Immune Response of Healthy Adults to the Ingested Probiotic Lactobacillus casei Shirota.

Daily ingestion of a probiotic drink containing Lactobacillus casei Shirota (LcS; 1.3 × 1010 live cells) by healthy adults for (1) 4-week LcS, (2) 6-week discontinuation of LcS and (3) a final 4 weeks of LcS was investigated. There was a significant increase in expression of the T cell activation marker CD3+ CD69+ in ex vivo unstimulated blood cells at weeks 10 and 14, and there was a significant increase in the NK cell marker CD3+ CD16/56+ in ex vivo unstimulated blood cells at weeks 4, 10 and 14. Expression of the NK cell activation marker CD16/56+ CD69+ in ex vivo unstimulated blood cells was 62% higher at week 10 and 74% higher at week 14. Intracellular staining of IL-4 in ex vivo unstimulated and PMA-/ionomycin-stimulated CD3+ ß7+ integrin blood cells was significantly lower at weeks 10 and 14. Intracellular staining of IL-12 in ex vivo unstimulated and LPS-stimulated CD14+ blood cells was significantly lower at weeks 4, 10 and 14. Intracellular staining of TNF-α in LPS-stimulated CD14+ blood cells was significantly lower at weeks 4, 10 and 14. Mucosal salivary IFN-γ, IgA1 and IgA2 concentrations were significantly higher at week 14, but LcS did not affect systemic circulating influenza A-specific IgA or IgG and tetanus-specific IgG antibody levels. In addition to the decrease in CD3+ ß7+ integrin cell IL-4 and a reduced CD14+ cell pro-inflammatory cytokine profile, at week 14 increased expression of activation markers on circulating T cells and NK cells and higher mucosal salivary IgA1 and IgA2 concentration indicated a secondary boosting effect of LcS.

Increased central adiposity may not underlie the marked elevation of IL-6 in diabetes mellitus patients in South-West, Nigeria.

BACKGROUND: Chronic inflammation is linked to disorders of obesity, insulin resistance and DM2. This reflects as increase in proinflammatory cytokines including IL-6. In Nigeria, there is no study that has measured IL-6 in diabetics, in spite of having the highest number of diabetics in Africa. METHODS: The twenty-eight DM2 patients and 13 controls recruited for this study had their BP, BMI, waist circumference (WC) and waist-hip-ratio (WHR) measured. They also had fasting plasma IL-6, fasting plasma glucose, total cholesterol (TC), Triglyceride (Tg), high density lipoprotein cholesterol (HDL-C), urea, creatinine, aspartate transaminases (AST), alanine transferases (ALT), total protein (TP) and albumin determined. RESULTS: Mean age was 51.83 years ± 13.28, with diabetics significantly older than controls (56.61 yrs. ± 9.62 vs. 41.54 years ± 14.53) P < 0.05. The mean IL-6 in diabetics (194.77 pg/ml ± 166.16) was significantly higher than controls' (26.29 pg/ml ± 6.65) at p ≤ 0.01. No significant difference in mean BMI in diabetics and controls. But WC and WHR of diabetics (100.75 cm ± 18.47; 1.01 ± 0.14) were significant higher than in controls (88.77 cm ± 13.36; 0.88 ± 0.07) at p ≤ 0.05 (WC; p value 0.043) and p ≤ 0.01(WHR; p value 0.002). Among diabetics, there were significant correlations between IL-6 and Tg (p<0.01, r = 0.007**),IL-6 and LDL-C (p<0.05, r = 0.02*), IL-6 and AST (p<0.05, r = 0.041*r4) and IL-6 and ALT (p<0.01, r = 0.004**) CONCLUSION: Elevated IL-6 in DM2 patients in South West Nigeria correlates with liver transaminases and not increased markers of central adiposity.

Breast milk levels of zinc and omega-6 polyunsaturated fatty acids and growth of healthy Chinese infants.

AIM: To examine the concentrations of zinc and omega-6 polyunsaturated fatty acids (omega-6 PUFAs) in breast milk, the impact of zinc on omega-6 PUFA metabolism, and the growth rate of infants. METHODS: Forty-one mother-term infant pairs from a rural area of northern Beijing, China, who were 1 month (n = 18, group I) and 3 months (n = 23, group II) old and exclusively breastfed, were studied. The dietary records and the concentrations of zinc and omega-6 PUFAs in the milk of lactating women and the increase in weight and length of their infants during 1 and 3 postnatal months were analysed. RESULTS: The dietary intakes of mothers in the two groups were the same, i.e. high in carbohydrate and low in fat, protein and energy. The maternal zinc intake was 7.5mg/d and thus reached only 34.6% of the current Recommended Nutrient Intake (RNI). The levels of zinc and arachidonic acid (AA, C20:4 omega-6) in the milk of group I were significantly higher than those in group II. Furthermore, significant positive correlations were found between the concentrations of zinc and AA in the breast milk and between the level of milk AA and weight gain. CONCLUSION: Zinc may be a co-factor and essential for essential fatty acids (EFA) metabolism. Thus suboptimal zinc intake may cause EFA imbalance. Further studies of Chinese rural mother-infant pairs are necessary to determine whether zinc supplementation should be recommended when lactation exceeds 3 months.

Diet and gene expression: delta-5 and delta-6 desaturases in healthy Chinese and European subjects.

AIM: To compare the composition of fatty acids (FAs) in diet, and the expression of delta-6 desaturase (D6D) and delta-5 desaturase (D5D) genes in peripheral blood mononuclear cells (PBMCs) between Chinese and Europeans. METHODS: Three-day dietary records from 20 subjects from Beijing, China (n = 10) and Kent, UK (n = 10) were analysed. Expression of PBMC D6D and D5D genes of the subjects was determined using RT-PCR. RESULTS: The dietary intake of Chinese subjects contained less saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs), but more essential fatty acids (EFAs) than that of Europeans. Levels of expression of PBMC D6D and D5D genes of Chinese subjects were significantly lower than those of Europeans. A significant positive correlation was found between dietary intake of total SFAs and total MUFAs and expression of PBMC D6D and D5D genes, but a significant negative correlation between dietary intake of linoleic acid (LA) and alpha-linolenic acid (LNA) and the expression of PBMC D6D and D5D genes. CONCLUSION: Intake of high SFAs and MUFAs appears to increase expression of PBMC D6D and D5D genes, whilst high EFAs intake appears to decrease expression of PBMC D6D and D5D genes. A follow-up study of the expression of D6D and D5D genes in Chinese who live in European countries with high SFA and MUFA diets would be of interest.

Dietary fatty acid modulation of mucosally-induced tolerogenic immune responses.

Immunological unresponsiveness or hyporesponsiveness (tolerance) can be induced by feeding protein antigens to naive animals. Using a classical oral ovalbumin gut-induced tolerance protocol in BALB/c mice we investigated the effects of dietary n-6 and n-3 polyunsaturated fatty acids (PUFA) on high-and low-dose oral tolerance (and in non-tolerised animals, i.e. effects of antigen challenge alone) in relation to lymphoproliferative, cytokine and antibody responses. Fish oil rich in long-chain n-3 fatty acids decreased both T-helper (Th) 1- and Th2-like responses. In contrast, borage (Borago officinalis) oil rich in n-6 PUFA, of which gamma-linolenic acid is rapidly metabolised to longer-chain n-6 PUFA, increased Thl-like responses and decreased Th2-like responses, and possibly enhanced suppressor cell or Th3-like activity. These findings are in general agreement with other studies on the effects of long chain n-3 PUFA on immune system functions, and characterise important differences between long-chain n-3 and n-6 PUFA, defining more precisely and broadly the immunological regulatory mechanisms involved. They are also discussed in relation to autoimmune disease.

The protective effects of omega-6 fatty acids in experimental autoimmune encephalomyelitis (EAE) in relation to transforming growth factor-beta 1 (TGF-beta1) up-regulation and increased prostaglandin E2 (PGE2) production.

Polyunsaturated fatty acids are known to affect the immune response and administration of the omega-6 fatty acid linoleic acid has been reported to be beneficial in multiple sclerosis (MS) and EAE. In this study we have investigated the effects of oral feeding of plant lipid rich in the omega-6 fatty acid gamma-linolenic acid from Borago officinalis on acute and relapse disease and the immune response in EAE using SJL mice. EAE was induced by an encephalitogenic peptide (92-106) of myelin oligodendrocyte glycoprotein (MOG), and mice were fed the plant lipid daily from 7 days after EAE induction to assess the effects on acute disease and from day 25 to assess the effects on disease relapse. The clinical incidence and histological manifestations of acute EAE, and the clinical relapse phase of chronic relapsing EAE (CREAE) were markedly inhibited by omega-6 fatty acid feeding. A significant increase in the production of TGF-beta1 in response to concanavalin A (Con A) at day 13 and a significant increase in TGF-beta1 and PGE2 to Con A, PPD and MOG peptide (92-106) at day 21 were detected in spleen mononuclear cells from fatty acid-fed mice. There was no difference in interferon-gamma, IL-4 and IL-2 production between the fatty acid-fed and control groups. Significantly higher TGF-beta mRNA expression was found in the spleens of omega-6 fatty acid-fed mice at day 21. There were no differences in spleen cell proliferative response to Con A, PPD and MOG peptide (92-106). Biochemical analysis of spleen cell membrane fatty acids revealed significant increases in the eicosanoid precursor fatty acids dihomo-gamma-linolenic acid and arachidonic acid in response to gamma-linolenic acid feeding, indicating rapid metabolism to longer chain omega-6 fatty acids. These results show that oral feeding of gamma-linolenic acid-rich plant lipid markedly affects the disease course of acute EAE and CREAE and is associated with an increase in cell membrane long chain omega-6 fatty acids, production of PGE2 and gene transcription and, on activation, secretion of TGF-beta1.

Plasma vitamin A and E in preterm babies fed on breast milk or formula milk with or without long-chain polyunsaturated fatty acids.

Plasma vitamin A and E, the antioxidant nutrients copper and zinc, and magnesium were investigated in preterm babies. They were fed on their own mother's breast milk, or a formula with, or without, AA and DHA. Vitamin A (2.4 mg/d) and E (15 mg/d) supplements were also given. Vitamin A and E levels of most of the babies were sub-optimal at birth. The mean concentrations of vitamin E increased in all the groups by the expected date of delivery (EDD) (p < 0.001). Those fed on their mother's breast milk had the highest value compared with the other groups (p < 0.001). There was an increase in the mean level of vitamin A (p > 0.05) and copper (p < 0.05) and a decrease in zinc (p < 0.05) between birth and EDD. Concentrations of the two vitamins were not different (p > 0.05) between the babies fed on the formula with, and without, AA and DHA. It is concluded that the amount of AA and DHA incorporated in the formula milk did not adversely influence the plasma vitamin A and E of the babies.

Dietary n-6 and n-3 fatty acids in immunity and autoimmune disease.

Clearly there is much evidence to show that under well-controlled laboratory and dietary conditions fatty acid intake can have profound effects on animal models of autoimmune disease. Studies in human autoimmune disease have been less dramatic; however, human trials have been subject to uncontrolled dietary and genetic backgrounds, infection and other environmental influences, and basic trial designs have been inadequate. The impact of dietary fatty acids on animal autoimmune disease models appears to depend on the animal model and the type and amount of fatty acids fed. Diets low in fat, essential fatty acid-deficient, or high in n-3 fatty acids from fish oils increase the survival and reduce disease severity in spontaneous autoantibody-mediated disease, whilst linoleic acid-rich diets appear to increase disease severity. In experimentally-induced T-cell-mediated autoimmune disease, essential fatty acid-deficient diets or diets supplemented with n-3 fatty acids appear to augment disease, whereas n-6 fatty acids prevent or reduce the severity. In contrast, in both T-cell and antibody-mediated auto-immune disease the desaturated and elongated metabolites of linoleic acid are protective. Suppression of autoantibody and T lymphocyte proliferation, apoptosis of autoreactive lymphocytes, and reduced pro-inflammatory cytokine production by high-dose fish oils are all likely mechanisms by which n-3 fatty acids ameliorate autoimmune disease. However, these could be undesirable long-term effects of high-dose fish oil which may compromise host immunity. The protective mechanism(s) of n-6 fatty acids in T-cell- mediated autoimmune disease are less clear, but may include dihomo-gamma-linolenic acid- and arachidonic acid-sensitive immunoregulatory circuits such as Th1 responses, TGF beta 1-mediated effects and Th3-like responses. It is often claimed that n-6 fatty acids promote autoimmune and inflammatory disease based on results obtained with linoleic acid only. It should be appreciated that linoleic acid does not reflect the functions of dihomo-gamma-linolenic and arachidonic acid, and that the endogenous rate of conversion of linoleic to arachidonic acid is slow (Hassam et al. 1975, 1977; Phylactos et al. 1994; Harbige et al. 1995). In addition to effects of dietary fatty acids on immunoregulation, inflammation as a consequence of immune activation in autoimmune disease may also be an important mechanism of action whereby dietary fatty acids modulate disease activity. In conclusion, regulation of gene expression, signal transduction pathways, production of eicosanoids and cytokines, and the action of antioxidant enzymes are all mechanisms by which dietary n-6 and n-3 fatty acids may exert effects on the immune system and autoimmune disease. Probably the most significant of these mechanisms in relation to our current understanding of immunoregulation and inflammation would appear to be via fatty acid effects on cytokines. The amount, type and balance of dietary fatty acids and associated antioxidant nutrients appear to impact on the immune system to produce immune-deviation or immunosuppressive effects, and to reduce immune-mediated inflammation which will in turn affect the susceptibility to, or severity of, autoimmune disease.

Characterization of the cellular and cytokine response in the central nervous system following Semliki Forest virus infection.

Cytokines are important mediators in the pathogenesis of central nervous system (CNS) inflammatory diseases including multiple sclerosis (MS), experimental allergic encephalomyelitis (EAE), viral encephalitis and virus induced demyelinating diseases. We have used immunohistochemical techniques to characterize the mononuclear cell infiltrate and cytokine profiles in the CNS following infection of mice with the demyelinating A7(74) strain of Semliki Forest virus (SFV), an important viral model of MS. Mononuclear cell infiltrates in the CNS, first observed at 3 days and maximal during clearance of infectious virus, were comprised predominantly of CD8+ lymphocytes. F4/80+ macrophage/microglia and CD45/B220+ B lymphocytes were most numerous during the subsequent phase of demyelination. CD4+ T-lymphocytes were observed at low levels throughout infection. By immunostaining MHC class I, IL-1beta , IL-3 and TGF beta1 were constitutively expressed in normal mice and were upregulated following infection. MHC class II, IL-1alpha, IL-2, IL-2R, TNF-alpha and IL-6 were strongly upregulated in the CNS of SFV-infected mice and mice with chronic relapsing EAE. The spatial and temporal distribution of these cytokines during the course of disease was analysed. Whereas IL-1alpha, IL-1beta, IL-10, and TGF beta1 were observed on day 3 following infection GMCSF, IL-2 and TNF alpha were first apparent at day 7 when the cellular infiltration in the CNS was most intense. In contrast IFN gamma and IL-6 were first observed on day 10 prior to the demyelination phase of disease. Cytokines in the lesions of demyelination suggest a role in the pathogeneisis of myelin damage. Based on cytokine profiles no clear bias of either a Th1 or Th2 response was observed in the CNS during infection.

Nutrition and immunity with emphasis on infection and autoimmune disease.

Nutrition and nutritional status can have profound effects on immune functions, resistance to infection and autoimmunity in man and other animals. Nutrients enhance or depress immune function depending on the nutrient and level of its intake. Protein-energy malnutrition and vitamin A deficiency are strongly associated with impaired immunity and infectious disease. The essential role vitamin A plays in infection and maintenance of mucosal surfaces has long been known. Recent evidence shows that T-cell subpopulations, cytokines and antibody subclasses are all affected by vitamin A. In animal studies supplementation with vitamin E protects against infection and is linked to stimulatory effects on the immune system. In man vitamin E and other anti-oxidants increase the number of CD4+ cells. Dietary lipids and zinc have a substantial impact on autoimmunity from protective to potentiation of immuno-pathological processes in animals. There is considerable potential to modify human autoimmune disease by manipulation of lipid nutrition. Deficiency of pyridoxine induces atrophy of lymphoid organs, marked reduction in lymphocyte numbers, impairs antibody responses and IL-2 production. Dietary copper is important in the prevention of infection in some animal species and T-cell function is defective under deficiency states due to an inability to produce IL-2. Selenium has been linked to viral infection, enhanced T-cell functions and TNF beta induced increase in natural killer cell activity. Understanding the molecular and cellular immunological mechanisms involved in nutrient-immune interactions will increase our applications for nutrition of the immune system in health and in disease

Prevention of experimental autoimmune encephalomyelitis in Lewis rats by a novel fungal source of gamma-linolenic acid.

The effects of oral administration of linoleic- and gamma-linolenic-acid-rich oils on the clinical and histopathological manifestations of experimental autoimmune encephalomyelitis (EAE) were investigated in Lewis rats 7 d post-inoculation. gamma-Linolenic-acid-rich fungal (Mucor javanicus) oil at 500 mg/kg body weight abrogated clinical and histological signs of EAE although at doses of 200 and 1000 mg/kg body weight it was only effective in delaying the onset of clinical disease. Linoleic-acid-rich safflower-seed (Carthamus tinctorius) oil at 500, 750 and 1000 mg/kg body weight decreased the severity of clinical EAE disease in a dose-dependent manner. The effects in healthy animals of orally administered gamma-linolenic-acid-rich fungal oil (500 mg/kg body weight) and linoleic-acid-rich safflower-seed oil (1000 mg/kg body weight) on splenic lymphocyte proliferative responses to the T-cell mitogen concanavalin-A (Con A), membrane fatty acid composition and lymphocyte sub-sets were also studied. Both treatments enhanced the T-cell proliferative response to Con A. There was no significant effect on the proportion of splenic CD8+ or CD4+ lymphocytes. Compositional studies on splenic phosphoglyceride fatty acids of oil-treated animals suggest the above responses were associated with increases in spleen dihomo-gamma-linolenic and arachidonic acids.

Vitamin A and related essential nutrients in cord blood: relationships with anthropometric measurements at birth.

Following the advice given by the Department of Health to women who are, or may become pregnant, not to eat liver and liver products because of the risk of vitamin A toxicity, the concentrations of vitamins A and E, and copper, magnesium and zinc in cord blood were investigated. The study was conducted in Hackney, an inner city area of London. Esters of vitamin A were not detected in any of the samples, indicating that there was no biochemical evidence of a risk of toxicity. Indeed, vitamin A correlated significantly with birthweight, head circumference, length, and gestation period. There was also a significant positive relationship between zinc and birthweight. In contrast, copper showed a negative correlation with birthweight and head circumference. Vitamin E and magnesium were not associated with any of the anthropometric measurements, although magnesium showed an increasing trend with birthweight. The data suggest that most of the mothers of the subjects studied may have been marginal with respect to vitamins A and E and zinc. In those with low birthweight babies. a higher intake would have improved their nutritional status and possibly the outcome of their pregnancy. For these low-income mothers, liver and liver products are the cheapest and the best source of vitamins A and E, haem iron, B vitamins and several other essential nutrients; hence the advice of the Department of Health may have been misplaced.

Essential fatty acids alter the activity of manganese-superoxide dismutase in rat heart.

The effects of oil-derived dietary essential fatty acids on the activities of mitchondrial Mn-SOD (manganese-superoxide dismutase) and cytosolic cupric zinc-superoxide dismutase (Cu/Zn-SOD) were investigated in rat heart. A control group of rats was fed a stock diet for 29 d, and a second group was fed on a fat-free diet. Three other groups were fed fat-free diets that were supplemented with (i) borage oil, which is rich in linoleic (18:2n-6) and gamma-linolenic (18:3n-6) acids, (ii) fungal oil, which is rich in gamma-linolenic, but low in linoleic acid, or (iii) evening primrose oil, which is rich in linoleic acid and low in gamma-linolenic acid. An increase in the percentage composition of arachidonic acid (20:4n-6) in both the choline and ethanolamine phospholipids, together with a decrease in linoleic acid in ethanolamine phospholipids, were found in heart membranes after feeding the rats with diets containing borage oil or fungal oil as compared to those fed the stock diet. The respective activities of Mn-SOD in rats fed the borage or fungal oil diets were also significantly higher than in rats fed the stock diet alone. No change in cytosolic Cn/Zn-SOD activity was observed. Dietary supply of linoleic acid-rich evening primrose oil resulted in an increased proportion of choline phospholipid linoleic acid without any changes in arachidonic acid content or in the activity of Mn-SOD. By contrast, a reduction in the activity of Mn-SOD was detected in rats fed a fat-free diet.(ABSTRACT TRUNCATED AT 250 WORDS)

The importance of energy and nutrient supply in human brain evolution.

Current evolutionary theories do not adequately address the question of how the human brain evolved to be larger and more sophisticated than that of other primates. The human brain/body weight ratio is 4-5 times higher than in primates and, relative to the rest of the body, requires up to 10 times as much energy as in other land-based mammals. Human brain evolution must therefore have required a stable food supply providing a reliable source of both high dietary energy and a cluster of 'brain-specific' nutrients over a long period of time. These nutrient and energy requirements are available in the marine and shore-based food chain but are difficult if not impossible to obtain in the terrestrial food chain. We suggest that marine and estuarine ecosystems provided hominids with the appropriate stimulus to develop a relatively large brain. This occurred in conjunction with the evolution of other uniquely human features, particularly relative hairlessness, bipedalism and abundant neonatal subcutaneous fat. Invertebrates, molluscs, small or slow-moving fish, and marine algae would have provided a stable, abundant supply of energy, long chain polyunsaturates and other nutrients essential for the brain and would have done so with comparatively little mammalian competition. The land-water interface would thus have allowed the hominid brain to develop sufficient neurological complexity to enable sophisticated tool and behaviour patterns to evolve in humans as a natural sequel to such a biochemical and environmental stimulus.

A review of the aetiology of multiple sclerosis: an ecological approach.

Multiple sclerosis (MS) is a disease of unknown aetiology characterized by myelin destruction. Approaches to the aetiology of the condition have noted its differential geographic and demographic distribution; a tentative and inconsistent link with infectious agents; a genetic susceptibility; and a tendency to track with dietary lipids. This review proposes a multidisciplinary approach to the aetiology of MS, emphasizing the interlocking and interdependent nature of current aetiological arguments. We suggest that the most profitable interpretation of current empirical data on the aetiology of MS can be accomplished through this kind of analysis.

The effect of dietary change on in vitro erythrocyte haemolysis, skin lesions and alopecia in common marmosets (Callithrix jacchus).

1. The effect of dietary change on common marmosets (Callithrix jacchus) exhibiting high in vitro erythrocyte haemolysis, skin lesions and alopecia was investigated. 2. The experimental diet was low in lipid, high in vitamin E and was devoid of n-3 long chain polyunsaturated fatty acids. 3. There were significant increases in the concentrations of plasma alpha-tocopherol (vitamin E) (P less than 0.05), all-trans retinol (vitamin A) (P less than 0.001) and total lipid (P less than 0.025), and a reduction in fibrinogen (P less than 0.02) concentration and neutrophil count (P less than 0.02). 4. Moreover, the mean in vitro erythrocyte haemolysis declined from an initial value of 64% to less than 2%, and there was a regression of skin lesions and alopecia with complete fur regrowth. 5. The amelioration of the conditions appear to be due to high circulating vitamin E which provided protection against oxidative stress, and the correction of membrane n-6:n-3 balance.