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The effects of pregnancy on the mouse thymic epithelium.

Changes in the murine thymus during pregnancy were studied using immunocytochemistry with monoclonal antibodies against thymic epithelial, neuroendocrine, and thymulin-producing cells, fibroblasts, blood vessels and connective tissue components. Extensive alterations occur in mid-pregnancy. The medulla was greatly enlarged in the involuted thymus, and there were greater numbers of epithelial cells. These epithelial cells had an altered distribution forming large structures surrounding spherical masses of mononuclear cells, lacked epithelial cells and often contained a central blood vessel with fibroblasts and connective tissue. We have called these structures 'medullary epithelial rings' (MERs). To our knowledge these structures have not been described before. Late in pregnancy the loss of the central mononuclear cells leaves collapsed structures in a smaller medulla that nevertheless retains many epithelial cells. In virgins and early-pregnancy, there are cortical channels free of epithelial cells that are very infrequent later in pregnancy. This may reflect the loss of steroid-sensitive thymocytes from the cortex. The influence of sex-steroids, neurological impulses and immune activity in causing the changes are discussed, as are the possible consequences in pregnancy of a reduced, thymocyte-depleted cortex and an enlarged medulla that shows great complexity and activity.

Animals↗

Suppression of acute experimental allergic encephalomyelitis by synthetic serum thymic factor. Clinical, histopathological, and immunohistochemical studies.

Acute experimental allergic encephalomyelitis (EAE) was induced in Hartley guinea pigs and Lewis rats, which were then treated with synthetic serum thymic factor (FTS). When a dose of 30 micrograms/100 g body weight of FTS was subcutaneously administered to the animals on days--1 (before inoculation), 4, 9 and 15 intermittently, clinical symptoms of acute EAE were suppressed. Histopathological evaluation showed that the severity of EAE in FTS-treated guinea pigs was less than in untreated guinea pigs. Immunohistochemical examination showed that the numbers of OX6+, W3/25+, W3/13+ and OX19+ cells in FTS-treated rats were less than in untreated rats and that the number of OX8+ cells in FTS-treated rats was greater than in untreated rats. These findings suggest that FTS induced OX8+ cells in inflammatory lesions and suppressed inflammation in acute EAE.

Animals↗

Purification of thymocyte growth peptide (TGP) from sheep thymus. Relationship to FTS/thymulin.

Thymocyte growth peptide (TGP) initiates DNA synthesis in immature thymocytes and has previously been characterized as an acidic peptide isolated from calf thymus. We now report the isolation of TGP from sheep thymus and show it to be a nonapeptide with a large N-terminal blocking moiety characterized by high UV absorbance. The amino acid composition is identical to FTS, consisting of 2 Gly, 2 Ser, 2 Glx, 1 Ala, 1 Lys, 1 Asx. In contrast to FTS, TGP is acidic with an apparent isoelectric point of 4.2 and a high UV absorbance at 270-280 nm. Reverse phase chromatography of TGP at an acidic pH results in a change of the molecule and the appearance of two new compounds TGP-A and TGP-B, both with less than 50% of the original TGP activity. Full activity could be restored by the addition of ZnCl2 to TGP-A. Both TGP-A and B have some amino acid composition and high UV absorbance as native TGP. We propose that TGP consists of a non-peptide moiety bound to the N-terminal of the nonapeptide Glu-Ala-Lys-Ser-Gln-Gly-Gly-Ser-Asn and that the active molecule is stabilized by Zn2+.

Amino Acid Sequence↗

Thymic factor-induced reduction of pulmonary metastases in mice with FSA-1 fibrosarcoma.

The biological activities of two thymic factors, serum thymic factor thymulin normally present in serum and thymosin alpha-1 (Ta-1) extracted from the thymus gland, have been studied. The effects of the factors on the growth of pulmonary metastases and survival of mice were evaluated in pathogen-free C3H/fSed males. Mice were injected i.v. with the single cell suspension of the syngeneic methylcholanthrene-induced fibrosarcoma. The treatment with thymulin and Ta-1 started two days after injection of 5 x 10(4) to 2 x 10(5) tumor cells per mouse. Different doses of the thymic factors were administered S.C. in sets of 5 daily injections through a period of 2 or 3 weeks. Numbers of tumor colonies in the lung were determined two weeks after the cell injection. Treatment with 0.1 micrograms Ta-1 per injection through the period of two or three weeks, prolonged the survival of tumor-injected mice. Similar effects were observed in mice treated with 0.01 microgram thymulin per injection. Numbers of tumor colonies in lungs of these mice two weeks after the cell injection were also reduced in comparison with saline-treated controls. These findings correlated with prolonged survival time of identically treated mice. The effectiveness of thymic factors in reducing tumor growth was dependent on the tumour load. In addition, the effects induced by Ta-1 persisted longer than observed in thymulin-treated mice. Mice challenged 150 days after the primary tumor cell injection and treatment with Ta-1 demonstrated increased resistance to tumor, while mice treated with other factors behaved as saline-treated controls. The results indicate that both factors exert beneficial effects against tumor growth, although mode of action for each factor may be different.

Animals↗

Thyroid and thymic endocrine function and survival in severely traumatized patients with or without head injury.

OBJECTIVE: Functional links among the brain, endocrine and immune system have been described previously. An impairment of both immunological defence mechanisms and thyroid hormone turnover was present in trauma conditions. An investigation on the relevance of thymulin and thyroid hormones in multiple trauma patients with or without head injury has been performed. The role of these hormones as predictive factors for patients outcome was also evaluated. DESIGN: Plasma thymulin levels and plasma thyroid hormone concentrations were tested in multiple trauma patients 24 h after admission to the Intensive Care Unit (ICU) and again after 5 and 10 days. SETTING: Department of Immunology Ctr. INRCA, IInd ICU, S. Matteo Hospital Pavia and ICU "Umberto I" Hospital, Ancona. PATIENTS: 45 patients were evaluated including 14 multiple trauma patients without head injury and 31 multiple trauma patients with head injury at various level of coma, graded according to the Glascow Coma Score (GCS). INTERVENTIONS: Routine protocol interventions were performed in all head injured patients. MEASUREMENTS AND RESULTS: Thymulin and triiodothyronine (T3) levels were reduced, and reverse triiodothyronine (rT3) increased in all traumatized patients, but multiple trauma patients with head injury and GCS < or = 5 had the lowest levels of thymulin and T3 and the highest levels of rT3. No difference in plasma thyroxine (T4) and thyrotropin (TSH) levels was observed among injured patients. The analysis of predictive factors for the outcome has assigned to thymulin the highest score (29.6%) compared with the score for T3 (19.3%) and rT3 (26.3%). The total relative risk (delta %) calculated on the basis of T3 or rT3 rises significantly when thymulin relative risk is added. CONCLUSIONS: Thymulin is markedly reduced in multiple trauma patients with head injury and it represents a predictive factor for the outcome better than the one deriving from the single measurements restricted to thyroid hormones.

Accidental Falls↗

In vivo administration of serum thymic factor (FTS) prevents EMC-D virus-induced diabetes and myocarditis in BALB/cAJcl mice.

The effect of serum thymic factor (FTS) on the D-variant of encephalomyocarditis (EMC-D) virus-induced diabetes and myocarditis in BALB/cAJcl mice was investigated. Mice pretreated with 50 or 10 micrograms of FTS were infected with 10 or 10(3) PFU of EMC-D virus. In the mice inoculated with 10 PFU of virus, 40% developed diabetes on post-infection day (PID) 14, whereas those treated with FTS (50 micrograms/administration) on day 2 and 1 before infection did not develop diabetes. FTS (10 micrograms)-pretreated mice developed diabetes. In histological observation, FTS non-treated mice which developed diabetes showed severe necrosis and inflammation of mononuclear cells in the islets of Langerhans and myocardia on 19 PID. Mice pretreated with 50 micrograms of FTS, however, manifested mild islet degeneration without any myocardial inflammation. Furthermore, in FTS non-treated mice, immunohistological staining showed a loss of insulin granules. This loss was markedly reversed and insulin granules remained largely intact in FTS-pretreated mice. Viral titers in pancreas of FTS-pretreated mice approximated well to those of non-treated mice on PID 4, 7 and 19. In mice inoculated with higher titer of EMC-D virus (10(3) PFU), however, 50 micrograms of FTS pretreatment did not change the course of these acute pathological developments (diabetes and myocarditis observed from PID 4).

Animals↗

Have we underestimated the importance of the thymus in man?

Recent immunological research has concentrated on the complex and subtle interactions between T cells, B cells and accessory cells. In these studies, little attention has been given to the adult thymus gland. Modern textbooks of disease and anatomy all stress that the gland undergoes fatty involution with age in man but omit reference to the statements here and there in the literature that the gland is active and produces lymphocytes throughout life. To suggest that the bone marrow, which also builds up fat throughout life, is atrophic and not important to adult man would deny all modern hematological concepts. Yet few people today take a parallel view of the thymus except perhaps those investigating aging and thymic hormones. In both of these areas of research it is obvious that the thymus must be active throughout life for continued good health. This brief review urges that a thorough understanding of the vital importance of the thymus in adult life is now needed. From it could emerge a new philosophy on the treatment of immune diseases in both the young (SCID and AIDS patients) and in the aged (autoimmune conditions and cancers) and it would aid our treatment of patients recovering from illness and from many drug treatments.

Acquired Immunodeficiency Syndrome↗

Serum thymic factor as a radioprotective agent promoting survival after X-irradiation.

Serum thymic factor (FTS, zinc-free thymulin) protected mice from death after whole-body X-irradiation. It was significantly radioprotective even when administered after irradiation, but it was more effective when administered both before and after irradiation. The protective effect appears to be due to the enhancement of hematologic recovery in the animals.

Animals↗

Impaired peripheral zinc metabolism in patients with senile dementia of probable Alzheimer's type as shown by low plasma concentrations of thymulin.

Plasma concentrations of a zinc carrier peptide, namely thymulin, were measured according to a bioassay in young donors, healthy elderly, and patients with senile dementia of Alzheimer's type (SDAT). Thymulin is a hormone released by thymic epithelial cells and its biological activity on cells of immune system is dependent on the presence of one molecule of zinc bound to the peptide. Plasma from different subjects were fractionated by gel filtration to yield protein-bound thymulin and free thymulin. The biological activity of the peptide was then assessed in the two different fractions. The activity of protein-bound thymulin was higher in young donors than in elderly or SDAT patients, being the lowest in SDAT. Addition of zinc ions to plasma fractions increased the thymulin activity of samples from elderly and SDAT patients to levels observed in young donors. Thymulin activity in free thymulin fractions was lower in the elderly than in the young and was practically undetectable in SDAT patients. The addition of zinc ions normalized the activity of thymulin in these fractions from both the elderly and SDAT patients. These findings confirm the presence of an altered zinc status in the elderly and suggest that an impaired zinc metabolism may be present in SDAT patients.

Adult↗

Thymopentin structure and relation to thymic factors.

High-performance liquid chromatography (HPLC) was used to evaluate the purity and batch-to-batch consistency of four commercially available thymic extracts and thymopentin, the synthetic pentapeptide corresponding to the biologically active region of the thymic hormone thymopoietin. The thymus extracts were all extremely heterogeneous, differing one from the other. Additionally, they showed high batch-to-batch variation. In contrast, thymopentin was homogeneous and this homogeneity was consistent in different batches.

Animals↗

Thymulin, a zinc-dependent hormone.

Thymulin (formerly called FTS) is a well defined nonapeptide hormone produced by thymic epithelial cells. Its biological activity and antigenicity depend upon the presence of the metal zinc in the molecule. This pharmacologically active metallopeptide induces the differentiation of T-cells and enhances several functions of the various T-cell subsets in normal or partially thymus-deficient recipients. Its effect on suppressor T-cells is, so far, the most remarkable and should be the first to find useful clinical applications. The peptide is a natural hormone, available in synthetic form. It is not toxic and one may foresee its clinical use as one of the major immunoregulatory agents in the near future.

Animals↗

Structural and conformational analysis of metal-containing peptides by one- and two-dimensional NMR spectroscopy. The case of thymulin.

Thymulin (formerly called FTS) is a well-defined nonapeptide hormone produced by thymic epithelial cells. Its biological activity and antigenicity depend on the presence of the metal zinc in the molecule. The interaction between this metal ion and thymulin has been investigated by means of one- and two-dimensional NMR experiments. These experiments were performed in dimethyl-d6 sulfoxide solution and in aqueous medium with different metal:peptide ratios. The results are compared with those obtained for complexes of thymulin with other metal ions (Cu2+ and Al3+) and for the [Ala4]- and [Ala8]-analogs in terms of biological activity. These comparative studies suggest that the 1:1 complex is the only conformation recognized by the antibodies. From the NOESY data, a spatial conformation has been proposed for this complex. This conformation should be the physiological one and could lead to a better insight into the conformation requirements at receptor sites.

Amino Acid Sequence↗

Thymulin, zinc and insulin-like growth factor-I (IGF-I) activity before and during recombinant growth hormone (rec-GH) therapy in children and adults with GH deficiency.

Plasma thymulin (active and total) levels; IGF-I and zinc concentrations were evaluated in 9 children and in 8 adults with GH-deficiency (GHD) before and after 3-6 months of recombinant-GH treatment. Before therapy, GH deficient children had lower plasma active thymulin levels (1.0 +/- 0.3 log-2), not due to a peripheral defect in zinc saturation since plasma zinc levels were within the normal range, and total thymulin levels (1.3 +/- 0.3 log-2) than in the age-matched control group. GH therapy significantly increased active thymulin (3rd month: 3.0 +/- 0.2 log-2, 6th month: 4.0 +/- 0.2 log-2), total thymulin (3rd month: 3.3 +/- 0.3 log-2, 6th month: 4.3 +/- 0.2 log-2) and IGF-I levels (3rd month: 283.3 +/- 7.2 micrograms/L, 6th month: 411.2 +/- 44.2 micrograms/L, vs basal: 144.3 +/- 11.5 micrograms/L); at the 6th month of therapy, thymulin levels (active and total) were comparable to those found in controls. A positive correlation existed between zinc and plasma IGF-I levels (r = 0.66, p < 0.05). In adults with GHD, plasma active (1.9 +/- 0.3 log-2) and total thymulin levels (3.9 +/- 0.1 log-2), significantly lower (p < 0.01 and 0.05, respectively) than in controls before treatment, increased after GH therapy (active thymulin, 3rd month: 3.0 +/- 0.2 log-2, 6th month: 4.4 +/- 0.3 log-2; total thymulin, 3rd month: 3.9 +/- 0.3 log-2, 6th month: 4.7 +/- 0.2 log-2), being at 6th month of therapy no more different from the values recorded in the age-matched control group. In conclusion, children and adults with GHD have a marked impairment of the thymic endocrine activity, which can be restored by six months of GH treatment. The effects of GH on thymic functions may be mediated by IGF-I, through the modulation of zinc turnover, suggesting the possible existence of an interplay among GH, zinc, IGF-I and thymulin both in children and adults with GHD.

Adolescent↗

Neuroendocrine-thymus interactions. I. In vitro modulation of thymic factor secretion by thyroid hormones.

Several in vivo experimental and clinical studies suggest that the production of thymic hormones, such as thymulin (Zn-FTS), is modulated by thyroid hormones. It was not determined in these studies, however whether such modulation is exerted directly on the thymic epithelial cells which synthesize and secrete thymic hormones. In order to discriminate between direct and indirect modulation, the effect of thyroid hormones on the in vitro production of thymulin by whole thymic organ culture, as detected by the rosette inhibition assay, has been investigated. Donors of thymuses were young 6N-propyl-2 thiouracil (PTU)-treated hypothyroid Balb/c mice and normal littermates. Thymuses from hypothyroid mice were shown to produce concentrations in vitro nearly undetectable of thymic hormone, when compared to thymuses from normal mice. The in vitro addition of triiodothyronine (T3) caused a complete recovery of the thymic hormone production by thymuses from hypothyroid mice and an increased synthesis even by normal thymuses over control values. The complete blockade of in vitro thymic hormone production with cycloheximide, which inhibits mRNA and protein synthesis but not thyroid hormone permissive actions, suggests that the T3 induced increment of thymic hormone level in the supernatant is due to de novo synthesis. Furthermore, the number of thymulin-producing cells, as detected by immunofluorescence using a specific antithymulin monoclonal antibody, which is quite low in thymuses from hypothyroid mice, is completely regained after in vitro incubation with T3. These findings support the idea that the modulation of thyroid hormones on thymic endocrine activity is directly exerted at thymic level.

Animals↗

Prolactin, thymulin and zinc in chronic hemodialysis: effect of renal transplant.

The interrelationships between PRL, thymulin and Zn, were studied in 25 patients with chronic renal failure (CRF) undergoing kidney transplantation and immunosuppressed with cyclosporine A (CsA). The possible role of serum PRL levels in predicting allograft rejection was also investigated. Before the kidney transplant serum PRL levels were significantly higher than in normals (mean +/- SE, 28.3 +/- 7.1 vs 7.5 +/- 0.6 micrograms/l, p less than 0.001) and their response to TRH (200 micrograms iv) was impaired (mean delta % at peak, 45.4 +/- 9.5 vs + 641 +/- 47.5, p less than 0.001). After kidney transplantation a dramatic decrease in serum PRL concentrations was observed in all patients, followed by a slight upward trend in the following two weeks, while TRH test administered on 3rd, 7th and 14th day, induced a progressive increase in serum PRL responses (delta % at peak, 201 +/- 43.3, 220 +/- 37.1 and 305 +/- 15.5, respectively). No difference in serum PRL patterns was observed between patients with (8 cases) and without (17 cases) clinical features and kidney fine needle biopsies suggestive of rejection. Basal serum Zn levels of patients with CRF (18.1 +/- 0.6 mumol/l) were similar to those observed in normals (17.7 +/- 0.2 mumol/l) and without any correlation with serum PRL levels. A decrement in serum Zn was recorded during CsA infusion and on the first day after the surgery, followed by a slight and slow upward trend. Basal serum thymulin titers were low [2.92 +/- 0.18 (1/log2)], were further reduced after CsA infusion [1.68 +/- 0.15 (1/log2)] and returned to the pretransplant levels in the two weeks after grafting.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Plasma thymulin and nerve growth factor levels in patients with primary open angle glaucoma and elevated intraocular pressure.

BACKGROUND: The objective was to measure the plasma concentrations of thymulin and nerve growth factor (NGF) in a group of patients with primary open angle glaucoma (POAG) and compare them with age- and sex-matched normal controls. METHODS: Twenty-eight patients newly diagnosed with POAG who were not undergoing treatment were compared with the same number of age- and sex-matched healthy controls. Blood samples were drawn into heparinized tubes and plasma samples were collected for the determination of the concentrations of thymulin and NGF, using specific enzyme-linked immunosorbent assay (ELISA). The Student's t test was used to perform the necessary statistical analysis of the results. RESULTS: Seventeen women and 11 men were enrolled in each of the two groups (study and control), with a mean age of 63.7 (SD 10.3) years in the former and 63.3 (SD 9.6) years in the latter. There was a highly significant (p<0.001) elevation in the thymulin levels in POAG patients compared with the control group. However, no significant difference was observed when comparing the plasma NGF levels. CONCLUSION: This is the first report to measure plasma thymulin levels in glaucoma patients. The significant results point the possible role of this immunomodulator in the pathogenesis of primary open angle glaucoma. The potential role of NGF seems to be less likely. These findings warrant further investigation.

Enzyme-Linked Immunosorbent Assay↗

Rat thymic epithelial cells in vitro and in situ: characterization by immunocytochemistry and morphology.

A new method for the long-term culture of pure rat thymic epithelial cells was established. The cultures were characterized by immunocytochemistry, electron microscopy and proliferation assays. Non-epithelial thymic cells were eliminated with a reliable and reproducible pre-plating method, by differential trypsin treatment of the cultures and by addition of horse serum to the culture medium instead of fetal calf serum. The final cultures contained more than 95% pure epithelial cells as evidenced by immunostaining for cytokeratin. Ultrastructural studies indicated that these cells are physiologically active epithelial cells with tonofilaments, desmosomes and filopods. The subsets of the thymic epithelial cells in vitro were investigated by comparing their staining pattern with that obtained in situ using several subtype-selective antibodies. Thymic epithelial cells in vitro showed a preferential expression of subcapsular/perivascular and medullary markers. Only few cultivated cells were of cortical origin. In the first to the fourth subcultures, some cells were immunopositive for the thymus hormone/factor thymulin. The proliferation of thymic epithelial cells was stimulated by horse serum and to a lesser extend by fetal calf serum. The adenylate cyclase activators isoproterenol and forskolin, and the glucocorticoid cortisol inhibited the proliferation.

Animals↗