Cellular and humoral adjuvant activity of lectins isolated from Korean mistletoe (Viscum album colaratum)

The adjuvant effect of lectins (KML-C) isolated from Korean mistletoe (Viscum album coloratum) on induction of humoral and cellular immune responses against keyhole limpet hemocyanine (KLH) was examined. When mice were immunized subcutaneously (s.c.) with KLH (20 micrograms/mouse) admixed with or without 50 ng/mouse of KML-C (KLH + KML-C), mice immunized with KLH + KML-C showed significantly higher antibody titers against KLH than those immunized with KLH alone, showing the highest titer 5 weeks after immunization. Furthermore, boost immunization with KLH + KML-C at 2-week interval elicited much higher activity than single immunization to enhance antibody responses against KLH. The assay for determining isotypes of antibodies revealed that KML-C augmented KLH-specific antibody titers of IgG1, IgG2a and IgG2b. The culture supernatants obtained from the splenocytes of mice treated with KLH + KML-C also showed a higher level of both KLH-specific Th-1 (IL-2 and IFN-gamma) and Th-2 type cytokine (IL-4). In an in vitro analysis of T lymphocyte proliferation to KLH on week 4, the splenocytes of mice treated with KLH + KML-C showed a significantly higher proliferating activity than those treated with KLH alone. In addition, mice immunized twice with KLH + KML-C and followed by intrafootpad (i.f.) injection of KLH (50 micrograms/site) 14 weeks after the primary immunization induced a higher delayed-type hypersensitivity (DTH) reaction than mice treated with KLH alone. These results suggest that KML-C is a potent immunoadjuvant to enhance cellular and humoral immune responses.     

鸡枞菌多糖对免疫抑制小鼠免疫功能的影响

目的探讨鸡枞菌多糖(TAP)对环磷酰胺所致免疫抑制小鼠免疫功能的影响。方法以黄芪多糖(APS)为阳性对照,用不同剂量的TAP对免疫抑制小鼠进行腹腔注射,检测其腹腔巨噬细胞吞噬功能,免疫器官指数、血清溶血素、T淋巴细胞亚群、细胞因子、脾淋巴细胞增殖等免疫指标。结果 TAP在5～20 g.L-1浓度范围内不同程度增强巨噬细胞吞噬功能及提高免疫器官指数,且有剂量依赖性;20 g.L-1TAP可明显降低免疫低下小鼠CD4+/CD8+比值,并使IL-2、IFN-γ水平分别上升312.3%和88.1%,同时降低IL-4水平;TAP能明显提高脾淋巴细胞增殖能力。结论 TAP可提高免疫抑制小鼠体液及细胞免疫功能,且效果优于APS。     

Immunotoxicity of paraquat after subacute exposure to mice

The immunotoxic effect of paraquat (PQ), a herbicide that has been used widely in agriculture was investigated using Balb/c mice. Paraquat was administered at doses of 1, 0.1, and 0.01 mg/kg for 21 days. Body weight, organ weight, cellularity of spleen, delayed type of hypersensitivity (DTH) response, plaque-forming cell (PFC) assay, hemagglutination titer (HA), quantitative hemolysis of SRBC (QHS) assay, spleen cell subtypes, cytokine production and lymphocyte proliferation assay were studied in various groups of animals. Results showed that high dose of PQ (1 mg/kg) could suppress both cellular and humoral activity of the immune system. PQ at medium dose (0.1 mg/kg) did not show any changes in organ weight, body weight and spleen cellularity but significantly decreased the proliferation response to PHA and the production of IFNγ. PQ at low dose (0.01 mg/kg) did not produce any significant changes in humoral or cellular responses of the immune system. In conclusion, paraquat at high dose has an inhibitory effect on the cell-mediated and humoral immunity. It seems that PQ has no adverse effects on mice immune system at low doses of 0.01 mg/kg, which is two times the PQ allowed daily intake (ADI) limit.     

Tumor inhibition and improved immunity in mice treated with flavone from Cirsium japonicum DC

The purpose of these experiments is to study the effects of flavone from Cirsium japonicum DC (FLCJ) on tumor activity and on the regulation of the immune response in mice with S180 and H22 tumors. Pectolinarin and 5, 7-dihydroxy-6, 4'-dimethoxyflavone are two major components of FLCJ. FLCJ, pectolinarin and 5, 7-dihydroxy-6, 4'-dimethoxyflavone can inhibit the growth of the implanted tumors S180 and H22 and promote cellular and humoral immune responses. FLCJ is stronger than Pectolinarin and 5, 7-dihydroxy-6, 4'-dimethoxyflavone in inhibiting tumors and promoting immune response significantly. Our results indicated that the tumor inhibition of FLCJ had close relation with its regulation of the immune response Furthermore, this tumor inhibition by FLCJ may activate the innate immune system, which would be expected as a potential anti-tumor drug.     

黄芪多糖对免疫功能影响的体外实验研究

目的探讨黄芪多糖对小鼠免疫功能的影响,为黄芪多糖的临床应用提供理论依据。方法取小鼠脾细胞,置于96孔细胞培养板中,设黄芪多糖组和空白对照组,WST-1法测细胞的增殖活性;ELISA测细胞因子IL-2、IFN-γ;流式细胞术（FCM）检测细胞表型。结果黄芪多糖组细胞增殖活性显著高于空白对照组（P〈0.01）;黄芪多糖组IL-2、IFN-γ的浓度随黄芪多糖浓度的增加而增多,在一定浓度范围内呈剂量依赖关系;黄芪多糖既能刺激B淋巴细胞增殖,也能刺激T淋巴细增殖。结论黄芪多糖可增强免疫功能,它不仅能提高体液免疫功能,而且能提高细胞免疫功能。     

Posintro™-HBsAg, a modified ISCOM including HBsAg, induces strong cellular and humoral responses

To improve the hepatitis B vaccines on the market new adjuvant systems have to substitute aluminium. In this study the hepatitis B surface antigen (HBsAg) was incorporated into a novel adjuvant system, the Posintro?, a modification of the traditional immune stimulatory complexes (ISCOMs). This new HBsAg vaccine formulation, Posintro?-HBsAg, was compared to two commercial hepatitis B vaccines including aluminium or monophosphoryl lipid A (MPL) and the two adjuvant systems MF59 and QS21 in their efficiency to prime both cellular and humoral immune responses. The Posintro?-HBsAg induced the strongest humoral response with high titers of HBsAg specific antibody, high number of antigen specific B-cells and a strong T helper 1 (Th1) antibody profile when compared to the other adjuvant formulations. The Posintro?-HBsAg was also a strong inducer of cellular immune responses with induction of delayed type hypersensitivity (DTH) reaction and CD4(+) T-cell proliferation. In addition, Posintro?-HBsAg was the only vaccine tested that also induced a strong cytotoxic T lymphocyte (CTL) response, with high levels of antigen specific CD8 T-cells secreting IFN-gamma mediating cytolytic activity. The results demonstrate that this novel experimental vaccine formulation, the Posintro?-HBsAg, is strongly immunogenic and can induce both class I and class II responses in experimental animals. This shows promise both for the protection against hepatitis B virus infection and as a potential therapeutic vaccine.     

肌力康饮对正常小鼠免疫功能的影响

目的:观察中药复方肌力康饮对正常小鼠免疫功能的影响。方法:小鼠随机分为6组,即正常对照组,地塞米松组(0.25mg/kg·d),环磷酰胺组(80mg/kg·d),肌力康饮低、中、高剂量组(2、4、8g/kg·d)。给药14天后,测定免疫器官指数来观察肌力康饮对免疫器官的影响;应用刀豆球蛋白A(ConA)、细菌脂多糖(LPS)诱导小鼠淋巴细胞增殖,观察肌力康饮对小鼠细胞免疫和体液免疫的影响。结果:与对照组比较,地塞米松、环磷酰胺均能显著降低小鼠胸腺和脾脏指数,对ConA、LPS所诱导小鼠脾T、B淋巴细胞增值有明显抑制作用(P<0.01);肌力康饮各剂量组胸腺和脾脏指数稍降低,但无显著性差异,中、高剂量组能显著增强小鼠脾T、B淋巴细胞增值能力(P<0.01)。结论:肌力康饮对正常免疫器官没有抑制作用,对细胞免疫和体液免疫有显著的增强作用。     

A nano silicon adjuvant enhances inactivated transmissible gastroenteritis vaccine through activation the Toll-like receptors and promotes humoral and cellular immune responses

Inactivated transmissible gastroenteritis virus (TGEV) vaccines are widely used in swine herds in China. These are limited, however, by the need to elicit both humoral and cellular immunity, as well as the efficiency of adjuvants. In this study, a 70-nm nano silicon particle was applied with inactivated TGEV vaccine in mice, and its immune-enhancing effects and mechanism of action investigated. We found that nano silicon applied with inactivated TGEV vaccine induced high antibody titers, increase IL-6, TNF-α and IFN-γ expression, and stimulate CD3+ T cell proliferation with a high CD4+/CD8+ T lymphocyte ratio. Nano silicon could quickly activate innate and adaptive immunity by stimulating Toll-like receptor signaling pathways, indicating that the nano silicon adjuvant enhanced long-term humoral and early cellular immune responses when combined with inactivated TGEV vaccine. Nano silicon could be considered for use as an antigen- carrier and adjuvant for veterinary vaccines.     

A synergistic decline in humoral and cellular immunity of diabetic mice on exposure to polluted air

It is clinically known that diabetic patients are more prone to infectious diseases, due to low immune status. Since, some of the common air pollutants are reported to suppress immune system, how exposure to artificially polluted air influences the immune responses in experimental diabetic mice was studied. A diabetic state was induced by alloxan and mice were exposed to artificially polluted air for 30 days. During the period of exposure, the humoral (antibody titer) and cellular (foot and swelling) immune responses to antigenic challenges with sheep RBC were investigated. The exposure to polluted air produced a significant decline in the immune responses in non-diabetic mice whereas a synergistic decline was observed in diabetic group. Since, daily oral treatment with vitamin E (150 mg/kg) significantly prevented the pollution induced immunosuppression, the involvement of free radicals is suggested.     

In vitro effects of an immunostimulating bacterial lysate on human lymphocyte function

MLBL is an oral immunostimulating vaccine consisting of bacterial standardized lysates obtained by mechanical lysis of different strains of Gram-positive and Gram-negative bacteria that can cause acute and chronic infections of the respiratory tract. Previous studies suggested a stimulating effect of MLBL both on humoral and cellular immune responses. In the present study, the in vitro effects of MLBL on human lymphocyte effector functions and its mechanisms of action were evaluated. The results show that the most remarkable effects of MLBL on the immune system are: i) activation of the IL-2 receptor (IL-2Ralpha) on different lymphocyte subsets (B, CD4+ T and CD8+ T cells) involved both in humoral and cellular immune responses; ii) induction of cytokine synthesis (IL-2, IL-10, IL-12, IFNgamma) in the immune competent cells that induce and regulate immune responses; iii) generation of CD4+ and CD8+ effector T cells. Overall, these results suggest that the therapeutic effect of MLBL on acute and recurrent infections of the respiratory tract is related to its ability to activate the responses of different subsets of immune competent cells both for humoral and cellular immunity. Moreover, these effects can be induced either by direct immune cell activation or through the generation and activation of immune effector cells.     

Immunosuppression by chronic exposure to N-nitrosodimethylamine (NDMA) in mice.

Immunosuppression of humoral and cellular responses following chronic oral exposure to 1, 5, 10, and 20 ppm N-nitrosodimethylamine (NDMA) was examined in CD-1 mice. Monitoring of cumulative mortality and the incidence of peritoneal ascites in animals showed an NDMA dose-related mortality and hepatotoxicity. No visible changes in immunological parameters were noted at the 1 ppm NDMA dose. Immunosuppression of immunoglobulin M (IgM) antibody response by NDMA to sheep red blood cells (SRBC) was time-related, dose-related, and could be reversed within 30 d by removal of the chemical from the drinking water. Cellular immune response, monitored by allogeneic stimulation of cells in mixed lymphocyte reaction (MLR), was markedly suppressed by 10 and 20 ppm NDMA. Thus, chronic exposure to NDMA, except for the low-hepatotoxic doses of nitrosamine, resulted in a marked and persistent immunosuppression of cellular and humoral responses in CD-1 mice. In conclusion, chronic exposure to the hepatotoxic (ascite-inducing) doses of NDMA suppressed humoral and cellular immunity. The persistent immunosuppression could be reversed after the removal of NDMA from the drinking water. Although no direct NDMA-related cancer was reported in humans, our data point to a potential epigenetic carcinogenicity of nitrosamines due to chronic immunosuppression.     

乙双吗啉对有丝分裂原诱导的小鼠淋巴细胞增殖反应的抑制作用

当浓度为1.56mg/L时,乙双吗啉就可明显抑制LPS诱导的体外小鼠淋巴细胞增殖反应,其作用随浓度的增高而加强。当此药浓度增大到6.25mg/L时,ConA和PHA诱导的体外小鼠淋巴细胞增殖反应才开始出现明显抑制。表明乙双吗啉对B细胞增殖反应的抑制作用强于对T细胞增殖反应。乙双吗啉对淋巴细胞无非特异细胞毒作用。     

Antigen levels and antibody titers after DNA vaccination

DNA vaccination generates strong cellular and humoral immunity in animal models. The mechanisms by which plasmid DNA uptake and expression after intramuscular injection lead to immune responses are not well understood. In particular, the importance of antigen expression levels on subsequent antibody immune responses has not been established. We found that a chemiluminescent assay for alkaline phosphatase allows measurement of antigen levels of secreted alkaline phosphatase (SEAP) in vivo after intramuscular injection of a wide range of plasmid doses. The mice produced antibodies to the alkaline phosphatase reporter gene and both antigen levels and antibody titers were measured over time. We found that the correlation between initial antigen level and antibody response was high (r = 0.74, p < 0.001) and remained high even after accounting for the dose of plasmid injected (r = 0.61, p < 0.001). The correlation between DNA dose and antibody titer was statistically significant (r = 0.53, p < 0.001) but was reduced to almost zero after we accounted for initial antigen levels.     

黄芪多糖对阿霉素治疗小鼠乳腺癌的增效减毒机制研究

目的：探讨黄芪多糖对阿霉素治疗乳腺癌的增效减毒作用机制,为乳腺癌的治疗提供帮助。方法：建立小鼠乳腺癌皮下移植肿瘤模型,44只荷瘤小鼠随机平分为4组,即对照组（生理盐水治疗）、DOX组（阿霉素治疗）、APS组（黄芪多糖治疗）和APS+DOX组（黄芪多糖和阿霉素联合治疗）;观察治疗后荷瘤小鼠肿瘤体积大小、体质量变化以及平均存活时间、各器官组织学变化;检测荷瘤小鼠治疗前后T淋巴细胞、应激反应因子以及肝肾功能变化。结果：经过联合治疗后,APS+DOX组荷瘤小鼠肿瘤体积比显著低于对照组、APS组和DOX组;同时APS+DOX组荷瘤小鼠平均存活时间明显长于对照组、APS组和DOX组;HE染色表明荷瘤小鼠经过黄芪多糖和阿霉素联合治疗后,各器官损伤程度低于DOX组和对照组;治疗后APS+DOX组荷瘤小鼠肝功能和肾功能各指标、GSH、SOD以及T淋巴细胞（CD3⁺、CD4⁺、CD4⁺/CD8⁺和NK）水平显著高于DOX组但显著低于APS组和对照组,APS+DOX组CD8⁺、MDA水平显著低于DOX组但显著高于APS组、对照组,数据比较差异存在统计学意义（P<0.05）。结论：黄芪多糖能够有效促进阿霉素对乳腺癌皮下移植肿瘤小鼠的治疗和平均存活时间的延长,同时能够提高荷瘤小鼠免疫功能、抑制氧化应激反应和改善荷瘤小鼠的肝肾功能,有利于促进乳腺癌的治疗。     

A predictive F344 rat immunotoxicology model: cellular parameters combined with humoral response to NP-CgammaG and KLH

The purpose of this study was to examine the predictive value of humoral and cellular immune parameters in determining the immunotoxic effects of the oral administration of azathioprine (AZA), cyclophosphamide (CY), or cyclosporin A (CsA) at doses of 25/17, 10, or 25 mg/kg per day, respectively, for 30 days in F344 female rats. The effect of these known immunosuppressive compounds on the immune response was assessed in a humoral model that consisted of the administration of nitrophenyl-chicken gamma globulin (NP-CgammaG) and keyhole limpet hemocyanin (KLH) antigens during immunosuppressive treatment and the measurement of resulting rat antigen-specific IgG and IgM, as well as total IgG, levels. Cellular assessment parameters were collected from the same groups of animals as the humoral parameters and included organ weights and cellularity, hematology, lymphocyte phenotype characteristics, spleen cell mitogen stimulation (T and B cell-dependent), splenic natural killer (NK) cell cytotoxicity, and bone marrow cellularity and lymphocyte phenotype differential. Although decreases in several of the cellular assay parameters were observed, the only functional assays to demonstrate a statistically significant immunosuppressive effect by all three immunosuppressive agents were the antigen-specific serum IgG levels. The primary (day 10; 15 days post-immunization) and secondary (day 25; 5 days post-rechallenge) nitrophenyl (NP) responses were significantly suppressed by > or =60%. The use of NP hapten provided consistent responses when analyzed with a sensitive, well developed, ELISA methodology. Absolute lymphocyte phenotyping and lymphocyte hematology were also predictive of T cell immunosuppression for all three compounds. The data presented herein suggests that these two parameters, NP-IgG humoral response and lymphocyte phenotyping, are sufficient for identifying immunosuppressive compounds.     

Phenyl isocyanate is a potent chemical sensitizer

Diisocyanates, as a chemical class, are recognized for their ability to cause respiratory and dermal sensitization. By contrast, monofunctional isocyanates have not been associated with cases of clinical sensitization. We investigated the immunologic activity of phenyl isocyanate (PI), an aromatic monoisocyanate, which is a trace constituent in commercial diphenyl methane diisocyanate (MDI) products, since it is has been reported to cause an ‘asthma-like’ syndrome in rats. The potency of PI as a contact sensitizer was assessed using the mouse ear swelling test. PI was found to be the most potent isocyanate tested yielding an SD₅₀ (dose predicted to sensitize 50% of the mice) of 0.04 μmol/kg, compared with SD₅₀ values of 0.5, 2.1, and 30.4 μmol/kg, for the diisocyanate sensitizers hexamethylene diisocyanate, MDI, and toluene diisocyanate (TDI), respectively. When tested for ability to stimulate humoral immune responses, antibody titers to PI were more than ten-fold greater than those induced by TDI. The mean hapten-specific IgC antibody titer to PI was 1.4 × 10⁴, compared with 1.3 × 10³ for TDI. The anti-PI IgG₁ anaphylactic antibody titer (1.2 × 10⁴) was significantly greater than the anti-TDI IgG₁ titer of 6.4 × 10₂. Hapten-specific IgE was not detected to either isocyanate. These results indicate that phenyl isocyanate is a potent inducer of both cellular and humoral immune responses. This activity may be a significant contribution to the sensitization potential of commercial products in which it is a trace contaminant.     

Effects of swainsonine on the humoral immune response of lipopolysaccharide

Effects of swainsonine (SW; 8α, β-indolizidine-1α, 2α, 8-triol from Locoweed) on the humoral immune responses of lipopolysaccharide (LPS) were studied in ICR mice. Mice were divided into 4 groups (10mice/group), and LPS was given to each mouse 1 hr after i.p. injection with 3.7 mg/kg of swainsonine, by i.p. injection twice a week for 14 days at a dose of 2 mg/kg. Humoral immune responses were evaluated by hemagglutination (HA) titer and splenic plaque forming cells (PFC). The results of this study were summarized as follows: Mice administrated each of LPS and SW showed significant enhancement of the weight ratios of spleen to body, HA titer, 2-mercaptoethanol-resistant HA (MER-HA) titer and PFC compared with those in controls. However, the LPS plus SW treatment decreased HA titer, MER-HA titer and PFC corresponding to humoral immunity, as compared with those in the mice treated with LPS alone. These findings indicated that LPS significantly enhanced humoral immune responses, but their enhancement effects were lowered somewhat by SW.     

不同制备工艺板蓝根多糖对小鼠脾淋巴细胞增殖作用的影响

目的研究板蓝根在不同制备工艺条件下得到的多糖对小鼠脾淋巴细胞增殖的影响。方法用不同浓度乙醇将板蓝根水提液进行分级沉淀,并结合二乙基氨基乙基纤维素离子交换层析柱,凝胶色谱柱分离纯化得到IP-Ⅰ、IP-Ⅱ、IP-Ⅲ…IP-Ⅷ8个粗多糖部分及IP-Ⅸ、IP-Ⅹ、IP-Ⅵ、IP-Ⅻ4个均一多糖部分。采用四甲基偶氮唑蓝法检测板蓝根多糖对小鼠脾细胞增殖反应的影响。结果板蓝根多糖各部分均具有促进淋巴细胞增殖作用,且存在一定的量效关系。结论板蓝根多糖对淋巴细胞的促增殖作用的最适剂量为IP-Ⅸ(0.125mg.mL-1)和IP-Ⅻ(0.125mg.mL-1)。     

Suppression of immune response associated with functions of B-cells, Th1- and Th2-lymphocytes and interleukins produced by these cells, and pharmacological correction of these disturbances upon acute intoxication with methanol

Experiments on Wistar rats showed that acute poisoning with methanol (0.75 LD50) leads to the suppression of cellular and humoral immune responses and decreases the blood concentration of interleukins (IL-2, IL-4) with an increase in the IL-2/IL-4 ratio. These facts indicate that a decrease in Th2 lymphocyte activity is more pronounced in comparison to that of Th1 cells. The immunomodulators mielopide and polyoxidon administered in a daily dose of 10 mg/kg for 4 days upon acute poisoning with methanol virtually completely restore the cellular and humoral immune responses, the activity of natural killers, and the synthesis of interleukins.     

干扰素在狂犬病疫苗中的佐剂效果研究

目的研究干扰素在狂犬病疫苗中的佐剂效果。方法将人用狂犬病疫苗与干扰素按一定比例混合,制成干扰素佐剂狂犬病疫苗。将干扰素佐剂狂犬病疫苗和无佐剂狂犬病疫苗分别免疫昆明小鼠,并于免疫前和免疫后4,7,14,30,60d时眶静脉采血,用快速免疫荧光灶抑制试验检测小鼠血清中和抗体;同时经腹腔免疫小鼠,第15天时取脾检测淋巴细胞转化率。结果干扰素佐剂狂犬病疫苗可促进免疫细胞增殖。与无佐剂狂犬病疫苗相比,干扰素佐剂狂犬病疫苗诱导产生中和抗体的时间早、滴度高。结论干扰素佐剂狂犬病疫苗具有良好的免疫学效果。