Quillaja saponin can modulate ovalbumin-induced IgE allergic responses through regulation of Th1/Th2 balance in a murine model

Quillaja saponin is the extract from the balk of a South American tree, and it is considered to modulate immunological responses. We hypothesized that Quillaja saponin may change allergy-associated cytokine profile and antigen-specific immune responses. The purpose of this study is to investigate whether Quillaja saponin can suppress ovalbumin (OVA)-induced IgE-mediated allergic responses through promoting a dominant Th1 immune response. The spleen cells from BALB/c mice, which were primed by OVA, were used for an in vitro challenge test. The level of total and OVA-specific IgE, IL-4, IFN-gamma, and IL-12 was determined by enzyme-linked immunosorbent assay (ELISA). BALB/c mice were orally administered with saponin for 35 days. The mice were immunized intraperitoneally with OVA on days 14 and 21. After intraperitoneal challenge with OVA on day 35, anaphylactic symptoms were monitored. Total and specific IgE and IgG, specific IgG1 and IgG2a, and histamine levels in serum were analyzed by ELISA. The increase of IL-12 and IFN-gamma levels was observed in the presence of Quillaja saponin, while the IL-4 level was decreased. Furthermore, Quillaja saponin suppressed total and OVA-specific IgE secretion in spleen cells. Balb/c mice that were orally administered Quillaja saponin exhibited lower total and OVA-specific IgE and OVA-specific IgG secretions, whereas total IgG levels remained unchanged. Suppression of OVA-specific IgG1 and an increase of OVA-IgG2a were observed in mice fed saponin. Quillaja saponin also decreased serum histamine levels and diminished anaphylactic symptoms. The present study indicates that Quillajasaponin can suppress allergen-specific IgE-mediated reactivity in a murine model of food allergy, which results from shifting from a Th2-dominated to a Th1-dominated immune response.     

Suppression of allergic reactions by dehulled adlay in association with the balance of TH1/TH2 cell responses

Dehulled adlay is known as a natural Chinese medicine having antiallergic activity, although its mechanism remains unclear. This study examined the effects of dehulled adlay on antigen-specific antibody and cytokine production. Mice were immunized three times with ovalbumin (OVA) in alum adjuvant. It was found that oral administration of dehulled adlay in mice suppressed the production of IgE against OVA antigen. Serum anti-OVA IgG(2a) antibody levels were significantly increased in mice after oral administration of dehulled adlay. Furthermore, the production of IL-2 by OVA-stimulated splenocytes was augmented in dehulled adlay-fed mice. Although dehulled adlay had no effect on the serum anti-OVA IgG(1) antibody levels, it had a great capacity to reduce IL-5 secretion by means of OVA-stimulated splenocytes. Hydrothermal processes, including steaming and extrusion cooking, did not change the capacity of dehulled adlay to suppress IgE production. Three fractions of dehulled alday, including methanolic extract, warm water extract, and residue, were obtained. The methanolic extract exhibited the greatest capacity to reduce anti-OVA IgE production. These results suggest that dehulled adlay has a modulating ability to shift the balance from Th2 to Th1 dominance in the T cell mediated immune system and may be beneficial for the treatment of allergic disorders.     

不同乳酸菌发酵甘薯渣产物组分分析

为提高甘薯淀粉加工副产物的高值化利用水平,本试验以植物乳杆菌(Lp)、干酪乳杆菌(Lc)、保加利亚乳杆菌(Lb)、戊糖片球菌(Pp)、嗜热链球菌(St)、商业植物乳杆菌(SZ)6种乳酸菌分别对甘薯渣进行发酵,系统分析不同发酵产物的pH值、总酸含量及营养功能成分,并采用灰色理论加权关联度对甘薯渣发酵产物进行综合营养评价,筛选适宜发酵甘薯渣的乳酸菌菌种。结果表明,6种乳酸菌的发酵能力存在显著差异,其中Lb组pH值最低(3.15),Lc组总酸含量最高(27.90);同时,不同乳酸菌对发酵产物中营养功能成分的影响存在显著差异,其中Lb组乳酸含量最高(11.60 mg·mL^-1),SZ组乙酸含量最高(66.99μg·mL^-1),Lb组可溶性膳食纤维含量最高(0.74 g·100 mL^-1),St组总酚含量最高(146.87μg GAE·mL^-1)。与未发酵样品相比,Lc组游离氨基酸总量提高了3.71倍;所有发酵产物中的Mg、Ca、Fe、Zn、Se等矿物质元素含量均显著提高。进一步通过灰色理论加权关联度分析发现...     

Effect of heat-inactivated kefir-isolated Lactobacillus kefiranofaciens M1 on preventing an allergic airway response in mice

In this study, we assessed the anti-asthmatic effects of heat-inactivated Lactobacillus kefiranofaciens M1 (HI-M1) and its fermented milk using different feeding procedures and at various dosage levels. The possible mechanisms whereby HI-M1 has anti-allergic asthmatic effects were also evaluated. Ovalbumin (OVA)-allergic asthma mice that have been orally administrated the HI-M1 samples showed strong inhibition of production of T helper cell (Th) 2 cytokines, pro-inflammatory cytokines, and Th17 cytokines in splenocytes and bronchoalveolar fluid compared to control mice. An increase in regulatory T cell population in splenocytes in the allergic asthma mice after oral administration of H1-M1 was also observed. In addition, all of the features of the asthmatic phenotype, including specific IgE production, airway inflammation, and development of airway hyperresponsiveness, were depressed in a dose-dependent manner by treatment. These findings support the possibility that oral feeding of H1-M1 may be an effective way of alleviating asthmatic symptoms in humans.     

Use of murine models to detect the allergenicity of genetically modified Lactococcus lactis NZ9000/pNZPNK

By introducing aprN into Lactococcus lactis NZ9000, the genetically modified L. lactis NZ9000/pNZPNK successfully expressed the nattokinase. The safety assessment of this novel strain was based on allergenicity of pepsin digestion stability and murine model serologic identity. Subjecting to the GM strain and host to pepsin digestion, the soluble fractions and cell debris were fast degraded completely. Feeding with ovalbumin resulted in significantly higher production of IgG1 and IgE as compared to that of L. lactis NZ9000/pNZPNK or L. lactis NZ9000. Further, the serum IgG2a level increased dose-dependently at week 2 and induced immune reaction toward Th1 pathway. Secretion of cytokines IL-4 and IL-10 fed with lactococci was significantly lower than that of the OVA group. L. lactis NZ9000/pNZPNK did not increase the proliferation of type 2 helper T cells in spleen or induce allergenicity in BALB/c mice. On the basis of the results, the new GM lactic acid bacterium is regarded as safe to use.     

Heat-killed cells of lactobacilli skew the immune response toward T helper 1 polarization in mouse splenocytes and dendritic cell-treated T cells

It is believed that probiotics play an important role for the health of the host, including modulation of immune responses. Most studies have focused on the immunomodulatory effects of viable cells of lactic acid bacteria; however, we investigated those of heat-killed cells of lactic acid bacteria in this study. We first observed the effects on immune functions via stimulating splenocytes with three heat-killed Lactobacillus strains. Furthermore, we also investigated the effect of mouse dendritic cells (DCs) treated with these heat-killed Lactobacillus strains on T cell responses. The results showed that these Lactobacillus strains were able to stimulate cell proliferation and interleukin (IL)-10, IL-12 p70, and interferon (IFN)-gamma production but not transforming growth factor (TGF)-beta in splenocytes. In addition, these heat-killed Lactobacillus strains also stimulated high-level secretion of IL-12 p70 in DCs and switched T cells to T helper (Th) 1 immune responses, as evidenced by the elevated secretion of IFN-gamma but not IL-5, IL-13, and TGF-beta. These results showed that lactobacilli play a potentially important role in modulating immune responses and allergic reactions.     

Effect of yak milk casein hydrolysate on TH1/TH2 cytokines production by murine spleen lymphocytes in vitro

Yak casein hydrolysate was derived from the enzymatic alcalase-hydrolysate of a typical northwestern China milk product called Qula. An in vitro study was conducted to examine their immunoregulatory effects on murine T cells, including Con-A-induced lymphoproliferation and splenocyte cell cycle, production, and messenger RNA (mRNA) expression of interleukin-2 (IL-2), interferon-gamma (IFN-gamma), and interleukin-4 (IL-4). The results showed that yak casein hydrolysate had lymphoproliferation activity on murine splenocytes and induced their cell cycle from the G1 to the S phase. It could increase Con-A-induced IL-2 and IFN-gamma production in spleen cells, but a very weak or no effect was observed in the absence of Con-A. The present study also showed that it could markedly increase the production and mRNA expression of IFN-gamma and IL-2, which are key cytokines for T helper 1 cell (Th1) cell development, in a dose-dependent manner. However, their effects on IL-4 secretion were not obvious, and the enhancement was much lower than that of IFN-gamma and IL-2. All of these demonstrated that yak casein hydrolysate could increase type 1 cytokine production, thereby shifting the Th1/T helper 2 cell (Th2) balance toward a Th1-dominant phenotype, which meant that yak casein hydrolysate could indeed not only modulate the differentiation of helper T cell but also has the capacity to modulate the Th1/Th2 balance. Therefore, yak casein hydrolysate may be useful for the treatment of cell-mediated immune diseases.     

Isolation and characterization of three novel peptides from casein hydrolysates that stimulate the growth of mixed cultures of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus

In this study, sodium caseinate hydrolysates produced by papain with strong growth-stimulating activity for Streptococcus thermophilus (St) and Lactobacillus delbrueckii subsp. bulgaricus (Lb) were obtained. A series of separation methods including ultrafiltration, macroporous adsorption resin chromatography, gel filtration chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC) were applied to isolate and purify the peptide(s), which were mainly responsible for the activity. Finally, three novel growth-stimulating peptides, H-2-A, F2-c, and F2-b, corresponding to amino acid residues 29-35 and 103-108 of bovine α(S2)-casein and 181-186 of bovine α(S1)-casein, respectively, were obtained. With supplementation of H-2-A, F2-b, or F2-c at a protein concentration of 0.3%, the biomass yield of these two lactic acid bacteria (LAB) was enhanced by 193.3, 166.7, or 151.7%, respectively. In addition, there were significant (p < 0.05) increases in viable counts of St and lactic acid production of LAB in the presence of the purified peptides.     

Antioxidative ability of lactic acid bacteria

Nineteen strains of lactic acid bacteria were investigated for antioxidative activity. These includedLactobacillus acidophilus B, E, N1, 4356, LA-1, and Farr; Lactobacillus bulgaricus 12 278, 448, 449, Lb, 1006, and 11 842; Streptococcus thermophilus 821, MC, 573, 3641, and 19 987; and Bifidobacterium longum B6 and 15 708. Intracellular cell-free extract of all strains demonstrated antioxidative activity with inhibition rates of ascorbate autoxidation in the range of 7-12%. Antioxidative mechanisms including metal ion chelating ability, scavenge of reactive oxygen species, enzyme inhibition, and reducing activity of intracellular cell-free extract of lactic acid bacteria were studied. S. thermophilus 821 had the highest metal ion chelating ability for Fe(2+), and B. longum 15 708 showed the highest Cu(2+) chelating ability among the 19 strains tested. All strains demonstrated reactive oxygen species scavenging ability. L. acidophilus E showed the highest hydroxyl radical scavenging ability, and B. longum B6 had the best hydrogen peroxide scavenging ability. Reducing activity was also found in all strains. Most of the strains tested demonstrated excellent reducing activity. B. longum B6 showed the highest reducing activity among the 19 strains tested. In enzyme inhibition, superoxide dismutase activity was not found in these 19 strains, and the activity of superoxide dismutase was not induced when metal ion Mn(2+), Fe(2+), or Cu(2+)Zn(2+) was present.     

Antiosteoporotic effects of Lactobacillus -fermented soy skim milk on bone mineral density and the microstructure of femoral bone in ovariectomized mice

Osteoporosis is a major skeletal disease associated with loss of estrogen in postmenopausal women. In this study, Lactobacillus paracasei subsp. paracasei NTU 101 (NTU 101F) and Lactobacillus plantarum NTU 102 (NTU 102F) were used as starters to ferment soy skim milk. This was then used as a nutritional supplement for 8 weeks to ovariectomized (OVX) mice. This study reveals that soy skim milk fermented with lactobacilli can increase the contents of aglycone isoflavones, soluble calcium, and vitamin D(3). The trabecular bone volumes and trabecular number of the distal femur in mice fed NTU 101F increased by a factor of 1.48 and 1.74 compared with the OVX group. The bone network density and thickness of the distal metaphyseal trabecular in mice fed NTU 101F and Fosamax was significantly greater than that of OVX mice. These results suggest that fermented soy skim milk can attenuate bone loss in OVX mice and lower the risk of osteoporosis.     

主成分分析法优选枸杞乳酸菌发酵饮品发酵剂

乳酸菌发酵作为果蔬汁的一种绿色加工技术,不仅可以赋予产品独特的风味,还可以转化其中的活性物质,提高产品的营养价值和保健功效。该研究以湖北杂交枸杞为原料,使用6种乳酸菌(植物乳杆菌、嗜热链球菌、嗜酸乳杆菌、鼠李糖乳杆菌、干酪乳杆菌及发酵乳杆菌)进行发酵,研究发酵前后枸杞果汁理化特性、主要活性成分及体外抗氧化变化,并利用主成分分析进行综合评价优选出理想的发酵菌株。结果表明,6种乳酸菌在枸杞果汁中生长良好,活菌数均能达到10.0 lg CFU/mL以上。发酵后的枸杞果汁中总糖和还原糖含量显著降低(P0.05),且植物乳杆菌和嗜热链球菌产酸能力更强,发酵后总酸含量达6.74、6.07g/kg。与未发酵枸杞果汁相比,经植物乳杆菌、嗜热链球菌、鼠李糖乳杆菌和发酵乳杆菌发酵的枸杞果汁中总酚含量增加了13.76%～28.07%,而嗜酸乳杆菌和干酪乳杆菌发酵后无显著性差异(P0.05)。6种乳酸菌发酵后枸杞果汁中总黄酮含量增加了55.80%～161.97%。发酵枸杞果汁的抗氧化活性与发酵前相比均有显著提高(P0.05)。基于主成分分析的综合评价函数显示经植物乳杆菌、发酵乳杆菌发酵的枸杞果汁品质更优,适宜作为开发枸杞高值化绿色加工饮品的发酵剂。     

自然发酵和接种发酵方法对白菜品质的影响(简报)

以自然发酵和人工接种发酵(Lactobacillus pentosus 和 Leuconostoc.mesenteroides 为发酵剂)加工的白菜为材料,对比了两种加工白菜亚硝酸盐含量随时间的变化以及有益微生物(乳酸菌)和有害微生物数量的变化,并对其制品做了感观分析.结果得出人工接种发酵白菜中亚硝酸盐含量在发酵整个过程均比自然发酵的低,且未出现自然发酵白菜初期出现的"亚硝峰"; 人工接种发酵白菜中肠杆菌及乳酸菌之外的需氧嗜温菌的数量较自然发酵的少;乳酸菌数量较自然发酵的多;人工接种发酵白菜感官分析优于自然发酵.说明人工接种发酵白菜的品质优于自然发酵加工的制品.     

Conjugated linoleic acid content and organoleptic attributes of fermented milk products produced with probiotic bacteria

The effect of probiotic bacteria on the formation of conjugated linoleic acid (CLA), microbial growth, and organoleptic attributes (acidity, texture, and flavor) of fermented milk products was determined. Four probiotic bacteria, Lactobacillus rhamnosus, Propionibacterium freudenreichii subsp. shermanii 56, P. freudenreichii subsp. shermanii 51, and P. freudenreichii subsp. freudenreichii 23, were evaluated individually or in coculture with traditional yogurt cultures (Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus salivarius subsp. thermophilus). The lipid source was hydrolyzed soy oil. L. rhamnosus, in coculture with yogurt culture, resulted in the highest content of CLA. Growth and CLA formation of propionibacteria were enhanced in the presence of yogurt cultures. Texture and flavor attributes of fermented milks produced with propionibacteria were significantly different than the fermented milks processed with yogurt cultures. The fermented milks processed with probiotic bacteria in coculture with yogurt cultures demonstrated similar acidity, texture, and flavor as the fermented milk produced with yogurt cultures.     

Immunoadjuvant activity, toxicity assays, and determination by UPLC/Q-TOF-MS of triterpenic saponins from Chenopodium quinoa seeds

The adjuvant activity of Chenopodium quinoa (quinoa) saponins on the humoral and cellular immune responses of mice subcutaneously immunized with ovalbumin (OVA) was evaluated. Two quinoa saponin fractions were obtained, FQ70 and FQ90, and 10 saponins were determined by UPLC/Q-TOF-MS. Mice were immunized subcutaneously with OVA alone or adjuvanted with Quil A (adjuvant control), FQ70, or FQ90. FQ70 and FQ90 significantly enhanced the amount of anti-OVA-specific antibodies in serum (IgG, IgG1, and IgG2b) in immunized mice. The adjuvant effect of FQ70 was significantly greater than that of FQ90. However, delayed type hypersensitivity responses were higher in mice immunized with OVA adjuvanted with FQ90 than mice treated with FQ70. Concanavalin A (Con A)-, lipopolysaccharide-, and OVA-stimulated splenocyte proliferation were measured, and FQ90 significantly enhanced the Con A-induced splenocyte proliferation. The results suggested that the two quinoa saponin fractions enhanced significantly the production of humoral and cellular immune responses to OVA in mice.     

Antioxidative effects of lactic acid bacteria on the colonic mucosa of iron-overloaded mice

The antioxidative effects of lactic acid bacteria on lipid peroxidation in the colonic mucosa were investigated. Among 49 strains of lactic acid bacteria, Streptococcus thermophilus YIT 2001 showed the highest inhibitory activity against lipid peroxidation in liposomes induced by ferrous iron. Feeding a diet containing 0.4% St. thermophilus YIT 2001 (2 x 10(8) colony-forming units per mouse per day) for 2 weeks caused a significant decrease of lipid peroxide (thiobarbituric acid reactive substance) in the colonic mucosa of iron-overloaded mice (0.07% Fe in the diet). The mucosal lipid peroxide level did not correlate with the soluble iron concentration of the cecal contents. Therefore, it is suggested that the antioxidative effect of St. thermophilus YIT 2001 in the colonic mucosa was not due to the removal of ferrous iron from the reaction system of lipid peroxidation.     

Immunomodulatory effects of heated ovomucoid-depleted egg white in a BALB/c mouse model of egg allergy

Oral immunotherapy (OIT) is a promising therapeutic approach for treating food allergy. The treatment with heated ovomucoid-depleted egg white (HOMEW) in egg-allergic patients is noteworthy; however, OIT protocols are still experimental, and a better knowledge of the underlying mechanism is required. The objective of this work was to investigate the immunomodulatory effects of HOMEW and characterize the underlying mechanism in a BALB/c mouse model of egg allergy. Mice were sensitized with EW and treated with HOMEW. Post treatment, mice were challenged with EW and euthanized for collecting blood and spleen. Markers of allergic clinical outcomes were measured as histamine concentration, serum antibody activity, and cytokine production from cultured splenocytes. Digestibility of HOMEW was assessed mimicking physiological conditions in vitro. The HOMEW demonstrated high digestibility. The treatment induced a marked increase of the Th1/Th2 ratio in the high-dose treatment group. Treated mice had significantly less histamine, EW-specific IgE, and IL-4 and more IFN-γ and IL-10. This study confirms the mechanisms involved in successful tolerance induction with OIT using HOMEW and allows understanding of the vital role of surrogate allergy markers involved in immune modulation.     

Modification of IgE binding during heat processing of the cow's milk allergen beta-lactoglobulin

The effect of heat treatment on the IgE binding ability of beta-lactoglobulin, as pure protein or in whole milk, was studied by inhibition of IgE antibody binding using FEIA-CAP inhibition. A slight but significant decreased IgE binding was seen between unheated and heat-treated beta-lactoglobulin solution at 74 degrees C (IC(50) = 2.03 and 3.59 microg/mL, respectively, p = 0.032). A more pronounced decrease was found at 90 degrees C with an IC(50) of 8.45 microg/mL (p = 0.014). The inhibition of IgE binding of milk after heat treatment at 90 degrees C was also significantly decreased (p = 0.007). However, at all heat treatments, a similar total amount of IgE antibodies could be inhibited at a sufficiently high concentration of beta-lactoglobulin. The inhibiting ability of beta-lactoglobulin was significantly impaired in some fermented acidified milk products such as yogurt as compared to that in nonfermented milk (p < 0.001). There was only a small difference of IgE binding between the native forms of genetic variants A and B.     

Development of a new method, based on a bioreactor coupled with an L-lactate biosensor, toward the determination of a nonspecific inhibition of L-lactic acid production during milk fermentation

The development and characteristics of a bioreactor employing bacteria (Streptococcus thermophilus) encapsulated in Ca-alginate beads coupled with an L-lactate biosensor are reported. The biosensor comprises a carbon paste electrode modified with enzymes HRP (horseradish peroxidase), LOD (lactate oxidase), and FcH (ferrocene) as redox mediator. The measurement of L-lactate is based on the signal produced by H(2)O(2), the product of the enzymatic oxidation of L-lactate by LOD. The detection of H(2)O(2) is performed at the electrode surface via HRP/FcH at low operating potential (-100mV vs Ag/AgCl). Optimization studies were performed using the bioreactor in conjunction with an L-lactate electrode operating in a flow injection system to assess the ability of encapsulated bacteria to ferment carbohydrate solutions. The possibility of using the developed method to assess the fermentation capability of milk samples was evaluated. Bronopol (2-bromo-2-nitro propane-1,3-diol) was chosen to simulate the effect of an inhibitory agent of milk fermentation. The obtained results indicated that the evaluation of the amount of L-lactate amount produced through the bioreactor could be used as a measure of inhibition of lactic acid production in milk samples.     

龙眼果浆复合乳酸菌发酵工艺优化

为筛选出适合龙眼发酵的复合乳酸菌,优化建立其发酵工艺,比较分析了7种不同的乳酸菌发酵龙眼的降糖能力和释放结合态酚类物质的能力,发现明串珠菌降糖能力最强,植物乳杆菌释放结合态酚能力最强。利用植物乳杆菌和明串珠菌复合发酵龙眼果肉,以游离酚质量分数和还原糖质量分数结合的综合模糊评判值为指标,通过Box-benhnken中心组合试验设计,确定复合乳酸菌发酵龙眼的最佳工艺条件。结果表明,各因素对复合乳酸菌发酵龙眼果浆的模糊评判值影响程度从强到弱依次为:发酵时间发酵温度接种量料液比。复合乳酸菌发酵龙眼的最佳工艺为:料液比为1∶7(g/m L),发酵温度为37℃,发酵时间为53 h,接种量为1.4%,菌种配比为1∶1。在最佳工艺条件下,发酵龙眼果浆的游离酚质量分数为(161.77±3.93)mg/(100 g),还原糖质量分数为(122.80±3.64)mg/g。该研究为开发低糖含量,且适合糖尿病、肥胖症等病人食用的龙眼乳酸菌发酵饮料新产品提供参考。