Rizal Technological University College of Arts and Sciences Department of Biology

A research paper in Immunology: Effects of Dietary Antioxidants on the Function of the Immune System

Submitted by: Jose, Louise Vironica D. CAS-02-501A

Submitted to: Professor Marita Geroy Immunology Instructor

Introduction

The ability to interact with the world and remain healthy is dependent to a large extent on the healthy functioning of the immune system, which is the first line defense of the human body, since it is responsible for fighting foreign invaders that enters the system of the human body, such as pathogenic bacteria and viruses; and also it destroys cells within the body when it shows the signs of being cancerous. The immune system could be an orchestra, because it contains many different instruments that works harmoniously together with one goal and that is to defend and protecting the human body from foreign substances that can cause damage to it. And, like an orchestra itself, the immune system has different components that are suppose to play their roles at the same time by cell-to-cell communication. There is a vast variety of components in regards to the immune system from the structural barriers to the specific messenger molecules to cause the action of the other components and also to stop the action. Thus, nutritionally supporting the immune system means supporting all the different components that comprises the whole orchestra. The bodys zillion or so cells always faces formidable threats that ranges from the lack of food to the infections caused by viruses, thereby making the immune system continuously fight off these foreign invaders.

Exogenous sources such as ozone, exposure to UV radiation via sunlight, and cigarette smoke, which stimulates the production of free-radicals. These free radicals may come in many shapes, sizes and chemical configurations; but even though they may differ in some aspect they also share a similarity, which is having a voracious appetite for electrons. Whereas, these free radicals steals electrons from any nearby substances that will yield them. Thus, the electron theft can radically alter the losers structure or function. Free radical damage can change the encoded amino acids that are encased in a DNA strand. This damage causes certain diseases such as heart attack, cancer and etcetera, since it results into the low-density lipoprotein molecule more likely to get trapped in an artery wall. Or it can alter a cells membrane, thereby changing the flow of what enters and leaves within the cell. Free radical molecules have also endogenous sources wherein they are generated during normal cellular metabolism and also made deliberately.

Free radicals are highly-reactive molecules containing one or more than unpaired election in the outer orbit such as oxygen free radicals are superoxide, hydroxyl, peroxyl (RO2), alkoxyl (RO), and hydroperoxyl (HO2) radicals; there are two nitrogen free radicals such as nitric oxide and nitrogen dioxide (NO2). Reactive oxygen species, also known as ROS, is collective term that includes not only oxygen-centered radicals but also some of which are, non-radical derivatives of oxygen such as superoxide anion (O2-) and hydroxyl radical (OH) and hydrogen peroxide (H2O2). In producing a hydroxyl radical, which the most reactive and damaging of the oxygen free radicals, H2O2 plays a major role, since it can easily breakdown particularly in the presence of transition metal ions such as iron (Fe2+)

Oxygen radicals exert critical actions such as signal transduction, gene transcription, and regulation of soluble guanylate cyclase activity in the cells. Also, nitrogen oxide (NO) is one of the most widespread signaling molecules and participates in virtually in every cellular and organ function in the body. Endothelial cells that produce physiologic levels of NO, that are essential in regulating the relaxation and proliferation of vascular smooth muscle cells, leukocyte adhesion, platelet aggregation, angiogenesis, thrombosis, vascular tone, and hemodynamics; it is also generation by activated macrophages that serves as an important mediator of the immune response. Free radicals induce oxidative stress, which is balanced by the bodys endogenous antioxidant systems with an input from the co-factors, and by the ingestion of exogenous antioxidants. Hence, as oxidants and inhibitors of enzymes that contains an iron-sulfur center, free radicals and other reactive species cause the oxidation of biomolecules (e.g., amino acids, proteins, lipids and DNA), that leads into cell injury, formation of cataracts and even death. Free radical production in the cells can significantly increased by certain toxic called redox -cycling compounds such as various drugs and CCl4. ROS are produced in vivo, since it is majorly produced by the mammalian mitochondria and enzymes is another endogenous source. As organisms have evolved antioxidant defense systems are either to prevent the generation of ROS or to intercept any that is being produced. They exist in both the aqueous and membrane compartments of the cells and can be enzymes or non-enzymes. Catalase and glutathione peroxidase are enzymes that can safely decompose peroxides, particularly H2O2 that are produced during the respiratory burst that involves in

microbial killing n phagocytic cells, while superoxide dismutase that intercepts or scavenges free radicals. Most free-radical scavengers are not enzymes, and many can be obtained through diet. -tocopherol is the most important in cell membranes, which is a major member of the vitamin E family. This molecule acts as a chain-breaking anti-oxidant, that intercepts lipid peroxyl radicals and in so terminating lipid peroxidation chain reactions. Another group of lipidsoluble compounds that can also act as antioxidants are the carotenoids, such as -carotene,

lycopene and lutein; these compounds can be found in highly-pigment fruits and vegetables (e.g., mangoes, tomatoes, spinach and etcetera,). The polyene structure of these compounds allows the molecules to quench, or to inactivate, singlet oxygen and free-radicals. Ascorbic acid, also known as vitamin C, that play a role in sparing vitamin E that regenerates -tocopherol from the oxidized tocopheroxyl radical. As viewed from the perspective of a two-faces with ROS as one face while antioxidants is the other, it has been thought that tipping the balance in favor of the ROS. Due to the fact, that ROS is a major contributor to several degenerative disorders such as cancer, chronic diseases, arthritis and cardiovascular diseases; also to the ageing process, in general. Strong associations between diets that are rich in antioxidant nutrients and a reduced incidence of cancer have been observed in numerous epidemiological studies, and it has been suggested that a boost to the bodys immune system by antioxidants might, at least may taken part in, account for this findings. It is probably crucial to attempt to keep the balance of ROS to antioxidants levels as possibly. Thus, ideally by the dietary means rather than by taking supplements, from as early age as possible, to prolong, if not prevent, the onset of many-age related disorders.

The immune system is particularly sensitive to oxidative stress. As the immune cells relies heavily on cell-cell communication, particularly via membrane- bound receptors for working more effectively. Cell membranes that are rich in polyunsaturated fatty acids which, if peroxidized, can lead to a loss of membrane integrity, altered membrane fluidity, and that could result into the alterations in intracellular signaling and cell function. In addition, the production of ROS by phagocytic immune cells can damage the cells themselves, if they are not sufficiently protected by antioxidants.

Terminologies

Angiogenesis The physiological process through which new blood vessels form from pre-existing vessels.

Cytokines Cells of the immune system communicate with one another by releasing and responding to chemical messengers

Dismutase is an enzyme that catalyzes a dismutation reaction

Dismutation reaction A disproportionation reaction, especially in a biological context, in which oxidized and reduced forms of a chemical species are produced simultaneously

Ewes A female sheep

Glucocorticoids A class of steroid hormones that bind to the glucocorticoid receptor, which is present in almost every vertebrate animal cell

Glutathione peroxidase (GPx) The general name of an enzyme family with peroxidase activity whose main biological role is to protect the organism from oxidative damage. The biochemical function of glutathione peroxidase is to reduce lipid hydroperoxides to their corresponding alcohols and to reduce free hydrogen peroxide to water.

Hydroperoxyl Also known as the perhydroxyl radical, is the protonated form of superoxide with the chemical formula HO2.

Hydroxyl radical HO, is the neutral form of the hydroxide ion. Hydroxyl radicals are highly reactive and consequently short-lived; however, they form an important part of radical chemistry.

Immunosuppressive Suppression of the immune response, as by drugs or radiation, in order to prevent the rejection of grafts or transplants or to control autoimmune diseases. Also called immunodepression.

Leukotriene B4 A leukotriene involved in inflammation. It is produced from leukocytes in response to inflammatory mediators and is able to induce the adhesion and activation of leukocytes on the endothelium, allowing them to bind to and cross it into the tissue. In neutrophils, it is also a potent chemoattractant, and is able to induce the formation ofreactive oxygen species and the release of lysosome enzymes by these cells. It is synthesized by leukotriene-A4 hydrolase from leukotriene A4.

Metallothionein S family of cysteine-rich, low molecular weight (MW ranging from 500 to

14000 Da) proteins. They are localized to the membrane of the Golgi apparatus. MTs have the capacity to bind both physiological (such as zinc, copper, selenium) andxenobiotic (such as cadmium, mercury, silver, arsenic) heavy metals through

the thiolgroup of its cysteine residues, which represents nearly the 30% of its amino acidicresidues.

Muscle proteolysis Proteolysis is the breakdown of proteins into smaller polypeptides or amino acids. In general, this occurs by the hydrolysis of the peptide, and is most commonly achieved by cellular enzymes called proteases, but may also occur by intramolecular digestion, as well as by non-enzymatic methods such as the action of mineral acids and heat.

Nitrogen free radicals An important cellular signaling molecule involved in many physiological and pathological processes. It is a powerful vasodilator with a short half-life of a few seconds in the blood Peroxidation A type of reaction in which oxygen atoms are formed leading to the production of peroxides. It is stimulated in the body by certain toxins and infections.

Selenoproteins Any protein that includes a selenocysteine (Se-Cys) amino acid residue. Among functionally characterized selenoproteins are five glutathione peroxidases (GPX) and three thioredoxin reductases, (TrxR/TXNRD) which both contain only one Se-Cys.

Soluble guanylate cyclase activity (sGC) The only known receptor for nitric oxide, NO. It is soluble, i.e. completely intracellular. Most notably, this enzyme is involved in vasodilation. In humans, it is encoded by the genes GUCY1A2, GUCY1A3, GUCY1B2 andGUCY1B3.

Xanthise oxidase is a form of xanthine oxidoreductase, a type of enzyme that generates reactive oxygen species.These enzymes catalyze theoxidation of hypoxanthine to xanthine and can further catalyze the oxidation of xanthine touric acid. These enzymes play an important role in the catabolism of purines in some species, including humans

List of Antioxidants:

Vitamin E. Alpha-tocopherol, the most widely available isomer, has the highest biopotency, or strongest effect in the body. Because it is fat-soluble (and can only dissolve in fats), alphatocopherol is in a unique position to safeguard cell membranes -- largely composed of fatty acids -- from damage by free radicals. Alpha-tocopherol also protects the fats in low-density lipoproteins (LDLs, or the "bad" cholesterol) from oxidation. Broccoli, carrots, chard, mustard and turnip greens, mangoes, nuts, papaya, pumpkin, red peppers, spinach, and sunflower seeds

Vitamin C, also known as ascorbic acid, is a water-soluble vitamin. As such, it scavenges free radicals that are in an aqueous (watery) environment, such as inside your cells. Vitamin C works synergistically with vitamin E to quench free radicals. Vitamin C also regenerates the reduced (stable) form of vitamin E. Berries, broccoli, Brussels sprouts, cantaloupe, cauliflower, grapefruit, honeydew, kale, kiwi, mangoes, nectarines, orange, papaya, red, green or yellow peppers, snow peas, sweet potato, strawberries, and tomatoes

Beta-carotene, also a water-soluble vitamin, is the most widely studied of the 600 carotenoids identified to date. It is thought to be the best quencher of singlet oxygen (an energized but uncharged form of oxygen that is toxic to cells). Beta-carotene is also especially excellent at scavenging free radicals in low oxygen concentration. Apricots, asparagus, beets, broccoli, cantaloupe, carrots, corn, green peppers, kale, mangoes, turnip and collard greens, nectarines, peaches, pink grapefruit, pumpkin, squash, spinach, sweet potato, tangerines, tomatoes, and watermelon

Selenium is a trace element. It is a mineral that we need to consume in only very small quantities, but without which we could not survive. It forms the active site of several antioxidant enzymes including glutathione peroxidase. Found in Brazil nuts, tuna, beef, poultry and fortified breads, and other grain products

Quercetin: a plant-based chemical (phytochemical) found in apples, onions, teas, red wines, and other foods; it fights inflammation and may help reduce allergies.

Luteolin: a flavonoid found in abundance in celery and green peppers; it also fights inflammation, and one study showed it may help protect against inflammatory brain conditions like Alzheimers.

Catechins: a type of flavonoid found in tea; catechins in tea may help reduce risk of heart disease, cancer, and Alzheimers disease.

Review of Related Literature A 1999s study that was conducted by David A. Hughes about the Effects of dietary antioxidants on the immune function of middle-aged adults and it was said that:

The immune system is highly reliant on accurate cell-cell communication for optimal function, and any damage to the signalling systems involved will result in an impaired immune responsiveness. Oxidant-mediated tissue injury is a particular hazard to the immune system, since phagocytic cells produce reactive oxygen species as part of the bodys defence against infection. Adequate amounts of neutralizing antioxidants are required, therefore, to prevent damage to theimmune cells themselves. Many antioxidants can be obtained directly from the diet (e.g. ascorbic acid, a-tocopherol, carotenoids and polyphenolic flavonoids) or require micronutrients as integral components (e.g. Se in the metalloenzyme glutathione peroxidase (EC 1.11.1.9)). Numerous epidemiological studies have found strong associations between diets rich in antioxidant nutrients and a reduced incidence of cancer, and it has been suggested that a boost to the bodys immune system by antioxidants might, at least in part, account for this. Although more striking effects have been observed in the elderly, there is also evidence that antioxidant nutrients can modify cell mediated immune responses in younger individuals. Indeed, it might be essential to have an adequate intake of antioxidant nutrients from an early age in order to help prevent the development of, or at least delay the onset of, several degenerative disorders. The present paper will review the effects of specific nutrients on immune function in young to middle-aged human subjects, focusing on the antioxidant vitamins C and E, and on Se. A further review, dealing more specifically with the effects of carotenoids on human immune function, will be presented at a forthcoming meeting of the Nutrition Society.

A European research on the function effects of dietary antioxidants in 2003, it has been concluded that:

The Antioxidant Hypothesis suggests that reducing agents prevent oxidative damage and thus increased levels will also reduce the risk of chronic disease. This lead to the suggestion that increased intake of dietary compounds, some of which are capable of acting as chemical antioxidants, would be of benefit by augmenting cellular defenses and protect components of the cell against oxidative damage and, ultimately disease. There is compelling epidemiological evidence linking consumption of diets rich in antioxidants, direct and indirect, with reduced risk of developing degenerative disease. However, the enormous focus on this field of research worldwide has generated data that both support and challenge the hypothesis. Observational studies in humans have demonstrated high intakes of foods rich in dietary antioxidants (i.e. fruit and vegetable, cereals and grains, and some specific oils and fats) and high plasma concentrations of these dietary antioxidants are associated with reduced morbidity and mortality. However, plasma antioxidant concentrations are a measure of intake, rather than a measure of concentration at target sites, and an association is not necessarily causal. Human intervention studies have shown no consistent benefit of increased intake of dietary antioxidants, either individually or in combination as isolates or foods, in the prevention of human diseases. Many arguments have been put forward to explain why these studies have failed to demonstrate an effect consistent with the epidemiology, but few have addressed the underlying and fundamental problems associated with complex nutritional trials.

Several researches in the National Research Center on Giza, Egypt conducted a study about the Effects of Dietary Antioxidants Supplementation on Cellular Immune Response and Evaluation of their Antimicrobial activity against some Enteric Pathogens in Goats (2013). That stated:

Oxidant- mediated tissue injury is a particular hazard to the immune system, since phagocytic cells produce reactive oxidants, as a part of the bodys defense against infectious agents. Therefore, adequate amounts of neutralizing antioxidants are required to prevent damage in the immune cells themselves [2].The results of the present study as shown in Table (4), indicated that BCS and multi-antioxidants significantly (P<0.05) enhanced lymphocyte proliferative responses.

Similarly, our recorded data demonstrated significant (P<o.o5??) elevation of phagocytic % and phagocytic index in both groups received antioxidants supplement compared with non- supplemented group. Many effects have been reported for the seeds of N. sativa and Zn- Met. With Vit. E and Se including their antioxidant roles and immune potentiating effects [2, 3]. This explained the improvement of cellular immune response in the two groups of animals received antioxidants in the current study.

Recently, Haman and Abou- Zeina [7] reported that ewes supplemented with vitamin E and Se has better antioxidants status and greater immunoglobulins. It was concluded that vitamin E and Se supplementation improve the status of these micronutrients and humoral immune response in buffalo calves.

Meanwhile, Abou- Zeina et al., [16] demonstrated that Zn, Se and Vit E administered to buffalo cows under heat stress condition, induced better reproduction performance and increase in immunoglobulin levels.

How Se and Vit E enhance the immune response is not fully understood. It is possible that Se as a co-factor of the GPx family of enzymes, either through a direct effect or via destruction of inhibitory peroxides, contributes to a better immune response [10]. As a constituent of selenoproteins, Se is needed for the proper function of neutrophils, macrophages, natural killer cells and T lymphocytes [5]. Mild Se deficiency appears to increase susceptibility to oxidant stress [50] and risk of infections. It was documented by many authors that deficiency of Se also affects the ability of the macrophages to release to leukotriene B4 which signals the white blood cells to aid in the destruction of harmful microorganisms [51]. The antioxidant activity of vitamin E in preventing lipid oxidation may be one of the mechanisms by which vitamin E enhances immunity particularly phagocytosis [11]. Besides, Vitamin E causes a reduction of glucocorticoids, which are known to be immunosuppressive. Researchers suggested that relatively higher levels of vitamin E supplementation may improve animal performance, which may be due to enhanced immunity p[15, 52].

Likewise, Zinc is crucial for normal development and function of cells mediating non-specific immunity such as neutrophils and natural killer cells [53]. Zinc also, regulates the expression in lymphocytes of metallothionein and metallothionein-like proteins with antioxidant activity. In vitro intracellular killing by macrophages is very sensitive to zinc deficiency adversely affects the secretion and functions of cytokines, the basic messengers of the immune system [13].

Conclusion:

Free radicals have been implicated in the etiology of varying large number of major diseases. In which, they can adversely alter many crucial biological molecules leading to loss of form and function. Such undesirable changes in the body can lead to diseased conditions. Antioxidants can protect the body against the damaged induced by the free radicals that acts in various levels, and it may also enhance the function of the immune response thereby, strengthening the components that of which make-up the immune system. Dietary and other components of pants form major sources of antioxidants.

The relation between free radicals, antioxidants and function of the various organs and organ systems is highly complex and the discovery of the redox- signaling has been a milestone in this crucial relationship. Recent research centers on various strategies to protect crucial tissues organs against oxidative damage induce by the free radicals. Many novel approaches are made and significant finding have come to light in the last few years. The higher intake of food with functional attributes including high level of antioxidant sin functional foods is one strategy that is gaining importance in advanced countries and is making its appearance.

Co-ordinate research involving biomedical scientists, nutritionist and physicians can make significant differences to human health in the coming decades. Research on free radicals and antioxidants involving theses is one such effort in the right direction.

References:

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