Age-related involution and terminal disorganization of the human thymus

The terminal involution pattern of the human thymus was studied based on autopsy cases (both sexes, age range 63-91 years). Large sections through the entire thymic fat body were examined with the help of both conventional histological and immunohistochemical techniques. The findings demonstrate that thymic atrophy in old humans (a) goes far beyond the degree of involution observed in small rodents; (b) results in a system of thin, branching, in part interrupted, non-keratinizing epithelial plates containing no typical Hassall bodies; (c) concerns all components of the thymus except fat tissue which progressively replaces original thymic structures; and (d) involves various types of disorganization of individual lobules with T and B lymphocytes often located outside rather than within epithelial remnants. Effects of low-level radiation on this final regression of the human thymus are unknown.     

Morphometrical analysis of the thymus from mice submitted to low temperature

Thymic involution is a slow physiological process than can be accelerated by some pathological or experimental conditions. In this work we kept male mice under low temperatures in order to observe whether or not thymic involution would be promoted. For this purpose, we carried out a histometrical analysis of the thymus and observed that there was a decrease in both cortical and medullary regions of the thymic lobules. These changes paralleled a loss in the absolute and relative weights of the thymus. It was concluded that low temperatures induce thymic involution even in nonhibernating animals.     

Analysis of the mitotic activity of the lymphoid cells of the thymus in relation to age]

The mitotic activity of lymphoid cells of the thymus was studied separately in the subcapsule zone of the cortical substance, cortical substance and medullary substance of newborn, 21-day, 3-, 4-month and 12-month-old mice. In addition to counting mitoses (MA%) histograms of the organ lobes were drawn with exact localization of mitoses of dividing cells. The greatest index of mitotically dividing cells was noted in the subcapsule zone of newborns and 21-day-old mice (5,2+/-0,9 and 4,7+/-0,5). After sex maturation the mitotic activity considerably decreased but in 3-, 4- and 12-month-old mice, the mitotic coefficient of lymphoid cells also retained rather high level (2,6+/-0,3 and 2,1+/-0,2) though processes of involution took place in their thymus.     

Occurrence of GABA-transaminase in the thymus gland of juvenile and aged rats

The occurrence and distribution of GABA-transaminase (GABA-t) activity were examined in the thymus of juvenile, adult and aged rats, using enzyme-histochemical and biochemical methods. Quantitative image analysis showed that specific GABA-t reactivity was localized in the wall of the arteries and, to a lesser extent, to the veins. Only a low activity could be observed in association with the subcapsular and medullary part of the parenchyma of the thymus. Many structures resembling nerve fibers also showed low positive reactivity. Biochemical results gave the following decreasing order of GABA-t activity: arteries, veins, whole thymus and parenchyma. Histoenzymatic staining and related values of quantitative analysis of images, are in agreement with the biochemical results; moreover, they demonstrated that the intensity of histoenzymatic staining for GABA-t in thymus of rats strongly decreases with age. GABA-t in thymic tissue is concentrated in blood vessels and particularly in the arteries. Therefore, our findings do not support the earlier assumptions that GABA-t is exclusively concentrated in cerebral vessels. Moreover, the decrease of GABA-t activity during age in thymic tissues may be related to the decrease of thymic microvessels as consequence of the thymic involution. On the contrary, the thymic macrovessels show an elevated staining in all ages with an apparent increase due to the thymic involution.     

Changes in the expression of the nerve growth factor receptors TrkA and p75LNGR in the rat thymus with ageing and increased nerve growth factor plasma levels

The nerve growth factor (NGF) receptors p75LNGR and TrkA are expressed by thymic epithelial cells. Presumably, the NGF-TrkA system is involved in the paracrine communication between thymic epithelial cells and thymocytes, whereas the functional role of p75LNGR is still unknown. The thymus of vertebrates undergoes age-related changes that in part depend on hormonal factors. In order to find out whether thymic epithelial cells are responsive to NGF during the whole lifespan of the rat, we studied NGF receptor expression in the thymus from birth to 2 years of age, using immunohistochemistry. Furthermore, to evaluate whether increased plasma levels of NGF affected the ageing process, either NGF or 4-methylcatechol (4MC), an inductor of NGF synthesis, was administered. Both TrkA and p75LNGR were expressed by a subpopulation of thymic epithelial cells during the whole age range studied and their expression peaked at around 3 months. TrkA was primarily found in subcortical and medullary epithelial cells, whereas p75LNGR was seen in a subpopulation of medullary cells. Cortical epithelial cells, neural crest-derived cells, other stromal cells and thymocytes were not immunoreactive for NGF receptors. Neither the administration of NGF nor the increased NGF plasma levels obtained after 4MC treatment seemed to affect the ageing of the thymus as assessed by morphological and immunohistochemical criteria, but this increase in NGF levels did produce a shift in the expression of p75LNGR from epithelial cells to ED1-positive macrophages in animals of 6 months and older. Present results indicate that the expression of p75LNGR and TrkA in the rat thymus undergoes age-dependent changes that parallel those of epithelial cells. NGF could therefore be important for thymus homeostasis, possibly acting on epithelial cells. Nevertheless, NGF did not seem to be able to prevent the involution of this organ, although it produced a switch in the expression of p75LNGR, the significance of which remains to be established.     

Status of the rat thymus after antenatal antigenic stimulation

Morphological reactions of the thymus to penetration of foreign antigens into the organism should be further investigated. In the work presented, morphometric, histological and histochemical methods have been applied to study specific thymic reactions of the rat fetuses to antenatal antigenic stimulation. The antigenic stimulation of the fetuses results in an increased thymic function. The changes observed in it resemble, in general, an accidental involution. A specific feature of the fetal thymic reaction is an essential enlargement of the medullary substance volume and appearance of the thymic bodies (Hassall's bodies) that demonstrates an accelerated maturation of the thymus.     

Neuropeptides of human thymus in normal and pathological conditions

Human thymus of healthy subjects and patients affected by thymoma-associated Myastenia Gravis were studied in order to visualize and compare the morphological distributive pattern of four neuropeptides: vasoactive intestinal peptide, substance P, neuropeptide Y, and neurotensin. Based on our observations, we formulated hypotheses on their relations in neuro-immunomodulation under physiological and pathophysiological conditions. Immuno-histochemical staining for neuropeptides was performed and morphological and morphometrical analyses were conducted on healthy and diseased thymus. In normal thymus, a specific distributive pattern was observed for the several neuropeptide-positive nerves in different thymus lobular zones. In particular substance P-positive fibers were observed in subcapsular zone, specifically located into parenchyma, where they represent the almost total amount of fibers; neurotensin-positive fibers were observed primarily located in parenchyma than perivascular site of several thymus lobular zones, and more abundant the cortico-medullary and medullary zones. Instead VIP- and NPY-positive fibers were widely distributed in perivascular and parenchymal sites of several thymus lobular zones. In thymoma, the distribution of neuropeptide-positive fibers was quantitatively reduced, while cells immunopositive to VIP and substance P were quantitatively increased and dispersed. Observation of the perivascular and parenchymal distribution of the analyzed neuropeptides suggests evidence that a regulatory function is performed by nerves and cells that secrete neuropeptide into the thymus. The alteration of neuropeptide patterns in thymoma suggests that these neurotransmitters play a role in autoimmune diseases such as Myastenia Gravis.     

Involution of the rat thymus in experimentally induced hypothyroidism

The thymus, as part of the immune-neuroendocrine axis, is greatly influenced by factors from most endocrine glands, especially the thyroid. Antithyroid drugs (carbimazole and methimazole) were used to induce hypothyroidism in rats. Histological and ultrastructural examination of the thymus showed progressive thymic involution after 4 weeks of drug treatment to the end of observations (7 weeks). The involution was characterised by increased thymocyte apoptosis and thymocyte phagocytosis by macrophages. This resulted in thymocyte depopulation, increases in numbers of interdigitating cells, alterations to mainly subcapsular and medullary epithelial cells, an apparent increase of mast cells and collagen in the capsule and septa, and increased numbers of B cells and plasma cells. Lymphoid cells immuno-reactive with MRC OX12 (which detects B cells) were observed within blood vessel walls, suggesting that they may have been moving in and out of the thymus. The administration of drugs causing hypothyroidism, therefore, also caused marked involution of the thymus.     

Morphologic changes in the thymus and lymphoid nodules of the appendix of the white rat subjected to increasing physical loads

By means of histological and morphometrical methods the thymus and appendage have been studied in male white rats subjected to a dosed physical loading (swimming). The physical loading is accompanied with essential changes in structure and cell composition of the immunogenic organs. The rate and character of the changes depend on the adaptability level of the animal's organism to the physical loading. At adaptation to the loading, the process of age involution of the thymus decelerates, amount of lymphoid nodules in the appendage increases, comparing the control parameters, contents of lymphocytes noticeably increase in all zones of the lymphoid nodules. When adaptation to the physical loading is not sufficient, the rate of the thymus involution sharply increases, while in the appendage the number of the lymphoid nodules decreases. However, in some animals at a sharp involution of the thymus, the changes in the appendage do not differ from the control ones.     

A morphometrical study of human fetal thymus

A morphometrical study was made of the thymus lobules in the cortical and medullary layers of male and female fetuses, 16 to 31 weeks old, divided into the age groups 16 to 19, 20 to 23, 24 to 27 and 28 to 31 weeks of intrauterine life. Results were correlated with body and thymus weights; the lobular, cortical and medullary volumes, as obtained using a histometrical method, were proportional to thymus, but not body weight increase. No differences were noted in the volumes and weights investigated as a function of fetal sex.     

Histology and histochemistry of the seal ovaries and the age-related changes in the ovaries of the Greenland seal]

Histological and histochemical methods were used to study the ovaries of Greenland seal (Pagophoca groenlandica) from birth-time up to 30 years of age and mature females of Phoca vitulina and Erignathus barbatus. The ovary of the new-born Greenland seal has fetal medullary substance which is a provisory endocrinous gland producing not only sex hormones but also corticosteron. In other species of seals the intestinal cells of the medullary substance are the equivalent of this gland. Within 3-4 weeks after birth the reduction of the fetal medullary substance is completed, it is substituted by the connective tissue and the ovary acquires its typical structure. The rest of the fetal medullary substance is in the depth of the cortex and near the infundibulum of the ovary as lipofuscincontaining cells. When the maturation period approaches, the process of the follicle atresia regularly changes: the epithelium dies quicker, and the multiplication of intestinal cells increases. The ovaries of seals are rich in interstitial cells. Their amount cyclically changes. The cells producing steroid hormones always well hydrolize AS naphthyl-phosphates, the reaction with glycerophosphate is more variable. The connective tissue is poor in acid mucopolysaccharides, its amorphous substance in the ovary cortex is rich in protein. Senile changes of the ovary are noticed in the seal beginning from 20 years of age.     

Substance P innervation of the rat thymus

Immunohistochemistry for substance P (SP) in the rat thymus revealed fine varicose neural profiles in specific regions of the thymus. Thymic SP innervation was abundant within the capsule and interlobular septa. The majority of SP+ nerve fibers within the septa were free of vascular association, although some fibers were associated with the vasculature deep within the septa. SP+ nerve fibers entered the thymic cortex from the septa and distributed among cortical thymocytes and mast cells. Along the corticomedullary junction, SP+ nerve fibers were found in association with the vasculature. The medullary region of the thymus received only a sparse innervation of SP+ fibers. In addition, SP+ nerve fibers coursed adjacent to OX-8+ cells and mast cells in the extrathymic connective tissue surrounding the thymus. The present study provides evidence that SP is present in nerve fibers in the thymus, and may be available to interact with thymocytes, mast calls, and other cells in the thymus, and affect their development and function.     

Changes in the structure and cell composition of the thymus as a result of dosed physical loads

By means of histological and morphometrical methods normal age involution of the thymus has been studied, as well as its changes under conditions of dosed physical loadings. The experiment has been performed in 92 non-inbred white male rats. At adaptation of the organism to the loadings, involution of the gland decelerates, and at an insufficient adaptation--accelerates. This is, probably, the cause of decreasing protective forces of the organism.     

Ultrastructural changes in the thymus of the turtle Mauremys caspica in relation to the seasonal cycle

Summary Changes in the ultrastructure of the thymus of the turtle Mauremys caspica, with special reference to its non-lymphoid components, were studied in relation to the seasonal cycle. The thymic cortex contains framework-forming epithelial-reticular cells and free macrophages, while the medulla includes, in addition, mature and presumptive pro-interdigitating cells. The ultrastructural features of these cells are generally similar to those described for non-lymphoid components of the mammalian thymus. The turtle thymus undergoes cortical involution in spring, with recovery periods in May–June and during autumn. A moderate involution occurs in winter. At the beginning of spring, cortical (but not medullary) epithelial-reticular cells show degenerative changes, probably related to high levels of circulating testosterone. In spring and autumn, mature interdigitating cells are absent, but macrophages, monocytes, and pro-interdigitating cells are found. During May–June, the cortical epithelial-reticular population recovers and macrophages, monocytes, and interdigitating cells are actively phagocytic. In summer, the epithelial-reticular cells in both cortex and medulla display normal ultrastructural features; mature and immature interdigitating cells are absent and some macrophages are detected occasionally. The results suggest that non-lymphoid components of the reptilian thymus can play a role in governing T-lymphocyte differentiation, and that the thymic cortex and medulla exhibit different cycles of seasonal activity.     

Cellular composition and ultrastructure of the thymus of the newborn mouse after immunization of pregnant females with homologous brain antigens

Immunization is performed with 20% of water-saline extract of medulla oblongata on the 6th, 8th, 10th days of pregnancy (1 g per 200 g body mass). In the thymus cortical substance of newborn rats no statistically significant difference in content of small lympocytes, lymphoblasts and large lymphocytes, mitotically deviding cells is revealed as compared to the normal. The part of middle lymphocytes decreases up to 5.7 +/- 0.7% (control--10.0 +/- 1.7%). The content of distroying cells and fagocytic macrophages is increasing. In cytoplasm of one macrophage several fagocyted degenerating cells with pyknotic nuclei and destructively altered organells are often present. In the interlobular connective tissue an increased amount of degenerating forms of mast cells is noted. In the thymus medullary substance small lymphocytes are growing in number. Certain changes in vessels of the microcirculatory bed are revealed.     

Age-related characteristics of the myeloarchitectonics of the peripheral nerves

By means of macromicroscopy and microscopy myeloarchitectonics of morphologically and functionally different somatic and visceral nerves (branches: mandibular nerve, cervical spinal nerves, inferior laryngeal nerve, hepatic plexus) have been studied in 11 age groups. During the prenatal period of ontogenesis asynchronism of myelogenesis is stated in various muscle branches of the nerves, dependent on formation of function in corresponding muscles and muscle groups. As demonstrate investigations on peculiarities of myelogenesis course in the somatic and visceral nerves studied, during the period of pre- and postnatal ontogenesis, its dynamics embraces three stages of myelogenesis, determined by G. B. Stovichek for visceral nerves: productive myelogenesis, stages of stabilization and involution. The stage of productive myelogenesis in the somatic nerves studied lasts up to the end of the adolescent period. Two phases are determined in it: the first lasts up to 2-3 years; the second--up to the end of the adolescent period and is characterized with a complete formation of the myelin fibers spectrum. In the visceral nerves studied increase of general amount of myelin fibers and their differentiation are completed simultaneously during the adolescent age. The stabilization stage of myeloarchitectonics of the nerves studied corresponds to the mature age (I and II periods) and the involution stage--to the elderly (and old) age.     

Developmental changes in the cellular composition of the chicken thymus

The cellular composition of the chicken thymus has been analyzed at different ages by using size distribution analysis in combination with preparative cell electrophoresis. The combination of these two physical methods was able to clearly resolve two major cellular subpopulations in the young chicken thymus and suggested the exsistence of a third one. Microscopically, all three cell types appeared to be small lymphocytes. Medium and large lymphocytes are not detected as distinct peaks by the settings used.The analysis revealed dramatic developmental changes in the cellular composition of the thymus. The adult chicken thymus, which is known to have practically no cortex, contained mainly one relatively large cell type. This cell type may, therefore, represent the medullary lymphocyte and may be active in graft-versus-host (G.v.H.) reactions. In the early postnatal thymus that is known to contain little graft-versus-host reactivity this larger cell type was not detectable. Instead, smaller cell types were found to be dominant. The developmental shift from smaller to larger cells was discontinuous. Before thymus involution at 16 weeks of age, smaller and larger cells were both found to be present and to have the same typical size and electrophoretic mobility that is characteristic for the postnatal or the adult chicken thymus, respectively. Size and electrophoretic mobility were therefore taken as markers indicating distinct cellular subpopulation in the thymus.     

Cyclosporine and the thymus: influence of irradiation and age on thymic immunopathology and recovery

With the proper experimental conditions, previous studies have demonstrated that syngeneic and autologous radiation chimeras treated with cyclosporine (CsA) routinely develop a syndrome resembling graft-vs-host disease (GVHD) after CsA is discontinued. The thymus is clearly important in the pathogenesis. Thymectomy prior to CsA prevents the development of syngeneic GVHD and the process can be adoptively transferred via thymocytes. The thymus, however, must be within the field of irradiation and the animal must be young. Here we examine how irradiation and advanced age influence the thymic immunopathologic changes induced by CsA and influence the recovery post-CsA. Young LEW rats, with or without pre-CsA mediastinal irradiation, demonstrate a marked involution of the thymic medulla with associated loss of medullary epithelium, Hassall's corpuscles, class II antigen expression, and maturation of thymocytes. While the control group underwent rapid and complete regeneration of the medulla post-CsA, however, the medullary changes in the irradiated group were prolonged or permanent. Most of these animals had changes of chronic GVHD. Older LEW rats had a more prominent medulla prior to CsA. In contrast to younger rats, the medulla did not show significant involution with CsA. While the Hassall's corpuscles disappeared, the medullae still had fusiform epithelium, dendritic cells, and class II antigen expression. Phenotype stains demonstrated many mature-appearing CD4+/CD8- lymphocytes. In light of evidence indicating the importance of the medullary microenvironment to the maintenance of self tolerance, the medullary effects of CsA are most likely essential to the development of autoimmunity. Young rats rapidly lose the ability to maintain tolerance. While unirradiated rats rapidly reestablish the proper microenvironment following CsA, irradiated rats have a prolonged loss. Older rats may resist the development of autoimmunity by retaining the medullary microenvironment.     

Central tolerance is impaired in the middle‐aged thymic environment

One of the earliest hallmarks of immune aging is thymus involution, which not only reduces the number of newly generated and exported T cells, but also alters the composition and organization of the thymus microenvironment. Thymic T‐cell export continues into adulthood, yet the impact of thymus involution on the quality of newly generated T‐cell clones is not well established. Notably, the number and proportion of medullary thymic epithelial cells (mTECs) and expression of tissue‐restricted antigens (TRAs) decline with age, suggesting the involuting thymus may not promote efficient central tolerance. Here, we demonstrate that the middle‐aged thymic environment does not support rapid motility of medullary thymocytes, potentially diminishing their ability to scan antigen presenting cells (APCs) that display the diverse self‐antigens that induce central tolerance. Consistent with this possibility, thymic slice assays reveal that the middle‐aged thymic environment does not support efficient negative selection or regulatory T‐cell (Treg) induction of thymocytes responsive to either TRAs or ubiquitous self‐antigens. This decline in central tolerance is not universal, but instead impacts lower‐avidity self‐antigens that are either less abundant or bind to TCRs with moderate affinities. Additionally, the decline in thymic tolerance by middle age is accompanied by both a reduction in mTECs and hematopoietic APC subsets that cooperate to drive central tolerance. Thus, age‐associated changes in the thymic environment result in impaired central tolerance against moderate‐avidity self‐antigens, potentially resulting in export of increasingly autoreactive naive T cells, with a deficit of Treg counterparts by middle age.     

Age and changes in the number of endocrine cells in the stomach and their role in senile atrophy of the gastric glands

In the gastric mucous membrane of 40 rats representing 4 age groups (pubertal, mature, old and very old age) per 10 animals in each, thickness of the mucous membrane has been measured by the argyrophil method of Grimelius and the argentaffin method of Masson - Hamperl and the number of the endocrine cells has been counted. The thickness of the mucous membrane and the length of its glands, as well as the number of argyrophil cells increase from the pubertal towards the mature age. From the mature up to the old age the number of argyrophil cells rises in the antral part, and in the acid-producing zone it significantly decreases. The number of argentaffin cells in the stomach does not change from the pubertal towards the old age. In the very old age, against the background of atrophy of gastric glands, the number of both argyrophil and argentaffin cells decreases in both parts of the stomach studied. The data obtained are discussed taking into consideration functional role of various types of endocrine cells, producing biologically active peptides and monoamines.