Thymosin Fraction Research Articles

Thymus peptides interacting with opiate receptors.

The substances displacing labelled ligands from opiate receptors of the rat brain membrane fraction were found in the thymosin fraction 3 and acetoacid extract of the thymus by the radioreceptor assay. Comparison of the displacing activity of acetoacid extracts of perfused and non-perfused thymus and peripheral blood as well as an estimation of the blood content in the thymus allowed to conclude that blood does not participate in the ability of thymus preparations to bind to opiate receptors. On the basis of the enzymatic treatment data one can conclude that opiate receptor ligands present in thymus preparations are of peptide nature. The value of their sodium shift suggests that those peptides are partial agonists of morphine. The possible role of opioid peptides in the thymic endocrine function is discussed.

Thymic hormones in radiation-induced immunodeficiency: I. Induction of mature interleukin 1 responsive cell in the thymus by thymosin fraction 5

The restorative effect of thymosin fraction 5 (TF5) on the thymus of γ-irradiated mice was examined. Four different mouse strains were used in this study since earlier work determined that the degree of response to TF5 is strain dependent. The responsiveness to comitogenic effect of interleukin 1 (IL-1) was used to measure the rate of recovery of immunocompetent cells in the thymus, since only more mature PNA −, Lyt-1 +-2 − medullary cells respond to this monokine. Contrary to several earlier reports that radioresistant cells repopulating the thymus within the first 10 days after irradiation are mature, corticosteroid resistant, immunocompetent cells, the thymic cells from irradiated mice in all strains used had greatly reduced responses to IL-1. Daily intraperitoneal injections of TF5 increased significantly the responses of thymic cells to IL-1 in 10- to 13-weeks-old C57Bl/KsJ, C57B1/6, C3H/HeJ, and DBA/1 mice. Older mice, 5 months or more in age, of DBA/1 strain did not respond to treatment with TF5. However, C3H/HeJ mice of the same age were highly responsive. In conclusion, (1) cells repopulating the thymus within 12 days after irradiation contain lower than normal fraction of mature IL-1 responsive cells, (2) thymic hormones increase the rate of recovery of immunocompetent cells in the thymus, and (3) the effect of thymic hormones is strain and age dependent.

Thymosin α1-induced modulation of cellular responses and functional T-cell subsets in mice with experimental autoimmune thyroiditis

The effects of Ta-1, a peptide constituent of thymosin fraction 5, were studied on murine autoimmune thyroiditis using two congenic strains of mice, B10.Br (Br) and B10.D2 (D2), which are sensitive and resistant to experimental autoimmune thyroiditis (EAT) induction, respectively. EAT was induced by either 2 weekly iv injections of mouse thyroglobulin with adjuvant lipopolysaccharide (LPS) or intradermal injection of thyroglobulin mixed with complete Freund's adjuvant (CFA). The criteria for induction and intensity of thyroiditis were the level of lymphoid infiltration in the thyroid gland and the titer of anti-thyroglobulin antibodies. Ta-1 was given in 5 or 10 daily sc injections in doses ranging from 0.0001 to 0.1 μg/injection. The injections were commenced at varying intervals from the 1st to the 4th week after immunization. T-Cell subsets in the spleens were determined 2 weeks after the first antigen injection and thyroid infiltration was determined 3 weeks later. Treatment with Ta-1 between the two antigen injections increased the level of thyroiditis in resistant mice, but had no effect in sensitive mice. Treatment for the first 2 weeks had similar effects in resistant mice, but also suppressed thyroiditis in the sensitive strain. Later treatments, during the 3rd and 4th weeks after immunization also revealed immunomodulating properties of Ta-1, with a suppressing effect on thyroiditis in sensitive mice and an enhancing effect in the resistant strain. Both effects of Ta-1 were dose dependent. The effects of Ta-1 on the individual phenotypes were also dose dependent. The dose of 0.01 μg greatly lowered the percentages of Lyt-2 +3 + cells in D2 mice and mildly increased the percentages in Br mice, but did not change the Lyt-1 + cell level in either strain. On the other hand, the dose of 0.001 μg greatly increased the percentage of Lyt-1 + cells in D2 mice and mildly decreased it in the Br strain, but did not alter the Lyt-2 +3 + cell subset in either strain. Thus, both doses of Ta-1 modulated Lyt-1 +/2 +3 + ratios, with each dose affecting a different T-cell subset. The changes in the response to thyroglobulin are apparently exerted through the regulation of the functional T-cell subset balance.

Mechanism of action of thymosin. II. Effects of aspirin and thymosin on enhancement of IL-2 production

Recent studies have shown that thymosin fraction 5 (TF5) enhances production of interleukin-2 (IL-2) by phytohemagglutinin (PHA)-stimulated normal human peripheral blood mononuclear leukocytes (PBL). In this study we sought to determine whether this effect of TF5 might be mediated via the cyclooxygenase or lipoxygenase pathways. Our studies demonstrate that aspirin, an inhibitor of the cyclooxygenase pathway, given in vivo, or added to cultures in vitro, results in two-fold increased IL-2 production by PHA-stimulated PBL. This increase is comparable to that seen when PBL are cultured in vitro with TF5. When aspirin and TF5 are added simultaneously to PBL in the presence of PHA, an additive response is seen. An inhibitor of the lipoxygenase pathway, 15-hydroxyeicosatetraenoic acid, did not significantly change IL-2 production by PBL or influence the enhancement by TF5. Augmentation of IL-2 production by aspirin and/or TF5 was prevented by monocyte depletion of the PBL population. These results are interpreted as demonstrating (a) that TF5 and aspirin augment, by distinct mechanisms, IL-2 production by normal human PBL, (b) that the effects of both of these agents are mediated directly or indirectly via a monocyte population and (c) that aspirin, in addition to its analgesic and anti-inflammatory properties, may act as a modulator of immunological responsiveness, either alone or in combination with other biological response modifiers such as thymosin.

Thymosin fraction 5 causes increased serum corticosterone in rodents in vivo.

In these studies it was found that i.p. injection of thymosin fraction 5 (TF5) caused a dose-dependent increase in serum corticosterone in male Swiss Webster mice and in male Wistar rats. The maximum responses were seen at 1 and 2 hr, respectively. There was no effect on serum corticosterone in mice when Thymosin alpha 1 (a 28 amino acid peptide isolated from TF5) was injected i.p. at doses up to 100 micrograms. The steroidogenic effects of TF5 were seen only when the basal levels of serum corticosterone were low (less than 80 ng/ml). In studies in which the baseline levels in the animal colony were elevated (greater than 80 ng/ml), there were no steroidogenic effects, or they were minimal. These results suggest that some component of TF5 may influence pituitary adrenal function.

The effects of thymus-derived peptides on hypothalamic LRF and pituitary gonadotropin content in prepubertal congenitally athymic nude mice and their normal heterozygous littermates

Congenitally athymic nude mice have recognized reproductive defects, accompanied by decreased pituitary gonadotropin and hypothalamic LRF concentrations at 3 weeks of age, compared to their normal heterozygous littermates. To determine if these hormonal changes are due to decreased secretion of thymic peptides, we measured pituitary concentrations of LH and FSH and hypothalamic LRF content at 21 days of age in 122 congenitally athymic nude ( nu/nu) mice and in 190 heterozygous ( nu/+) littermates administered daily injections of a crude thymic extract, semipurified thymosin fraction 5, or a synthetic thymic protein preparation. All animals receiving crude thymic protein extract, and some receiving either thymosin or synthetic thymic factor, had reduced levels of hypothalamic LRF. The changes in concentrations of gonadotropins were more variable. All mice receiving crude thymic extract (with reduced hypothalamic LRF content) appeared to have increased pituitary concentrations of LH, FSH, or both when compared to control animals, although statistical significance was not achieved in all groups. A similar but less marked trend toward increased pituitary gonadotropins was also noted in mice receiving thymic factor. These preliminary results suggest that thymus-derived peptides can affect hypothalamic-pituitary function and support our hypothesis that thymic hormones may play a role in programming the hypothalamic-pituitary axis in the prepubertal period in rodents.

Terminal differentiation of cord blood lymphocytes induced by thymosin fraction 5 and thymosin alpha 1.

Cord blood lymphocytes (CBL) have been shown to be functionally immature compared with normal circulating adult lymphocytes (NAL). Differentiation of T cells is associated with changes in surface antigenic markers and in the pattern of purine degradative enzymes. Previous studies have demonstrated that thymosin fraction 5 (TMS-F5) and thymosin alpha 1 (TMS-alpha 1) can induce in vitro differentiation of murine T-cell precursors and human thymocytes. We have investigated the effects of TMS-F5 and TMS-alpha 1 on the pattern of the purine degradative enzymes adenosine deaminase, purine nucleoside phosphorylase, and ecto-5'-nucleotidase (5'NT) of CBL and on the phenotypic markers from the OKT series 3, 4, 8 and 11. Other than a significantly reduced level of 5'NT activity (P less than 0.001) and an elevated percentage of OKT4+ cells (P less than 0.01), CBL demonstrated the same immunological and biochemical patterns as NAL. Incubation of CBL with TMS-F5 (150 micrograms/ml) and TMS-alpha 1 (1 microgram/ml) for 40 h caused a significant rise in 5'NT level and decrease of cells positive for OKT4, resulting in a pattern characteristic of NAL. Thus TMS-F5 might induce the terminal differentiation of CBL, and TMS-alpha 1 seemed to be the active component.

Thymus Lymphocytes in Rats During Induction of Bladder Tumors and Effects of Thymosin Fraction 5 In Vitro

Bladder tumors were induced in Wistar rats by oral administration of N-butyl-N-(4-hydroxybutyl)nitrosamine [(BBN) CAS: 3817-11-6], after which changes in bladder mucous membranes and in the response of thymus lymphocytes to concanavalin A (Con A) were studied. The response of thymus lymphocytes to Con A began to decrease from the 8th week after BBN administration, with changes such as hypertrophy or vascular formation in the bladder membrane. The response decreased markedly from the 15th week, when the tumors became visible. At about the 20th week, approximately 95% of the rats had bladder tumors. When thymus lymphocytes were pretreated with thymosin fraction 5 extracted from calf thymus, the response to Con A improved between the 8th and 20th weeks. When the thymus small-lymphocyte population was separated into lighter, intermediate, and heavier subpopulations by discontinuous density gradient centrifugation and an adhesion column, the effect of thymosin fraction 5 was much greater in the lighter subpopulation.

Mechanism of action of thymosin. I. Thymosin fraction 5 increases lymphokine production by mature murine T cells responding in a mixed lymphocyte reaction.

The effect of thymosin on the murine thymocyte mixed lymphocyte response was studied. Thymosin fraction 5 (TF5) caused a two- to threefold enhancement of the proliferative response and production of IL 2 when murine thymocytes were cultured with alloantigenic stimulator cells. Production of a second lymphokine, CSF, was increased up to sevenfold. The target cell for thymosin was a mature T cell, because the PNA- subpopulation of thymocytes, as well as peripheral lymph node lymphocytes, responded to culture with TF5 and alloantigen by enhanced proliferation and lymphokine production. The active component of TF5 appears to be one or more as yet unidentified peptides, because neither of the well-characterized TF5 component peptides, alpha 1 or beta 4, were active. After incubation with TF5 in primary culture, cells remaining after 10 to 14 days were increased both in number and in secondary response to alloantigen, as measured by lymphokine production. These results suggest that TF5 contains one or more biologically active components which can modulate mature T cell activity and lymphokine production, and which provide the basis for understanding some of the previously reported diverse effects of thymosin.

In vitro modulation of purine enzyme metabolism and lymphocyte surface marker expression by thymosin fraction 5 in homosexual males

Thymosin fraction 5 (Thymosin) has numerous immunoregulatory activities including modulation of enzymes involved in lymphocyte maturation. The effect of Thymosin on the purine metabolic enzymes adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and 5′ nucleotidase (5′ NT) in null and T-enriched peripheral blood lymphocytes from sexually active asymptomatic homosexual males (AS), patients with the AIDS-related symptom complex (ARC), and those with acquired immune deficiency syndrome (AIDS) was examined and compared to its effect on lymphocytes from healthy heterosexual controls. Mean ADA activity was significantly higher in null cells from fourteen AIDS patients than in five asymptomatic homosexuals, ten ARC patients, or 27 controls. Mean PNP activity was significantly elevated in null-enriched lymphocytes from ten ARC and fourteen AIDS patients compared to controls. No differences in these enzymes were found in T-enriched cells from any group. 5′NT was markedly decreased in both null and T lymphocytes in all homosexual groups relative to controls. Homosexuals had significantly elevated percentages of OKT10 positive and Ia positive lymphocytes compared to controls. Thymosin at an optimal concentration of 150 μg/ml caused significant decreases in mean ADA and PNP activity in null lymphocytes from ARC + AIDS patients along with a significant decrease in the percentage of OKT10 positive lymphocytes. No phenotypic changes were seen in AS or control lymphocytes. The data suggest that Thymosin has a maturational effect in vitro on immature T cells from symptomatic homosexuals.

16] Thymosin β4

Publisher Summary This chapter provides an overview of β4. Thymosin β4 is a polypeptide isolated from thymosin fraction 5 (F5) and fraction 5A (5A). This peptide exhibits important activities in the regulation and differentiation of thymus-dependent lymphocytes. It induces expression of TdT activity in vivo 2 and in vitro. 3 It also inhibits the migration of macrophages, and exerts biological effects on the hypothalamus and pituitary. The chapter further provides purification procedure, thymosin β4 is present in both thymosin F5 and F5A. However, its relative concentration is higher in F5A than in F5. This peptide can be purified either by conventional column technique or by HPLC. Conventional column techniques are used for large-scale purification, while the HPLC procedure is used when the starting material available is limited. In addition, the chapter also explains purification by HPLC from thymosin F5A or tissue extracts.

Thymic peptide hormones: Basic properties and clinical applications in cancer

The manuscript will provide an in-depth and critical review of the nomenclature, biochemistry, biological properties, and a summary of published and on-going clinical trials with all reported thymic preparations, including both partially purified thymic factors (e.g., thymosin fraction 5, thymostimulin) as well as purified and synthesized thymic peptides (e.g., thymosin alpha 1, thymulin). Particular emphasis will be placed on which thymic peptides should be categorized as true hormones. In addition, the comparative biochemistry and biological activity in animals will be summarized and contrasted for all the currently available thymic factors. The effects, in vitro of thymic factors, on peripheral blood lymphocytes isolated from normal donors and patients with primary immunodeficiency disorders, autoimmune disorders, and neoplastic disorders will also be reviewed. Finally, a detailed critical summary of the clinical trials performed with each of the thymic preparations will be presented with an emphasis on treatment of patients with cancer.

Thymosins, Lymphokines, and the Immunology of Aging

Recent data point to a significant role for thymosins, lymphokines, and other soluble mediators in the senescence of the immune response that occurs with aging. In recent years, considerable progress has been made in the isolation and physiochemical characterization of several of these soluble mediators. We are now beginning to define the mechanisms by which these molecules regulate and mediate immune responses. In this paper we review the properties of the best characterized thymic hormones and lymphokines and focus on the role of the endocrine thymus in modulating immune responses. Of particular interest is the recent observation that thymosin fraction 5 can enhance production of interleukin-2 (IL-2) and colony-stimulating factor (CSF), and that IL-2 production, but not CSF production, is selectively diminished in aging mice. Several of the products of the immune system also can act as neuroactive immunotransmitters and modulate a number of neuroendocrine responses. Current studies point to an important role for these molecules in modulating neuroendocrine function, suggesting a broader role for the endocrine thymus in the aging process.

313 Effect of interleukin-2 and thymosin fraction V on the expression of Tac antigen and lymphocyte transformation in AIDS

313 Effect of interleukin-2 and thymosin fraction V on the expression of Tac antigen and lymphocyte transformation in AIDS

A new immunomodulator, LF 1695 — I. in vitro effects on the T lymphocyte lineage in man

A new immunomodulator, LF 1695, was analyzed in vitro for its capacity to induce T cell markers (HTLA and OKT3, 4 and 8 antigens), to enhance the mitogen-induced lymphocyte proliferation, and to increase the Con A-induced suppressor activity. The inductive activity was compared with that of thymic hormone (thymosin fraction V). LF 1695 was capable of significantly augmenting the percentages of HTLA + cells (10 – 15 %) and of OKT 3 +,T4 +, or T8 + cells (13 – 28 %) in human bone marrow prothymocytes. Maximum induction was observed at the concentration of 0.5 μg/ml. In addition, LF 1695 significantly augmented the proliferation of human peripheral blood lymphocytes stimulated by mitogens (Concanavalin A, phytohemagglutinin, pokeweed mitogen and phorbol myristate acetate). LF 1695 also increased the Con A-induced suppressor activity of human lymphocytes. These data indicate that LF 1695 is active both on T cell precursors and on more mature T cells.

13] Thymic hormones: An overview

Publisher Summary This chapter provides an overview of thymic hormones. The thymus gland is a bilobed lymphoid organ located in the chest immediately beneath the breastbone. The involution of the thymus is among the first noticed indications of aging. A vital part of the process by which the thymus works occurs via a hormonal mechanism. A partially purified thymic extract termed thymosin fraction is been demonstrated to be effective in reconstituting immune functions in thymic-deprived or immuno deprived animals as well as in humans with primary immunodeficiency diseases and in immunosuppressed cancer patients. In addition to thymosin fraction 5, a number of preparations with thymic hormone-like activity are isolated from thymus tissue, blood and thymic epithelial supernatants. Several of the newly described thymosin peptides have been found to bear high degree of homology with either thymosin a α 1 and β 4 . The most important contribution of thymic hormone research resides in its application to the clinical management of thymus-dependent diseases.

Immunomodulation and therapeutic characterization of thymosin fraction five.

In this report we describe the characterization of the immunomodulatory efficiency and therapeutic properties of thymosin fraction five (F5). We consistently observed the immunostimulation of T-cell activity in assays of allogeneic mixed lymphocyte response (MLR) and the development of cytotoxic effector cells in an allogeneic mixed lymphocyte tumor response-cell-mediated cytotoxicity assay (MLTR-CMC). No induction of suppressor cell activity was observed. Thymosin F5 also acted successfully as an adjuvant when admixed with irradiated tumor cells. We were unable to demonstrate either NK cell or macrophage activation by thymosin F5. Therapeutic protocols using thymosin F5 and directed against pre-existing experimental and spontaneous metastases, had a significant immunotherapeutic potential.

Autoimmune hemolytic anemia as a manifestation of T-suppressor-cell deficiency

This report describes two children in whom autoimmune hemolytic anemia was the initial clinical manifestation of an underlying T-cell deficiency. Further investigation revealed a profound deficiency of T suppressor cells in both children as detected by monoclonal T-cell antibody (OKT8) and/or functional assays. In vitro incubation of their lymphocytes with cultured thymus epithelium or thymic factors induced T suppressor cells. In vivo treatment with cultured thymus epithelium or calf thymosin fraction 5 resulted in increased T-suppressor-cell numbers and/or function in both and possibly decreased hemolytic activity in one. These results suggest that the autoimmune process in some patients with autoimmune hemolytic anemia is associated with T-suppressor-cell deficiency and that in vivo therapy with agents that modulate T-cell function may be of therapeutic value.

Influenza and aging: Age-related changes and the effects of thymosin on the antibody response to influenza vaccine

Despite massive immunization programs, influenza remains a major cause of morbidity and mortality for elderly people. This may occur because immune senescent recipients may respond to vaccination with inadequate antibody production. We measured antibody response to the trivalent 1983-1984 influenza vaccine in young and elderly volunteers and found a significantly reduced response in the latter. The age-associated decreased antibody production was also observed in lymphocyte cultures in which specific antiinfluenza antibody synthesis was measured. In these cultures, however, the addition of a thymic hormone preparation (either thymosin fraction 5 or thymosin alpha 1) was shown to enhance specific antibody synthesis to a greater extent in the cultures established from the elderly volunteers. If this in vitro observation of thymosin induced increased antibody production reflects what might occur in a clinical trial in which elderly subjects receive thymosin coincident with vaccine, greater protection against influenza infection may result.

Maturational impairment of thymic lymphocytes in streptozotocin-induced diabetes in rats

Streptozotocin (STZ)-induced diabetes in rats was associated with marked decreases in thymus weight and the number of thymic lymphocytes. Histologically, the cortical lymphocytes which were present near the cortico-medullary junction in the thymus seemed to be reduced selectively in the STZ-induced diabetes. Rosette-forming cells, which bind to guinea pig erythrocytes in the presence of fetal calf serum, were also significantly decreased. Insulin treatment allayed these intrathymic changes. Preincubation of thymic lymphocytes from diabetic rats with thymosin fraction 5 significantly enhanced the percentage of rosette-forming cells to near the control level. These results suggest that a maturational impairment of thymus cortical lymphocytes may be caused in STZ-induced diabetes with hypoinsulinemia and it may be intimately related to reductions in thymus weight and the number of thymic lymphocytes.