Stefan R. Bornstein

Plasma catecholamines in patients with Addison's disease

BACKGROUND AND OBJECTIVE TWO endocrine tissues are present within the adrenal gland: the steroid producing cortical cells and the catecholamine producing chromaffin cells. Glucocorticolds occur in high concentrations in the adrenal medulla. In vitro, glucocorticolds have been shown to induce the enzyme phenyl‐N‐methyl‐transferase which is necessary for the production of adrenaline in adrenal medullary cells. The purpose of this study was to evaluate the possible significance of a local glucocorticold effect on adrenomedullary function. DESIGN Plasma catecholamine levels were measured in patients with autoimmune Addisons disease where local production of corticosterolds is deficient In the presence of intact chromaffin tissue.

SYMPATHOADRENAL REGULATION OF ADRENOCORTICAL STEROIDOGENESIS

Sympathoadrenal regulation of adrenocortical steroidogenesis was studied on a physiological, cellular, molecular and morphological level. The effects of nerve activation and of epinephrine (EPI) on adrenal corticosteroid release were compared in isolated perfused pig adrenals with preserved nerve supply. Splanchnic nerve activation as well as perfusion with EPI provoked a significant release of cortisol, aldosterone and androstenedione. In cultured bovine adrenocortical cells steroid secretion and accumulation of P450scc, P450(17) alpha, P450c21 and P450(11) beta mRNAs were studied after stimulation with EPI with or without propranolol or phentolamine. Incubation with EPI stimulated steroidogenesis and increased the levels of all four P450-mRNAs. The beta-adrenergic antagonist propranolol totally blocked the effects of EPI while the alpha-antagonist phentolamine had no effect. Using immunohistochemistry, adrenals were studied morphologically. The contact zones of the two cell types were investigated on an electron microscopical level. Cortical and medullary cells were closely interwoven with cortical and chromaffin cells in direct apposition, providing the possibility for paracrine interactions. It is concluded that the release of corticosteroids can be stimulated through the sympatho-adrenal system. The stimulatory action of EPI upon adrenal steroid formation and accumulation of all four P450-mRNAs requires beta-adrenergic receptors. Taking into consideration the close colocalization of cortical and medullary tissue, this stimulation may be mediated by chromaffin cells in a paracrine manner.

Macrophages within the human adrenal gland.

There is increasing evidence for an immune-adrenal interaction in which macrophages may play an important role. However, few data are available with respect to a human intra-adrenal macrophage system. In this study, we have investigated the density, distribution and phenotype of human adrenal macrophages using monoclonal antibodies. Macrophages are localized in all zones of the adrenal gland. These cells exhibit the phenotype of the phagocytotic macrophage compartment (CD11c+, KiM8+). At the ultrastructural level, macrophages are frequently attached to the endothelial wall, but also lie in direct contact with cortical and chromaffin cells. This investigation reveals the cellular basis for the possible role of macrophages in the local immune-neuroendocrine axis.

Morphological and functional studies of the paracrine interaction between cortex and medulla in the adrenal gland

Within the last years it has become evident that besides the hypothalamo‐pituitary‐adrenal axis, extrapituitary mechanisms exist that regulate the activity of the adrenal cortex. In this context, intra‐adrenal regulatory mechanisms play an important role. Several secretory products from adrenomedullary cells are able to influence adrenocortical steroidogenesis. Since the main blood flow within the adrenal is directed centripetally from the cortex to the medulla, chromaffin cells should act on cortical cells in a paracrine manner. The morphological prerequisite for this regulatory pathway is seen in the close apposition of the two tissues. Within the mammalian adrenal, the two endocrine tissues are interwoven to an astonishing degree with cortical cells located within the medulla and vice versa. It is concluded from morphological and functional studies that paracrine interactions between cortex and medulla play an important role in the regulation of adrenocortical steroidogenesis. Microsc. Res. Tech. 36:520–533, 1997.

Cellular communication in the neuro-adrenocortical axis: role of vasoactive intestinal polypeptide (VIP)

It is well established now that adrenocortical function, besides being regulated through systemic factors, is influenced by intra-adrenal mechanisms. In this context paracrine influences between the sympathoadrenal system and the adrenal cortex play an important role. As a prerequisite for these interactions, adrenal medullary cells and cortical cells are highly interwoven as revealed by immunohistochemistry. The potential role of VIP in the regulation of human adrenal steroidogenesis was now investigated in human adrenal cells in primary culture. The primary cultures contained both, cortical and chromaffin cells which were found to be in close cellular contact as revealed by immunocytochemistry. VIP enhanced cortisol secretion from adrenal cells in a dose-dependent manner with a maximal effect at 10(-7) M. VIP stimulated the release of dehydroepiandrosterone (DHEA), testosterone, androstenedione, and aldosterone significantly. The addition of propranolol, a beta-adrenergic antagonist, to the incubation medium attenuated VIP-induced corticosteroid secretion. It is concluded that VIP is a paracrine messenger in the human adrenal that could regulate adrenocortical function at least in part via catecholamines released from the medulla.

Cellular immune-endocrine interaction in adrenocortical tissues

The detection of important immunocompetence‐like features on endocrine steroid cells raises questions about direct intercellular communication between the adrenal and immune systems. This article summarizes our recent work and new data on immune–adrenal interactions.

A New View on Immune-Adrenal Interactions: Role for Fas and Fas Ligand?

The hypothalamic-pituitary-adrenal axis and the immune system interact in a bidirectional manner providing the basis for the regulation of the immune response due to a pathogenic stimulus. This interplay is commonly believed to be based on the action of hormones or cytokines, respectively. Since it has been detected that adrenocortical cells offer immunological properties such as expression of MHC class II antigens and/or CD95 (Fas antigen) and its ligand, the question has to be raised whether direct intercellular communication between immune cells and ‘immunocompetent’ endocrine cells contributes to the complexity of immunoregulation. Here we discuss the possible reciprocal relevance of physiological and pathological adrenal changes, as well as T-cell-mediated immune response for immune and/or adrenal pathology during disease.

Angiotensin II regulates both adrenocortical and adrenomedullary function in isolated perfused pig adrenals

The effect of angiotensin II (ANG II) on all four zones of the adrenal gland was studied in preparations of isolated perfused porcine adrenals. The experimental design offered the possibility to analyze directly the actions of ANG II while preserving the structure of the gland. ANG II stimulated aldosterone, cortisol, and androstenedione release in a dose-dependent manner. At a final ANG II concentration of 10(-8) M aldosterone increased from 0.7 +/- 0.05 to 3.4 +/- 0.9 ng/ml, cortisol from 50 +/- 5 to 430 +/- 60 micrograms/l, and androstenedione from 1.4 +/- 0.2 to 4.4 +/- 0.8 ng/ml. In addition, ANG II provoked a release of adrenaline from 4.1 +/- 0.6 to 27.5 +/- 0.5 micrograms/ml and of noradrenaline from 5.5 +/- 1.1 to 36.0 +/- 8.7 micrograms/ml. Our results show that secretion of both adrenocortical steroids and adrenomedullary catecholamines can be evoked by ANG II. ANG II seems to influence not only the function of the zona glomerulosa but the function of the entire adrenal gland.

Expression of interleukin-1 in human pheochromocytoma

Cytokines and in particular interleukin 1 (IL-1) play a role in the micro-environment of various tumors. In addition, it is well established that IL-1 is a neurotrophic, angiogenetic and fibrogenetic factor. Using in situ hybridization we analyzed the expression of endogenous IL-1 in human pheochromocytoma (PCC). With the help of specific antibodies to B- and T-cells, macrophages, and neuroendocrine cell antigens, we characterized the distribution and localization of various cell types in human PCC. The combination of immunohistochemistry and in situ hybridization was eminently suited to defining the exact cellular source of IL-1 expression. In situ hybridization and immunostaining revealed that IL-1 mRNA was located primarily in the tumor cell itself. IL-1 may therefore constitute an important paracrine/autocrine factor in neuroendocrine tumors.

Evaluation of apoptotic parameters in normal and neoplastic human adrenal

The migration and proliferation of adrenocortical cells is accompanied by mechanisms of cellular knock-out. We compared the programmed cell death of normal and malignant adrenocortical tissues on the basis of apoptotic rates by the nonradioactive in situ end-labelling of DNA-fragments, immunohistochemistry against PCNA, CD95 and ultrastructural analysis. The highest labelling index (LI) was detectable in the outermost zones of the adrenal cortex of normal adrenals. Average LI in normal adrenal cortex was 20% whereas only 2% was detectable in adrenocortical neoplasms. MHC class II, which was previously shown to be involved in programmed cell death in lymphocyte populations (1), was detectable in normal and benign but not in malignant adrenocortical neoplastic cells. In conclusion, the analysis of apoptosis provides new aspects of normal adrenal zonation and allows the differentiation between normal and neoplastic adrenal cortex although the differentiation between malignant and benign neoplasms requires further markers.