of anabolic steroids can have significant effects on immune responses and extrapituitary production of corticotropin. Furthermore, the mouse model should provide an effective means by which to study other deleterious effects of anabolic steroid abuse in humans.     

Modulation of humoral immune responses by endogenous opioids

The effects of opioid agonists and antagonists were investigated on humoral immune mechanisms in mice and rats. Opioid agonists like morphine, Leu-enkephalin, and Met-enkephalin, enhanced antigen-induced histamine release from mixed peritoneal cells of rats in vitro; this enhancement was effectively antagonized by naloxone, an opioid antagonist. Naloxone, per se, decreased anaphylactic mortality in doses of 10 mg/kg, while it increased mortality in a dose of 1 mg/kg. Reduced IgE antibody titer, measured by passive cutaneous anaphylaxis, decreased hemagglutination titer to sheep red blood cells, blocked histamine release from mixed peritoneal cells of rats in vitro induced by antigen, but had no significant effect when histamine release was induced by compound 48/80. Thus, it appears that endogenous opioids are involved in humoral immune responses.     

Modulation of host immune responses by helminth glycans

Summary:  Parasitic infections regulate/alter host immune responses. Among parasitic infections, helminth infection often leads to systemic immune suppression or anergy. Helminth infection or helminth extracts drive CD4⁺ T-helper (Th) cell responses towards Th2 type and activate antigen-presenting cells (APCs) such that these cells express an anti-inflammatory phenotype. Among the myriad molecules present on or secreted by helminth parasites, glycans have been shown to be key in inducing Th2-type and anti-inflammatory immune responses. The majority of studies on immune modulatory helminth glycans have focused on Lacto-N-fucopentaose III and Lewis^X. When presented as glycol-conjugates, with multiple copies of the sugars conjugated to a carrier molecule, these compounds activate APCs, inducing an alternative activation pattern, whose phenotypic profile is substantially different than that seen using pro-inflammatory activators such as lipopolysaccharide. Though the mechanism of APC activation by LNFPIII/Lewis^X glycoconjugates has not been fully elucidated, it involves C-type lectin ligation on the surface of APCs, with subsequent antagonism of Toll-like receptor signaling. In this article, we discuss the APC surface receptors known to play roles in LNFPIII/Lewis^X induced alternative activation of APCs. We also discuss what is currently known regarding downstream signaling pathways, closing with a discussion of future research directions for this field of investigation including the potential use of immune modulatory glycans as vaccine adjuvants and anti-inflammatory therapeutics.     

Induction of immune responses in mice and pigs by oral administration of classical swine fever virus E2 protein expressed in rice calli

Classical swine fever (CSF), caused by the CSF virus (CSFV), is a highly contagious disease in pigs. In Korea, vaccination using a live-attenuated strain (LOM strain) has been used to control the disease. However, parenteral vaccination using a live-attenuated strain still faces a number of problems related to storage, cost, injection stress, and differentiation of CSFV infected and vaccinated pigs. Therefore, two kinds of new candidates for oral vaccination have been developed based on the translation of the E2 gene of the SW03 strain, which was isolated from an outbreak of CSF in 2002 in Korea, in transgenic rice calli (TRCs) from Oriza sativa L. cv. Dongjin to express a recombinant E2 protein (rE2-TRCs). The expression of the recombinant E2 protein (rE2) in rE2-TRCs was confirmed using Northern blot, SDS-PAGE, and Western blot analysis. Immune responses to the rE2-TRC in mice and pigs were investigated after oral administration. The administration of rE2-TRCs increased E2-specific antibodies titers and antibody-secreting cells when compared to animals receiving the vector alone (p < 0.05 and p < 0.01). In addition, mice receiving rE2-TRCs had a higher level of CD8+ lymphocytes and Th1 cytokine immune responses to purified rE2 (prE2) in vitro than the controls (p < 0.05 and p < 0.01). Pigs receiving rE2-TRCs also showed an increase in IL-8, CCL2, and the CD8+ subpopulation in response to stimulation with prE2. These results suggest that oral administration of rE2-TRCs can induce E2-specific immune responses.     

Modulation of the immune responses against SRBC after oestriol treatment in mice.

Delayed-type hypersensitivity (DTH), primary direct and indirect PFC, memory antibody response and suppressor cell induction against sheep red blood cells (SRBC) have been examined in oestriol (E3) pretreated mice. The results showed that DTH and primary direct and indirect PFC responses were suppressed by E3 treatment. These suppressive effects could, however, be overcome when oestrogenized mice were given supra-optimal doses of SRBC for each response. On the other hand, the memory antibody response was markedly enhanced when E3 was given prior to the primary antigen stimulation. Induction of the suppressor cells for the antibody response seemed not to be affected by E3 treatment, but the characterization of the suppressor cells revealed that those obtained from E3 treated mice were surface immunoglobulin positive (sIg+) cells whereas those from control mice were not. These results were discussed in terms of the altered antigen distribution due to the activated phagocytic activity of the reticuloendothelial system (RES) after E3 treatment.     

Modulation of cancer patients' immune responses by anti-idiotypic antibodies

The immunomodulatory role of anti-idiotypic antibodies (Ab2) in patients with gastrointestinal cancer has been demonstrated in two types of clinical trials. In the first, cancer patients were treated with a monoclonal antibody (MAb) defining a tumor-associated antigen (Ag). MAb administration initiated an idiotypic network as demonstrated by the induction of both Ab2 and anti-anti-idiotypic antibodies (Ab3) in the treated patients. The Ab3 bound to tumor cells and isolated tumor Ag with the same specificity as the Mab (Ab1) at the beginning of the idiotypic cascade. A beneficial role of Ab3 is postulated for patients showing delayed clinical responses to MAb therapy. In a recent trial, patients with advanced colorectal cancer responded to immunization with Ab2 that functionally mimicked in vitro and in vivo (animals) a gastrointestinal tumor-associated Ag by developing highly specific Ab3 with anti-tumor binding reactivities. Thus, Ab2 are promising agents in immunotherapy approaches to cancer. These studies suggest an immunoregulatory role for Ab2 in cancer patients. Modulation of cellular immune responses by Ab2 in cancer patients will be an important consideration in future studies.     

外源质粒DNA经胃肠道途径对小鼠免疫基因的调控作用

目的:研究外源质粒DNA经胃肠道途径对小鼠免疫基因表达的调控作用。方法:给BALB/c小鼠灌胃质粒pcDNA3200μg,分别在灌胃后4h和18h分离脾脏,提取脾脏总RNA。利用Affymetrix基因表达谱芯片对灌胃质粒pcD-NA3后的BALB/c小鼠脾脏进行基因表达谱研究。采用Genmmapp和MAPPFinder软件进行功能聚类分析。结果:灌胃质粒pcDNA3后,大量与免疫应答相关的基因发生上调,这些基因包括转录因子、细胞因子和细胞因子受体、主要组织相容性基因、蛋白酶体基因、补体分子基因和细胞凋亡相关基因;下调的基因主要是免疫球蛋白基因。MAPPFinder分析结果显示大量免疫应答过程发生上调。结论:外源质粒DNA通过胃肠道途径可调控大量免疫基因的表达,对相关基因及免疫应答过程的研究有助于在分子水平上深入了解外源质粒DNA的胃肠道作用机制。     

Effects of oral administration of Lactobacillus casei on antitumor responses induced by tumor resection in mice

The effects of an oral administration of BLP, a preparation of viable Lactobacillus casei YIT 9018, upon tumor growth and the mitogenic responses of splenocytes from tumor-bearing mice were studied. BALB/c mice were insufficiently pre-immunized by resecting a Colon 26 tumor mass (primary tumor) grown for 5 days intradermally. Thereafter, mice were rechallenged by injecting Colon 26 tumor (secondary tumor) into the hind footpad. The secondary tumor grew progressively in control mice, but was markedly suppressed by oral administration with BLP at a dose of 100 or 200 mg/kg/day for 7 consecutive days. The suppression was a primary tumor-specific response. Viable L. casei, not heat-killed L. casei, could suppress the secondary tumor.Although the lymphoproliferative responses of splenocytes from the secondary tumor-bearing mice with T-cell mitogens (concanavalin A and phytohaemagglutinin) and cytokines (interleukin-1 and interleukin-2) were lower than those of normal mice, this suppression of mitogenic responses under tumor-bearing conditions was abolished by oral administration with BLP.Thus we concluded that oral BLP potentiated systemic immune responses that modified T-cell functions in tumor-bearing mice.     

Modulation of the adoptive immune response in mice by histamine

The immunodulatory actions of histamine in mice were examined by a combinedin vitro/in vivo approach.Spleen cells from mice were incubated between 2 and 24 hours with histamine (10^–12–10^–3 M) under conditions which prevent a change of the free histamine concentration. The cells were subsequently transferred to sublethally irradiated syngenic mice in order to measure the adoptive IgM response. Only stimulatory effects of histamine were found at NMRI mice. However, both stimulatory and inhibitory actions were observed at different histamine concentrations if mice of the strains AB or XVII were used. The graft versus host reaction was measured after transfer of histamine treated spleen cells (strain XVII) to neonatal F₁ (XVII×B10.LP) hybrid mice and revealed both suppressive and stimulatory effects of histamine at different concentrations. A maximal expression of the immunomodulatory effects of histamine was found after 8 hours of preincubation with the donor cells. The action of selective histamine antagonists and cell separation experiments indicated that the effect of histamine on the adoptive IgM response was mediated by H₂-receptors on spleen T-cells. Summarizing, the results indicate that low histamine concentrations elicit bidirectional immunodulatory effects in mice which vary considerably among different strains.