Differential expression of histamine receptors in the bladder wall tissues of patients with bladder pain syndrome/interstitial cystitis - significance in the responsiveness to antihistamine treatment and disease symptoms.

BACKGROUND: Activation of mast cells plays an important role in the pathogenesis of bladder pain syndrome/interstitial cystitis (BPS/IC). Histamine, a mast cell-derived mediators, induced inflammation and hypersensitivity of the bladder. The present study investigated the expressions of histamine receptors in the bladder wall tissues of patients with BPS/IC, and its association with the effectiveness of antihistamine therapy and disease symptoms. METHODS: Bladder tissues were collected from 69 BPS/IC patients and 10 control female patients. The expression of H3R in BPS/IC was further examined in an independent cohort of 10 female patients with BPS/IC and another 10 age-matched female patients. Immunohistochemistry, Western blotting, and quantitative RT-PCR were performed to quantify the expressions of histamine receptors. Statistical analyses of the correlation of histamine receptor expression with antihistamine therapy outcome and severity of disease symptoms were also performed. RESULTS: The expression of four histamine receptors was significantly elevated in BPS/IC (H1R, P < 0.001; H2R, P = 0.031; H3R, P = 0.008; H4R, P = 0.048). Western blotting revealed that H3R were significantly reduced in the patients, whereas the mRNA levels of H3R were significantly increased. The patients were further divided into antihistamine responders (n = 38) and nonresponders (n = 22). No significant correlation was found in the expression of histamine receptors between responder and nonresponder groups. However, significant correlations between OLS and H1R (P = 0.003) and H3R (P = 0.045) were found. CONCLUSION: The present study showed that expression of all the 4 histamine receptors were elevated in BPS/IC. There were no statistical significant correlations between the expression levels of the four different histamine receptors and the treatment outcome of antihistamine therapy (amtitriptyline or cimetidine).

Histamine intolerance and dietary management: A complete review

BACKGROUND: Present in several types of food, bioactive amines are described as organic bases of low molecular weight, which constitute a potential health risk. An awareness of amine levels in foods today is therefore important in relation to food safety and patient care. This review aims to emphasise the need to unify the information on the content of biogenic amines in foods and prevent patients' misunderstanding. METHODS: Selective literature search for relevant publications in PubMed and other scientific data bases combined with further data from the World Wide Web on histamine and other amines content in foods. RESULTS: Available reference sources do not reflect a homogeneous consensus, and the variation between foods makes it impossible for dieticians to accurately estimate amines content to correctly advise patients. CONCLUSIONS: To achieve the goal of collecting reliable information, all methods and tools used in analytical studies should be standardised and information exposed to patients should be verified

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BODIPY(®) FL histamine as a new modality for quantitative detection of histamine receptor upregulation upon IgE sensitization in murine bone marrow-derived mast cells.

Flow cytometry is one of the most widely used methods for the qualitative and quantitative analysis of cell surface expressed proteins by making use of fluorescent specific antibodies. Lacking an antibody validated for flow cytometry, an alternative approach for labeling cell surface receptors is the use of fluorescently tagged ligands. In this study, histamine H4 receptor transfected Chinese hamster ovary cells and murine bone marrow-derived mast cells (mBMMCs) were selected for studying the possibility of staining individual histamine receptors using BODIPY(®) FL histamine and selective antagonists. Flow cytometric measurements and supporting calculations showed that BODIPY FL histamine is suitable tool for quantitating cell surface histamine receptors. The binding, and competitive inhibition of this fluorescent ligand were characterized, which were in good agreement with a semi-empirical model constructed from fundamental protein-binding relationships. Using this method it was shown for the first time that even though mature mBMMCs express H2R and H4R to the same extent, immunoglobulin E sensitization results in H4R upregulation only, while the surface expression of H2R remains unchanged.

Human inflammatory dendritic epidermal cells express a functional histamine H4 receptor.

Expression of histamine H(4) receptor (H(4)R) on leukocytes suggests an immunomodulatory role of this receptor. Here we investigated the expression and function of H(4)R on human inflammatory dendritic epidermal cells (IDECs). H(4)R is expressed by IDEC of the skin. On monocyte-derived IDECs (Mo-IDECs), H(4)R is also expressed and upregulated by IFN-gamma. Functionally, histamine and H(4)R agonists clobenpropit and 4-methylhistamine downregulated the production of the Th2-linked chemokine CCL2 and the Th1 cytokine IL-12 on Mo-IDEC, whereas agonists for the other histamine receptors did not. An H(4)R-selective antagonist (JNJ7777120) blocked the effect of H(4)R agonists. Downregulation of CCL2 also led to a decreased migration of monocytes. Thus, IDEC express a functionally active H(4)R, which upon stimulation leads to downregulation of CCL2 and IL-12. This might have implications for the treatment of atopic dermatitis, since H(4)R agonists may have beneficial effects in downregulating inflammation.

The role of histamine in human mammary carcinogenesis: H3 and H4 receptors as potential therapeutic targets for breast cancer treatment.

There is increasing evidence that describes a histamine role in normal and cancer cell proliferation. To better understand the importance of histamine in breast cancer development, the expression of histamine H3 (H3R) and H4 (H4R) receptors and their association with proliferating cell nuclear antigen (PCNA), histidine decarboxylase (HDC) and histamine content were explored in mammary biopsies. Additionally, we investigated whether H3R and H4R were implicated in the biological responses triggered by histamine in MDA-MB-231 breast cancer cells. The expression levels of H3R, H4R, PCNA, HDC and histamine content were determined by immunohistochemistry in 40 benign and malignant lesions. MDA-MB-231 cells proliferation (clonogenic assay and BrdU incorporation) and cell cycle distribution (flow cytometry) were evaluated upon treatment with histamine, H3R and H4R agonists and antagonists. Apoptosis was determined by Annexin staining and TUNEL assay. Cell migration was assessed by transwell system. Results indicate that H3R was detected in 67% (10/15) of benign lesions and in almost all carcinomas (24/25), being the level of its expression significantly higher in carcinomas (p = 0.0016). The non-tumoral breast tissue surrounding carcinomas revealed a lower H3R expression compared to the tumor cells. Only 13% (2/15) of the benign lesions expressed H4R compared to 44% (11/25) of the carcinomas. Interestingly, H3R expression was correlated in carcinomas with the expression of HDC and PCNA (p < 0.0001), and also histamine content (p = 0.0229). Accordingly, histamine increased MDA-MB-231 cells proliferation and also migration via H3R. In contrast, activation of H4R inhibited proliferation and this effect was associated with an arrest in the G(0)/G(1) phase of the cell cycle and an induction of apoptosis. Present findings demonstrate the presence of H3R and H4R in human mammary tissue and suggest that H3R may be involved in the regulation of breast cancer growth and progression representing a novel molecular target for new therapeutic approach.

The CML-related oncoprotein BCR/ABL induces expression of histidine decarboxylase (HDC) and the synthesis of histamine in leukemic cells.

Basophil numbers are typically elevated in chronic myeloid leukemia (CML) and increase during disease progression. Histamine is an essential mediator and marker of basophils and is highly up-regulated in CML. We examined the biochemical basis of histamine synthesis in CML cells. The CML-specific oncoprotein BCR/ABL was found to promote expression of histidine decarboxylase (HDC) and synthesis of histamine in Ba/F3 cells. Moreover, the BCR/ABL tyrosine kinase inhibitors imatinib (STI571) and nilotinib (AMN107) decreased histamine levels and HDC mRNA expression in BCR/ABL-transformed Ba/F3 cells, in the CML-derived basophil cell line KU812, and in primary CML cells. Synthesis of histamine was found to be restricted to the basophil compartment of the CML clone and to depend on signaling through the PI3-kinase pathway. CML cells also expressed histamine receptors (HRs), including HR-1, HR-2, HR-4, and histamine-binding CYP450 isoenzymes which also serve as targets of HR antagonists. The HR-1 antagonists loratadine and terfenadine, which bind to CYP450, were found to counteract proliferation of CML cells, whereas no growth inhibition was observed with the HR-1 antagonist fexofenadine which is not targeted or metabolized by CYP450. Moreover, DPPE, an inhibitor of histamine-binding CYP450 isoenzymes, produced growth inhibition in CML cells. Together, these data show that BCR/ABL promotes histamine production in CML cells and that certain HR-targeting drugs exert antileukemic effects on CML cells.

Selective expression of histamine receptors H1R, H2R, and H4R, but not H3R, in the human intestinal tract.

BACKGROUND AND AIMS: Histamine is known as a regulator of gastrointestinal functions, such as gastric acid production, intestinal motility, and mucosal ion secretion. Most of this knowledge has been obtained from animal studies. In contrast, in humans, expression and distribution of histamine receptors (HR) within the human gastrointestinal tract are unclear. METHODS: We analysed HR expression in human gastrointestinal tissue specimens by quantitative reverse transcription-polymerase chain reaction and immunostaining. RESULTS: We found that H1R, H2R, and H4R mRNA were expressed throughout the gastrointestinal tract, while H3R mRNA was absent. No significant differences in the distribution of HR were found between different anatomical sites (duodenum, ileum, colon, sigma, and rectum). Immunostaining of neurones and nerve fibres revealed that H3R was absent in the human enteric nervous system; however, H1R and H2R were found on ganglion cells of the myenteric plexus. Epithelial cells also expressed H1R, H2R and, to some extent, H4R. Intestinal fibroblasts exclusively expressed H1R while the muscular layers of human intestine stained positive for both H1R and H2R. Immune cells expressed mRNA and protein for H1R, H2R, and low levels of H4R. Analysis of endoscopic biopsies from patients with food allergy and irritable bowel syndrome revealed significantly elevated H1R and H2R mRNA levels compared with controls. CONCLUSIONS: We have demonstrated that H1R, H2R and, to some extent, H4R, are expressed in the human gastrointestinal tract, while H3R is absent, and we found that HR expression was altered in patients with gastrointestinal diseases.

Epithelial distribution of neural receptors in the guinea pig small intestine.

Neural and paracrine agents, such as dopamine, epinephrine, and histamine, affect intestinal epithelial function, but it is unclear if these agents act on receptors directly at the enterocyte level. The cellular localization and villus-crypt distribution of adrenergic, dopamine, and histamine receptors within the intestinal epithelium is obscure and needs to be identified. Single cell populations of villus or crypt epithelial cells were isolated from the jejunum of adult guinea pigs. Enterocytes were separated from intraepithelial lymphocytes by flow cytometry and specific binding was determined using fluorescent probes. Alpha1-adrenergic receptors were located on villus and crypt intraepithelial lymphocytes and enterocytes. Beta-adrenergic receptors were found on villus and crypt enterocytes. Dopamine receptors were found on all cell types examined, whereas histamine receptors were not detected (<10% for each cell population). These studies demonstrated that (1) receptors for epinephrine and dopamine exist on epithelial cells of the guinea pig jejunum, (2) beta-adrenergic receptors are found primarily on villus and crypt enterocytes and (3) intraepithelial lymphocytes contain alpha1-adrenergic, but have few beta-adrenergic, receptors. The presence of neural receptors suggests that these agents are acting, at least in part, at the enterocyte or intraepithelial lymphocyte levels to modulate intestinal and immune function.

Immunohistochemical localization of histamine receptors in rat cochlea.

OBJECTIVE: Histamine may have physiologic functions in the inner ear. The locations of histamine receptors, however, have not yet been identified in the mammalian cochlea. The aim of this study was to investigate the localization of histamine receptor subtypes (H1, H2, and H3 receptors) in rat cochlea. METHODS: Immunohistochemistry was performed with antibodies specific for each of the histamine receptors (H1, H2, and H3). To identify the type I and II spiral ganglion cells in the cochlea, some cryostat sections were double stained with antibodies to both a histamine receptor and neurofilament 200 kD, which predominantly stains type II spiral ganglion cells in the cochlea. RESULTS: All H1, H2, and H3 receptor immunoreactive staining was limited to the spiral ganglion cells of the cochlea. Spiral ganglion cells with positive immunoreactivity to the neurofilament 200 kD antibody were stained only slightly by histamine H1, H2, and H3 receptor antibodies, indicating that histamine receptor immunoreactivity is specific to type I ganglion cells. CONCLUSIONS: These findings indicate that histamine receptors are present in the cochlea and support the hypothesis that histamine plays a physiologic role in the cochlea.

New concepts of histamine receptors and actions.

Histamine and antihistamines are so deeply woven into the fabric of allergic diseases that it is sometimes difficult to see how this field could advance beyond our current, potent histamine H1-receptor drugs. Investigations of other actions of histamine and the identification of H2, H3, and now H4 receptors have suddenly reignited the search for new mono- and multi-receptor-specific agonists and antagonists. There is great excitement due to preliminary findings that H3 receptors act as neural inhibitory autoreceptors, and H4 receptors might modulate immune cell functions.

Pterygial derived fibroblasts express functionally active histamine and epidermal growth factor receptors.

Pterygia are characterised by a fleshy outgrowth of altered conjunctival tissue over the cornea and are most common in tropical regions. Pterygial fibroblasts are characteristically distinct from normal conjunctival fibroblasts, and therefore the aim of this study was to determine the presence and functional significance of histamine and epidermal growth factor (EGF) receptors in these cells. Pterygial specimens were cultured in vitro and cellular outgrowths were phenotypically characterised as fibroblasts using vimentin and cytokeratin staining. Intracellular calcium mobilization was used to characterise the functional activity of histamine receptors on these cells. Maximal response was obtained with 100 microM histamine. However, lower concentrations of histamine also caused mobilization of calcium that were totally abolished by pre-incubation with H1 but not H2 or H3 receptor antagonists. EGF receptor was diffusely expressed over the cell surfaces. EGF stimulated receptor internalization, ERK protein phosphorylation and intracellular calcium mobilization. Therefore, fibroblasts derived from human pterygia express functionally active histamine and epidermal growth factor receptors. Controlled modification of either the receptors or the appropriate ligands could have beneficial effects in pterygia treatment.

Histamine regulates cytokine production in maturing dendritic cells, resulting in altered T cell polarization.

Atopic diseases such as allergy and asthma are characterized by increases in Th2 cells and serum IgE antibodies. The binding of allergens to IgE on mast cells triggers the release of several mediators, of which histamine is the most prevalent. Here we show that histamine, together with a maturation signal, acts directly upon immature dendritic cells (iDCs), profoundly altering their T cell polarizing capacity. We demonstrate that iDCs express two active histamine receptors, H1 and H2. Histamine did not significantly affect the LPS-driven maturation of iDCs with regard to phenotypic changes or capacity to prime naive T cells, but it dramatically altered the repertoire of cytokines and chemokines secreted by mature DCs. In particular, histamine, acting upon the H2 receptor for a short period of time, increased IL-10 production and reduced IL-12 secretion. As a result, histamine-matured DCs polarized naive CD4(+) T cells toward a Th2 phenotype, as compared with DCs that had matured in the absence of histamine. We propose that the Th2 cells favor IgE production, leading to increased histamine secretion by mast cells, thus creating a positive feedback loop that could contribute to the severity of atopic diseases.

Positron emission tomography receptor studies in epilepsy.

Investigations with specific PET ligands that bind to specific neuro-receptors give information on abnormalities of neurotransmission involved in the pathophysiology of the epilepsies. Data need to be interpreted in the light of optimal structural imaging. Objective voxel-based and region-based analyses, with correction of partial volume effect, are complementary. Central benzodiazepine (cBZR) and opioid receptors have been studied most. Reduced cBZR binding is commonly seen at an epileptic focus, in a more restricted distribution than an area of hypometabolism, and sometimes also in projection areas. In contrast to acquired lesions causing partial seizures, focal increases in cBZR binding have been demonstrated in malformations of cortical development and also in areas of brain that appear normal on MRI, indicating the widespread nature of the abnormalities. Focal abnormalities of cBZR are also commonly found in patients with partial seizures and normal MRI. It is not yet clear how useful these data will prove to be in presurgical evaluation. Mu- and delta-opioid receptors have been found to be increased in temporal neocortex overlying mesial temporal epileptic foci, but with different patterns of increase. Dynamic studies of the binding of 11C-diprenorphine to opioid receptors are possible using PET, and have implied the release of cerebral endogenous opioids at the time of serial absences and reflex seizures induced by reading. Other tracers, that have been applied less widely, label the enzyme monoamine oxidase type B and peripheral benzodiazepine and histamine H1 receptors.

Platelet derived growth factor-BB induced calcium transients in cultured human peritoneal mesothelial cells.

The peritoneal membrane is used as an artificial dialysis organ for patients with end-stage renal failure during continuous ambulatory peritoneal dialysis (CAPD). Resident peritoneal mast cells may influence the course of the inflammatory process during acute infection or prolonged exposure to dialysate in a paracrine fashion, with the release of preformed mediators, including platelet derived growth factor (PDGF), which may activate human peritoneal mesothelial cells (HPMC) and other peritoneal cells. PDGF receptor interactions have multiple intracellular effects, typified by cell proliferation, protein synthesis, cytoskeletal organization, extracellular matrix turnover, and angiogenesis. PDGF may mediate HPMC inflammatory response by stimulating cytokine release. The presence of transcripts for both PDGF-B polypeptide and PDGF-beta receptor in cultured HPMC was confirmed using a reverse transcriptase-polymerase chain reaction. To investigate the activity of PDGF-beta receptors in HPMC, the authors examined intracellular calcium (Ca2+(i)) mobilization in HPMC in response to PDGF-BB (100 ng/ml), histamine (1.0 mmol/L), and 4-brA23187 (1.0 micromol/L) using the calcium indicator, fura-2. HPMC calcium mobilization in response to PDGF was assessed in Krebs-Ringer saline, with and without added external calcium (Ca2+(ext)). HPMC responded to PDGF with a transient rise in Ca2+(i) (approximately 1.6-fold) that returned to an elevated resting value. In the absence of Ca2+(ext), PDGF produced a Ca2+(i) transient indicative of Ca2+ release from intracellular stores. These results suggest that HPMC have functional PDGF-beta receptors that may bind and respond to PDGF in both an autocrine and paracrine

Gastric carcinoid with histamine production, histamine transporter and expression of somatostatin receptors.

A case of sporadic, histamine-producing gastric carcinoid with liver metastases is reported. The patient was treated with somatostatin analogue (octreotide) combined with cortisone and blockade of histamine receptors prior to surgery, which included subtotal gastrectomy, excision of lymph node metastases and superficial liver metastases. Residual liver metastases were injected with ethanol. These interventions markedly reduced the urinary excretion of the main histamine metabolite (MelmAA). Eighteen months later combined immuno- and chemotherapy was initiated due to tumour progression and recurrent hormonal symptoms with good clinical results over 12 months. Scintigraphy, using 111In-DTPA-D-Phe1-octreotide, visualized somatostatin receptors (sstr) in primary tumour, lymph node metastases and liver metastases. The tissue/blood 111In concentration ratios of tumour biopsies were very high. Northern analyses confirmed expression of all subtypes of sstr1-5. Immunocytochemically, tumour cells were strongly positive for chromogranin A, histamine and vesicular monoamine transporter (VMAT) 2 (histamine transporter), but negative for VMAT 1, suggesting an origin from gastric enterochromaffin-like cells. In primary tumour cell cultures, histamine, 5-HTP and 5-HIAA, but not 5-HT, could be detected in conditioned culture medium, indicating a defective decarboxylation of the tryptamine precursor. This rare case of histamine-producing gastric carcinoid demonstrates that excellent symptom relief can be achieved despite disseminated disease, if active, multimodal treatment strategy is instituted. The presence of high numbers of sstr in tumour tissue also raises the possibility of receptor-guided radiotherapy.

Histamine mediates the muscle layer-specific responses in the isolated swine myometrium.

To clarify the role of histamine in uterine contractility, the effect of this biogenic amine on the myometrium of cyclic mature gilts was investigated by an isometric tension recording study in vitro. In addition, using crude membrane preparations isolated from the longitudinal (LM) and circular muscle (CM), the distribution of H1 histamine receptors was characterized by 3H-pyrilamine binding assay. Histamine caused a tetrodotoxin-resistant contractile response of LM and CM in Krebs solution, but LM (-logEC50 = 6.34) was more sensitive than CM (-logEC50 = 5.4). Pyrilamine decreased the excitatory response of histamine in both muscle layers. In pyrilamine-treated LM, a high concentration of histamine (1-30 microM) caused a slight inhibition of spontaneous contraction, and this inhibition was abolished by ranitidine. On the other hand, histamine did not cause any inhibition in the pyrilamine-treated CM preparations. Dimaprit (10-300 microM) concentration-dependently inhibited the spontaneous contraction of LM but not of CM. In the presence of pyrilamine and ranitidine, N alpha-methylhistamine, even at 10 microM, did not affect the spontaneous and electrical field stimulation (5Hz)-induced contraction of LM and CM layers. Specific 3H-pyrilamine binding sites were distributed heterogeneously in the swine myometrium. The maximum number of binding sites in LM (132.5 +/- 9.9 fmol/mg protein, n = 10) was 2.5 times higher than that in CM (52.2 +/- 3.2 fmol/mg protein, n = 6). These results indicate that there is a muscle layer-dependent difference of histamine-induced response in the swine myometrium. In the LM layer, histamine acts on both H1 and H2 histamine receptors, and causes contraction (via H1 receptors at a low concentration) or relaxation (via H2 receptors at a high concentration in the presence of pyrilamine). However, histamine causes only a contraction in the CM layer, likely the result of the absence of H2 histamine receptors. Histamine-induced contraction is conspicuous in the LM layer, because of the heterogeneous distribution of H1-receptors between LM and CM.

A preparative method for sequencing proteins and peptides: in situ gel staining with subsequent passive elution onto polyvinylidine difluoride membranes.

A preparative method for obtaining both N-terminal and internal peptide amino acid sequences from purified proteins is reported. The methodology reliably yields high fidelity signal from between 14 to 30 residues per purified protein or peptide, with low backgrounds on amino acid analysis. The procedure relies on the use of in situ staining of proteins during preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the utilisation of microconcentrators to repeatedly concentrate small amounts of proteins onto a small polyvinylidene difluoride (PVDF) disc until sufficient amounts have been adsorbed so as to give a strong sequencing signal. The protein elution and subsequent adsorption can be monitored visually with a dye and the final product, a PVDF disc with the adsorbed protein or peptide, can be directly inserted into the automated amino acid sequencer.

A novel fluorescein-histamine reagent to quantitate histamine receptor expression on leucocytes.

The expression of histamine receptors on the surface of rat lymph node cells was studied using a reagent made by directly coupling fluorescein isothiocyanate (FITC) to histamine. This approach contrasts with the use of previous reagents, made by coupling histamine and fluorescein separately to a protein carrier, which bind non-specifically to cells and cause staining unrelated to histamine receptor expression. The new reagent was used, in combination with a panel of monoclonal antibodies, for the dual staining of rat lymph node cells for two-colour flow cytometric analysis to investigate the distribution of histamine receptors on different leucocyte subsets. The majority of cells were stained by the FITC-histamine reagent and these constituted two distinct populations, those with the properties of small lymphocytes and a second population which included macrophages. Inhibition studies with the drugs mepyramine and cimetidine, which are antagonists of H1 and H2 receptors, respectively, showed that most lymphocytes possess H1 receptors while the macrophages have H2 receptors. It seems that macrophages have a higher number of histamine receptors than the majority of lymphocytes, but that they are of lower affinity.