Disorganization of the splenic microanatomy in ageing mice.

The precise mechanisms responsible for immunosenescence still remain to be determined, however, considering the evidence that disruption of the organization of primary and secondary lymphoid organs results in immunodeficiency, we propose that this could be involved in the decline of immune responses with age. Therefore, we investigated the integrity of the splenic microarchitecture in mice of increasing age and its reorganization following immune challenge in young and old mice. Several differences in the anatomy of the spleen with age in both the immune and stromal cells were observed. There is an age-related increase in the overall size of the white pulp, which occurs primarily within the T-cell zone and is mirrored by the enlargement of the T-cell stromal area, concurrent to the distinct boundary between T cells and B cells becoming less defined in older mice. In conjunction, there appears to be a loss of marginal zone macrophages, which is accompanied by an accumulation of fibroblasts in the spleens from older animals. Furthermore, whereas the reorganization of the white pulp is resolved after several days following antigenic challenge in young animals, it remains perturbed in older subjects. All these age-related changes within the spleen could potentially contribute to the age-dependent deficiencies in functional immunity.

Immunosenescence: a product of the environment?

The major function of the immune system is to provide protection against pathogens, in order to prevent infections and potential death. However, with increasing age the immune system undergoes alterations culminating in a progressive deterioration in the ability to respond to infection and vaccination. The precise mechanisms associated with immunosenescence have not been fully elucidated although extensive analyses have suggested that intrinsic defects within immune cells are potentially involved. Despite the stromal niche playing a critical role in the development and activation of immune cells, the role of extrinsic factors within the microenvironment in immunosenescence is less well understood. Moreover, emerging evidence suggests that the aged microenvironment contributes significantly to the age-associated decline of immune function and additionally may offer a potential target for rejuvenating the immune system. Indeed, rejuvenation strategies which have targeted the thymic stromal microenvironment have proved to be successful in recovering thymic function in the aged.

It's not all equal: a multiphasic theory of thymic involution.

Regression of the thymus is a key feature of immunosenescence, which coincides with a decrease in T cell output and contributes to the restriction of the T cell repertoire in the elderly, leading to increased susceptibility to illness and disease. However, the mechanisms involved in thymic involution are still not fully known. Although, it is often believed that thymic involution occurs during the onset of puberty, increasing data suggests alterations to the thymus happen much earlier in life. Therefore, the changes in the thymus and subsequent thymic function may not just be an ageing phenomenon. In this article, we propose that there are several, non-linear, phases to thymic atrophy, which are regulated by different mechanisms, including the familiar age-dependent thymic involution and a much earlier growth-dependent thymic involution.

The origin and implication of thymic involution.

Age-related regression of the thymus is associated with a decline in naïve T cell output which is thought to contribute to the reduction in T cell diversity in older individuals that is partially responsible for an increase in susceptibility and severity of infections, cancers and autoimmune diseases. Thymic involution is one of the most dramatic and ubiquitous changes in the ageing immune system, but the precise regulators remain anonymous. However, a picture is emerging, implicating extrinsic and intrinsic factors that may contribute towards age-associated thymic involution. In this review we assess the role of the thymic microenvironment as a possible target of thymic involution, question whether thymocyte development in the aged thymus is functional and explore why the thymus involutes.

BTN1A1, the mammary gland butyrophilin, and BTN2A2 are both inhibitors of T cell activation.

Butyrophilin (BTN) genes encode a set of related proteins. Studies in mice have shown that one of these, BTN1A1, is required for milk lipid secretion in lactation, whereas butyrophilin-like 2 is a coinhibitor of T cell activation. To understand these disparate roles of BTNs, we first compared the expression and functions of mouse Btn1a1 and Btn2a2. Btn1a1 transcripts were not restricted to lactating mammary tissue but were also found in virgin mammary tissue and, interestingly, spleen and thymus. In confirmation of this, BTN1A1 protein was detected in thymic epithelial cells. By contrast, Btn2a2 transcripts and protein were broadly expressed. Cell surface BTN2A2 protein, such as the B7 family molecule programmed death ligand 1, was upregulated upon activation of T cells. We next examined the potential of both BTN1A1 and BTN2A2 to interact with T cells. Recombinant Fc fusion proteins of murine BTN2A2 and, surprisingly BTN1A1, bound to activated T cells, suggesting the presence of one or more receptors on these cells. Immobilized BTN-Fc fusion proteins, but not MOG-Fc protein, inhibited the proliferation of CD4 and CD8 T cells activated by anti-CD3. BTN1A1 and BTN2A2 also inhibited T cell metabolism, IL-2, and IFN-gamma secretion. Inhibition of proliferation was not abrogated by exogenous IL-2 but could be overcome following costimulation with high levels of anti-CD28 Ab. These data are consistent with a coinhibitory role for mouse BTNs, including BTN1A1, the BTN expressed in the lactating mammary gland and on milk lipid droplets.

An evolutionary perspective on the mechanisms of immunosenescence.

There is an accumulating body of evidence that a decline in immune function with age is common to most if not all vertebrates. For instance, age-associated thymic involution seems to occur in all species that possess a thymus, indicating that this process is evolutionary ancient and conserved. The precise mechanisms regulating immunosenescence remain to be resolved, but much of what we do know is consistent with modern evolutionary theory. In this review, we assess our current knowledge from an evolutionary perspective on the occurrence of immunosenescence, we show that life history trade-offs play a key role and we highlight the possible advantages of the age-related decline in thymic function.

Neuropeptides and thymic hormones in the Xenopus thymus.

T-cell development is characterised by a complex series of events in the thymus, which results in the development of self-restricted immunocompetent lymphocytes. We have previously reported the expression of neuropeptides in the thymus of various species, highlighting the evolutionary importance of neuroendocrine interactions in thymocyte development. Despite the many physiological and functional similarities in their immune systems, no study has addressed the importance of neuropeptides and thymic hormones in T-cell development in Xenopus. Immunohistochemical analysis revealed that the neuropeptides substance P, neuropeptide Y, somatostatin, calcitonin gene related peptide, and vasoactive intestinal polypeptide and the thymic hormones thymosin alpha1, thymosin beta4, and thymopoietin are found in the Xenopus thymus. This was further corroborated by RT-PCR. Furthermore, double staining revealed that neuropeptides and thymic hormones are coexpressed within the epithelial cell component of the thymus. These results show that neuropeptides and thymic hormones are expressed in the thymus of Xenopus, and suggest that they are likely to play a role in T-cell development.

Phenotypical and morphological changes in the thymic microenvironment from ageing mice.

The thymus is crucial for T-cell output and the age-associated involution of this organ, is thought to have a major impact in the decline in immunity that is seen in later life. The mechanism that underlines thymic involution is not known, however, we have evidence to suggest that this is may be due to changes in the thymic microenvironment. To further test this hypothesis, we quantified the in situ changes to markers that identify cortical and medullary thymic epithelial cells. This analysis revealed an age-dependent decline in cortical and medullary markers together with an increase in Notch and Delta expression, in older mice, as judged by immunohistochemistry. This was accompanied by alterations of the archetypal staining patterns and three dimensional analysis revealed changes in the morphology of the thymic microenvironment. These studies suggest that there are age-associated alterations in the thymic microenvironment, which may therefore play a role in thymic involution.

Is thymocyte development functional in the aged?

T cells are an integral part of a functional immune system with the majority being produced in the thymus. Of all the changes related to immunosenescence, regression of the thymus is considered one of the most universally recognised alterations. Despite the reduction of thymic size, there is evidence to suggest that T cell output is still present into old age, albeit much diminished; leading to the assumption that thymocyte development is normal. However, current data suggests that recent thymic emigrant from the aged thymus are functionally less responsive, giving rise to the possibility that the generation of naïve T cell may be intrinsically impaired in the elderly. In light of these findings we discuss the evidence that suggest aged T cells may be flawed even before exiting to the periphery and could contribute to the age-associated decline in immune function.

Architectural changes in the thymus of aging mice.

Age-associated thymic involution is one of the most dramatic and ubiquitous changes in the immune system, although the precise mechanisms involved still remain obscured. Several hypotheses have been proposed incorporating extrinsic and intrinsic factors, however, changes in the thymic microenvironment itself is one of the least investigated. We therefore decided to undertake a detailed histological examination of the aging thymus in order to elucidate possible mechanisms of thymic atrophy. This investigation provides insight into the changes within the murine thymus with age, demonstrating a new approach to quantify protein expressional differences while preserving the thymic architecture. There is a decline in expression of thymic epithelial cell-specific makers and an increase in fibroblast content in the aging mouse thymus. This is concurrent with a disorganization of the thymic compartments, a morphological transformation within the epithelial cells and alterations of their archetypal staining patterns. Furthermore, this is linked to a rise in apoptotic cells and the novel finding of increased senescence in the thymus of older mice that appears to be colocalized in the epithelial compartment. These changes within the thymic epithelial cells may be in part accountable for thymic atrophy and responsible for the decline in T-cell output.

Functional analysis of neuropeptides in avian thymocyte development.

The function of lymphoid organs and immune cells is often modulated by peptides and hormones produced by the neuroendocrine and immune systems. We have previously reported the intrathymic expression of neuropeptides in the thymus of different species and that neuropeptides can influence murine thymocyte development in vitro. To further explore the evolutionary nature of neuroendocrine interactions in the thymus, we identified the expression of calcitonin-gene-related peptide, neuropeptide Y, somatostatin (SOM), substance P and vasointestinal polypeptide, as well as their receptors on chicken thymic epithelial cells (TEC) and thymocytes by immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR). All the studied neuropeptides and their receptors were found to be expressed in both TEC and thymocytes, suggesting that intrathymic neuroendocrine interactions may take place within the avian thymus. In order to elucidate whether such interactions play a role in avian thymocyte development, neuropeptides and their antagonists were added to embryonic thymus organ cultures and found to influence chicken thymopoiesis. In particular, an antagonist of SOM increased the proportion of double-positive thymocytes, while SOM itself appeared to inhibit the early stages of thymocyte development. Taken together, these data provide further evidence to suggest that neuropeptides play a conserved role in vertebrate thymocyte development.

Immunosenescence: emerging challenges for an ageing population.

It is now becoming apparent that the immune system undergoes age-associated alterations, which accumulate to produce a progressive deterioration in the ability to respond to infections and to develop immunity after vaccination, both of which are associated with a higher mortality rate in the elderly. Immunosenescence, defined as the changes in the immune system associated with age, has been gathering interest in the scientific and health-care sectors alike. The rise in its recognition is both pertinent and timely given the increasing average age and the corresponding failure to increase healthy life expectancy. This review attempts to highlight the age-dependent defects in the innate and adaptive immune systems. While discussing the mechanisms that contribute to immunosenescence, with emphasis on the extrinsic factors, particular attention will be focused on thymic involution. Finally, we illuminate potential therapies that could be employed to help us live a longer, fuller and healthier life.

Evolutionary conservation of neuropeptide expression in the thymus of different species.

Evidence suggests that the immune and neuroendocrine systems cross talk by sharing ligands and receptors. Hormones and neuropeptides produced by the neuroendocrine system often modulate the function of lymphoid organs and immune cells. We have previously reported the intrathymic expression of somatostatin (SOM) in the mouse and that several neuropeptides, most notably calcitonin-gene-related peptide (CGRP), neuropeptide Y (NPY), SOM and substance P (SP), can modulate thymocyte development. However, little is known about the intrathymic expression of these neuropeptides either in the mouse or in other species. Moreover, a comparative analysis of the expression of these molecules would highlight the evolutionary importance of intrathymic neuroendocrine interactions in T-cell development. We have studied the expression of different neuropeptides in the thymus of zebrafish, Xenopus, avians, rodent, porcine, equine and human by immunohistochemistry and reverse transcription-polymerase chain reaction. We found that CGRP, NPY, SOM, SP and vasointestinal polypeptide (VIP) are expressed in the thymus of all species investigated. The thymic location of many of these neuropeptides was conserved and appears to be within the stromal compartments. Interestingly, in the avian thymus the expression of CGRP, SOM and SP appears to change depending on the age of the tissue. These findings suggest that neuropeptides may play an important role in T-cell development and provide further evidence of cross talk between the immune and neuroendocrine systems.