QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lim, B. O. Articles by Fernandes, G. Search for Related Content PubMed PubMed Citation Articles by Lim, B. O. Articles by Fernandes, G.

---

Article

Dietary (n-6) and (n-3) Fatty Acids and Energy Restriction Modulate Mesenteric Lymph Node Lymphocyte Function in Autoimmune-Prone (NZB x NZW)F1 Mice¹

Beong O. Lim, Christopher A. Jolly, Kaliquz Zaman and Gabriel Fernandes²

Department of Medicine, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900

²To whom correspondence should be addressed.

	ABSTRACT

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
We previously showed that dietary fish oil (FO) and energy restriction (R) have beneficial anti-inflammatory properties in the peripheral blood and spleens of (NZB x NZW)F1 (B/W) lupus-prone mice. Furthermore, unsaturated fatty acids also were shown in the past to influence mesenteric lymph node (MLN) lymphocyte function in healthy young rats. The MLN play a pivotal role in mediating food allergy. To date, the effect of R on intestinal immunity is not well understood; therefore we determined the effect of diet on MLN lymphocyte function. Mice were given either free access to a 5 g/100 g corn oil (CO) or fish oil (FO) diet or the same corn oil (CR) or fish oil (FR) diets restricted to 60% of the intake of the control group. At the age of 4 (young) and 8 (old) mo, MLN lymphocytes were isolated and B- (CD19⁺) and T-lymphocyte subsets (CD4⁺ and CD8⁺) were determined by flow cytometry. Additional MLN lymphocytes were placed in culture with or without concanavalin A and culture supernatants collected after 72 h for cytokine and immunoglobulin (Ig) quantitation by ELISA. Aging significantly (P < 0.05) decreased both CD4⁺ and CD8⁺ T-lymphocytes. Spontaneous and activation-induced interleukin-4 (IL-4), IL-10, and interferon- secretion were greater while IL-2 was lower in CO-fed old mice compared to CO-fed young mice. In contrast, CR or FO alone partially blunted the age-dependent alterations in T-lymphocyte ratios including cytokine and Ig secretion, whereas the FR diet significantly (P < 0.005) normalized the accelerated aging effects on these immune variables. We show for the first time that FR is a far more potent anti-inflammatory therapy than either CR or FO alone in modulating MLN lymphocyte function.

KEY WORDS: • lymphocyte • energy restriction • fatty acid • cytokine • immunoglobulin • mice

	INTRODUCTION

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
Dietary fish oil (FO), rich in (n-3) fatty acids, possesses potent anti-inflammatory properties in human and rodent autoimmune disease models (Endres et al. 1993 , Fernandes et al. 1994 , Fernandes 1998 , James and Cleland 1997 , Kremer et al. 1995 , Lim et al. 1997 , Venkatraman et al. 1994 ). These properties likely involve changes in T-lymphocyte function, which indeed influences both the type and extent of immune response (Calder 1997 ). The T-lymphocyte exerts its modulatory properties by proliferating in response to stimulation and producing various cytokines. Cytokines derived from the T-lymphocyte are categorized as Th-1 or Th-2. The Th-1 cytokines including interleukin-2 (IL-2),³ interferon- (IFN-), and tumor necrosis factor (TNF-) influence cell-mediated immunity, while the Th-2 cytokines, IL-4, -5, -6, and -10, regulate humoral or antibody-mediated immunity. Recently, dietary (n-3) fatty acids were found to modulate lymphocyte proliferation, cytokine production, signal transduction, and gene expression in healthy humans (Endres et al. 1993 , Meydani et al. 1991 ) and rodents (Fernandes and Jolly 1998 , Fernandes et al. 1998 , Jolly et al. 1997 , Jolly et al. 1998 ). Furthermore, other studies reported that dietary lipids also modulate immunoglobulin (Ig) production in both spleen and mesenteric lymph node (MLN) lymphocytes from healthy rats (Hung et al. 1997 , Lim et al. 1995 , Lim et al. 1996 , Sugano et al. 1998 , Yamada et al. 1996 ). Specifically unsaturated fatty acids derived from vegetable oils enhance IgE production and inhibit IgG and IgM production by spleen or MLN lymphocytes (Yamada et al. 1996 ). Indeed, the MLN play a pivotal role in intestinal immunity, whereas IgE is the primary mediator of the Type I hypersensitivity reaction to food allergens (Sampson 1997 ).

Alterations in both cell-mediated and humoral immunity occur with age in both elderly humans and aged rodents, thereby increasing the risk of developing autoimmune disease and cancer as well as both viral and bacterial infections (Miller 1991 , Murasko and Goodnewardene 1990 ). Many of these age-related changes involve defects in T-lymphocyte function, including a diminished proliferative response to mitogenic stimulation [i.e., concanavalin A (Con A)] and subsequent production of IL-2, a potent polyclonal T-lymphocyte mitogen, and alterations in Th-2 cytokine production in the blood and spleen. Cytokines can act directly on B-lymphocytes, promoting polyclonal activation and subsequent synthesis of antibodies such as IgG, IgE, IgM, and IgA (Stevens et al. 1988 ). To date, the only known intervention to increase life span and abrogate some of the age-dependent immune abnormalities is energy restriction (R) (Birt et al. 1999 , Keenan et al. 1997 , Kim et al. 1997 , Weindruch and Walford 1988 ). Energy restriction (R) is defined as a 30–40% reduction in food consumption. Our previous studies showed that R extended the life span of MRL/l pr mice by inhibiting the development and expression of the lymphoproliferative syndrome (Fernandes and Good et al. 1984 ) and by delaying the onset of autoimmune kidney disease in (NZB x NZW)F1 (B/W) mice (Fernandes et al. 1976 ). We also reported increasing the life span of B/W mice by feeding (n-3) fatty acid-enriched diets without R when compared to corn oil (CO) diet (Fernandes et al. 1994 ). However, the effects of R on intestinal immunity are not well-understood. We recently reported that dietary (n-3) fatty acids, from FO, or R delay the onset of autoimmune kidney disease in B/W mice in association with reduced cytokine production in kidneys (Chandrasekar et al. 1995 , Chandrasekar and Fernandes 1994 ) and in peripheral blood T-lymphocytes (Jolly and Fernandes 1999 ). Since little information exists on the influence of R on intestinal immune function and, at least, in blood and spleen, R appears to have beneficial effects (Fernandes and Jolly 1998 , Jolly and Fernandes 1999 ), we therefore undertook to study the effect of R and FO in combination on MLN lymphocyte populations, and cytokines and Ig production in young and old B/W mice. The purpose of this study was to determine if MLN lymphocytes respond in a similar or different manner to peripheral blood and splenic lymphocytes when fed FO and/or R diets. Our previous results showed that the B/W MLN B-lymphocyte response is impaired relative to spleen lymphocytes or to the MLN of healthy mice (Jung et al. 1984 ).

	MATERIALS AND METHODS

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
Materials.

ConA (10 mg/L) was purchased from Pharmacia (Piscataway, NJ). Monoclonal antibodies and cytokine and Ig standards were purchased from PharMingen (San Jose, CA). All other chemicals were reagent grade or better from Sigma (St. Louis, MO).

Animal and diets.

Four-week-old female B/W mice were obtained from the Jackson Laboratory (Bar Harbor, ME). The mice were housed five per microisolator cage in a pathogen-free environment at 25°C and maintained on a 12-h light-dark cycle. Following 1-wk acclimation, two groups of mice were adapted to a 40% energy-restricted diet. For 1 wk, the groups were given 10% less food relative to the free access group followed by another week of 20% restriction. Then five mice per group were maintained on either free access to a CO or FO diet or were given the corn oil diet (CR) or fish oil diet (FR) with intake restricted by 40%. Mice ingested all of the required amounts of essential nutrients from the diets. Vitamin and mineral content was not increased in the CR or FR groups to maintain the same nutrient intake per gram body weight. The diets were prepared according to the recommendation of the American Institute of Nutrition (AIN 76 diet) (77). Body weight was recorded weekly and food intake daily as previously described (Troyer et al. 1997 ). Beginning at 4 mo of age, proteinuria was measured weekly to assess disease severity (i.e., nephritis). Prior to killing, the mice were anesthetized, and peripheral blood was collected by retroorbital bleeding for the analysis of lymphocyte populations and cytokine production (Jolly and Fernandes 1999 ). All animal-care techniques were performed within the guidelines approved by the institutional animal care and use committee.

Lymphocyte culture.

Mice were killed at the age of either 4 (young) or 8 (old) mo and MLN were aseptically removed. The MLN_S were disrupted between two frosted glass slides into 3 mL of RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100,000 U/L penicillin, 100 mg/L streptomycin, 10 µmol/L 2-mercaptoethanol, and 100 mmol/L L-glutamine (RPMI complete) (Lim et al. 1997 ). The MLN lymphocytes were then passed through a wire-mesh filter to obtain a single-cell suspension; viability (95%) was determined by trypan blue exclusion; then the cells were resuspended at 2 x 10⁹ cells/L in RPMI complete. Lymphocytes were cultured at 2 x 10⁵ cells per well in 96-well microtiter plates in the presence or absence of 10 mg/L ConA for 72 h. Culture supernatants were collected and stored at -80°C for future analysis.

Lymphocyte population analysis.

Single-cell suspensions of MLN lymphocytes were incubated at 37°C for 30 min to remove fibroblasts and then centrifuged at 1500 x g for 30 min. The cells were rinsed three times with RPMI 1640 complete and then resuspended in PBS containing 10% FBS. MLN lymphocyte subsets were determined by an FACScan flow cytometer (Becton Dickinson, Mountain View, CA) with a direct staining immunofluorescence technique as previously described (Fernandes 1999 ). T-lymphocyte subsets (CD4⁺ and CD8⁺) and B-lymphocyte (CD19⁺) proportions were determined by placing 2.5 x 10⁵ lymphocytes, in RPMI 1640 medium (containing 2% FBS), in 12 x 75 mm tubes containing either CD4 fluorescein isothiocyanate, CD8 phycoerythrin (PE) or CD19 PE conjugated monoclonal antibodies and counting 20,000 cells (lymphocytes) per sample. Relative proportions of CD4, CD8, and CD19 lymphocytes were determined. The data represent the means ± SEM of five determinations.

Measurement of cytokine and Ig concentration.

Supernatants from 72 h Con A-activated MLN lymphocyte cultures were obtained. Cytokines (IL-2, IL-4, IL-5, IL-10, and IFN-) were measured by ELISA using cytokine-specific capture and detection monoclonal antibodies as previously described (Fernandes 1999 ). IgE, IgA, IgG₁, IgG_2a, IgG_2b, IgG₃, and IgM content in the culture supernatants was measured by ELISA as described previously (Lim et al. 1997 ).

Statistical analyses.

Data were analyzed by three-way ANOVA for the main effects and interactions of age, energy level, and lipid source. Individual means were separated using Duncan’s new multiple-range test to evaluate significant differences (Duncan 1955 ). The significance of Con A stimulation was determined using a paired t-test. Differences of P < 0.05 were considered significant.

	RESULTS

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
Body weights and proteinuria.

The body weights of the young (4 mo) and old (8 mo) CO and FO fully fed diet groups did not differ with dietary lipid source (44 ± 4 vs. 45 ± 5 g, respectively, for the young and 40 ± 5 and 46 ± 6 g, respectively, for the old). Furthermore, the body weights of the young and old CR and FR diet groups did not differ with dietary lipid source (28 ± 3 vs. 32 ± 1 g, respectively, for the young and 32 ± 3 and 31 ± 1 g, respectively, for the old). Therefore, the restricted groups weighed ca. 30 to 35% less than their counterparts given free access to food. Dietary fat source did not have a significant effect on mouse weight. All the mice given free access to CO were 3+ for proteinuria (Table 1 ) at 8 mo. The proteinuria for the FO group ranged from 1+ to 3+ while the CR and FR groups had trace or undetectable proteinuria. Similarly, all young mice had trace proteinuria (data not shown).

Relative to the CO group, CR and FR moderately reduced CD4⁺ T-lymphocyte proportions in young mice (Fig. 1 , upper panel P < 0.05). On the other hand, the proportion of CD8⁺ lymphocytes was significantly (P < 0.001) higher (17%), in CR than in CO mice, while FO and FR reduced the proportions (13%) of CD8⁺ T-lymphocytes in young mice (Fig. 1 , middle panel). Age in the CO-fed mice significantly (P < 0.001) reduced the relative proportions of CD4⁺ and CD8⁺ T-lymphocytes (65 and 52%, respectively), relative to that of the young CO controls. CR partially prevented the age-associated loss of CD4⁺ proportions, while FO had no effect. On the other hand, FR significantly (P < 0.001) maintained CD4⁺ lymphocyte proportions at 87% of that of young CO controls. Similar to the CD4⁺ lymphocytes, the age-associated decrease in CD8⁺ proportions was partially normalized by CR (88% of young control levels), while FO had no effect. Interestingly, in the FR group the age-dependent decline in CD8⁺ proportions was not observed.

View larger version (29K): [in this window] [in a new window]   Figure 1. Diet reverses the age effects on lymphocyte populations in mesenteric lymph node lymphocytes of lupus-prone mice. (NZB x NZW) F1 mice were given free access to either a corn oil (CO) or fish oil (FO) diet or were given 40% less food from the CO (CR) or FO (FR) diets. At 4 (young) and 8 (old) mo of age, mesenteric lymph node lymphocytes were isolated and analyzed by flow cytometry for the relative percentage of CD4+ (A), CD8+ (B) T-lymphocyte subsets and CD4+ to CD8+ ratio (C). Values are means ± SE, n = 5. Values within an age group that have different letters are significantly different, P < 0.05. nd; not detectable.

Both IL-2 and IFN- were spontaneously produced in young mice with free access to CO (Table 2 ). CR and FO significantly (P < 0.001) augmented IL-2 secretion by 100% while FR was only partially effective (25% increase). In contrast, diet did not significantly influence the spontaneous production of IFN- in young mice. As would be predicted, age significantly (P < 0.001) decreased spontaneous IL-2 production in mice given free access to CO and in CR mice, while in all diet groups, age significantly (P < 0.001) increased IFN- production. Activation of MLN cells augmented (P < 0.001) IL-2 secretion in young mice with free access to CO. This augmentation was less dramatic in young mice with CR and free access to FO and was greatest in FR mice. Aged mice also responded to Con A by producing higher levels of IL-2, albeit to a lesser extent than in young mice. Both CR and FR resulted in significantly (P < 0.001) less IL-2 secretion relative to the old CO-fed mice. Interestingly, FR in aged mice yielded the highest level of IL-2 production, which was similar to the level seen in young mice with free access to CO following activation. Activation also significantly (P < 0.001) enhanced IFN- secretion in young mice with free access to CO, which was significantly (P < 0.001) reduced by CR, FO and FR. With age, Con A-induced IFN- production in the CO group was significantly higher relative to their young counterparts. CR, FO and FR reduced activation induced IFN- levels to that of their young counterparts.

In young mice, FO and FR resulted in significantly (P < 0.001) higher B-lymphocyte proportions (24%) when compared to the young CO and CR groups. Age significant P < 0.001) increased (32%) B-lymphocytes in the CO group when compared to CO-fed young mice. While both CR and FO partially blunted the age effect (Fig. 2 ), FR completely normalized B-lymphocyte proportions when compared to their young counterparts. The only significant effect of diet on spontaneous IgE production was in the old group, where CR, FO and FR equally significantly (P < 0.001) reduced (28%) IgE production (Fig. 3 ). Activation-induced IgE production was less (35%) in the old vs. young CO group. Diet significantly (P < 0.001) reduced the IgE production (25 and 32%) for CR and FO, respectively, while FR completely blunted the response. Table 4 illustrates the effects of diet and age on spontaneous Ig production from MLN lymphocytes. All Ig (IgA, IgG₁, IgG_2a, IgG_2b, IgG₃, IgM) levels were significantly (P < 0.001) lower in the old than in the young CO-fed mice. CR was better than FO at reducing spontaneous IgA production in young mice while the opposite was true for IgG₁ and IgG_2a. Interestingly, FO produced significantly (P < 0.001) higher IgG_2b and IgM in young mice. With the exception of IgM, FR was the best at suppressing spontaneous Ig production, regardless of age. Diet did not significantly influence spontaneous IgM production in old mice. FR and CR and to a lesser extent FO significantly (P < 0.001) reduced spontaneous IgA, IgG₁, IgG₃ or IgM production in old mice. On the other hand, CR was more effective at suppressing IgG_2a and IgG_2b relative to FO in old mice.

View larger version (33K): [in this window] [in a new window]   Figure 2. Effect of diet and age on B-lymphocyte proportions in mesenteric lymph nodes of lupus-prone mice. B/W mice were given free access to either a corn oil (CO) or fish oil (FO) diet or were given 40% less food from the CO (CR) or FO (FR) diets. At 4 (young) and 8 (old) mo of age, mesenteric lymph node lymphocytes were isolated and analyzed by flow cytometry for the relative percentage of CD19+ B-lymphocytes. Values are means ± SE, n = 5. a–d Values without the same superscript letter are significantly different, P < 0.05. The main effect P values: age < 0.0001; amt < 0.0037; oil < 0.2200: Interactions: age x amt. < 0.0001; age x oil < 0.001; amt. x oil < 0.5362. Values within an age group that have different letters are significantly different at P < 0.05.

View larger version (35K): [in this window] [in a new window]   Figure 3. Spontaneous and Concanavalin A (Con A) induced immunoglobulin E (IgE) production in mesenteric lymph node lymphocytes of lupus-prone-mice is altered by diet and age. B/W mice were given free access to either a corn oil (CO) or fish oil (FO) diet or were given 40% less of the CO (CR) or FO (FR) diets. At 4 (young) and 8 (old) mo of age, mesenteric lymph node (MLN) lymphocytes were isolated. MLN lymphocytes (2 x 109 cells/L) were cultured for 72 h in the absence (A) or presence (B) of 10 mg/L of Con A then IgE levels in the culture supernatants determined by ELISA. Values are means ± SE, n = 5. Values within an age group that have different letters are significantly different, P < 0.05. The main effect P values for unstimulated: age < 0.2355; amt. < 0.0408; oil < 0.6897: Interactions: age x amt. < 0.0244; age x oil < 0.081; amt. x oil < 0.1047. The main effect P values for stimulated: age < 0.0001; amt. < 0.0001; oil < 0.2200: Interactions: age x amt. < 0.0935; age x oil < 0.001; amt. x oil < 0.2832.

	DISCUSSION

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

Aging reduced the relative proportions of both CD4⁺ and CD8⁺ T-lymphocytes in MLN, which is consistent with what we observed in the peripheral blood (Jolly and Fernandes 1999 ). Interestingly, only CR or FR was able to normalize CD4⁺ and CD8⁺ T-lymphocytes, indicating the effect to be mediated by R alone. This is most likely due to maintenance of thymic function (i.e., decrease in thymic atrophy, which occurs with age, allowing for the appropriate proportions of T-lymphocytes to be maintained). Indeed, we and others previously showed that thymic involution in aged mice is decreased by R. In young mice, FO and FR marginally suppressed CD8⁺ T-lymphocyte proportions while diet only moderately influenced CD4⁺ proportions. Similarly, Yaqoob et al. (1994) did not find a significant effect of dietary FO in young rats.

The reduction in spontaneous and activation-induced IL-2 secretion seen in aged mice is a hallmark of aging in both human and rodent models (Jung et al. 1982 , Miller 1991 ). This is most likely due to an increase in the proportions of memory (having been previously exposed to antigen) relative to naïve (no previous antigenic exposure) T-lymphocytes. Interestingly, only FR increased activation-induced IL-2 secretion while CR and FO brought spontaneous secretion to detectable levels, suggesting an increase in the relative proportions of naïve vs. memory T-lymphocytes. The increase in naïve T-lymphocytes is most likely due to the maintenance of active thymic function. In contrast, both spontaneous and Con A-induced IFN-, IL-4, IL-5 and IL-10 secretion were greater in aged mice. This is typical of what is commonly observed in lupus-prone mice in that the Th-1 cytokine, IFN-, plays a critical role in propagating autoimmune kidney disease (Haas et al. 1998 ) while the Th-2 cytokines IL-4, IL-5 and IL-10 (Kalechman et al. 1997 ) are critical in initiating kidney inflammation. The Th-2 cytokines induce the polyclonal activation and expansion of B-lymphocytes, which enhances antibody production. Indeed, we observed increased B-lymphocyte proportions in old CO-fed mice relative to the young group. The relevant antibodies in lupus would be the autoantibodies as well as dsDNA antibodies. Therefore, by suppressing both Th-1 and Th-2 cytokine secretion via dietary intervention, both the initiation and propagation of autoimmune glomerulonephritis can be delayed.

Age also decreased Ig production in MLN. This is consistent with our previous results showing that the MLN, unlike the spleen in B/W mice, has defective B-lymphocyte function as determined by decreased plaque-forming cells (Jung et al. 1984 ). The most dramatic effects of diet could be observed following Con A activation. In general, dietary treatment further down-regulated Ig production, with FR being the most effective. It is important to note that diet dramatically suppressed activation-induced IgE, IgA, IgG₁, IgG_2b and IgG₃ secretion while IgM and IgG_2a were only marginally affected in both young and old mice. IgE plays a major role in mediating food allergy while IgA and IgM are thought to suppress food allergy by preventing the binding of antigen to the intestinal mucosa. Therefore, IgE reduction by diet treatment while IgM is relatively unaffected would decrease the IgE-to-IgM ratio, which may indeed help to reduce food hypersensitivity. The suppression of Ig production by diet treatment is consistent with the reduced relative proportions of B-lymphocytes in CR-, FO- and FR-fed old mice. Previous results have shown that the addition of unsaturated fatty acids in vitro enhances Con A-induced IgE production in MLN lymphocytes (Yamada et al. 1996 ). This discrepancy is most likely related to oxidation status. Hung et al. (1997) found that the enhanced IgE production by polyunsaturated fatty acid (PUFA) in vitro could be blocked by the addition of an antioxidant (-tocopherol). We have previously shown that FO increases antioxidant enzyme activity (Fernandes et al. 1996 , Reddy and Fernandes 1999 ), suggesting that effects related to fatty acid oxidation may be blunted which may explain IgE secretion does not increase in FO- and FR-fed mice.

FR was the most effective dietary treatment, but there were some unique qualities of FO and CR. For example, both FO and CR influenced spontaneous Ig production to varying degrees in an isotype-specific manner in young and old mice. However, CR was consistently more effective than FO at suppressing activation-induced Ig production regardless of isotype examined. FO was far more effective at suppressing activation-induced IFN- secretion when compared to CR in young and old mice. In contrast, CR was more effective in down-regulating IL-10 production relative to FO-fed mice. Therefore, it would appear that CR was more effective at suppressing the antibody-mediated (via IL-10) aspect of autoimmune kidney disease while FO may be more effective at reducing the cell-mediated (via IFN-) arm.

In this murine model, it is difficult to determine whether the effects of the diets are related to disease and/or aging. However, it should be pointed out that like the data presented herein, aged, healthy long-lived mice also have reduced IL-2 and IFN- secretion by lymphocytes and CR maintains IL-2 and IFN- secretion in the aged mice. Furthermore, our previous results showed that IL-2 secretion by splenic lymphocytes from aged lupus-prone mice is reduced. Thus, we chose this animal model because its life span is 1 y while that of healthy mice is 3 y. The data in this paper have implications for a wide range of diseases. For example, CR and FR, and to a lesser extent FO, reduced IL-4, IL-5 and IL-10 secretion by aged MLN lymphocytes, suggesting that these diets would reduce autoantibody production and could be valuable for preventing the development of autoimmune diseases. Of particular importance is the observation that diet reduced age-dependent elevations of IgE secretion, which may help to prevent the development of allergies, especially food allergies. Two other key points should be made with respect to the influence of CR and FO on Ig production. First, both CR and FR reduced IgA secretions in unstimulated and Con A-stimulated lymphocytes from young mice which may be beneficial in the prevention of IgA-related nephropathies. Second, the ability of CR and FR to reduce IgG_2a secretions in unstimulated and Con A-stimulated lymphocytes from young mice has direct application to this animal model. The IgG_2a isotype is thought to be the primary autoimmune antibody to deposit in the kidney, resulting in nephritis.

In summary, we show for the first time that dietary CR and FO can partially normalize age-associated alterations in MLN T- and B-lymphocyte proportions and function (cytokine and Ig secretion, respectively) while FR appears to be by far the most effective. Specifically, the reduced IgE-to-IgM ratios in FR-fed mice may indicate a protective effect of diet on the type I food hypersensitivity response as well. This study however warrants further new investigations into the potential use of CR, FO or FR as therapeutic means of managing immune disorders including food allergy.

	FOOTNOTES

Supported in part by: F32 AG05826;

Supported in part by: RO1 AG14541 and AG13693

³ Abbreviations used: B/W, (NZB x NZW)F1 mice; CO, corn oil; Con A, Concanavalin A; CR, corn oil with restriction (mice were fed a 40% energy-restricted diet containing 5% CO); FBS, fetal bovine serum; FO, fish oil; FR, fish oil with restriction (mice were fed a 40% energy-restricted diet containing 4.5% FO plus 0.5% CO); IFN-, interferon-; Ig, immunoglobulin; IL, interleukin; IL-2, interleukin-2; MLN, mesenteric lymph node; PE, phycoerythrin; PUFA, polyunsaturated fatty acids; R, energy restriction; Th-1 or Th-2, T-lymphocyte; TNF-, tumor necrosis factor-.

Manuscript received July 28, 1999. Initial review completed August 25, 1999. Revision accepted February 24, 2000.

	REFERENCES

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 

1. American Institute of Nutrition Report of the American Institute of Nutrition ad hoc committee on standards for nutritional studies J. Nutr 1977;107:1340-1348

2. Birt D. F., Yaktine A., Duysen E. Glucocorticoid mediation of dietary energy restriction inhibition of mouse skin carcinogenesis. J. Nutr. 1999;129:571S-574S

3. Calder P. C. n-3 Polyunsaturated fatty acids and cytokine production in health and disease. Ann. Nutrient Metabolism 1997;41:203-234

4. Chandrasekar B., Fernandes G. Decreased proinflammatory cytokines and increased antioxidant enzymes gene expression by n-3 lipids in murine lupus nephritis. Biochem. Biophys. Res. Commun. 1994;200:893-898[Medline]

5. Chandrasekar B., Troyer D. A., Venkatraman J. T., Fernandes G. Dietary omega-3 lipids delay the onset and progression of autoimmune lupus nephritis by inhibiting transforming growth factor ß mRNA and protein expression. J. Autoimmun. 1995;8:381-393[Medline]

6. Duncan D. B. Multiple range and multiple F test. Biometrics 1955;11:1-42

7. Endres S., Meydani S. N., Ghorbani R., Schindler R., Dinarello C. A. Dietary supplementation with n-3 fatty acids suppresses interleukin-2 production and monnuclear cell proliferation. J. Leuk. Biol. 1993;54:599-603[Abstract]

8. Fernandes G. Dietary lipids and risk of autoimmune disease. Clin. Immunol. Immunopathol. 1994;72:193-197[Medline]

9. Fernandes G. Omega-3 fatty acids and immune function including apoptosis. Kremer J. M. eds. Medicinal Fatty Acids in Inflammation 1998:73-90 Birkhauser Publishing Ltd Basel, Switzerland.

10. Fernandes G. Methods for the study of immune cells in aging. Methods in Aging Research 1999:549-567 CRC Press LLC Salem, MA.

11. Fernandes G., Bysani C., Venkatraman J. T., Tomar V., Zhao W. Increased TGF-ß and decreased oncogene expression by -3 fatty acids in the spleen delays onset of autoimmune disease in B/W mice. J. Immunol. 1994;152:5979-5987[Abstract]

12. Fernandes G., Chandrasekar B., Luan X., Troyer D. A. Modulation of antioxidant enzymes and programmed cell death by n-3 fatty acids. Lipids 1996;31:S91-S96

13. Fernandes G., Good R. A. Inhibition by restricted-calorie diet of lympho-proliferative disease and renal damage in MRL/lpr mice. Proc. Natl. Acad. Sci. USA 1984;81:6144-6148[Abstract/Free Full Text]

14. Fernandes G., Jolly C. A. Nutrition and Autoimmune Disease. Nutr. Rev. 1998;56:S161-S169[Medline]

15. Fernandes G., Troyer D. A., Jolly C. A. The effects of dietary lipids on gene expression and apoptosis. Proc. Nutr. Soc. 1998;57:543-550[Medline]

16. Fernandes G., Yunis E. J., Good R. A. Influence of diet on survival of mice. Proc. Natl. Acad. Sci. USA 1976;73:1279-1283[Abstract/Free Full Text]

17. Haas C., Ryffel B., Le Hir M. INF- receptor deletion prevents autoantibody production and glomerulonephritis in lupus-prone (NZB x NZW)F1 Mice. J. Immunol. 1998;160(8):3713-3718[Abstract/Free Full Text]

18. Hung P., Yamada K., Lim B. O., Mori M., Yuki T., Sugano M. Effect of unsaturated fatty acids and -tocopherol on immunoglobulin levels in culture medium of rat mesenteric lymph node and spleen lymphocytes. J. Biochem. 1997;121:1054-1060[Abstract/Free Full Text]

19. James M. J., Cleland L. G. Dietary n-3 fatty acids and therapy for rheumatoid arthritis. Semin. Arthritis. Rheum. 1997;27:85-97[Medline]

20. Jolly C. A., Chapkin R. S., McMurray D. N. Dietary (n-3) polyunsaturated fatty acids suppress murine lymphoproliferation, interleukin-2 secretion, and the formation of diacylglycerol and ceramide. J. Nutr. 1997;127:37-43[Abstract/Free Full Text]

21. Jolly C. A., Fernandes G. Diet modulates Th-1 and Th-2 cytokine production in the peripheral blood of lupus-prone mice. J. Clin. Immunol. 1999;19(3):171-177

22. Jolly C. A., McMurray D. N., Chapkin R. S. Effect of dietary n-3 fatty acids on interleukin-2 and interleukin-2 receptor alpha expression in activated murine lymphocytes. Prost. Leuk. Essen. Fatty Acids 1998;58(4):289-293

23. Jung L. K. L., Good R. A., Fernandes G. In vitro immune response of cells of various lymphoid tissues in (NZB x NZW)F1 mice: Evidence for abnormality of the mesenteric lymph node cells. J. Immunol. 1984;132(3):1265-1270[Abstract]

24. Jung L. K. L., Palladino M. A., Calvano S., Mark D. A., Good R. A., Fernandes G. Effect of calorie restriction on the production and responsiveness to interleukin 2 in (NZB x NZW)F1 mice. Clin. Immunol. Immunopathol. 1982;25:295-301[Medline]

25. Kalechman Y., Gafter U., Da J. P., Albeck M., Alarcon-Segovia D., Sredni B. Delay in the onset of systemic lupus erythematosus following treatment with the immunomodulator AS101: association with IL-10 inhibition and increase in INF- levels. J. Immunol. 1997;159(6):2658-2667[Abstract]

26. Keenan K. P., Ballam G. C., Dixit R., Soper K. A., Laroque P., Mattson B. A., Adams S. P., Coleman J. B. The effects of diet, overfeeding and moderate dietary restriction on Sprague-Dawley rat survival, disease and toxicology. J. Nutr. 1997;127:851S-856S

27. Kim M. J., Aiken J. M., Havighurst T., Hollander J., Ripple M. O., Weindruch R. Adult-onset energy restriction of rhesus monkeys attenuates oxidative stress-induced cytokine expression by peripheral blood mononuclear cells. J. Nutr. 1997;127:2293-2301[Abstract/Free Full Text]

28. Kremer J. M., Lawrence D. A., Petrillo G. F., Litts L. L., Mullaly P. M., Rynes R. L., Stocker R. P., Parhami N., Greenstein N. S., Fuchs B. R., Mathur A., Robinson D. R., Sperling R. I., Bigaouette J. Effects of high-dose fish oil on rheumatoid arthritis after stopping nonsteroidal antiinflammatory drugs. Arthritis Rheum 1995;38:1107-1114[Medline]

29. Lim B. O., Yamada K., Hung P., Watanabe T., Taniguchi S., Sugano M. Effect of n-3 polyunsaturated fatty acids and lectins on immunoglobulin production by spleen lymphocytes of Sprague-Dawley rats. Biosci. Biotechnol. Biochem. 1996;60(6):1025-1027[Medline]

30. Lim B. O., Yamada K., Nonaka M., Kuramoto Y., Hung P., Sugano M. Dietary fibers modulate indices of intestinal immune function in rats. J. Nutr. 1997;127:663-667[Abstract/Free Full Text]

31. Lim B. O., Yamada K., Yoshimura K., Watanabe T., Hung P., Taniguchi S., Sugano M. Free bile acids inhibit IgE production by mouse spleen lymphocytes stimulated by lipopolysaccharide and interleukins. Biosci. Biotechnol. Biochem. 1995;59(4):624-627[Medline]

32. Meydani S. N., Endres S., Woods M. M., Goldin B. R., Soo C., Morrill-Labrode A., Dinarello C. A., Gorbach S. L. Oral (n-3) fatty acid supplementation suppresses cytokine production and lymphocyte proliferation: comparison between young and older women. J. Nutr. 1991;121:547-555

33. Miller R. A. Aging and immune function. Int. Rev. Cytol. 1991;124:187-215(Review)[Medline]

34. Murasko D. M., Goodnewardene I. M. T-cell function in aging: mechanisms of decline. Annu. Rev. Gerontol. Geriatri. 1990;10:71-96

35. Reddy Avula C. P., Fernandes G. Modulation of antioxidant enzymes and apoptosis in mice by dietary lipids and treadmill exercise. J. Clin. Immunol. 1999;19:35-44[Medline]

36. Sampson H. A. Food Allergy. J. Am. Med. Assoc. 1997;278(22):1888-1894(Review)[Abstract/Free Full Text]

37. Stevens T. L., Bossie A., Sanders V. M., Fernandez-Botran R., Coffman R. L., Mosmann T. R., Vietta E. S. Regulation of antibody isoype secretion by subsets of antigen-specific helper T cells. Nature 1988;334(6179):255-258[Medline]

38. Sugano M., Tsujita A., Yamasaki M., Noguchi M., Yamada K. Conjugated linoleic acid modulates tissue levels of chemical mediators and immunoglobulins in rats. Lipids 1998;33:521-527[Medline]

39. Troyer D. A., Chandrasekar B., Barnes J. L., Fernandes G. Calorie restriction decreases platelet-derived growth factor (PDGF)-A and thrombin receptor mRNA expression in autoimmune murine lupus nephritis. Clin. Exp. Immunol. 1997;108:58-62[Medline]

40. Troyer D. A., Chandrasekar B., Thinnes T., Stone A., Loskutoff D. J., Fernandes G. Effects of energy intake on type I plasminogen activator inhibitor levels in glomeruli of lupus-prone B/W mice. Am. J. Pathol. 1995;146:111-120[Abstract]

41. Venkatraman J. T., Chandrasekar B., Kim J. D., Fernandes G. Effects of n-3 and n-6 fatty acids on the activities and expression of hepatic antioxidant enzymes in autoimmune-prone NZB x NZW F1 mice. Lipids 1994;29(8):561-568[Medline]

42. Weindruch R., Walford R. L. Dietary restriction: Effects on biological parameters. Weindruch R. eds. The Retardation of Aging and Disease by Dietary Restriction 1988:179-191 Charles C. Thomas Springfield, IL.

43. Yamada K., Hung P., Yoshimura K., Taniguchi S., Lim B. O., Sugano M. Effect of unsaturated fatty acids and antioxidants on immunoglobulin production by mesenteric lymph node lymphocytes of Sprague-Dawley rats. J. Biochem. 1996;120:138-144[Abstract/Free Full Text]

44. Yaqoob P., Newsholme E. A., Calder P. C. The effect of dietary lipid manipulation on rat lymphocyte subsets and proliferation. Immunol 1994;82:603-610[Medline]

This article has been cited by other articles:

				C. A. Jolly Dietary Restriction and Immune Function J. Nutr., August 1, 2004; 134(8): 1853 - 1856. [Abstract] [Full Text] [PDF]

				J. Bassaganya-Riera, R. M. Pogranichniy, S. C. Jobgen, P. G. Halbur, K.-J. Yoon, M. O'Shea, I. Mohede, and R. Hontecillas Conjugated Linoleic Acid Ameliorates Viral Infectivity in a Pig Model of Virally Induced Immunosuppression J. Nutr., October 1, 2003; 133(10): 3204 - 3214. [Abstract] [Full Text] [PDF]

				R. Hontecillas, M. J. Wannemeulher, D. R. Zimmerman, D. L. Hutto, J. H. Wilson, D. U. Ahn, and J. Bassaganya-Riera Nutritional Regulation of Porcine Bacterial-Induced Colitis by Conjugated Linoleic Acid J. Nutr., July 1, 2002; 132(7): 2019 - 2027. [Abstract] [Full Text] [PDF]

				C. A. Jolly, A. Muthukumar, C. P. R. Avula, D. Troyer, and G. Fernandes Life Span Is Prolonged in Food-Restricted Autoimmune-Prone (NZB NZW)F(1) Mice Fed a Diet Enriched with (n-3) Fatty Acids J. Nutr., October 1, 2001; 131(10): 2753 - 2760. [Abstract] [Full Text] [PDF]

				M Ehrenfeld, M Blank, Y Shoenfeld, and M Hidvegi AVEMAR (a new benzoquinone-containing natural product) administration interferes with the Th2 response in experimental SLE and promotes amelioration of the disease Lupus, September 1, 2001; 10(9): 622 - 627. [Abstract] [PDF]

				J. Bassaganya-Riera, R. Hontecillas, D. R. Zimmerman, and M. J. Wannemuehler Dietary Conjugated Linoleic Acid Modulates Phenotype and Effector Functions of Porcine CD8+ Lymphocytes J. Nutr., September 1, 2001; 131(9): 2370 - 2377. [Abstract] [Full Text] [PDF]

				A Leiba, H Amital, M E Gershwin, and Y Shoenfeld Diet and lupus Lupus, March 1, 2001; 10(3): 246 - 248. [Abstract] [PDF]

This Article Abstract Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lim, B. O. Articles by Fernandes, G. Search for Related Content PubMed PubMed Citation Articles by Lim, B. O. Articles by Fernandes, G.