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B L Spangelo

Publications and source records attributed to B L Spangelo.

At least 37 records · Page 2Linked to original sources
PubMed1990

Production of interleukin-6 by anterior pituitary cells is stimulated by increased intracellular adenosine 3',5'-monophosphate and vasoactive intestinal peptide.

Interleukin-6 (IL-6) is an inflammatory cytokine that is produced by a variety of cells and tissues. We recently demonstrated that IL-6 is produced by anterior pituitary cells in response to the bacterial endotoxin lipopolysaccharide and phorbol diester in vitro. Lipopolysaccharide (0.01-100 ng/ml) increased, whereas dexamethasone (0.1-100 nM) decreased, IL-6 production by anterior pituitary cells in vitro as measured by the 7TD1 cell growth factor assay. In addition, we now report that IL-6 production by anterior pituitary cells is stimulated by agents that elevate intracellular cAMP concentrations. Exposure of anterior pituitary cells to (Bu)2cAMP (0.01-10 mM), prostaglandin E2 (1.0-1000 nM), forskolin (50-1000 nM), or cholera toxin (0.25-250 ng/ml) for 6 h resulted in concentration-related increases in the production of IL-6, which, in the cases of forskolin and cholera toxin, correlated well with increased intracellular cAMP concentrations. Vasoactive intestinal peptide (1-1000 nM), which stimulates adenylate cyclase activity in the anterior pituitary, caused a concentration-related enhancement of IL-6 production that was unaffected in the presence of 10-100 nM somatostatin. In contrast, GH-releasing factor had no effect on IL-6 production. These data suggest that anterior pituitary cells produce IL-6 in response to increased intracellular cAMP, and that the neuropeptide vasoactive intestinal peptide may act to regulate IL-6 production.

Animals↗
PubMed1990

Endotoxin-induced release of interleukin-6 from rat medial basal hypothalami.

Interleukin-6 (IL-6) is an inflammatory cytokine that stimulates T-cell activation and B-cell differentiation. We recently reported that picomolar concentrations of IL-6 stimulated PRL, GH, and LH release in vitro. These data suggested that IL-6 may function as a hypothalamic releasing factor for anterior pituitary hormones. Medial basal hypothalami (MBH) were incubated for 60-90 min in Krebs-Ringer bicarbonate buffer supplemented with 0.025% BSA, and the conditioned medium was assayed for IL-6 concentrations by the 7TD1 cell growth factor assay. It was found that MBH released IL-6 in vitro. Although depolarizing concentrations of K+ (56 mM) did not increase IL-6 release, somatostatin release from the MBH was increased significantly. The bacterial endotoxin lipopolysaccharide (LPS; 1-100 ng/ml) induced significant increases in IL-6 release from the MBH. The presence of IL-6 in the hypothalamus suggested a possible role for this cytokine in the regulation of neuropeptide release; however, the release of somatostatin was not affected by 20 ng/ml IL-6. Comparison studies of neural and neuroendocrine tissues revealed that the anterior and posterior pituitaries released larger amounts of bioactive IL-6 than the MBH or parietal cortex during a 4-h incubation; induction of IL-6 release by endotoxin occurred only in the anterior pituitary and hypothalamus. IL-6 mRNA was detectable in the MBH and anterior pituitary tissue after a 4-h incubation; however, no IL-6 mRNA was detectable in freshly isolated tissues. LPS (100 ng/ml) and (Bu)2cAMP (1 mM) increased IL-6 mRNA accumulation in and IL-6 release from the MBH and anterior pituitary. These data suggest that the MBH synthesizes and releases IL-6 via a nonneuronal source in vitro.

Animals↗
PubMed1989

Thymic stromal elements contain an anterior pituitary hormone-stimulating activity.

Conditioned medium from thymic reticular monolayers displayed time-dependent accumulations of a concentration-responsive pituitary hormone-releasing activity that has been named thymic neuroendocrine-releasing factor (TNRF). Dopamine blocked and somatostatin (SRIF) attenuated TNRF-induced prolactin (PRL) release. Conversely, SRIF had no effect on TNRF-induced growth hormone (GH) release. TNRF potentiated thyrotropin-releasing hormone (TRH)-stimulated PRL release and was additive to the effects of GH-releasing hormone (GHRH) on GH release. Anterior pituitary cells perifused with TNRF responded with immediate, sustained and reversible increases in hormone release. Partial purification revealed this activity to be greater than 10,000 in molecular weight. These data suggest that the thymus may affect pituitary function.

Animals↗
PubMed1989

Interleukin-6 stimulates anterior pituitary hormone release in vitro.

Interleukin-6 (IL-6), a cytokine produced by inflammatory reactions, was found to stimulate PRL, GH and LH release from anterior pituitary cells at concentrations similar to those which affected lymphocyte mitogenesis. Perifused pituitary cells responded to IL-6 with prompt increases in hormone release that declined rapidly following cessation of exposure. Dopamine (DA) attenuated IL6-induced PRL release. In addition, IL-6 potentiated both GHRF- and TRH-induced hormone release without an affect on intracellular cAMP. These data demonstrate a new biological activity for IL-6 and provide evidence for immune system regulation of anterior pituitary hormone release.

Animals↗
PubMed1988

Angiotensin II increases pituitary cell prolactin release and arachidonate liberation.

The effects of angiotensin II (AII) on prolactin release and arachidonate liberation were studied in anterior pituitary cells preincubated with [3H]arachidonate to label the cellular phospholipids. AII increased prolactin release and [3H]arachidonate liberation over similar concentration ranges with the dynamics of these two events proving identical. Dopamine attenuated both prolactin release and [3H]arachidonate liberation. The diacylglycerol lipase inhibitor RHC80267 decreased AII-stimulated prolactin release and arachidonate liberation. Further evidence that AII-induced release of arachidonate is mediated by a diacylglycerol lipase is suggested by the finding that AII increased [14C]stearate liberation from cells prelabeled with the fatty acid. Although arachidonate itself may have some role in prolactin secretion, it is likely that arachidonate metabolites are more directly involved because BW755c and AA861, inhibitors of arachidonate metabolite formation, increased AII-stimulated arachidonate liberation, but decreased prolactin release.

Angiotensin II↗
PubMed1988

A possible role for lipoxygenase and epoxygenase arachidonate metabolites in prolactin release from pituitary cells.

We studied the effects of selected leukotrienes and hydroxyeicosatetraenoic acids (HETEs) on prolactin release from primary cultures of female rats anterior pituitary cells. Leukotrienes B4, C4, and D4 had no effect on basal prolactin release; however, they did enhance prolactin release that was stimulated by 1 or 5 nM thyrotropin-releasing hormone (TRH). Leukotriene C4 also enhanced prolactin release that was induced by phorbol myristate acetate (a protein kinase C activator) by maitotoxin (a calcium uptake stimulator), and by angiotensin II. 5-HETE, 12-HETE, and 15-HETE stimulated basal prolactin release at high concentrations (1 microM and greater), and 5-HETE and 12-HETE enhanced TRH- and angiotensin II-induced prolactin release at lower (nanomolar) concentrations as well. In order to determine the role of endogenous arachidonate metabolites in prolactin release, pituitary cell cultures were exposed to selected inhibitors of the 5-lipoxygenase enzyme, which metabolizes arachidonate to leukotrienes and 5-HETE, and to those of the epoxygenase enzyme, which metabolizes arachidonate to epoxyeicosatrienoic acids. These inhibitors decreased basal and secretagogue-induced prolactin release. In additional experiments, it was determined that TRH enhances the liberation from pituitary cells of arachidonate metabolites with high-performance liquid chromatography elution profiles similar to those of leukotriene C4 and omega-OH-leukotriene B4 (a metabolite of leukotriene B4) and the HETEs. Therefore, the production of leukotrienes, HETEs, and epoxyeicosatrienoic acids may be necessary for the normal release of prolactin.

Animals↗
PubMed1988

Characterization of the MMQ cell, a prolactin-secreting clonal cell line that is responsive to dopamine.

Although dopamine inhibits PRL release from the normal anterior pituitary lactotroph, a conclusive demonstration of the mechanisms involved in this response has been impeded by the presence of other cell types in the anterior pituitary. To circumvent this problem, we have isolated a clonal cell line, designated MMQ, from the 7315a rat pituitary tumor. The MMQ cell is an exemplary model for our use because it only secretes PRL. Our studies show that dopamine inhibits secretagogue-induced PRL release from these cells. In addition, dopamine decreases the intracellular cAMP concentration in MMQ cells that have been exposed to forskolin, cholera toxin, or vasoactive intestinal polypeptide, each a stimulator of cAMP generation. This inhibition is, in turn, reversed by the dopamine antagonist haloperidol and by pertussis toxin, an inactivator of the GTP-binding coupling protein. Dopamine also decreases the uptake and fractional efflux of 45Ca2+ by MMQ cells that have been exposed to the calcium channel activator maitotoxin. It seems, therefore, that dopamine decreases PRL release from MMQ cells at least in part by decreasing intracellular cAMP levels and calcium uptake. In additional experiments, we have found that MMQ cells are responsive to somatostatin, estrogen, progesterone, and acetylcholine, but not to TRH, angiotensin II, neurotensin, or bombesin. Furthermore, these cells possess a functional protein kinase-C system, as evidenced by the increase in PRL release and decrease in stimulated intracellular cAMP levels that occur in response to treatment with phorbol diesters. We suggest that the MMQ cell line will prove a useful model system for study of the biochemical effects of dopamine and other factors that modify PRL release.

Calcium↗
PubMed1987

Thymosin fraction 5 stimulates the release of prolactin from cultured GH3 cells.

Thymosin fraction 5, a bovine thymus preparation, has recently been implicated in the regulation of neuroendocrine function. The purpose of the present study was to investigate the effects of thymosin fraction 5 treatment upon the GH3 rat pituitary cell line. Thymosin fraction 5 stimulated prolactin (PRL) release from these cells in a dose and time dependent manner. These results suggest that a product of the endocrine thymus may regulate the release of PRL.

Dose-Response Relationship, Drug↗
PubMed1987

Immunoglobulins of Lambert-Eaton myasthenic syndrome inhibit rat pituitary hormone release.

We speculated that IgG from patients with Lambert-Eaton myasthenic syndrome (LES) would inhibit calcium-dependent hormone secretion in rat anterior pituitary (AP) cells. Primary cultures of normal AP cells were continuously exposed to crude IgG from an LES patient or from healthy control subjects, and the incubation media were assayed for prolactin (PRL) or growth hormone (GH). The LES IgG caused a time- and concentration-dependent reduction of PRL and GH release compared with control IgG over 0.5 to 48 hours using concentrations of 0.01 to 4.0 mg/ml. The calcium-channel activator maitotoxin stimulated 45Ca2+ uptake and PRL release from AP cells under control conditions, and the LES IgG significantly reduced both actions of maitotoxin. Thus LES IgG appears to modulate an AP antigenic site, perhaps representing an integral component of voltage-gated calcium channels that may share properties with similar presynaptic elements at the neuromuscular junction.

Animals↗
PubMed1987

Stimulation of in vivo antibody production and concanavalin-A-induced mouse spleen cell mitogenesis by prolactin.

Regulation of the immune system by neuroendocrine hormones is receiving increased attention. Prolactin, a hormone normally associated with lactation, has been shown recently to reconstitute immunosuppressed hypophysectomized rats. The present studies demonstrate that prolactin administration to normal mice results in a biphasic stimulation of antibody production to sheep red blood cells. While 100 and 200 micrograms bovine prolactin/animal stimulated antibody production, 400 micrograms had no effect. Potentiation of lectin-induced T-cell mitogenesis by prolactin was also biphasic. As the concentration of prolactin increased the incorporation of [3H]thymidine into the cells first increased and then decreased. Decreasing serum prolactin levels with the dopaminergic agonist bromocriptine resulted in a reduction of antibody titers to sheep erythrocytes and a modulation of thymic weight. These data show that prolactin can stimulate the immune system in a biphasic manner and that a reduction in the basal levels of this hormone results in an attenuated immune response.

Animals↗
PubMed1987

Thymosin fraction 5 stimulates prolactin and growth hormone release from anterior pituitary cells in vitro.

Thymosin fraction 5 (TF5) is a partially purified extract of bovine thymus containing 40-60 peptides. In addition to its well documented immunopotentiating effects, TF5 reportedly modulates the secretion of some hypothalamic peptides and pituitary hormones. In this study, TF5 (10-100 micrograms/ml) stimulated PRL release from normal, MtTW15, and 7315a cells and GH release from normal and MtTW15 cells, but had no apparent effect on LH release. No changes in intracellular cAMP or cGMP levels could be correlated with these responses. Stimulation of PRL release from perifused normal anterior pituitary cells was rapid, sustained, and concentration related. Although it had no apparent effect on normal prelabeled anterior pituitary cells with respect to 45Ca2+ efflux, the calcium channel blocker D-600 inhibited TF5-mediated hormone release from these cells. Additive increases in TRH-stimulated PRL release and GRF-stimulated GH release by TF5 suggested independent mechanisms of action. Dopamine (500 nM) blocked TF5-stimulated PRL release, but somatostatin (10-100 nM) had no effect on TF5-stimulated PRL or GH release. TF5 failed to affect either basal or TRH-induced polyphosphoinositide hydrolysis. Perifused normal anterior pituitary cells prelabeled with [3H]arachidonate responded to TF5 treatment with a liberation of radioactive arachidonate and/or its metabolites. BW755c, an inhibitor of all known catabolic pathways of arachidonic acid, blocked the ability of TF5 to stimulate PRL and GH release. Reversed phase HPLC separation of TF5 into five fractions resulted in two fractions that exhibited hormone-releasing activity. These data suggest that TF5 stimulates pituitary hormone release through a mechanism different from that ascribed to TRH or GRF. The stimulus-secretion coupling mechanism involves neither polyphosphoinositide hydrolysis nor cAMP generation, but appears to be dependent on the generation of arachidonate metabolites.

Animals↗
PubMed1985

Evidence that thymosins and other biologic response modifiers can function as neuroactive immunotransmitters.

An increasing amount of data supports the hypothesis that there are bidirectional circuits between the central nervous system (CNS) and the immune system. Soluble products that appear to transmit information from the immune compartment to the CNS include thymosins, lymphokines, and certain complement proteins. Opioid peptides, adrenocorticotropic hormone (ACTH), and thyroid-stimulating hormone (TSH) are additional products of lymphocytes that may function in immunomodulatory neuroendocrine circuits. It is proposed that the term "immunotransmitter" be used to describe molecules that are produced predominantly by cells that comprise the immune system but that transmit specific signals and information to neurons and other cell types. Examples would include thymosin alpha 1 and beta 4, lymphocyte-derived ACTH, TSH, and beta-endorphin, interleukin 1, interferon as well as certain other lymphokines and cytokines. The evidence that certain thymosin peptides can serve as immunotransmitters by modulating the hypothalamic-pituitary-adrenal and gonadal axes will be discussed.

Adrenocorticotropic Hormone↗
PubMed1984

Immunochemical studies on thymosin: radioimmunoassay of thymosin beta 4.

Thymosin beta 4, a peptide with hormonal-like properties first isolated from the thymus gland, can be measured in serum using a newly described radioimmunoassay. The radioimmunoassay utilizes an antibody raised in rabbits against synthetic thymosin beta 4 conjugated by glutaraldehyde to keyhole limpet hemocyanin. A 125I-tyrosine-C13 analogue of the biologically active C-terminal fragment is used as the radioactive tracer. The radioimmunoassay is sensitive in the nanogram range and no cross-reactivity with common serum proteins is demonstrable. High performance liquid chromatography of serum samples indicates that two thymosin beta 4 cross-reactive species are present in human serum. Levels in serum range from 450 to 1100 ng/ml and decline with age.

Amino Acid Sequence↗
PubMed1983

Isolation of thymosin alpha 1 from thymosin fraction 5 of different species by high-performance liquid chromatography.

High-performance liquid chromatography (muBondapak C18 column with 0.05% trifluoroacetic acid in acetonitrile as solvent system) was used to isolate thymosin alpha 1 (alpha 1) from thymosin fraction 5 (f5) of various species (calf, pig, sheep and mouse). Each of the f5 preparations gave a protein peak similar in retention time to bovine thymosin alpha 1. This peak coincided with the immunoreactive peak determined by a radioimmunoassay for alpha 1. Chromatographic analysis of fresh thymus tissue extracts using a high-performance liquid chromatographic similar system did not reveal a detectable protein peak or immunoreactive peak at the alpha 1 position. Our results suggest that alpha 1 may be synthesized in a precursor form in animal tissues.

Animals↗
PubMedDate not supplied

Thymosin stimulates interleukin-6 production from rat spleen cells in vitro.

Thymosin fraction 5 (TF5) is a partially purified preparation of bovine thymus that affects the differentiation and function of T-cells in vitro. Interleukin-6 (IL-6) is a pleiotropic cytokine that induces terminal maturation of B-cells and T-cell activation and differentiation. Although TF5 had previously been shown to stimulate the production of a number of lymphokines, its effects on IL-6 were not known. In this study we determined the effect of TF5 on IL-6 production from rat spleen cells in vitro. TF5 (100 micrograms/ml) stimulated IL-6 production from splenocytes (0.75-3.0 x 10(5) cells/well) in the presence of 0.008-0.2 micrograms/well of the T-cell mitogen concanavalin-A (con-A) by 10-20 fold during a 72 h incubation period. Dose-response studies demonstrated that 10 micrograms/ml of TF5 was the lowest concentration capable of enhancing IL-6 production. The ability of TF5 to stimulate IL-6 production in the presence of con-A could be demonstrated within 24 h of incubation; longer incubation periods (48-72 h) correlated with further enhancements of IL-6 production. Partial purification of the IL-6-inducing activity from TF5 resulted in three subfractions possessing activity in the presence of con-A (MB2, MB3, MB7) and one in the absence of con-A (MB2). The previously characterized thymosin peptides T alpha 1 and T beta 4 had no effect on IL-6 production in the absence or presence of mitogen. This study reports a new biological activity for TF5 and suggests that a novel constituent of TF5 may enhance the production of IL-6 from spleen cells.

Animals↗