Impacts of Intralipid on Nanodrug Abraxane Therapy and on the Innate Immune System.

A major obstacle to nanodrugs-mediated cancer therapy is their rapid uptake by the reticuloendothelial system that decreases the systemic exposure of the nanodrugs to tumors and also increases toxicities. Intralipid has been shown to reduce nano-oxaliplatin-mediated toxicity while improving bioavailability. Here, we have found that Intralipid reduces the cytotoxicity of paclitaxel for human monocytic cells, but not for breast, lung, or pancreatic cancer cells. Intralipid also promotes the polarization of macrophages to the anti-cancer M1-like phenotype. Using a xenograft breast cancer mouse model, we have found that Intralipid pre-treatment significantly increases the amount of paclitaxel reaching the tumor and promotes tumor apoptosis. The combination of Intralipid with half the standard clinical dose of Abraxane reduces the tumor growth rate as effectively as the standard clinical dose. Our findings suggest that pre-treatment of Intralipid has the potential to be a powerful agent to enhance the tumor cytotoxic effects of Abraxane and to reduce its off-target toxicities.

Soybean oil containing ginseng saponins as adjuvants promotes production of cytokines and enhances immune responses to foot-and-mouth disease vaccine.

In the present study, the adjuvant effect of soybean oil containing ginseng root saponins (SO-GS-R) on the immune response to foot-and-mouth disease vaccine (FMDV) in mice was investigated. When immunized with FMDV antigen emulsified in an SO-GS-R formulation, mice generated remarkably higher serum antibody and cytokine responses than mice immunized with FMDV antigen alone. To elucidate the mechanisms underlying the adjuvant effect of SO-GS-R, we measured cytokines in serum and muscle tissue after intramuscular injection of SO-GS-R. The results showed that injection of SO-GS-R significantly increased the levels of IL-1ß, IL-5, IL-6, G-CSF, KC, MCP-1, MIP-1α, and MIP-1ß in both serum and muscle. These results suggested that SO-GS-R recruits neutrophils, eosinophils, T cells and macrophages, causing immune cell recruitment at the injection site, driving antigen-presenting cells to actively participate in the onset of immunity, and amplifying the immune responses. Considering its adjuvant activity and plant-derived properties, SO-GS-R should be further studied for its adjuvant effect on vaccines used in food animals.

Increased intake of vegetable oil rich in n-6 PUFA enhances allergic symptoms and prevents oral tolerance induction in whey-allergic mice.

Increased intake of vegetable oils rich in n-6 PUFA, including soyabean oil, has been associated with an increase in allergic disease. The present study aimed to determine the effect of an increasing dose of dietary vegetable oil on allergic outcomes in mice. To study this, mice received a 7 v. 10 % soyabean oil diet before and during oral sensitisation with whey or whey hyperimmune serum transfer. Another group of mice received partial whey hydrolysate (pWH) while being fed the diets before oral sensitisation. The acute allergic skin response, serum Ig level, mouse mast cell protease-1 (mMCP-1) concentration and/or splenic T-cell percentages were determined upon whey challenge. When the diets were provided before and during oral sensitisation, the acute allergic skin response was increased in mice fed the 10 % soyabean oil diet compared with the 7 % soyabean oil diet. Whey IgE and IgG1 levels remained unaltered, whereas mMCP-1 levels increased in mice fed the 10 % soyabean oil diet. Furthermore, allergic symptoms were increased in naive mice fed the 10 % soyabean oil diet and sensitised with whey hyperimmune serum. In addition to enhancing the mast cell response, the 10 % soyabean oil diet increased the percentage of activated Th1 and Th2 cells as well as increased the ratios of Th2:regulatory T cells and Th2:Th1 when compared with the 7 % soyabean oil diet. Oral tolerance induction by pWH was abrogated in mice fed the 10 % soyabean oil diet compared with those fed the 7 % soyabean oil diet during pretreatment with pWH. In conclusion, increased intake of soyabean oil rich in n-6 PUFA suppresses tolerance induction by pWH and enhances the severity of the allergic effector response in whey-allergic mice. Dietary vegetable oils rich in n-6 PUFA may enhance the susceptibility to develop or sustain food allergy.

[Allergic enterocolitis with protein-losing enteropathy in a 5-month-old girl: a case report].

We report the case of a 5-month-old female infant who had allergic enterocolitis with protein-losing enteropathy and had low birth weight. Until the age of 4 months, she was fed only breast milk and did not experience any related problems. When she was 5-months-old, she was admitted to our hospital with the chief complaints of vomiting and diarrhea. She had whole body edema, thoracic and abdominal ascites, and bloody stools. Laboratory examinations showed the presence of hypoalbuminemia, hypogammaglobulinemia, and an elevated CRP level. Subsequently, she was fed an elemental diet with enteral tube feeding, and her condition gradually improved. However, 2 weeks later, her symptoms reappeared, and therefore, she was admitted to the National Center for Child Health and Development. She underwent upper and colon endoscopies and was found out to have multiple inflammatory lesions in the area extending from the cecum to the rectum. The biopsy findings showed features of eosinophilic enterocolitis and she was diagnosed with allergic eosinophilic enterocolitis. Subsequently, an elemental formula that does not include soybean was used for feeding the infant. The symptoms resolved and her condition improved; currently, she is well and is showing normal development. We have reported this case because allergic enterocolitis is becoming a topic of concern.

Influence of the slow infusion of a soybean oil emulsion on plasma cytokines and ex vivo T cell proliferation after an esophagectomy.

BACKGROUND: Lipid emulsions have been suggested to reduce immune responses, particularly in severely stressed patients. The authors investigated the influence of the slow intravenous infusion of a soybean oil-based lipid emulsion on some immune parameters in patients who had undergone an esophagectomy for esophageal cancer. METHODS: Thirty-two patients who had undergone an esophagectomy were randomly divided into a lipid emulsion (LPD)-treated group and a control group. All patients received parenteral feeding with a glucose-based solution. Patients in the LPD group received 100 mL of a 20% soybean oil emulsion for 7 days after the esophagectomy in addition to the glucose-based feeding. A slow infusion rate (0.09-0.12 g/kg/h) was adopted to take account of the intrinsic degradation of infused lipids. Immune responses were measured based on lymphocyte proliferation and serum concentrations of monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). The authors also measured levels of rapid turnover proteins (ie, transferrin, prealbumin, and retinol-binding protein). RESULTS: Phytohemagglutinin- and concanavalin A-stimulated lymphocyte proliferation significantly decreased after the esophagectomy, but no significant difference was seen between the LPD and control groups. No significant difference in changes in plasma concentrations of MCP-1, IL-6 and TNF-α occurred between the 2 groups either. Plasma concentrations of rapid turnover proteins did not differ between the groups. CONCLUSIONS: These results indicate that the lipid emulsion did not affect the immune parameters measured in patients who had undergone an esophagectomy when administered at a slow rate.

Induction of immune tolerance in mice with a novel mucosal nanoemulsion adjuvant and self-antigen.

AIM: The aim of this study was to investigate the impact of a novel nanoemulsion (NE) adjuvant, a soybean oil emulsion, on autoimmune response. To this end, we used murine thyroglobulin (mTg)-induced experimental autoimmune thyroiditis in mice as a study model. MATERIALS & METHODS: Mice received NE or NE + mTg by nasal delivery. At 1 week after the second nasal delivery of NE with or without mTg, all mice were immunized with mTg and lipopolysaccharides to induce experimental autoimmune thyroiditis. RESULTS: Compared with controls, mTg-NE-treated mice had much more antigens accumulated in the nasal passage and thymus and developed a milder form of thyroiditis. This was accompanied by an increase in IL-10, IL-17 and reduced IFN-γ. The production of anti-mTg antibodies was significantly decreased in mTg-NE-treated mice. The percentage of Tregs in cervical lymph nodes was higher in mTg-NE-treated mice than NE-treated mice. Furthermore, Foxp3 and TGF-ß levels were prominently enhanced in mTg-NE-treated mice. CONCLUSION: This study indicates that a low dose of mTg in NE can significantly enhance antigen uptake and Tregs, resulting in inhibition of experimental autoimmune thyroiditis development.

Simple extraction method of non-allergenic intact soybean oil bodies that are thermally stable in an aqueous medium.

This study supplied a simple extraction method for intact soybean oil body (ISOB) and examined the heating effect on ISOB. ISOB, which just contained its intrinsic protein (oleosin), could be obtained by pH 11 extraction (50000g, 45 min). ISOB suspension was dialyzed to deionized water (1:3600) and named DISOB. DISOB aggregated at pH 5.7, but NaCl pre-addition (5-500 mM) made ISOB disperse well at pH 5.7. The heating (30, 40, 50, 60, 70, 80, and 90 degrees C and boiling water baths, 30 min) did not affect the particle size distributions of ISOB. The pH and CaCl(2) effects on DISOB and its surface hydrophobicity were also not affected by heating (>95 degrees C, 5 min). Both unheated and heated ISOB were bound to native soybean protein but were not bound to the heat-denatured one. Thus, it was suggested that ISOB changed little by heating. This study was meaningful in two aspects: (1) pH 11 extraction removed beta-conglycinin, glycinin, and allergenic proteins (such as Gly m Bd 30K), and the obtained ISOB had good stability in an aqueous medium. (2) Heating could denature the contamination allergenic proteins.

Analytic investigations on protein content in refined seed oils: implications in food allergy.

BACKGROUND: A number of scientific reports have investigated the possible implications of refined seed oils in allergic reactions, resulting in conflicting points of view. Also the total amount of residual proteins after refinement is still a matter of debate. Nevertheless, seed oils are now blamed as possible cause of allergic reactions. OBJECTIVE: To determine the true amount of proteins after oil refinement and to shed new lights on allergenic properties of refined seed oils. METHODS: We optimized a protein extraction procedure on several commercial refined seed oils. Both colorimetric and amino acid analysis were used to measure residual protein content. SDS-PAGE was also used for characterizations of protein band patterns. Sensitized child patients sera were tested by Western blot on PAGE-resolved proteins. RESULTS: Our extraction method proved to be effective and reproducible. Amino acid analysis resulted more accurate in determining the protein content with respect to colorimetric methods, indicating a higher protein content than that previously reported. IgE responsive residual proteins were found in peanut oil extracts. CONCLUSIONS: Our preliminary data suggest that fully refined seed oils should be taken into account in the context of allergic reactions and would benefit of further toxicological studies.

Immune modulation by parenteral lipid emulsions.

Total parenteral nutrition is the final option for nutritional support of patients with severe intestinal failure. Lipid emulsions constitute the main source of fuel calories and fatty acids (FAs) in parenteral nutrition formulations. However, adverse effects on patient outcomes have been attributed to the use of lipids, mostly in relation to impaired immune defenses and altered inflammatory responses. Over the years, this issue has remained in the limelight, also because technical advances have provided no safeguard against the most daunting problems, ie, infectious complications. Nevertheless, numerous investigations have failed to produce a clear picture of the immunologic characteristics of the most commonly used soybean oil-derived lipid emulsions, although their high content of n-6 polyunsaturated FAs (PUFAs) has been considered a drawback because of their proinflammatory potential. This concern initiated the development of emulsions in which part of the n-6 FA component is replaced by less bioactive FAs, such as coconut oil (rich in medium-chain saturated FAs) or olive oil (rich in the n-9 monounsaturated FA oleic acid). Another approach has been to use fish oil (rich in n-3 PUFA), the FAs of which have biological activities different from those of n-6 PUFAs. Recent studies on the modulation of host defenses and inflammation by fish-oil emulsions have yielded consistent data, which indicate that these emulsions may provide a tool to beneficially alter the course of immune-mediated conditions. Although most of these lipids have not yet become available on the US market, this review synthesizes available information on immunologic characteristics of the different lipids that currently can be applied via parenteral nutrition support.

Parenteral N-3 fatty acids modulate inflammatory and immune response in rats undergoing total gastrectomy.

This study investigated the effect of n-3 fatty acid (FA)-containing parenteral nutrition on the circulatory lymphocyte subpopulation, intracellular cytokine and leukocyte adhesion molecule expression, and phagocytic activity in rats undergoing total gastrectomy. Normal rats with internal jugular catheters were assigned to normal control (NC) and two experimental groups and received total parenteral nutrition (TPN). At the same time, a total gastrectomy was performed in the experimental groups, whereas the NC group underwent a sham operation. The TPN solutions were isonitrogenous and identical in nutrient compositions except for differences in fat emulsion contents. The NC and one of the experimental groups received a soybean oil emulsion (SO), and the other experimental group received 50% soybean oil and 50% fish oil emulsion (FO). Half of the rats in each respective group were sacrificed 1 or 3 days after surgery or the sham operation to examine their immune response. The results showed that the FO group had a higher CD4 proportion and CD4/CD8 ratio than those of the SO and NC groups postoperatively. The phagocytic activity of peritoneal macrophages was higher in the FO group than in the NC group, but no difference was found between the SO and NC groups 3 days after surgery. The intracellular interferon (IFN)-gamma distribution in the FO group was higher than that of the SO group on postoperative days. Leukocyte adhesion molecule expressions and peritoneal monocyte chemotactic protein-1 levels were lower in the FO group than in the SO group on postoperative day 3. These results suggest that parenterally infused FO did not result in immunosuppression. In addition, FO administration promotes lymphocyte Th1 cytokine production, enhances peritoneal macrophage phagocytic activity, and reduces leukocyte adhesion molecule expression in rats with total gastrectomy.

Anti-soya antibodies and trilucent breast implants.

In light of the media attention surrounding the withdrawal of Trilucent breast implants in March of 1999, we undertook a study of 20 women with these implants to compare the level of serum antibodies to soya oil and protein with that of a control group. Two control groups were chosen: 20 women without breast implants and 20 women with silicone-containing breast implants. Serum antibodies to soya protein were detected with an enzyme-linked immunoabsorbent assay technique, because direct assay to soya oil is not possible owing to insolubility of the soya oil component. No antibody formation was detected in the patients with Trilucent breast implants or in either of the control groups, lending support to the view that these implants are unlikely to give rise to a systemic immune response.

Enhancing effect of dietary oil emulsions on immune responses to protein antigens fed to mice.

Repeated intragastric administration of beta-lactoglobulin (beta-Lg) with emulsified soybean oil elicited an antigen-specific, systemic humoral immune response in different strains of mice. The antibody response was enhanced as the dose of oil was increased and the particle size of emulsions was decreased. Feeding of aqueous beta-Lg could induce the antibody response only when emulsified oil was fed almost simultaneously. However, the emulsion-driven humoral immune response was not observed when mice were treated with anti-CD40 ligand antibody or in athymic mice. It is likely that the intestinal coexistence of emulsified oil with dietary antigens modulates the immune system to crucially support B cell response. A practical application of the present results to the prevention of cow's milk protein sensitization in infants is proposed.

Antigenicity of the proteins in soy lecithin and soy oil in soybean allergy.

BACKGROUND: Soy lecithin and soy oil are usually produced from the hexane extract of soybean. Some of the soybean proteins are included in the extract and are therefore present in small amounts in both soy lecithin and soy oil. The antigenicity of the proteins present in defatted soybean has been studied with respect to soybean allergy, but the antigenicity of those found in the extract is yet to be investigated. OBJECTIVE: The antigenicity of soy lecithin and soy oil proteins with regard to soybean allergy were investigated. METHODS: The proteins present in soy lecithin and soy oil were determined according to already established method and analysed by SDS-PAGE. The IgE- and IgG4-binding abilities of the soy lecithin proteins were investigated by immunoblotting with sera from 30 soybean-sensitive patients, including seven with a positive challenge test. Immunoblotting of soy oil proteins was performed with the sera from some of these patients. RESULTS: In 100 g of sample, the soy lecithin and soy oil contained 2.8 mg and 1.4-4.0 microg of proteins, respectively. The results of SDS-PAGE demonstrated the presence of only three proteins, with molecular weights of about 58-67 kDa in soy oil, and suggested that soy lecithin also contains these proteins. The soy lecithin also contained many proteins besides these. In the soy lecithin, the detection rate of only one protein, with a molecular weight of 31 kDa, by the serum IgE of patients was significantly different compared with controls (detection rate: 40%). The proteins with molecular weights of 58-67 kDa rarely bound to serum IgE. Only one of the patients who presented a positive challenge test had IgE antibodies to soy lecithin proteins. IgG4-binding proteins were found only rarely in soy lecithin. Neither the IgE nor the IgG4 present in the patients' sera reacted to any soy oil protein. CONCLUSION: Proteins present in soy lecithin and soy oil have little antigenicity with regard to soybean allergy.

Kinetics of cell-mediated cytotoxicity in mice fed diets of various fat contents.

We previously reported lower mitogen-induced blastogenic and cytotoxic activity of splenocytes from C3H/OUJ female mice fed 20% soybean oil (SBO) in their diet compared to those fed 5% SBO. The present study examined the kinetics of cell-mediated cytotoxicity using the same animal model and dietary treatments. Kinetic parameters were determined by analyzing the lytic efficiency of splenocytes cultured for various times with several concentrations of radiolabeled P815 mastocytoma cells. The apparent avidity constant (K1/2) of the reaction was not changed by dietary SBO intake (1.0 +/- 0.2 x 10(5) cells for 20% SBO versus 1.3 +/- 0.3 x 10(5) cells for 5% SBO). However, the maximum velocity (Vmax) of the reaction for splenocytes from mice fed 20% SBO was significantly lower than that for splenocytes from mice fed 5% SBO (1.4 +/- 0.2 x 10(4) cells/h for 20% SBO versus 2.3 +/- 0.4 x 10(4) cells/h for 5% SBO, p less than 0.05). The evidence indicates that the rate of target cell lysis, but not the avidity of the lymphocytes for the target cell antigen, was altered by increasing dietary SBO concentration.