Lymphocyte Growth Hormone Research Articles

Expression of lymphocyte-derived growth hormone (GH) and GH-releasing hormone receptors in aging rats

In the present study, we show that higher levels of lymphocyte GH are expressed in spleen cells from aging animals compared to young animals. Further, leukocytes from primary and secondary immune tissues and splenic T and B cells from aging rats all express higher levels of GHRH receptors compared to younger animals. Bone marrow and splenic T cells express the highest levels of GHRH receptor in aging animals. Spleen cells from aging animals showed no significant change in proliferation or GH induction after treatment with GHRH. Taken together, the data for the first time show alterations in GH synthesis and expression of the GHRH receptor on cells of the immune system that may play a role in the immune response in aging.

Acute circuit-resistance exercise increases expression of lymphocyte agouti-related protein in young women

Exercise-induced leukocytosis and lymphocytosis is accompanied by up-regulation and down-regulation of hundreds of genes in white blood cells (WBCs). Agouti-related protein (AgRP) is an orexigenic peptide secreted predominantly from the arcuate nucleus in the hypothalamus. AgRP affects feeding behavior and plays a role in energy and glucose homeostasis and adiposity. The purpose of the study was to determine effects of circuit resistance exercise (CRE) (9 exercises, 25 s per exercise) at different intensities on peripheral blood lymphocyte (PBL) AgRP mRNA expression and its concentrations in lymphocytes and plasma. Twenty-five young female college students were randomly divided into five groups: control, 40% 1-repetition maximum (1-RM), 60% 1-RM, 80% 1-RM and combined (40 + 60 + 80% 1-RM) loads. Peripheral blood mononuclear cells were isolated by a lymphocyte density gradient centrifugation method for AgRP mRNA expression. Lymphocyte ATP, glycogen, AgRP, growth hormone (GH), and plasma AgRP, GH and glucose concentrations were measured. CRE increased AgRP mRNA lymphocyte expression significantly (P < 0.0001) at all intensities. A higher and significant (P < 0.01) increase was found in the 60% 1-RM group when compared with the other groups. The CRE-induced lymphocyte AgRP expression was accompanied by elevations in plasma AgRP, glucose and GH levels as well as higher WBCs, lymphocytes and neutrophil counts. Lymphocyte AgRP and GH concentrations were significantly reduced (P < 0.05). Lymphocyte ATP content was unchanged and glycogen was reduced in the combined group but not in the other groups. Data indicate that AgRP mRNA is expressed in PBLs and that CRE increases its expression. Data also reveal that the expression of AgRP was accompanied with higher plasma AgRP and GH concentrations. Findings suggest that AgRP may provide an important signal in the immune environment and that the lymphocyte may be considered as an extra-hypothalamic source of plasma AgRP following exercise stress.

Expression of insulin-like growth factor-1 and insulin-like growth factor-1 receptors in EL4 lymphoma cells overexpressing growth hormone

Almost all of the previous studies with growth hormone (GH) have been done with exogenously supplied GH and, therefore, involve actions of the hormone through its receptor. However, the actions of endogenous or lymphocyte GH are still unclear. In a previous study, we showed that overexpression of GH (GHo) in a lymphoid cell line resulted in protection of the cells to apoptosis mediated by nitric oxide (NO). In the present study, we show that the protection from apoptosis could be transferred to control cells with culture fluids obtained from GHo cells and blocked by antibodies to the insulin-like growth factor-1 (IGF-1) or antibodies to the IGF-1-receptor (IGF-1R). Northern and Western blot analysis detected significantly higher levels of IGF-1 in cells overexpressing GH. An increase in the expression of the IGF-1R in GHo cells was also detected by Western blot analysis, 125I-IGF-1 binding and analysis of IGF-1R promoter luciferase constructs. Transfection of GHo cells with a dominant negative IGF-1R mutant construct blocked the generation of NO and activation of Akt seen in GHo cells compared to vector alone control EL4 cells. The results suggest that one of the consequences of the overexpression of GH, in cells lacking the GH receptor, is an increase in the expression of IGF-1 and the IGF-1R which mediate the protection of EL4 lymphoma cells from apoptosis.

The Inhibition of Superoxide Production in EL4 Lymphoma Cells Overexpressing Growth Hormone

A substantial body of research exists to support the production of growth hormone by cells of the immune system. However, the function and mechanism of action of lymphocyte-derived growth hormone remain largely unelucidated. Since, it has been found that exogenous growth hormone (GH) primes neutrophils for the production of reactive oxygen intermediates (ROI) and in particular superoxide we investigated the role of GH on the production of in T cells. Furthermore, we examined whether endogenous and exogenous GH act similarly. Our studies show that overexpression of GH in EL4, a T-cell lymphoma cell line, results in a decrease in the production of compared to control cells, as detected using the fluorescent dye, dihydroethidium. production in control cells was not affected by treatment with inhibitors of xanthine oxidase or a non-specific NADPH-oxidase inhibitor. However, treatment with diallyl sulfide, an inhibitor of cytochrome P450 2E1 mimicked the reduction in production seen in cells overexpressing GH. Although no significant change could be detected in CYP2E1 protein levels, CYP2E1 activity was found to be greater in control EL4 than in cells overexpressing GH. Both the decrease in production and the lower CYP2E1 activity in GH overexpressing cells could be abrogated by treatment with NG-monomethyl-L-arginine, an inhibitor of nitric oxide synthase. The overexpression of GH protects cells from apoptosis induced by isoniazid, a CYP2E1 inducer, suggesting a role for nitric oxide as a mediator in the regulation of xenobiotic metabolism and apoptosis-protection by lymphocyte GH.

The production of nitric oxide in EL4 lymphoma cells overexpressing growth hormone

Growth hormone (GH) is produced by immunocompetent cells and has been implicated in the regulation of a multiplicity of functions in the immune system involved in growth and activation. However, the actions of endogenous or lymphocyte GH and its contribution to immune reactivity when compared with those of serum or exogenous GH are still unclear. In the present study, we overexpressed lymphocyte GH in EL4 lymphoma cells, which lack the GH receptor (GHR), to determine the role of endogenous GH in nitric oxide (NO) production and response to genotoxic stress. Western blot analysis demonstrated that the levels of GH increased approximately 40% in cells overexpressing GH (GHo) when compared with cells with vector alone. The results also show a substantial increase in NO production in cells overexpressing GH that could be blocked by N G-monomethyl- l-arginine ( l-NMMA), an l-arginine analogue that competitively inhibits all three isoforms of nitric oxide synthase (NOS). No evidence was obtained to support an increase in peroxynitrite in cells overexpressing GH. Overexpression of GH increased NOS activity, inducible nitric oxide synthase (iNOS) promoter activity, and iNOS protein expression, whereas endothelial nitric oxide synthase and neuronal nitric oxide synthase protein levels were essentially unchanged. In addition, cells overexpressing GH showed increased arginine transport ability and intracellular arginase activity when compared with control cells. GH overexpression appeared to protect cells from the toxic effects of the DNA alkylating agent methyl methanesulfonate. This possibility was suggested by maintenance of the mitochondrial transmembrane potential in cells overexpressing GH when compared with control cells that could be blocked by l-NMMA. Taken together, the data support the notion that lymphocyte GH, independently of the GH receptor, may play a key role in the survival of lymphocytes exposed to stressful stimuli via the production of NO.

Human lymphocyte growth hormone stimulates interferon gamma production and is inhibited by cortisol and norepinephrine

Lymphocyte growth hormone (L-GH) is distributed throughout the human immune system; however, its biologic role has not been defined. In order to clarify this issue, we determined if Candida, commonly used as a recall antigen, and IL-12, an important monocyte cytokine that stimulates the TH1 cytokine pathway, could stimulate L-GH mRNA synthesis in cultured human peripheral blood mononuclear cells (PBMCs). Both Candida and IL-12 produced significant ( p<0.01) stimulation of L-GH mRNA synthesis and the TH1 cytokine, gamma interferon (IFN-γ). These effects on IFN-γ production were significantly ( p<0.001) inhibited by a GH antagonist, suggesting that L-GH was acting by an autocrine or paracrine mechanism to enhance IFN-γ production in these serum-free cultured cells. Concentrations of norepinephrine and cortisol achievable in humans during stress were able to significantly ( p<0.001) decrease L-GH synthesis in the PBMCs. They also diminished IFN-γ significantly ( p<0.001), but not the TH2 cytokine, IL-10, in the supernatants of the cultured PBMLs. These studies suggest that L-GH plays a role in cellular immune function mediated via the TH-1 pathway.

Growth Hormone and Pit-1 expression in bovine fetal lymphoid cells

Bovine fetal lymphoid cells were examined for growth hormone (GH) and the transcription factor Pit-1/GHF-1 mRNA. GH and Pit-1/GHF-1 transcripts were detected in thymocytes and splenocytes from fetuses at 60, 90, 120, and 270 d of gestation using reverse transcription-polymerase chain reaction (RT-PCR). Northern analysis indicated that the lymphoid GH mRNA was approximately 350 nucleotides larger than in the pituitary. RT-PCR analysis demonstrated that the coding regions as well as 3′ untranslated regions of the lymphocyte GH and pituitary transcripts were the same. Analysis of the 5′-untranslated region of the lymphocyte GH mRNA showed that transcription began upstream from the start site in the pituitary gland, suggesting differences in regulation in these tissues. Fetal thymocytes and splenocytes expressed Pit-1/GHF-1 mRNA; however, they contained only the 2.5-kb transcript. The GH and Pit-1/GHF-1 mRNA in fetal lymphoid cells supports the hypothesis that lymphocyte-derived GH may function as an autocrine and/or paracrine factor during the development and maturation of the bovine fetal immune system.

Cloning and nucleotide sequencing of rat lymphocyte growth hormone cDNA.

The messenger RNA for rat growth hormone (GH) was isolated from rat spleen lymphocytes and cloned as a cDNA after PCR. Four clones, including part of the prehormone sequence and the full-length mature GH sequence, were obtained and sequenced. The sequence data revealed that rat lymphocyte GH was identical to that reported for pituitary GH. The results do not support the expression by alternate splicing of the 20-kD variant described in the pituitary. We also cloned the promoter of lymphocyte GH and analyzed its nucleotide sequence including 300 base pairs of the 5'-flanking region. The promoter sequence we obtained did not exactly match that reported in the literature because of reading compressions on the gel. However, parellel sequencing of thymus, spleen and pituitary GH promoter sequences gave identical patterns of compressions in the gel. The results suggest that the sequence in all the tissues was the same. Collectively, these studies demonstrate that lymphocytes express authentic GH mRNA and suggest that GH gene expression may be regulated in immune tissue by similar transcriptional factors to those described in the pituitary.

Effect of the Administration of Growth-Hormone-Producing Lymphocytes on Weight Gain and Immune Function in Dwarf Mice

Our previous work has shown that cells of the immune system produce a growth hormone (GH) molecule similar to that secreted by the pituitary. In the present studies, we evaluated the possibility that normal spleen cells producing GH transferred to dwarf mice could stimulate their growth. The results showed that normal spleen cells alone or spleen cells treated with growth-hormone-releasing hormone (GHRH) did not appear to significantly stimulate the growth of dwarf mice. Spleen cells activated in vitro with concanavalin A or lipopolysaccharide and then transferred to dwarf mice, or thymus cells alone, were also without effect, whereas GH alone stimulated growth as expected. Serum levels of insulin-like growth factor-I (IGF-I) and IGF-I-liver RNA were undetectable in control dwarf mice and dwarf mice receiving spleen cells, whereas serum levels of IGF-I increased after treatment of dwarf mice with GH. The immune system of dwarf mice receiving spleen cells, however, was significantly altered. Spleen cells from dwarf animals showed enhanced immunoglobulin, interleukin (IL)-6, IL-2, and interferon-gamma production whereas no significant change was apparent in natural killer cell activity. Despite the absence of the pit-1 protein in dwarf mice, their spleen and thymus cells retained the ability to produce almost as much lymphocyte GH as normal. Overall, the findings support the idea that the pit-1 protein in lymphocytes of dwarf mice may not be obligatory for the expression of lymphocyte GH.(ABSTRACT TRUNCATED AT 250 WORDS)

1α,25-Dihydroxyvitamin D 3 and a novel vitamin D analogue MC 903 are potent inhibitors of human interleukin 1 in vitro

1α,25-dihydroxycholecalciferol (1,25(OH) 2D 3) inhibits the lymphocyte growth hormone, interleukin 2. Since its production is dependent upon interleukin 1 (IL-1) produced by antigen-presenting cells, we tested five vitamin D 3 analogues for effects on the production and function of human natural and recombinant IL-1. The production was not affected, but 1,25(OH) 2D 3 (>10 −11M) and a synthetic derivative MC 903 (> = 10 −10M) inhibited the proliferation of mouse thymocytes to IL-1. The vitamins failed to affect the cytotoxic activity of tumor necrosis factor. 1,25(OH) 2D 3 may play a physiological immunomodulatory role as a selective inhibitor of the function of IL-1, and MC 903 may prove clinically useful in this regard because of its limited calcium metabolic activity.

Growth hormone receptors in lymphocytes of growth hormone-deficient children

Human growth hormone was injected intravenously into 18 growth hormone-deficient children and growth hormone binding sites in lymphocytes were investigated. Fresh circulating lymphocytes had a low initial value for the binding of growth hormone to solubilized receptors (3.45 ± 1.46%) but after growth hormone injection, the binding rapidly increased to 14.8 ± 4.2% at 2 1 2 h and 8.7 ± 1.8% at 5 h. The sharp increase in binding is due to increase in the number of binding sites. Two control children who received chorionic gonadotropin had no change in lymphocyte growth hormone receptors. The methodological differences between the present study and previous attempts to identify human growth receptors in lymphocytes were (1) lymphocytes were separated and disrupted with Triton X-100 as quickly as possible (to avoid error from receptor leaking out of the cell) and (2) the receptors were assayed at 2 1 2 h after growth hormone administration (previous studies were 12–24 h later). One possible explanation for the data is that growth hormone receptor from liver is taken up by lymphocytes and rapidly released again, thus, contributing to the hormonal receptor economy in humans.