A fetal sheep liver extract reverses age-related increments in spontaneous and induced cytokine production by indirect environmental effects.

BALB/c, DBA/2 and C57BL/6 mice of different ages (ranging from 8 to 110 weeks of age) were used as spleen cell donors to assay cytokine production from ConA activated spleen and Peyer's Patch (PP) lymphocytes. As reported in an earlier publication, there was an age-related decline in IL-2 production in all strains, with a general increase in IL-4 and IL-10 production with age, this being particularly marked for PP cell preparations. Similar conclusions were reached from independent analysis of CD44hi and CD44lo cell populations in these groups (memory vs. naive cells, respectively). Interestingly, IL-6 production was dramatically increased (some 4-5-fold in the different strains) and significantly increased levels of IL-6 were detected in the serum of aged mice. A previously described sheep fetal liver extract was able to reverse, to varying degrees, these cytokine changes associated with aging. Interestingly, when cells from aged mice were adoptively transferred to lethally irradiated young (8 week) recipients, the cytokine production phenotype of cells harvested from recipient mice 3 weeks later was that of the aged donor, unless recipients were treated continually with extract. Treatment of the donor alone produced minimal changes in cytokine production 3 weeks following adoptive transfer. The effect of extract was reversed in treated aged mice by concomitant daily intravenous infusion of the competitive inhibitor of nitric oxide synthesis (NG-monomethyl-L-arginine (NMMA)), which also decreased the increased serum nitrate levels in mice treated with extract. Our data suggest an important role for reactive nitrogen products, themselves induced by fetal liver extract, in age-associated changes in cytokine production.

An antibody specific for interleukin-6 reverses age-associated changes in spontaneous and induced cytokine production in mice.

A number of quantitative and qualitative changes in the pattern of cytokine production have been reported to accompany the process of ageing in laboratory animals and in human populations, including an increase in serum levels of interleukin-1 (IL-1) and IL-6, as well as increased concanavalin A (ConA)-stimulated production of IL-4, IL-10 and transforming growth factor-beta (TGF-beta), and decreased production of IL-2 from cultured spleen cells. Increased IL-1 and IL-6 production is a feature of splenic adherent cells and peritoneal exudate cells taken from aged mice and stimulated with lipopolysaccharide in vitro. We have asked whether the altered production of lymphocyte-derived cytokines (IL-4, IL-2, TGF-beta) is itself a function of a primary alteration in IL-1/IL-6 production (from macrophage/monocytes) by infusing monoclonal antibodies to these cytokines prior to harvesting cells from aged mice and stimulating the cells in vitro. Anti-IL-6, but not anti-IL-1, reversed the age-associated alteration in lymphocyte cytokine production. The general pattern of cytokine production in aged mice is of a type-2 cytokine type, and thus these data are consistent with the idea that increased production of IL-6 in aged animals is causally implicated in this age-associated polarization to type-2 cytokine production.

Dietary fat composition and age affect synaptosomal and retinal phospholipid fatty acid composition in C57BL/6 mice.

The purpose of this study was (i) to determine whether dietary fat-induced differences in neural and retinal membranes occur when dietary fat treatment is implemented in aged animals and (ii) to characterize the effect of long-term differences in dietary fat on neural and retinal membrane composition. For the first objective, young (six-week-old) and old (95-week-old) mice were randomly assigned to beef tallow (TAL) or soybean oil (SBO) diets for eight weeks. For the second objective, young (four-week-old) mice consumed either TAL or SBO diets for 99 weeks. Young and old mice challenged with a change in dietary fat for an eight-week period showed both diet and age effects on neural and retinal phospholipid fatty acid composition (P < 0.05). In addition, significant diet by age interactions were evident. In mice that consumed TAL and SBO diets throughout their life, only retinal phosphatidylethanolamine (PE) 18:2n-6 and neural phosphatidylserine 22:5n-6, PE 18:2n-6 and phosphatidylcholine 18:2n-6 differed between dietary treatments (P < 0.05). Neither the unsaturation index nor the n-6/n-3 ratio was affected by diet. Neural and retinal phospholipid fatty acid composition were responsive to changes in dietary fat even when the treatment was implemented beyond developmental or post-weanling stages. In contrast, when mice consumed TAL or SBO diets throughout their life, fewer differences in phospholipid fatty acid composition were detected, suggesting that the effect of the dietary treatment was mitigated by aging.

Aging. Polymorphism, compartmentalization and environmental impact.

The proportion of elderly in our population is steadily increasing and so is the need to provide sophisticated health care. We must intensify research which provides results, leading to the design of preventive medicine, before the increased proportion of aged causes a crisis in our health care and social systems. The potential impact of such research represents the best and most cost-effective means of preparing for the future, and providing directions for a better quality of life with reduced chronic and debilitating illness for the elderly. Indeed, prevention appears to be the only approach able to lower the enormous economic burden of the cost of geriatric medicine [1,2]. There are many precedents in medical research for preventive measures being much more cost-effective than therapeutic means: one of them is immunization against poliomyelitis as an alternative to development of improved models of iron lungs.

Age-related changes in interleukin production in BALB/cNNia and SJL/J mice and their modification after administration of foreign macromolecules.

In vitro production of Interleukin-2, -3, -4 and -10 by activated lymphocytes of BALB/cNNia and SJL/J was studied. While IL-2 production in BALB/c mice remains constant throughout the life span of the animals (8-113 weeks), an increase in production from stimulated SJL cells was seen. Age-related increases in IL-3 and IL-4 production occur between young and middle age (8-60 weeks) in both strains. Some organ differences in quantity of lymphokine produced were seen; the direction of age-related changes was the same for lymphocytes of spleen and MLN. The exceptional feature of BALB/cNNia was the relative stabilization of the levels of interleukin production, as animals approach old age. BALB/cNNia and SJL, which are at the two opposite extremes of lifespan, differ also in their response to molecular interventions: in BALB/cNNia fetal sheep liver extract and hemocyanin increase the output of interleukins. This is in striking contrast to the effects observed in older SJL mice in which the extract reduced the output of IL-3 and IL-4 by old animals, whereas hemocyanin increased the output of IL-2 and IL-3 at all ages tested.

Age-related changes in hepatic and splenic insulin receptors and serum insulin and glucose levels in inbred mice.

Inbred mice of strains A/J, DBA/1J, and SJL/J were housed and aged in our animal colony, and parameters of carbohydrate metabolism were assessed at various ages. The patterns of age-related change were both organ- and strain-specific. Age-related changes in two of the strains were associated with relative carbohydrate intolerance. Common to all three strains was a biphasic pattern of change in hepatic insulin receptor number, with a decrease in early life and a return to earlier levels late in life. In both A/J and DBA/1J mice, there was a sharp increase in serum insulin level (twofold to 9.7-fold) that corresponded to the decrease in hepatic insulin receptors and was associated with hyperglycemia; no significant change in serum insulin or glucose levels was seen in SJL/J mice, despite a similar biphasic pattern in hepatic insulin receptor concentration. Age-related changes in splenic insulin receptors resembled changes in the liver in A/J and SJL/J mice, ie, there were synchronous biphasic age-related patterns. This was not the case in the spleens of DBA/1J mice, in which we did not observe age-related changes. There was no change in insulin receptor affinity with age, nor was there any difference in affinity between tissues or mouse strains. The pattern of change in hepatic insulin receptors and serum insulin levels was more complex than has been previously recognized. We do not know the mechanisms responsible for this complex pattern, but it must involve at least two discrete age-related events.(ABSTRACT TRUNCATED AT 250 WORDS)

Polymorphism of age-related changes in the antibody response to the hapten phosphorylcholine.

Aging is accompanied by changes in the immune system that occur at different levels and at different periods of time. We have studied age-related changes in isotype and idiotype of the antibody response to hapten phosphorylcholine (PC) in C57BL/6, and A mice and in the congenic MRL/Mp(-)+/+ and MRL/Mp-1pr/1pr strains. Three groups, representing young, middle and old age were immunized with PC-keyhole limpet hemocyanin. Total anti-PC antibody and the contribution of each isotype and of the T15 idiotype were assessed in the initial and late response. Some features of the antibody-response were similar in all the strains tested, e.g. the largest quantity of anti-PC antibody is formed in middle age and IgM is dominant in the initial response. However, remarkable differences occur in the isotype and idiotype predominance. Particularly, congenic MRL/Mp strains, prone to autoimmune disease, express the T15 idiotype only at low levels, even though IgM, which normally expresses this idiotype, is produced in large amounts. Furthermore, the late (memory) response of the MRL/Mp strains is dominated by IgG2b rather than IgG1, which is the predominant isotype in mice of long-lived strains. We conclude from these results that the number of T helper cells, involved in isotype regulation decreases with age and that there is a genetic variation, i.e., polymorphism in the ability to express T15-idiotype producing subtypes.

Age-related polymorphism of thymus subpopulations in inbred mice.

There are striking age-related changes in the demography of thymus lymphocytes, i.e. in thymus-cell subpopulations of BALB/c and SJL mice; these changes occur in the proportion of cells, identified by various markers, and by the membrane density of these markers. The thymuses of both strains undergo an age-related increase in the proportion of CD4+ CD8- cells and decrease in CD4+ CD8+ cells. Age-related changes in cells that are Pgp-1+ also show marked strain differences: Pgp-1+ cells increase in SJL, but not in BALB/c thymuses. In both strains, cells with high density of Pgp-1 appear in later life, though this is more marked in the thymus of SJL, which also shows a higher relative density at an advanced age, than do BALB/c mice. Furthermore, the per cent of cells with high density of Pgp-1 is larger in thymuses of SJL than in BALB/c mice. The percentage of CD45+ thymocytes remains unchanged, as animals age. Thymocyte-membrane densities of CD-45 undergo age-related increases in both SJL and BALB/c. The per cent of cells with high density of CD-45 is similar in both strains. Individual variations in relative size of subpopulations in SJL mice of the same age are greater in old than in young mice; this increase in heterogeneity is manifested by increase in standard deviation. Corresponding significant changes have not been observed in BALB/c or C57BL/6 mice. Thus, we have detected an intrastrain variation which may reflect age-related effects of the impact of stochastic events.

The origins and early years of IUIS.

The International Union of Immunological Societies (IUIS) was born of the global linkages between scientists involved in an exciting intellectual journey of discovery and search for a conceptual framework for regulation, specificity and memory of the immune response. Starting in the 1950s, a dozen of us discussed the need for an international organization; we founded it in 1969.

Influence of genotype on diet-induced changes in membrane phosphatidylcholine and phosphatidylethanolamine composition of splenocytes, liver nuclear envelope and liver mitochondria.

Inbred congenic mice of strains MRL/Mp-lpr/lpr (lpr/lpr) and MRL/Mp(-)+/+ (+/+) were fed nutritionally adequate semipurified diets containing 20% (w/w) fat and differing in linoleic acid content. Levels of linoleic acid (18:2n-6) and arachidonic acid (20:4n-6) in phospholipids of splenocytes, liver mitochondria and liver nuclear envelopes were determined. Membranes of lpr/lpr mice exhibited significantly lower levels of 18:2n-6 and 20:4n-6 in phospholipids compared with the +/+ strain. The high linoleic acid diet increased incorporation of 18:2n-6 and 20:4n-6 in most phospholipid fractions of these membranes. These observations indicate that genotype as well as dietary 18:2n-6 content significantly influenced incorporation of 18:2n-6 and 20:4n-6 into membrane phospholipids. The results also suggest that membrane compositional abnormalities found in the lpr/lpr mice, which develop lymphoma and age faster than +/+ mice, are not restricted to the immune system but also extend to other organs. Differences observed in phospholipid fatty acid composition in splenocytes and liver subcellular membranes for mice fed diets differing in linoleic acid content suggest that the early expression of the lpr gene resulting in progression of autoimmunity may be delayed through dietary manipulation.

Age-related decline in striatal D2-dopamine receptors is subject to 'economic correction'.

We have previously reported a correlation between the number (Bmax) of striatal D2-dopamine receptors in youth and the magnitude of the decrease to the 40th to 60th week of age. This correlation was observed in five inbred strains of mice, which differ over a 2-fold range in youthful Bmax. To determine the extent to which this correlation can predict changes in strains, other than those so far examined, we measured the binding of [3H]spiperone to striatal membranes in two additional strains (MRL/Mp-++ and DBA/2NNia), in one strain previously tested, C57BL/6, but now maintained in pathogen-free conditions (C57BL/6NNia), and in hybrids (C6D2F1) of C57BL/6NNia and DBA/2NNia mice. The results were as expected from the correlation observed with other strains; that is, the magnitude of decline in Bmax with age is correlated with the density of receptors in youth. To test the stability of these age-related changes, we examined the effect of feeding diets with high (4.5) and low (0.2) polyunsaturated to saturated fatty acid (P/S) ratios to SJL/J and MRL/Mp-++ mice and found both receptor density and affinity and their age-related change to be independent of the dietary P/S ratio. In conclusion, our data are consistent with 'economic correction', i.e. with a direct correlation between youthful quantity of striatal D2-dopamine receptors and subsequent extent of decrease with aging.

Polymorphism of age-related changes in interleukin (IL) production: differential changes of T helper subpopulations, synthesizing IL 2, IL 3 and IL 4.

Inbred mice were examined for strain differences in age-associated changes in the capacity to synthesize interleukin (IL), i.e. IL 2, IL 3 and IL 4 after stimulation with phorbol myristate acetate (PMA) and calcium ionophore (A23187). Production of IL 2 remains constant in A/J, DBA/1 and DBA/2 mice and decreases with age in one of the strains examined (C57BL/6J). Strain differences in age-associated change of synthesis did not show the relation between youthful synthetic capacity and magnitude of later decrease ("economic correction") which is observed in several other systems. This difference between different types of polymorphisms is attributed to an age-associated defect in intrinsic capacity to synthesize IL 2 which may occur in only one of the tested strains, C57BL/6J. In contrast to IL 2 production, the quantities of IL 3 and IL 4 increase progressively, with advancing age, in mice of the three strains tested. T cells from old mice contain a greater frequency of cells producing IL 3, than do those of young mice. In addition, synthesis of IL 3 is induced at a relatively lower concentration of PMA in cells from old animals and this may be a consequence of different signal requirements of the two subsets of the T helper cells, but also a change in intrinsic properties of these cells. Since IL 3 and IL 4 production, but not IL 2 production, increases with age, it is reasonable to conclude that this reflects an expansion of a T helper cell population which secretes IL 3 and IL 4, but not IL 2, presumably TH2.

Fine specificity and isotype expression of anti-PC antibodies in BALB/c, MRL/Mp-lpr/lpr, and -(+)/+ mice at different ages.

The MRL/Mp congenic mouse strains develop autoimmune disease with age. We have investigated age- and autoimmune-related changes in fine specificity, isotype spectra and T15 idiotype expression of the anti-phosphorylcholine (PC) response in BALB/c, MRL/Mp- + and -lpr congenic mice and in (BALB/c x MRL/Mp-lpr) F1 hybrids. Two groups of anti-PC antibodies with distinct fine specificity are elicited in the memory response. Group I antibodies recognize the PC moiety and express the T15 idiotype. Antibodies of group II are specific for phenyl-phosphorylcholine and are found predominantly in the memory response. In the MRL/Mp-lpr and - + strains only a minor population of antibodies expresses the T15 idiotype at all ages. However, a third group of antibodies was observed which binds to PC-coated proteins and to Diplococcus pneumoniae R-36A. This binding was not inhibited by PC-chloride and appeared mainly in the memory response at old age. The isotype distribution among anti-PC antibodies was similar in all strains analysed. In the initial response primarily mu, gamma 3 and gamma 1 isotypes were produced, while in the memory response gamma 1 was dominant. Thus autoimmune defects and ageing result in altered anti-PC antibody and idiotype profile, probably related to altered states in both the T- and B-cell compartments.

Strain polymorphism in progression of aging: changes in CD4, CD8 bearing subpopulations.

Polymorphism of age-related changes in CD4 (L3T4) and CD8 (Lyt-2) determinants of spleen and thymus cells was assessed by fluorescence-activated flow cytometry. Cells from mice ranging from 5 weeks to greater than 2 years of age were examined. There is little age-related change in the proportion of CD4+ CD8- splenocytes in A, C57BL/6, DBA/1, DBA/2, and SJL mice (slopes 0.04, 0.06, 0.08, 0.17 and 0.17, respectively, when age in weeks was plotted against % of positive cells). Changes in the composition of the thymus are much more profound: CD4+ CD8+ cells of SJL mice decrease from 70% to less than 10% as the animals age from 5 to 69 weeks (slope -1.03), and in DBA/2 mice from 5 to 110 weeks (slope -0.88). While this decrease in CD4+ CD8+ cells occurs, there is a compensatory increase in CD4+ CD8- and CD4- CD8+ cells; this is a shift in the relative proportion of subpopulations rather than an increase in absolute cell numbers of a particular subpopulation. In contrast to the age-related changes of SJL and DBA/2 mice, there is relatively little change in the proportion of CD4+ CD8+ thymus cells in mice of strains C57BL/6, DBA/1 and A (slopes -0.03, -0.14 and -0.15, respectively).

Age-related changes in contact photosensitivity differ among mouse strains.

There are differences among mouse strains in the age-related changes in reactivity to the contact photosensitizer tetrachlorosalicylanilide (TCSA). We found a tendency to lower reactions in older mice, with some strains showing declines from an early age (BALB/cJ, MRL/MpJ +/+, MRL/MpJ lpr/lpr and SJL/J). Others had increasing reactions until about 30-50 weeks of age before declining (DBA/1J, C3H/HeJ, and A/J) and one strain (C57BL/6J) had increased reactivity with age. There are also differences in the role of cyclophosphamide-sensitive T-suppressor cells in these age-related changes. In some mouse strains, BALB/cJ, C57BL/6J, A/J, DBA/1J and C3H/HeJ, age-related changes in reactivity to TCSA are independent of changes in cyclophosphamide-sensitive suppressor cells. In other strains, MRL/MpJ +/+, MRL/MpJ lpr/lpr and SJL/J, the development of cyclophosphamide-sensitive suppressor cells is responsible for the initial, though not later, stages of the age-related decline in reactivity.

Economic correction; quantity and age-related rate of change.

Correlation between the relative magnitude of activity or concentration in youth and the relative magnitude of age-related decrease occurs in several systems. We have observed this correlation in density of D2-dopamine receptors of the striatal membrane, activity of lymphocyte activated killer cells, relative density of CD8 on thymocytes, augmenting activity of T splenocytes and mRNA, coding for IL-1 in Langerhans cells. We have suggested that this correlation should be considered in the context of balanced investment in lifespan and reproductive efficiency, that it may be the result of feedback regulation and have designated it as "economic correction".

Dietary fat influences electric membrane properties of neurons in cell culture.

1. SJL/J mice were maintained on semipurified diets which differed in the ratio of polyunsaturated/saturated fatty acid content (P/S). Exposure was from conception and was maintained for periods ranging from 6 to 34 weeks. 2. Neural cell cultures were prepared from dorsal root ganglia (DRG). After 6 and 20 days of culture, neuronal electric membrane properties were determined quantitatively by intracellular recording. 3. A number of significant differences were observed for the two dietary conditions. DRG from mice on the low-P/s diet had an increase in the rate of fall of both phases of repolarization which, in conjunction with the reduced action potential overshoot, led to a reduced action potential duration. This shift to shorter-duration action potentials was accompanied by a shift to more monophasic falling phases. The low-P/S neurons also exhibited a decreased afterhyperpolarization, decreased specific membrane resistance, and decreased membrane electrical time constant compared to high-P/S neurons. 4. It was concluded that the P/S ratio in the diet can have a significant effect on the electric properties of neurons. The high-P/S neurons tended to have action potentials with biphasic repolarizations and longer durations. In contrast, the low-P/S neurons tended to have action potentials with monophasic repolarizations and shorter durations. Moreover, the known ionic dependence of these two types of action potentials suggested that the low-P/S diet resulted in action potentials with a more exclusive Na dependence, while the high-P/S diet resulted in action potentials with both Na and Ca dependence.

Immunosuppression after injection of Friend virus into C57BL/6 mice of different ages.

Injection with Friend virus (FV) causes immunosuppression in young and old C57BL/6 mice, i.e. it occurs whether or not the virus replicates very briefly or for a long period. There are only minor age-related differences in the extent of immunosuppression, except that suppression appears to persist somewhat longer in old than in young animals.