Christopher A. Jolly

Calorie restriction decreases proinflammatory cytokines and polymeric Ig receptor expression in the submandibular glands of autoimmune prone (NZB x NZW)F1 mice

Calorie restriction or fish oil (enriched in n-3 fatty acids) supplementation ameliorates glomerulonephritis and Sj¨ogrens syndrome lesions in (NZB × NZW)F1(B/W) mice. Enhanced proinflammatory cytokine expression and deposition of immune complexes are the important pathological events in the development of Sj¨ogrens syndrome. In the present study, we have examined the effect of calorie restriction and fish oil supplementation on the expression of key inflammatory cytokines [gamma interferon (INF-γ), interleukin-10 (IL-10), and IL-12] and polymeric immunoglobulin (Ig) receptor (pIgR) (receptor for IgA and IgM) and the secretion of Ig in the submandibular glands (SMG) of B/W mice. Weanling B/W mice were fed either ad libitum (AL) or calorie restricted (CR) (40% less calories than AL) diet supplemented with 5% corn oil (CO) or 5% fish oil (FO) until 4 or 9 months of age. The SMGs were removed and a portion of the tissue used for semiquantitive determinations of IFN-γ, IL-10, IL-12 (p40), and pIgR mRNA. The remaining SMG tissue was fragmented and cultured for 7 days and the culture supernatants assayed for IgA, IgM, and IgG2a levels by enzyme-linked immunosorbent assay. Results revealed a significant increase in the expression of IFN-γ, IL-10, and IL-12 mRNA with age in AL fed mice, whereas CR fed mice maintained their levels to near those seen in young animals regardless of the dietary fat. PIgR mRNA expression also remained unaltered in CR animals irrespective of age and dietary fat, while it was found significantly increased in AL fed mice. CR significantly inhibited the elevated levels of IgA and IgG2a seen in aged mice. Interestingly, CR also influenced the Ig level in young animals. In summary, these results indicate that amelioration of autoimmune disease by CR in B/W mice is possibly mediated by the lowered mRNA expression of IFN-γ, IL-10, IL-12, and pIgR and the reduced Ig secretion.

Diet modulates Th-1 and Th-2 cytokine production in the peripheral blood of lupus-prone mice

Calorie restriction or fish oil extends life span. To investigate the potential mechanism(s) involved, young (4-month) and old (9-month) NZB × NZW(F1) mice were fed 5% (w/w) corn oil (CA) or fish oil (FA) ad libitum or 40% restricted (CR and FR, respectively). Peripheral blood T-lymphocytes were analyzed for Th-1 (IL-2, IFN-γrpar; and Th-2 (IL-5, IL-10) production. CR and FA partially blunted while FR completely abolished the decline in aged CD4+ T lymphocytes. In contrast, both CR and FR abolished the decline in CD8+ T lymphocytes with age, while FA had no effect. In aged mice, both CR and FR blunted the increase in Th-1 (IL-2, IFN-γrpar; cytokine production, while FA was only partially effective. Only FR completely blunted the age-related increase in Th-2 (IL-5, IL-10) cytokine production. These data suggest that FR delays the onset of autoimmune kidney disease by suppressing both Th-1 and Th-2 cytokine production.

THE EFFECTS OF DIETARY LIPIDS ON GENE EXPRESSION AND APOPTOSIS

The beneficial effects of dietary FO with respect to autoimmune disease, CVD and some types of cancer are well established. Studies conducted over the last 10-15 years have established the potent effects of FO on gene expression in the previously mentioned diseases. The effects of dietary FO appear to be selective in nature, with the expression of individual genes simultaneously being increased, decreased or completely unaffected. In order to elucidate the molecular mechanism(s) involved, recent studies have focused on analysing the effects of the long-chain polyunsaturated n-3 fatty acids EPA and DHA which are highly enriched in FO and thought to be the primary mediators of its biological activity. Indeed, it has been found that EPA and DHA appear to both directly and indirectly modulate gene expression in vivo, depending on the gene examined. The direct effects of EPA and DHA are most probably mediated by their ability to bind to positive and/or negative regulatory transcription factors, while the indirect effects appear to be mediated through alterations in the generation of intracellular lipid second messengers (e.g. diacylglycerol and ceramide). Future studies need to be focused on further elucidation of the inter- and intracellular signalling events mediated by dietary n-3 fatty acids. Understanding the molecular mechanism(s) modified by dietary FO will ultimately lead to improved dietary strategies to aid in the prevention of autoimmune disease, CVD and/or certain types of cancer.

Calorie restriction modulates Th-1 and Th-2 cytokine-induced immunoglobulin secretion in young and old C57BL/6 cultured submandibular glands

Immunoglobulin production by the salivary gland plays an important role in oral and upper respiratory tract immunity. Age and/or disease may compromise salivary gland function. In order to gain insight into the role of calorie restriction (CR) on immunoglobulin (Ig) production, we determined the effect of ad libitum (AL) feeding and CR in young (3 months) and old (18–24 months) C57BL/6 mouse submandibular glands (SM). The SM tissues were fragmented and cultured in the absence (control) or presence of either Th-1 cytokines, such as interleukin-2 (IL-2) and interferon-γ (IFN-γ), or Th-2 cytokines, e.g. IL-4 and IL-5, for seven days. Culture supernatants were then analyzed for immunoglobulin A (IgA), IgM, and IgG2a levels by ELISA. Aging increased basal (control) IgA and IgM production by 3.1- and 3.7-fold, respectively, in AL mice. CR prevented the age-dependent rise of both IgA and IgM, maintaining levels equal to those of young AL mice. Interestingly, age resulted in a decrease of Th-1 cytokine-induced IgA and IgM, and increased IgG2a secretion in AL mice, while Th-2 cytokines did not appear to have an age effect. In general, CR suppressed Ig production induced by both Th-1 and Th-2 cytokines in young mice. In contrast, CR in old mice resulted in enhanced IgA and IgM production to levels similar to those in their young counterparts, while IgG2a was predominantly suppressed by Th-1 and not Th-2 cytokines. The data presented herein show, for the first time, the ability of CR to offset age-induced changes in submandibular gland Ig production, which may play a role in maintaining mucosal immune function, including proper oral health.

Protein-Energy Malnutrition and Infectious Disease

The immune system plays a leading role in fighting off the constant bombardment of our bodies by invading pathogenic organisms, such as bacteria, fungi, viruses, toxins, and allergic compounds. Also, the immune system is intimately linked to the quality and quantity of nutrient intake (1,2). The host is defense-regulated and maintained by two branches of the immune system. One is cell-mediated immunity, carried out primarily by the CD4+ Th-1 and CD8+ T lymphocytes, which plays a pivotal role in cytotoxic responses against malignant cells and cells infected with intracellular bacteria and viruses. The other branch is humoral (antibody-mediated) immunity, of which the B lymphocyte plays a dominant role, to ward off and/or destroy extracellular pathogens. The T lymphocytes may also influence humoral immunity, as the Th-2 CD4+ T lymphocyte subset secretes a wide variety of antibody-inducing cytokines. The Th-1 CD4+ T lymphocyte produces primarily interleukin-2 (IL-2) and interferon gamma (IFN-y) and the Th-2 population produces primarily IL-4, -5, -6, and -10. Antibody isotype production (IgA, IgE, IgG, etc.) is dependent on the kind of cytokine produced to fight invading antigen assaults. Unfortunately, this vast array of defense mechanisms is not impregnable. Primary causes of a breakdown in these lines of defense (i.e., serious breakdown of our immune system) is either chronic or acute under nutrition and malnutrition.

Reduced food consumption increases water intake and modulates renal Aquaporin-1 and-2 expression in autoimmune prone mice

Aquaporin-1(AQP1) and AQP2 are members of the aquaporin family of cell membrane water channel transport proteins and have been implicated in the regulation of renal water excretion. We have previously shown that calorie restriction (CR) relative to ad libitum (AL) feeding extends lifespan and delays the onset of autoimmune kidney disease in lupus-prone (NZBxNZW)F1 (B/W) mice. To determine if AQP1 and/or AQP2 expression is influenced by CR, mice were fed an AL or CR (40% less food) diet until 4 (young) or 9 (old) months of age when mice were sacrificed. Kidneys were removed and the expression of AQP1 and AQP2 was determined at the protein and mRNA levels using western blotting and RT-PCR respectively. While age did not significantly increase AQP1 expression in the AL groups, CR did increase both the protein (1.4-fold) and mRNA (2.4-fold) levels. In old mice, AQP1 expression was higher (1.8-fold) in CR compared to the AL group while CR had no effect in young mice. In contrast, AQP2 showed an age related decrease (55%) in the AL groups and an increase in the protein (8.4-fold) and mRNA (1.7-fold) levels in the CR groups. Relative to AL, CR decreased AQP2 expression at the protein (90%) and mRNA (50%) levels in the young mice while an increase at the protein (2.9-fold) and mRNA (1.9-fold) levels was evident in the old mice. Interestingly, a significant increase in water intake per gram body weight was found in both young and old CR fed mice when compared to their AL counterparts which may contribute to the prevention of autoimmune disease with age and differences in longevity. These data show, for the first time, significant age and diet influences in renal AQP1 and AQP2 expression at both protein and mRNA levels in lupus-prone mice.