Nutrient supplementation strategy improves cell concentration and longevity, monoclonal antibody production and lactate metabolism of Chinese hamster ovary cells.

A careful selection of culture mediums and feeds has become necessary to maximize yields of recombinant proteins during bioprocesses of mammalian cells. Supplements contain a variety of concentrate nutrients, and their beneficial effects vary according to recombinant cell lines. In this study, the effects of PowerFeed A on growth kinetics, productivity and cellular metabolism were evaluated for two Chinese hamster ovary cell lines producing a monoclonal antibody in a batch culture. Supplemented cultures increased integral viable cell density of CRL-12444 and CRL-12445 cells by 2.4 and 1.6 times through extension of culture time at which viability was above 90% in 72 and 36 h, respectively, and increment of maximal cell concentration in 3.25 × 106 cells/ml (69%) for CRL-12445 cells. Product titer augmented 1.9 and 2.5 times for CRL-12444 and CRL-12445 cells, respectively, without changes in growth rate and specific productivity. Feed supplementation also stimulated full consumption of glucose and free glutamine and reduced 10 times lactate accumulation, while ammonium, sodium and potassium remained at similar concentrations at the end of the culture. About 44% of calcium, mainly provided by feed, was consumed by both cell lines. Maximization of cellular growth, viability and protein titer through feeding encourages extending its use to other cell lines and exploring novel combinations with other basal mediums or feeds. A thorough investigation of its impact on protein quality and the molecular mechanisms behind these effects will allow designing effective feeds and strategies to rationally optimize protein production in the biomanufacturing industry.

[Vitamin D and infectious diseases]. / Vitamine D et infections.

Vitamin D plays a role in the synthesis of antibacterial peptids and in autophagy. Several studies have shown that low levels of vitamin D are associated with the susceptibility and the severity of acute infections on one hand, and with an unfavorable outcome of some chronic infections (such as HIV infection). Vitamin D supplementation improves response to treatment of some viral (such as chronic hepatitis C infection) or bacterial infections (such as pulmonar tuberculosis). Vitamin D supplementation demonstrated no benefit in reducing the incidence of pulmonary infections. The target level of vitamin D to be reached after supplementation is not known yet.

Presurgical stress management improves postoperative immune function in men with prostate cancer undergoing radical prostatectomy.

OBJECTIVE: To assess whether stress management (SM) improved immune outcomes in men undergoing surgery for prostate cancer. METHODS: A total of 159 men were assigned randomly to a two-session presurgical SM intervention, a two-session supportive attention (SA) group, or a standard care (SC) group. Men in the SM group discussed their concerns about the upcoming surgery and were taught diaphragmatic breathing, guided imagery; they had an imaginal exposure to the day of surgery and learned adaptive coping skills. Men in the SA group discussed their concerns about the upcoming surgery and had a semistructured medical interview. Blood samples were collected at baseline (1 month before surgery) and 48 hours after surgery. Measures of mood (Profile of Mood States) were collected at baseline, 1 week pre surgery, and the morning of surgery. RESULTS: Men in the SM group had significantly higher levels of natural killer cell cytotoxicity (p = .04) and higher levels of circulating proinflammatory cytokines (interleukin [IL]-12p70, p = .02; IL-1ß, p = .02; tumor necrosis factor-α, p = .05) 48 hours post surgery than men in the SA group and higher levels of natural killer cell cytotoxicity (p = 0.02) and IL-1ß (p = .05) than men in the SC group. Immune parameters increased for the SM group and decreased or stayed the same for the SA and SC groups. The SM group had significantly lower Profile of Mood States scores than the SC group (p = .006), with no other group differences between SA and SC groups. Changes in mood were not associated with immune outcomes. CONCLUSIONS: The finding that SM leads to decreased presurgical mood-disturbance and increased immune parameters after surgery reveals the potential psychological and biological benefits of presurgical SM.

Effects of feeding polyphenols from pomegranate extract on health, growth, nutrient digestion, and immunocompetence of calves.

Objectives were to determine effects of feeding pomegranate extract (POMx) rich in polyphenols on performance, health, nutrient digestion, and immunocompetence of calves in the first 70 d of age. Holstein calves (n=67), at 2+/-1 d of age (d 0=birth day) were randomly assigned to 0 (control), 5 (POMx5), or 10 g/d (POMx10) of pomegranate extract containing 16.9% gallic acid equivalent (GAE) to result in intakes of 0, 850 and 1,700 mg of GAE/d or an average of approximately 0, 15, and 30 mg of GAE/kg of body weight (BW) per day. All calves received colostrum during the first 24 h, pasteurized milk thereafter until 61 d of age, and grain was fed ad libitum for the first 70 d of age. Calves were housed in individual hutches, and grain intake, attitude and fecal scores, incidence and duration of health disorders, and treatments for health problems were evaluated daily. Body weight was measured on 2 consecutive days at 2, 30, and 70 d of age and averaged for each measurement. Concentrations of glucose and 3-hydroxybutyrate were measured in plasma. Nutrient digestion was measured using total fecal collection during a 3-d period. Neutrophil phagocytic and killing activities and antibody response to immunization with ovalbumin were measured. Peripheral blood mononuclear cells were cultured and cytokine production measured. Feeding POMx had no effect on intake or BW gain in the first 30 d of age, but after 30 d of age, both grain dry matter intake and BW gain decreased with increasing addition of POMx, which resulted in calves that were 1.8 and 4.3 kg lighter at 70 d of age for POMx5 and POMx10, respectively, compared with controls. Feeding POMx did not influence dry matter, organic matter, or starch digestibility, but it reduced crude protein and fat digestion. Plasma concentrations of glucose and 3-hydroxybutyrate were similar among treatments throughout the first 70 d of age. Measures of calf health such as fecal and attitude scores, risk of fever, and rectal temperature were not altered by treatments. Similarly, neutrophil phagocytic and killing activities did not differ among treatments. On the contrary, feeding POMx increased synthesis of interferon-gamma and interleukin-4 by peripheral blood mononuclear cells and improved total immunoglobulin G responses to ovalbumin vaccination. These results suggest that feeding POMx top-dressed onto the grain suppresses intake of grain and digestibility of fat and protein, likely because of the high tannin content. Nevertheless, polyphenols from POMx enhanced mitogen-induced cytokine production and response to vaccination, which might benefit immune competence of calves and potentially health. Additional studies are warranted to minimize the effect of POMx on intake and digestibility and to better understand the mechanisms by which polyphenols improve immune response of calves.

Selected vitamins and trace elements support immune function by strengthening epithelial barriers and cellular and humoral immune responses.

Adequate intakes of micronutrients are required for the immune system to function efficiently. Micronutrient deficiency suppresses immunity by affecting innate, T cell mediated and adaptive antibody responses, leading to dysregulation of the balanced host response. This situation increases susceptibility to infections, with increased morbidity and mortality. In turn, infections aggravate micronutrient deficiencies by reducing nutrient intake, increasing losses, and interfering with utilization by altering metabolic pathways. Insufficient intake of micronutrients occurs in people with eating disorders, in smokers (active and passive), in individuals with chronic alcohol abuse, in certain diseases, during pregnancy and lactation, and in the elderly. This paper summarises the roles of selected vitamins and trace elements in immune function. Micronutrients contribute to the body's natural defences on three levels by supporting physical barriers (skin/mucosa), cellular immunity and antibody production. Vitamins A, C, E and the trace element zinc assist in enhancing the skin barrier function. The vitamins A, B6, B12, C, D, E and folic acid and the trace elements iron, zinc, copper and selenium work in synergy to support the protective activities of the immune cells. Finally, all these micronutrients, with the exception of vitamin C and iron, are essential for antibody production. Overall, inadequate intake and status of these vitamins and trace elements may lead to suppressed immunity, which predisposes to infections and aggravates malnutrition. Therefore, supplementation with these selected micronutrients can support the body's natural defence system by enhancing all three levels of immunity.

Effects of a Taiji and Qigong intervention on the antibody response to influenza vaccine in older adults.

Previous studies have suggested that Taiji practice may improve immune function. This study was intended to examine whether 5 months of moderate Taiji and Qigong (TQ) practice could improve the immune response to influenza vaccine in older adults. Fifty older adults (mean age 77.2 +/- 1.3 years) participated in this study (TQ N = 27; wait-list control [CON] N = 23). Baseline pre-vaccine blood samples were collected. All subjects then received the 2003-2004 influenza vaccine during the first week of the intervention. Post-vaccine blood samples were collected 3, 6 and 20 weeks post-intervention for analysis of anti-influenza hemagglutination inhibition (HI) titers. We found a significant (p < 0.05) increase in the magnitude and duration of the antibody response to influenza vaccine in TQ participants when compared to CON. The vaccination resulted in a 173, 130, and 109% increase in HI titer at 3, 6, and 20 weeks post-vaccine, respectively, in the TQ group compared to 58, 54, and 10% in CON. There was a significant between group difference at 3 and 20 weeks post-vaccine and at 20 weeks the TQ group had significantly higher titers compared to the pre-vaccine time point, whereas the CON group did not. A higher percentage of TQ subjects also responded to the influenza A strains with a protective (> 40HI) antibody response (37% TQ vs. 20% CON for the H1N1 strain and 56% TQ vs. 45% CON for the H3N2 strain), but the differences between groups were not statistically significant. Traditional TQ practice improves the antibody response to influenza vaccine in older adults, but further study is needed to determine whether the enhanced response is sufficient to provide definitive protection from influenza infection.

Advances in basic and clinical immunology in 2006.

This article reviews the progress in the field of basic and clinical immunology in 2006, focusing on the articles published in the Journal. The role of Toll-like receptors in the immune response was explored in detail in several articles. The knowledge gained in these investigations is being used to develop strategies that enhance the immunogenicity of vaccines to prevent infectious diseases and to have an immunomodulatory effect on allergic diseases. Other components of the innate immunity reported on were the recognition of allergens with lipid-derived motifs by CD1d-restricted T cells and the role of dendritic cells in the development of an allergic response. More than 120 primary immunodeficiencies were defined at a molecular level, and biological agents such as TNF-alpha antagonists and IFN-alpha were shown to have therapeutic use. New anti-HIV drugs that block cell entry were proven to be effective, thus offering alternative therapies to respond to the development of multidrug-resistant HIV strains. The modern understanding of immunologic concepts is helping to elucidate the mechanisms of defense against viruses, bacteria, and parasites; as a result, strategies to improve management and prevention continue to emerge.

Update: effects of antioxidant and non-antioxidant vitamin supplementation on immune function.

The purpose of this manuscript is to review the impact of supplementation with vitamins E and C, carotenoids, and the B vitamins on parameters of innate and adaptive immune function as reported from clinical trials in humans. There is evidence to support causal effects of supplementation with vitamins E and C and the carotenoids singly and in combination on selected aspects of immunity, including the functional capacity of innate immune cells, lymphocyte proliferation, and the delayed-type hypersensitivity (DTH) response. Controlled intervention trials of B vitamin-containing multivitamin supplements suggest beneficial effects on immune parameters and clinical outcomes in HIV-positive individuals.

Active immunization against ghrelin decreases weight gain and alters plasma concentrations of growth hormone in growing pigs.

Ghrelin has been implicated in the control of food intake and in the long-term regulation of body weight. We theorize that preventing the ability of ghrelin to interact with its receptors, would eventually lead to decreased appetite and thereby decrease body weight gain. To test our hypothesis, pigs were actively immunized against ghrelin. Ghrelin((1-10)) was conjugated to BSA and emulsified in Freund's incomplete adjuvant and diethylaminoethyl-dextran. Primary immunization was given at 19 weeks of age (WOA), with booster immunizations given 20 and 40 days after primary immunization. Body weight (BW) and plasma samples were collected weekly beginning at 19 WOA, and feed intake was measured daily. Fourteen days after primary immunization, the percentage of bound (125)I-ghrelin in plasma from immunized pigs was increased compared with control animals (P<0.001). Voluntary feed intake was decreased more than 15% in animals that were actively immunized against ghrelin compared with controls. By the end of the experiment, immunized pigs weighed 10% less than control animals (P<0.1). Concentrations of GH were increased (P<0.05) in immunized pigs. Apoptosis was not observed in post-mortem samples obtained from the fundic region of the stomach. Our observations suggest that immunization against ghrelin induces mild anorexia. This procedure could potentially be used as a treatment to control caloric intake and obesity.

Antioxidant protection, carotenoids and the costs of immune challenge in greenfinches.

Costs accompanying immune challenges are believed to play an important role in life-history trade-offs and warranting the honesty of signal traits. We performed an experiment in captive greenfinches (Carduelis chloris L.) in order to test whether and how humoral immune challenge with non-pathogenic antigen [sheep red blood cells (SRBC)] affects parameters of individual condition including intensity of coccidian infection, estimates of total antioxidant protection, plasma carotenoids and ability to mount a cell-mediated immune response. We also asked whether the potential costs of immune challenge can be alleviated by dietary carotenoid supplementation. None of the treatments affected intensity of coccidiosis. Humoral immune challenge suppressed the cell-mediated response to phytohemagglutinin (PHA), suggesting a trade-off between the uses of different arms of the immune system. Immune challenge reduced body-mass gain, but only among the carotenoid-depleted birds, indicating that certain somatic costs associated with immune system activation can be alleviated by carotenoids. No evidence for oxidative stress-induced immunopathological damages could be found because immune activation did not affect total antioxidant protection or carotenoid levels. Carotenoid supplementation inclined birds to fattening, indicating that lutein interfered with lipid metabolism. Altogether, our results support the hypotheses of biological importance of carotenoids and exemplify the overwhelming complexity of their integrated ecophysiological functions.

Effects of dietary supplementation with clostridium butyricum on the growth performance and humoral immune response in Miichthys miiuy.

The effects of dietary supplementation with Clostridium butyricum on growth performance and humoral immune response in Miichthys miiuy were evaluated. One hundred and fifty Miichthys miiuy weighing approximately 200-260 g were divided into five groups and reared in 15 tanks with closed circuiting culture system. The animals were fed 5 diets: basal diet only (control) or supplemented of the basal diet with C. butyricum at doses of 10(3) (CB1), 10(5) (CB2), 10(7) (CB3) or 10(9) (CB4) CFU/g. Compared with the control, the serum phenoloxidase activity was significantly increased by the supplementation (P<0.05), acid phosphatases activity was increased significantly (P<0.05) at the doses of 10(9) CFU/g. Serum lysozyme activity peaked at dose of 10(7) CFU/g and in the skin mucus at dose of 10(9) CFU/g. Immunoglobulin M level in the serum and skin mucus was increased except at dose of 10(3) CFU/g (P<0.05). The growth at the dose of 10(9) CFU/g was higher than that of the control (P<0.05). It is concluded that supplementation of C. butyricum can mediate the humoral immune responses and improve the growth performance in Miichthys miiuy.

Effect of vitamin and trace mineral withdrawal from finisher diets on growth performance and immunocompetence of broiler chickens.

The present study was carried out to examine the effect of vitamin (V) or trace mineral (TM) mix removal from finisher diet (42-56 d of age) on performance and immunocompetence of broilers. The dietary treatments were: (1) the basal diet with no supplemental V or TM; (2) the basal diet fortified with V mix only; (3) the basal diet fortified with TM mix only; and (4) the basal diet fortified with V and TM mix. At 47 d, four birds from each replicate (20/treatment) were injected with Newcastle disease (ND) antigen and blood samples were collected just before and 9 d after immunisation. The results showed that V and TM mix withdrawal from finisher diet did not impair either weight gain or feed conversion efficiency from d 42 to 56. Feed intake was significantly increased when both mixes were omitted from the diet. Dietary treatments had no effect on carcase yield or proportion of breast and thigh meat and abdominal fat deposition. Packed cell volume (PCV) significantly decreased as a consequence of removal of V, TM and V+TM. Neither heterophil:lymphocyte ratio (H:L ratio), haemagglutination inhibition (HI) nor total antibody titre measured by ELISA were influenced by removal of V or TM supplements. In conclusion, the results from this study suggest that 14-d withdrawal of V or TM mix did not influence the immunocompetence of broilers.

It takes nerve to tell T and B cells what to do.

The existence of an association between the brain and immunity has been documented. Data show that the nervous and immune systems communicate with one another to maintain immune homeostasis. Activated immune cells secrete cytokines that influence central nervous system activity, which in turn, activates output through the peripheral nervous system to regulate the level of immune cell activity and the subsequent magnitude of an immune response. In this review, we will focus our presentation and discussion on the findings that indicate a regulatory role for the peripheral sympathetic nervous system in modulating the level of cytokine and antibody produced during an immune response. Data will be discussed from studies involving the stimulation of the beta2 adrenergic receptor expressed on CD4+ T cells and B cells by norepinephrine or selective agonists. We will also discuss how dysregulation of this line of communication between the nervous and immune systems might contribute to disease development and progression.

Long-term effects of early-life dietary L-carnitine on lymphoid organs and immune responses in Leghorn-type chickens.

The long-term effects of supplementing Leghorn-type chickens with dietary L-carnitine after hatching for 4 weeks on growth, lymphoid organ weights, humoral and cell-mediated immune responses were assessed in a 12-week study. A commercial starter feed supplemented with 0 (control), 100 (LC) or 1,000 (HC) mg L-carnitine/kg was offered to day-old ISA Brown cockerels for 4 weeks, then all birds were given a commercial pullet grower feed for another 8 weeks. No differences (p > 0.05) in growth rates, feed intake or feed utilisation efficiency existed among the dietary treatments throughout the study. Compared with control birds, HC-fed birds had a lower (p < 0.05) thymus weight relative to bodyweight (BW) at Week 4, but a higher (p < 0.05) relative thymus weight at Week 12. A higher (p < 0.05) serum primary antibody response to sheep red blood cells (SRBC) in HC-fed birds than in the other two groups was also detected at Week 12. Relative spleen or bursa weights and cutaneous responses of toe webs to phytohaemagglutimin (PHA) (an in vivo indicator of cell-mediated immune responses) did not differ (p > 0.05) among dietary treatments at any time. It is concluded that a short-term supply of dietary L-carnitine to a conventional commercial feed after hatching enhanced subsequent humoral immunity in Leghorn-type chickens. Further study is warranted to confirm such a long-term effect.

Enhancement of cellular and humoral immunity following embryonic exposure to melatonin in turkeys (Meleagris gallopavo).

Two experiments were performed to determine the effect of in ovo melatonin supplementation on the ontogeny of immunity in the Large White turkey poult. Different levels of melatonin were injected into the air cell of the egg 4 days prior to hatch. In Experiment 1, turkey embryos received 3 ml of solution containing 200, 100, 50, 25, 10, or 1 microg/ml of melatonin. The hatchability at each dose was determined and compared to vehicle-injected controls. In Experiment 2, only poults from melatonin treatments in Experiment 1 that resulted in normal hatchability (10 and 1 microg/ml) were used. Lymphoproliferative responses to phytohemagglutinin (PHA-P) and primary antibody responses to Chukar red blood cells (CRBC) were determine at five time intervals: 0, 1, 7, 14, and 21 days post-hatch. At each of these times, including 28 days post-hatch, treatment effects on body weights were determined. At 28 days post-hatch, bursal, thymic, and splenic weights were obtained. In ovo melatonin administration significantly accelerated (P0.05) the development of cell-mediated (PHA-P) and humoral (CRBC) immune responses, and these responses were significantly elevated above vehicle-injected controls through 21 days post-hatch. No effect was observed on bursal, thymic, splenic or body weights. These data suggest that embryonic exposure to melatonin enhances post-hatch immune development and responsiveness.

Effects of arginine supplementation on the humoral and innate immune response of older people.

OBJECTIVE: To evaluate whether oral supplementation with arginine affects the humoral and innate immune response after vaccination against Streptococcus pneumoniae in a group of people aged 60 y and older, free-living in the community. DESIGN: A randomized controlled trial with one supplemented group and one control group. SETTING: Older persons living in the community. SUBJECTS: A total of 29 adults aged 60 y and older. INTERVENTIONS: The older people were randomized into two groups, one with arginine supplementation (15 g/day) for 4 weeks after pneumococcal vaccine. The control group received only the vaccine. Anthropometric measurements and immune system function parameters: neutrophil chemotaxis and phagocytosis, natural killer cell activity, determination of serum pneumococcal polysaccharide antibodies and serum C3 and C4. RESULTS: Neutrophil phagocytosis and the serum concentration of complement (C3 and C4) did not differ between groups. IgG antibodies against pneumococcal polysaccharide serotypes 1, 5 and 6B increased in both groups. The following parameters increased in the arginine-supplemented group compared to the nonsupplemented group: neutrophil chemotaxis (34 vs 19 units of migration, P = 0.002), natural killer cell cytotoxicity (23.3 vs 13.4 10 M/Ul 40%, P = 0.011) and IgG against antigen 5 (12.3 vs 6.2 mug/ml, P = 0.044). CONCLUSIONS: This study suggests that, after the pneumococcal vaccine, the intake of arginine increased neutrophil chemotaxis, natural killer cytotoxicity and serum concentration of IgG against antigen 5 in older people. These results suggest that arginine supplementation may enhance the immune response elicited by the pneumococcal vaccine in older people. SPONSORSHIP: Supported in part by CAPES and FAEPA.