Zinc-dependent low thymic hormone level in type I diabetes.

Zinc is required for optimal functioning of the immune system. It was recently reported that one of the best-known thymic hormones responsible for the maturation and differentiation of the thymus-derived T-lymphocyte line, i.e., serum thymic factor (STF), is biologically active only when bound to zinc ions; in this form it has been called thymulin (Zn-STF). Because low serum and tissue zinc values have been reported to occur in diabetic conditions, and because defects of T-lymphocyte-dependent functions are also present in diabetic patients, even metabolically well-controlled diabetic patients, we investigated the serum level of zinc and the plasma level of both active Zn-STF and inactive STF thymic hormones in 15 young patients suffering from type I (insulin-dependent) diabetes. Serum zinc levels were significantly reduced in diabetic conditions and did not correlate with the degree of metabolic compensation measured by glycosylated hemoglobin. In diabetes, the active form of thymulin is strongly reduced, whereas the inactive form is abnormally elevated. In vitro zinc addition to diabetic plasma samples also induces zinc saturation of inactive thymic hormone molecules: the total thymic hormone measured in these experimental conditions shows values in diabetic patients comparable with those observed in healthy age-matched individuals, suggesting that low thymulin levels recorded in diabetic conditions are due not to a thymic failure in synthesizing and secreting thymic hormone but to a peripheral defect in zinc saturation of the hormone molecules. The zinc-dependent failure of thymic hormone, present even in fairly compensated diabetic conditions, might account for the apparent insulin-independent immunological abnormalities associated with type I diabetes.

Effect of oral zinc administration on prolactin and thymulin circulating levels in patients with chronic renal failure.

High serum PRL and low zinc (Zn) levels are common findings in patients with chronic renal failure (CRF); in such patients serum Zn concentrations have been reported to be inversely correlated to serum PRL levels. Moreover, Zn regulates both thymus growth and the biological activity of the thymic hormone thymulin, and PRL-thymic interrelationships have been described. To determine whether hypozincemia alters serum PRL and plasma thymulin concentrations in CRF, 9 men with CRF treated by chronic hemodialysis were given 400 mg/day Zn sulfate, orally (4.96 meq/day Zn), for 6 months. Before treatment, serum PRL levels were significantly higher (P less than 0.001) in these patients than in normal men [mean, 28.7 +/- 20.7 (+/-SD) vs. 7.5 +/- 3.7 micrograms/L], and their serum PRL response to TRH (200 micrograms, iv) was impaired (mean maximal percent increase, 38.2 +/- 10.9 vs. 641 +/- 335; P less than 0.001). The plasma Zn-bound bioactive thymulin titer (1.3 +/- 0.7 1/log2), total thymulin titer (Zn-bound plus Zn-unbound forms, 2.1 +/- 0.8 1/log2), and serum Zn (13.1 +/- 2.4 mumol/L) were lower (P less than 0.001) in men with CRF than in normal men. Zn therapy did not induce any significant change in basal and TRH-stimulated serum PRL levels, while serum Zn levels significantly increased, reaching the normal range after the first week of treatment (17.8 +/- 6.3 mumol/L). Plasma total thymulin increased rapidly, reaching normal levels after 1 week, but Zn-bound thymulin increased modestly during the first month of treatment and more after 3 and 6 months of treatment. There was no age-related difference in plasma thymulin levels during therapy. We conclude that oral Zn administration in patients with CRF significantly increases both total and Zn-bound thymulin, but does not modify basal and TRH-stimulated serum PRL levels. The observation that Zn supplementation markedly increased plasma thymulin levels in uremic patients suggests that Zn is a potent stimulus for thymic hormone synthesis, and it can reverse the age-related diminution of thymic activity in CRF patients.

Involution of thymic epithelium and low serum thymulin bioactivity in weanling mice subjected to severe food intake restriction or severe protein deficiency.

The volume of epithelium in the cortex and in the medulla of the thymus was compared in four groups of weanling male and female CBA/J mice. Well-nourished controls (C), food intake restricted (R), and animals given a low-protein diet ad libitum (LP) were fed from 23 to 37 days of age. Baseline controls (B) were studied at 23 days of age. Epithelial volume fraction was estimated for each group by point-counting morphometry on electron micrographs. Other mice were used to obtain group mean estimates of thymic index (mg/g live weight) and volume fraction of cortex and medulla (light microscope-level point-counting morphometry). Cortical and medullary epithelial volumes were calculated for each animal examined by electron microscopy by obtaining the live weight and applying, in sequence, the group mean thymic index, an assumed thymic density of 1.0 mm3/mg, the group mean cortical or medullary volume fraction, and the measured cortical or medullary volume fraction for that animal. Serum thymulin bioactivity was also measured in C, R, and LP mice. The results reveal thymic epithelial involution in the two most common rodent models of malnutrition, and suggest that this may contribute to the low serum thymulin levels found in malnourished experimental animals and humans.

Thymulin (Zn-FTS) activity in protein-energy malnutrition: new evidence for interaction between malnutrition and infection on thymic function.

The combined effects of protein-energy malnutrition (PEM) and infection on thymic function evaluated by specific plasma thymulin activity were studied in Senegalese children: 29 hospitalized in Dakar for severe malnutrition and various diseases; 9 infected without sign of severe PEM, living in Dakar; 13 apparently healthy, uninfected, living in Dakar; and 7 apparently healthy, uninfected, living in Paris. Most of the free-living children in Dakar suffered from mild to moderate PEM. The specific thymulin activity (total plasma activity minus the activity recorded after adsorption of the plasma with a monoclonal antithymulin antibody) was almost undetectable in the infected children and was normal only in the children living in Paris. Such activity might be decreased by moderate and severe PEM and severe malnutrition may not be the only underlying cause of depressed level of thymulin in malnourished children from the Third World. Concurrent infections are important factors.

Neuroendocrine regulation of thymus hormones: hypothalamic dependence of "facteur thymique serique" level.

The influence of the hypothalamus extract (HE) on the blood level of the "facteur thymique serique" (FTS) was studied. Hypothalami collected from young mice were pooled, homogenated in saline, and centrifuged; finally, the supernatant injected in old or thymectomized mice with no detectable levels of FTS. In the old mice the treatment results in a reappearance of this circulating thymic hormone; in contrast, in adult thymectomized animals, the HE injection was not able to induce FTS activity. When HE donors were pretreated with thymosin fraction 3, known to contain FTS among other active peptides, the capacity of such a hypothalamus preparation to induce reappearance of FTS in old animals is greatly diminished. The data presented here suggest that the capacity of the thymus to secrete FTS depends on a hypothalamic factor, and therefore the absence of this thymic hormone in the aged reflects a failure of the thymus linked to its impaired neurologic control. On the other hand it seems evident that a feedback system operates in order to regulate the release of this hypothalamus stimulatory factor.

Circulating thymulin and thymosin-alpha 1 activity in pediatric acquired immune deficiency syndrome: in vivo and in vitro studies.

Twenty-five children with acquired immune deficiency syndrome (AIDS) or AIDS-related complex had a characteristic pattern of T cell deficiency. Abnormally low plasma thymulin levels preceded the development of peripheral blood T cell abnormalities. In contrast to patients with congenital T cell deficiencies, our patients had elevated serum levels of thymosin-alpha 1. Treatment with thymosin fraction 5 in three children with AIDS resulted in only transient clinical and immunologic improvement.

The wasted mutant mouse. II. Immunological abnormalities in a mouse described as a model of ataxia-telangiectasia.

Ataxia-telangiectasia (AT) is a complex multiparametric disease associating oculocutaneous telangiectasias, cerebellar ataxia, elevated chromosomal aberration frequency and varied degrees of immunodeficiency. Recently a wasted mutant mouse (wst) has been described as an animal model of AT. We have looked in the wasted mutants for the presence of immune and endocrine abnormalities characteristic of AT. In contrast to the T cell immunodeficiency in AT, wasted mutants had a marked hypoplasia of all lymphoid organs, which affected both T and B lymphocyte subsets. The marked thymic atrophy appearing at the final stage of their disease did not modify the endocrine function of the thymic epithelium which produced normal levels of the thymic hormone thymulin. Although in vitro interleukin 2 (IL-2) production by splenic T cells in response to Con A was markedly diminished, these mice presented normal T and B cell proliferative responses to mitogens. Finally, no significant increase in serum alpha-fetoprotein level (a typical marker of AT) was found throughout the course of the disease. Although by many aspects, i.e. neurological disorder, chromosomal aberrations and early death, wasted mice presented similarities with human AT, major discrepancies in the typical features of immune abnormalities were found between the mouse model and the human disease.

Thymic hormone-containing cells. VII. Adrenals and gonads control the in vivo secretion of thymulin and its plasmatic inhibitor.

The influence of adrenals and gonads on the intrathymic production and the circulating level of thymulin was evaluated in young adult mice. Adrenalectomy (Adx) and gonadectomy (Cx) induce a temporary decrease of thymulin serum level. One simultaneously notes, as a compensatory phenomenon, an increase in the thymic content of the hormone-producing cells. The decrease of serum thymulin levels after Adx and Cx is at least partially due to the appearance of low m.w. thymulin-inhibitory molecules. The fact that thymectomy prevents the appearance of these inhibitors suggests that the effects of Adx and Cx could be explained by a negative control by sex hormones of the synthesis or activity of thymulin inhibitors produced or controlled by the thymus. Specific hormone replacement therapy of castrated/adrenalectomized animals normalized thymulin serum level and thymic content. Such correction was also spontaneously observed after 4 mo, suggesting that other mechanisms (e.g., an influence of the hypothalamus-hypophysis axis) might be involved in the endocrine control of thymic hormone secretion.

Low circulating thymulin-like activity in children with AIDS and AIDS-related complex.

Thymic secretory function was assessed by determining levels of circulating thymulin-like activity in plasma of 21 pediatric patients infected with the HTLV-III/LAV retrovirus. All the patients had serum antibodies against p41 antigens of HTLV-III on Western blot analyses. In accordance with the latest definition established by the Centers for Disease Control, 14 patients had the acquired immunodeficiency syndrome (AIDS) and the remaining 7 were classified as having AIDS-related complex. Their ages ranged from 1 to 7 years, with 10 being less than 1 year of age. Circulating thymulin activity, normally highest in healthy children under 15 years of age, was undetectable in 11 patients and below normal range for age in the remaining. OKT4/OKT8 ratios of T-cell subsets in peripheral blood were below normal in the majority of patients. Our findings suggest that thymic epithelial injury may be an early event in HTLV-III/LAV-related disease and may precede the development of clinical and/or immunologic aberrations.

A zinc-dependent epitope on the molecule of thymulin, a thymic hormone.

Thymulin is a nonapeptide hormone produced by thymic epithelial cells. Its biological activity is strictly dependent on the presence of the metal zinc in the molecule. Antithymulin monoclonal antibodies have been produced against either the synthetic (AS1) or the natural intraepithelial (AE1) molecule. These monoclonal antibodies were screened for their abilities to inhibit the zinc-dependent biological activity of the hormone and were shown to bind to thymic epithelial cells. By using biological and immunofluorescence assays, the two antibodies were shown to recognize exclusively the zinc-coupled thymulin molecule. Other antithymulin antibodies screened by RIA or ELISA (using a zinc-deprived substrate) recognized a zinc-independent epitope on the thymulin molecule. These data indicate the existence of a zinc-specific conformation on the thymulin molecule. They are in agreement with NMR studies showing that the zinc-containing hormone has a unique structure.

Selective improvement of thymus and some T cell dysfunctions in NZB mice by in utero thymulin treatment.

NZB mice were treated during gestation with thymulin, a thymus-secreted, zinc-associated nonapeptide. Control pregnant NZB mice received either zinc alone or saline alone. Offspring from all three groups of NZB mothers, and age-matched DBA/2 mice, were tested for the following immunologic parameters: thymulin serum levels at 2 and 5 wk of age; splenic anti-sheep red blood cell (anti-SRBC) plaque-forming cell (PFC) numbers after immunization at birth or at 2 wk of age; anti-human gamma-globulin (anti-HGG) antibody titers after immunization at 2 wk of age, with or without prior tolerance induction at birth with deaggregated HGG; spontaneous IgM serum levels at 2 and 5 wk of age; spontaneous splenic anti-trinitrophenyl (anti-TNP) PFC numbers at 2 wk of age. As compared with DBA/2 mice, young NZB mice exhibited low circulating thymulin titers, high antibody responses to SRBC and to HGG, resistance to tolerance induction by deaggregated HGG, increased spontaneous IgM serum levels, and increased spontaneous anti-TNP PFC numbers. However, marked reductions in anti-SRBC and anti-HGG antibody production, both thymus-dependent responses, were observed in the young NZB offspring of thymulin-treated mothers as compared with NZB controls born from zinc- or saline-treated mothers. A delay in the postnatal decrease of serum thymulin levels was also noted in the offspring of thymulin-treated mothers. Interestingly, these effects of in utero thymulin treatment tended to become more pronounced with advancing age during the postnatal period. Conversely, IgM serum levels, spontaneous anti-TNP PFC and sensitivity to tolerance induction were not affected by thymulin treatment during fetal life. Taken together, the data suggest that in utero exposure to pharmacologic concentrations of thymulin induces a persistent and selective improvement of some thymus and T cell dysfunctions but has no effect on intrinsic B cell abnormalities of NZB mice.

Thymulin (Zn-facteur thymique serique) activity in anorexia nervosa patients.

Thymulin (or FTS-Zn) a well-defined thymic hormone was studied in fifteen female patients hospitalized for anorexia nervosa. The circulating hormone was measured together with the plasma levels of thyroid hormones, cortisol and zinc. Thymulin activity determined by the rosette assay was significantly reduced in the anorexia nervosa patients compared to sex- and age-matched healthy control subjects. The patients were characterized by very depressed plasma levels of triiodothyronine (T3) but exhibited normal concentrations of thyroxine (T4), thyroxine-binding globulin (TBG), cortisol and zinc. The distribution of their peripheral lymphocyte cells into several subsets was not affected. The observed decrease of thymulin activity in this illness might be the consequence of thymic atrophy secondary to malnutrition and/or hormonal disturbances. Our results suggested that the fall in thymulin level might explain the variability of cellular immune responses in anorexia nervosa patients and occurrence of energy when their weight loss is far advanced.

[Changes in the thymic factor level in rats with Jensen sarcoma and modification of the rate of tumor growth by administration of thymic hormone]. / Veränderungen des Thymusfaktorspiegels bei Ratten mit Jensen-Sarkom und Beeinflussung der Tumorangehrate durch Applikation von Thymushormon.

Rats were differently treated after transplantation of cells of Jensen sarcoma to test the influence of Doparkin (increase of the cAMP level), morbilli vaccine (interferon inductor) and local radio-therapy alone or in combination for a decrease of the rate of growing tumors. The lowest rate of growing tumors was received by combining the agents with a local radio-therapy. The analysis of the thymic factor level showed in the animals with a tumor a significant reduction in comparison to normal animals respectively to animals without tumor formation in consequence of the therapy. An application of the thymic factor alone respectively in combination with Doparkin and morbilli vaccine leads to a clear decrease of the rate of growing tumors.