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Int Arch Allergy Immunol 2014;163:11-19

Histamine Downregulates the Th1-Associated Chemokine IP-10 in Monocytes and Myeloid Dendritic Cells

Glatzer F.a · Mommert S.a · Köther B.a · Gschwandtner M.a, d · Stark H.b, c · Werfel T.a · Gutzmer R.a
aDivision of Immunodermatology and Allergy Research, Department of Dermatology and Allergy, Hannover Medical School, Hannover, bZAFES/CMP/ICNF, Biocenter, Institute of Pharmaceutical Chemistry, Johann Wolfgang Goethe University, Frankfurt am Main, and cInstitute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University, Düsseldorf, Germany, dResearch Division of Biology and Pathobiology of the Skin, Department of Dermatology, Medical University of Vienna, Vienna, Austria
email Corresponding Author


 goto top of outline Key Words

  • CXCL10
  • Histamine
  • Histamine receptors
  • IP-10
  • Monocytes
  • Myeloid dendritic cells

 goto top of outline Abstract

Background: Histamine is an important mediator of allergic diseases. It modulates the cytokine expression of various subtypes of antigen-presenting cells by four known receptors, H1R-H4R. The effects of histamine on myeloid dendritic cells (mDC) are unclear. Methods: Monocytes and mDC were isolated from human PBMC. Histamine receptor expression was evaluated by real-time PCR. Cells were stimulated with histamine and histamine receptor ligands, and restimulated with polyinosinic-polycytidylic acid (poly I:C), and supernatants were analyzed by protein array and ELISA. Results: Monocytes and mDC express H1R and H2R without significant differences between the two cell types, whereas H4R mRNA was significantly higher in mDC compared with monocytes and H3R mRNA was not detected in any cell type. Prestimulation with histamine caused a significant decrease in poly I:C-induced expression of interferon-γ-induced protein (IP-10) in mDC and monocytes. Stimulation with specific H1R, H2R and H4R agonists and antagonists showed that the observed effect was mediated via H2R and H4R in monocytes and mDC. Conclusion: Monocytes and mDC have similar histamine receptor repertoires with regard to H1R, H2R and H3R, but H4R expression is higher on mDC. Histamine stimulation shows similar functional effects on both cell types, i.e., downregulation of TLR3-induced IP-10 production. This might be a new mechanism how histamine fosters a Th2 milieu.

© 2013 S. Karger AG, Basel

goto top of outline Introduction

Histamine is a major mediator in allergic diseases and inflammatory processes, and it is involved in several physiological functions, e.g. differentiation and proliferation of cells, embryonic development, hematopoiesis and wound healing [1]. It plays an important role in acute inflammatory and immediate-type hypersensitivity responses as well as in chronic inflammation and delayed-type allergic reactions. Histamine concentrations are increased at sites of allergic reactions and inflammation, such as allergic asthma, atopic dermatitis (AD) or allergic contact dermatitis [2]. T helper cells are important for the initiation and regulation of immune responses. They can be divided into Th1 and Th2 cells, which are characterized by the production of IFN-γ and interleukin (IL)-4, respectively. Th1 cells play a role in delayed-type hypersensitivity and cytotoxic responses, whereas Th2 cells are associated with the regulation of allergic diseases by activating B cells and by inducing the production of IgE and IgG [3]. Th1 and Th2 cells show different histamine receptor (HR) expression patterns and histamine interferes with the Th1/Th2 balance, e.g. via a reduction in Th1 cell proliferation and an enhanced secretion of IL-10 in T cells. Histamine also modulates the development of allergies by upregulation of Th2 cell proliferation and increased production of Th2 cytokines IL-4, IL-5, IL-10 and IL-13 [3,4]. Furthermore, histamine influences the Th1/Th2 balance indirectly by acting on dendritic cells (DC), e.g. by blocking the production of the Th1-priming cytokine IL-12 in DC [5,6]. As the most potent antigen-presenting cells (APC), DC play an important role in the human immune system and can be found in nearly all peripheral tissues [7]. They represent an important link between adaptive and innate immune responses and are involved in inflammatory and allergic disorders, e.g. allergic airway inflammation [8], psoriasis [9] or rheumatoid arthritis [10]. The main function of these cells is their interaction with T cells: DC trigger the proliferation of specific T cells and they influence the type of T cell responses [11]. Various subtypes of DC are known. One of the main subsets are myeloid DC (mDC), characterized by CD11c+ and CD123-, which pick up antigens in the periphery and trigger immunity after migration to lymphoid organs. Depending on the antigen and other factors, mDC are capable to induce a Th1 or a Th2 cytokine pattern [12].

Whereas effects of histamine on monocytes and monocyte-derived DC (MoDC) have been investigated in previous studies [5,13], there are no data on the effects of histamine on human mDC. Therefore, we compared histamine receptor expression and function between primary mDC and monocytes. First, we studied the expression of the four histamine receptors at mRNA level and showed that mDC and monocytes express similar repertoires for H1R, H2R and H3R, whereas H4R expression is higher on mDC compared with monocytes. Next, we performed a protein array to evaluate functional effects of histamine on the expression of cytokines and chemokines. Finally, we analyzed the newly observed downregulation of the Th1-associated chemokine IP-10 by histamine in more detail. We observed similar functional effects towards histamine stimulation with regard to IP-10 regulation. Thus, our data show the similarity of monocytes and mDC with regard to histamine as well as a new mechanism that fosters a Th2 milieu induced by histamine.


goto top of outline Materials and Methods

goto top of outline Isolation and Culture of Peripheral Blood Mononuclear Cells, Monocytes and mDC

Peripheral blood mononuclear cells (PBMC) were isolated from buffy coats from anonymous healthy donors obtained from the local blood bank. PBMC were separated by density centrifugation on Lymphoprep (Fresenius Kabi Norge AS, Oslo, Norway) and erythrocytes were removed by Gey's lysis. Monocytes and mDC [characterized by CD1 (BDCA-1)+ and CD141 (BDCA-3)+; CD11chigh, CD123low, CD32+, CD64+ and FcεR1+] were isolated from PBMC by negative isolation using magnetic cell sorting (MACS; Miltenyi Biotech Inc., Bergisch-Gladbach, Germany). 1 × 108 PBMC were used per isolation and the procedure was executed on ice or in the fridge. Cells were washed with MACS buffer and incubated with FcR blocking reagent and mDC antibody-biotin cocktail for 10 min. After centrifugation and two washing steps with MACS buffer, cells were subsequently incubated with anti-biotin MicroBeads for 15 min and separated by magnetic field. The unlabeled cells passing the column contained the desired mDC. Isolated mDC had a purity of at least 90% based on the expression of CD1, CD11c and CD141, and a lack of expression of CD3, CD14, CD16 and CD19 (fig. 2b). Monocytes and mDC were cultured for the stimulation time in RPMI 1640 medium supplemented with 12 mM HEPES, 2 mML-glutamine, 100 mg/ml penicillin/streptomycin, 5% v/v fetal calf serum (PAN-Biotech, all other components from Biochrom, Berlin, Germany) at 37°C in a humidified atmosphere containing 5% CO2.

goto top of outline Proteome Analysis

Isolated mDC (5 × 104) were pretreated with 10-5M histamine (agonist for all histamine receptors; Alk-Scherax, Wedel, Germany) and stimulated with 10-5M polyinosinic-polycytidylic acid (poly I:C; Sigma-Aldrich, St. Louis, Mo., USA). Supernatants were collected after 24 h and five different independent experiments were pooled. This supernatant pool was analyzed with the Proteome Profiler™ human cytokine array panel A (R&D Systems, Minneapolis, Minn., USA). The probes were incubated for 1 h with a detection antibody cocktail and then the nitrocellulose membrane, containing 36 different anti-cytokine or anti-chemokine antibodies printed in duplicate, was incubated with 500 µl of the supernatants overnight at 4°C. Immunoreactivity was detected by chemiluminescence according to the manufacturer's instructions (Pierce, Thermo Fisher Scientific, Rockford, Ill., USA) and the intensity of the dot plots was evaluated by densitometric analysis with ChemiImager Software (Genetic Technologies, Miami, Fla., USA).

goto top of outline mRNA Isolation, Reverse Transcription and Quantitative Real-Time PCR

Monocytes and mDC were pretreated with histamine overnight and subsequently stimulated with 10-5M poly I:C. Cells were harvested and total RNA was isolated using a Quick-RNA™ MiniPrep kit including additional DNase digestion with DNase I (both ZymoResearch, Orange, Calif., USA). Reverse transcription was performed using a Quantitect reverse transcription kit (Qiagen, Hilden, Germany). Quantification of IP-10, the four histamine receptors and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was performed on a LightCycler (Roche Molecular Biochemicals, Risch, Switzerland) using SYBR Green with QuantiTect primer assays (IP-10: QT01003065; H1R: QT00199857, H3R: QT00210861, and GAPDH: QT01192646) according to the manufacturer's protocol (Qiagen) or for H2R and H4R with designed primers (H2R: 5′-TACCAGCTGTCCTGCAAGTG-3′ and 5′-CCCCAGGTGGATAGACAGAA-3′, and H4R: 5′-TGCTAGGAAATGCTTTGGTC-3′ and 5′-GCGTGTGAGGGATGTACAAA-3′) [14] synthesized by Tib Molbiol (Berlin, Germany), under standard cycler conditions (annealing temperature 55°C). Melting curve analysis was performed from 60 to 90°C with a ramp rate of 20°C/s. For visualization of the amplified PCR products, agarose gel electrophoresis (2% agarose; Roth, Karlsruhe, Germany) was performed. Relative gene expression data were analyzed using an external calibrator for calculation and using the Relative Quantification Software (Roche Molecular Biochemicals).

goto top of outline Enzyme-Linked Immunosorbent Assay

Protein expression of IP-10 was analyzed after 24 h by ELISA. 0.5 to 1 × 105 mDC and 3 × 105 monocytes were seeded per well in 96-well plates and were preincubated overnight with histamine (Alk-Scherax) or one of the histamine receptor agonists (H1R agonist 2-pyridylethylamine and H2R agonist amthamine; both Tocris Bioscience, Bristol, UK; the H4R agonist ST1006 was synthesized by Holger Stark, Goethe University, Frankfurt am Main, Germany, as published previously [15]). Afterwards cells were stimulated for 24 h with 10-5M poly I:C, then cell-free supernatants were harvested and analyzed for IP-10 content using a commercially available ELISA according to the manufacturer's protocol (IP-10: R&D Systems). The plate was read at a wavelength of 450 nm on a FluoStar plate reader (BMG Lab Technologies GmbH, Jena, Germany). For blocking experiments, cells were pretreated for 30 min with one of the three different antagonists before histamine was added (H1R antagonist levocetirizine; UCB, Anderlecht/Brüssel, Belgium; H2R antagonist ranitidine; Biomol, Hamburg, Germany, and H4R antagonist JNJ7777120; Sigma Aldrich).

goto top of outline Statistical Analysis

Data are expressed as means ± SEM. A Wilcoxon matched pair test (fig. 1 and 3a mDC) or paired t test (fig. 3, 4, 5, 6) was applied to determine statistically significant differences; a value of p < 0.05 was considered statistically significant. GraphPad Prism® (version 5; GraphPad Software, San Diego, Calif., USA) was used for statistical analysis.

Fig. 1. mDC and monocytes express the histamine receptors H1R, H2R and H4R at mRNA level. a Representative real-time LightCycler PCR melting peaks and agarose gel bands with the correct size of the amplified PCR products (H1R = 116 bp, H2R = 239 bp, H3R = 150 bp, H4R = 130 bp and GAPDH = 119 bp) of histamine receptors and GAPDH of 7 independent experiments are presented. As positive control for H3R amplification H3R-transfected HEK cells were used. b Both cell types were isolated from PBMC and mRNA expression for H1R, H2R, H4R and GAPDH was analyzed by LightCycler PCR. The relative amounts of the different receptors in monocytes and mDC were determined to a calibrator and normalized to GAPDH. mDC expressed less H1R mRNA, equal amounts of H2R mRNA and significantly more H4R mRNA compared to monocytes. Results of 9 (H1R and H4R) or 12 (H2R) independent experiments are shown. * p < 0.05 (Wilcoxon matched pair test).

Fig. 3. The expression of IP-10 occurs in a concentration- and time-dependent manner. ns = Nonstimulated. a Monocytes and mDC were pretreated overnight with different concentrations of histamine (Hista; 10-5-10-8M) and then stimulated for 24 h with 10-5M poly I:C. The content of IP-10 in supernatants was detected by ELISA (means and SEM of 6 independent experiments). Histamine significantly and dose-dependently decreased IP-10 release from human monocytes (mean ± SEM at 24 h of poly I:C = 2,119 ± 570 pg/ml, range 606-4,211 pg/ml) and mDC (mean ± SEM at 24 h of poly I:C = 4,302 ± 1,127 pg/ml, range 1,509-9,142 pg/ml) in response to poly I:C. b Monocytes were pretreated with 10-5M histamine overnight and then stimulated for different time points with 10-5M poly I:C. mRNA expression was detected by RT-PCR and peaked after 4 h (mean ± SEM at 4 h of poly I:C = 257 ± 120 relative mRNA expression, range 6-714; mean ± SEM at 4 h of poly I:C + 10-5M histamine = 122 ± 89 relative mRNA expression, range 3-552). Expression of IP-10 at protein level was detected in supernatants by ELISA (means and SEM of 6 independent experiments are shown) and peaked after 24 h (mean ± SEM at 24 h of poly I:C = 826 ± 155 pg/ml, range 425-1,431 pg/ml). * p < 0.05, ** p < 0.01 poly I:C vs. poly I:C + histamine.

Fig. 4. Different histamine (Hista) receptor agonists mimic the histamine effect of downregulating IP-10 protein production in monocytes (a) and mDC (b). Monocytes and mDC were pretreated overnight with 10-5M histamine or a specific histamine receptor agonist and then stimulated for 24 h with 10-5M poly I:C. The content of IP-10 in supernatants was detected by ELISA [means and SEM of 8 (mDC) or 10 (monocyte) independent experiments are shown]. a In monocytes, both the H2R agonist amthamine (Amtha) and the H4R agonist ST1006 mimics the histamine effect on the IP-10 expression (mean ± SEM at 24 h of poly I:C = 2,273 ± 449 pg/ml, range 738-5,038 pg/ml). b In mDC, H2R (Amtha) and H4R (ST1006) agonists imitate the histamine effect of reduced IP-10 protein expression (mean ± SEM at 24 h of poly I:C = 2,710 ± 715 pg/ml, range 682-6,376 pg/ml). * p < 0.05, ** p < 0.01. ns = Nonstimulated.

Fig. 5. Different histamine receptor antagonists blocked the downregulation of IP-10 protein by histamine (Hista) in monocytes (a) and mDC (b). Monocytes and mDC were pretreated for 30 min with 10-5M of the specific antagonist and subsequently incubated overnight with 10-5M histamine. The next day, cells were stimulated for 24 h with 10-5M poly I:C. The content of IP-10 in supernatants was detected by ELISA (mean and SEM of 7 independent experiments are shown). ns = Nonstimulated. a In monocytes, both antagonists for H2R (ranitidine) and for H4R (JNJ7777120) reverse the histamine effect of reduced IP-10 protein expression (mean ± SEM at 24 h of poly I:C = 2,565 ± 613 pg/ml, range 738-5,038 pg/ml). b In mDC, only the H4R antagonist JNJ7777120 can reverse the effect of histamine (mean ± SEM at 24 h of poly I:C = 2,180 ± 746 pg/ml, range 599-6,144 pg/ml). * p < 0.05, ** p < 0.01.

Fig. 6. H1R is not involved in the regulation of poly I:C-induced IP-10 expression. Monocytes (a) and mDC (b) were treated in the presence or absence of the H1R antagonist levocetirizine (Levo) with histamine (Hista) or the H1R-specific agonist 2-pyridylethylamine (2-Pyr) overnight. Then cells were stimulated for 24 h with 10-5M poly I:C. The content of IP-10 in supernatants was detected by ELISA (mean and SEM of 8-13 independent experiments are shown). The results show no significant involvement of H1R in monocytes (mean ± SEM at 24 h of poly I:C = 2,893 ± 573 pg/ml, range 283-5,594 pg/ml) and mDC (mean ± SEM at 24 h of poly I:C = 1,789 ± 549 pg/ml, range 305-4,508 pg/ml; * p < 0.05, ** p < 0.01). ns = Nonstimulated.


goto top of outline Results

goto top of outline H1R, H2R and H4R Expression on mDC and Monocytes

First we analyzed the expression of the four different HR in monocytes and mDC. Real-time PCR was performed and the amplified products were analyzed by LightCycler melting curves and agarose gel electrophoresis, where sharp bands at the expected sizes could be demonstrated (fig. 1a). The majority of monocytes expressed more H1R receptor compared with mDC, but the experiments were inconsistent and the difference not statistically significant (fig. 1b). H2R mRNA was detected at similar amounts and no mRNA for H3R was detected in both cell types. With respect to H4R, we observed a significantly higher expression pattern on mDC compared with monocytes (fig. 1b).

goto top of outline Effects of Histamine on Chemokine and Cytokine Expression on mDC

To assess the effects of histamine on mDC, we performed a proteome array. We stimulated mDC with poly I:C or with poly I:C and histamine. We pooled the cell-free supernatants from five different cell preparations. Proteome analysis revealed the regulation of different cytokines and chemokines and we observed upregulation of the poly I:C-induced CXCL1 expression (poly I:C + histamine results in 120% densitometry compared to stimulation with poly I:C only) and a downregulation of poly I:C-induced expression of TNF-α (76%), CCL1 (78%), IL-6 (46%) and IP-10 (63%) following preincubation with histamine by densitometry (fig. 2a).

Fig. 2. Histamine downregulates the expression of IP-10 in mDC. a mDC were cultured overnight in the presence or absence of 10-5 M histamine and then stimulated with 10-5M poly I:C for 24 h. Supernatants of five different cell stimulations were pooled. The expression of 36 different chemokines and cytokines in this pool was analyzed by protein array. The dot plots show downregulation of IP-10 in supernatants from cells stimulated with poly I:C and histamine compared with samples stimulated with poly I:C only. In addition to IP-10, we also detected effects on other mediators such as TNF-α, CXCL1, CCL1 and IL-6. b Representative dot plot of purity of mDC is shown in comparison to isotype staining. mDC were gated by cell surface staining of CD1, CD11c and CD141, and for linage staining CD3, CD14, CD16 and CD19 were used.

goto top of outline Inhibitory Effect of Histamine on Poly I:C-Induced Expression of IP-10

To confirm our observations from the protein array and to gain a deeper insight, we analyzed the effect of different histamine concentrations (10-5-10-8M). As shown in figure 3a in mDC, histamine concentrations from 10-6 to 10-8M led to a significant downregulation of IP-10. Also, in monocytes, it was possible to inhibit the production of IP-10 by histamine concentrations from 10-5 to 10-7M (fig. 3a).

To analyze the time dependence of the poly I:C-induced IP-10 expression, monocytes were preincubated with histamine and incubated for different time periods with poly I:C. IP-10 mRNA was detected after 2, 4, 6 and 8 h and maximal mRNA expression was reached after 4 h. Subsequently, we analyzed the IP-10 protein in supernatants between 2 and 24 h. Protein expression peaked at 24 h. At all measured time points, we could observe a significant downregulation of IP-10 by histamine at the protein level, while at the mRNA level a difference was only detected for the first three time points between 2 and 6 h (fig. 3b).

goto top of outline Effect of H2R and H4R on Poly I:C-Induced Expression of IP-10 on Monocytes and mDC

To analyze which receptor is at the functional level responsible for the observed effects of histamine, we performed experiments with specific agonists and antagonists. We demonstrated that amthamine (H2R agonist) and ST1006 (H4R agonist) mimic the histamine effect on IP-10 expression in both monocytes and mDC (fig. 4). To verify these observations, we incubated the cells for 30 min with specific receptor antagonists before they were stimulated with histamine. In monocytes, blockade of H2R with ranitidine as well as of H4R with JNJ7777120 abolished the histamine effect. However, in mDC the histamine effect could be reversed only by blocking H4R with JNJ7777120 (fig. 5). As both cell types also express H1R, we performed experiments with a specific H1R agonist and antagonist. With these ligands, no effects were observed in both monocytes and mDC (fig. 6). Taken together, the results with agonists and antagonists showed that IP-10 expression is regulated via H2R and H4R stimulation in these two cell types.


goto top of outline Discussion

In the immune system, histamine plays a role as an important mediator of inflammation and allergic diseases, and the development of some infections and allergic reactions is associated with a substantial production of histamine. It is also known that histamine modulates the function and activity of different cells of the immune system, including T lymphocytes and APC. Therefore, we focused our study on the expression pattern of histamine receptors and histamine effects on mDC, a major subpopulation of APC. Monocytes and mDC are human blood cell subpopulations and have the same myeloid descent from bone marrow stem cells, so they are related with each other [16]. Since mDC represent a rather small fraction of around 0.5% of PBMC [17], most previous investigations used in vitro generated MoDC as a model to determine the effects of histamine on the expression and function on mDC [5,18,19,20]. MoDC have differentiated from monocytes in vitro using a cytokine cocktail, most commonly IL-4 and granulocyte/macrophage colony-stimulating factor, and incubation results in so-called immature DC with high antigen-processing capacity and poor capability for T cell activation [21,22]. To eliminate possible effects of this in vitro generation and to get a more realistic view of the natural conditions, we isolated mDC directly from human blood by negative magnetic separation from PBMC. These cells represent the natural mDC population and were compared in our experiments to monocytes from the same donors. In earlier studies, other subtypes of DC and monocytes expressed the histamine receptors H1R, H2R and H4R [23]. The results of our present study demonstrated that mDC and monocytes also expressed three different histamine receptors at mRNA level (H1R, H2R and H4R). Comparing monocytes and mDC, in most donors, monocytes expressed more H1R than mDC. The amounts of H2R were not different between the two cell types. This is consistent with our previous finding that compared to monocytes MoDC downregulate H1R mRNA but not H2R mRNA [13]. In contrast, we were unable to detect H3R mRNA in both cell types, although other studies reported H3R expression on monocytes and MoDC [5,18]. However, H3R is a presynaptic autoreceptor which is mainly present in the peripheral and central nervous system. The expression of H4R mRNA was significantly higher in mDC than monocytes. This is consistent with our previous observation that compared to monocytes in vitro MoDC upregulate H4R during their differentiation [5].

Histamine can modulate T helper cell responses, influence antigen presentation capacity and enhance proinflammatory cytokine production [6]. Histamine is a factor for DC polarization but not for DC maturation [20,24], and mediates its influence on the Th1/Th2 balance through several effects on different cell types and through involvement of different histamine receptors. DC are often located in areas with high amounts of histamine [25]. Various mechanisms have been described how DC can influence Th1/Th2 polarization. One mechanism is the production of cytokines, such as IL-12, priming a Th1 response. In case IL-12 production is blocked, e.g. by histamine, DC prime a Th2 response [5,26,27]. McIlroy et al. [20] showed also an influence of histamine on the Th1/Th2 balance by an upregulation of the Th2 attractants CCL17 and CCL22 and the downregulation of IFN-γ-induced IP-10 in immature human MoDC through involvement of H2R. In other cell types, we were able to show that H2R and H4R stimulation led to downregulation of cytokine and chemokine production, e.g. of TNF-α, CXCL8 and IFN-α in plasmacytoid DC and of IL-27 in monocytes [28,29]. van der Pouw Kraan et al. [30] demonstrated that histamine inhibits the production of IL-12 via stimulation of H2R in monocytes. Also, in whole blood cultures, histamine dose-dependently increases IL-10 expression and significantly decreases IL-12 after lipopolysaccharide treatment via H2R stimulation [31]. Another mechanism for the influence of Th1/Th2 polarization is the DC type. Subtypes of DC (DC1 and DC2) have been characterized that stimulate either a Th1 or Th2 response, respectively [32]. DC1 have a high expression of H1R and histamine acts as a stimulus for increased antigen presentation capacity, enhanced production of proinflammatory cytokines and Th1 priming activity. In DC2, histamine plays a suppressive role on antigen-presenting ability, increases IL-10 production and induces Th2 cells via H2R [6]. Taken together, histamine is a potent mediator that primes DC to induce Th2 polarization [24,32].

To evaluate the functional role of histamine on mDC, we used a protein array to study the effects of histamine on cytokine and chemokine expression on mDC. We observed a regulation of the expression of CXCL1, TNF-α, CCL1, IL-6 and IP-10 through histamine. The expression of the C-X-C motif chemokine 10 (CXCL10), also called IP-10, is associated with inflammation. This chemokine is released from different cell types, e.g. monocytes, DC, endothelial and epithelial cells and keratinocytes, in response to IFN-γ [33]. CXCL chemokines primarily attract Th1 lymphocytes and neutrophils, and CXC receptors are found predominantly on Th1 lymphocytes [34]. Twenty to 40% of circulating Th1 lymphocytes express CXC receptors, whereas Th2 lymphocytes express predominately CC chemokine receptors [35]. Due to the important role of IP-10 in inflammation and as a key chemokine for the recruitment of Th1 lymphocytes into tissue and the association of IP-10 with inflammatory and allergic diseases, including allergic contact dermatitis, allergic pulmonary inflammation and AD [36,37,38,39], we chose IP-10 as a target for further investigations. For the first time, we describe an influence of histamine on poly I:C-induced expression of IP-10 in natural untouched mDC. Our results fit well in the picture that histamine exerts anti-inflammatory effects and promotes a Th2 milieu via H2R and H4R.

H2R and H4R might be new therapeutic targets also in allergic diseases such as AD. Potential antagonists of H2R and H4R mediate their therapeutic potential through alleviation of Th1 responses by targeting the expression of relevant cytokines such as IP-10 or IL-12. Thus, H2R and H4R antagonists might be useful for the treatment of allergic and inflammatory disorders with Th2 predominance, such as AD.

Since IP-10 is a Th1 chemokine and plays an important role in numerous diseases, first studies evaluating IP-10 as therapeutic target are in progress. Fife et al. [40] showed that the application of an anti-IP-10 antibody resulted in a decreased clinical and histological manifestation of experimental autoimmune encephalomyelitis. Blocking of IP-10 and CXCR3 reduced inflammatory colitis in mice [41].

In summary, our study demonstrates that mDC as another APC subpopulation express H1R, H2R and H4R. In addition, the histamine-induced downregulation of IP-10 via H2R and H4R might affect the regulation of the Th1/Th2 balance. These results improve the understanding of immunological functions of histamine and suggest that specific receptor antagonists might be candidates for pharmacological treatment to inhibit the Th2 shift in inflammatory and allergic disorders.


goto top of outline Acknowledgments

We acknowledge the support from the Deutsche Forschungsgemeinschaft (German Research Foundation) by grant Gu434/5-2 and the European Community COST Action BM0806 (Recent advances in histamine H4 receptor research).

 goto top of outline References
  1. Akdis CA, Jutel M, Akdis M: Regulatory effects of histamine and histamine receptor expression in human allergic immune responses. Chem Immunol Allergy 2008;94:67-82.
  2. Gutzmer R, Gschwandtner M, Rossbach K, Mommert S, Werfel T, Kietzmann M, Baeumer W: Pathogenetic and therapeutic implications of the histamine H4 receptor in inflammatory skin diseases and pruritus. Front Biosci (Schol Ed) 2011;3:985-994.
  3. Packard KA, Khan MM: Effects of histamine on Th1/Th2 cytokine balance. Int Immunopharmacol 2003;3:909-920.
  4. Osna N, Elliott K, Khan MM: Regulation of interleukin-10 secretion by histamine in Th2 cells and splenocytes. Int Immunopharmacol 2001;1:85-96.
  5. Gutzmer R, Diestel C, Mommert S, Kother B, Stark H, Wittmann M, Werfel T: Histamine H4 receptor stimulation suppresses IL-12p70 production and mediates chemotaxis in human monocyte-derived dendritic cells. J Immunol 2005;174:5224-5232.

    External Resources

  6. Mazzoni A, Young HA, Spitzer JH, Visintin A, Segal DM: Histamine regulates cytokine production in maturing dendritic cells, resulting in altered T cell polarization. J Clin Invest 2001;108:1865-1873.
  7. Banchereau J, Steinman RM: Dendritic cells and the control of immunity. Nature 1998;392:245-252.
  8. van Rijt LS, van Kessel CH, Boogaard I, Lambrecht BN: Respiratory viral infections and asthma pathogenesis: a critical role for dendritic cells? J Clin Virol 2005;34:161-169.
  9. Nestle FO, Turka LA, Nickoloff BJ: Characterization of dermal dendritic cells in psoriasis. Autostimulation of T lymphocytes and induction of Th1 type cytokines. J Clin Invest 1994;94:202-209.
  10. Thomas R, Davis LS, Lipsky PE: Rheumatoid synovium is enriched in mature antigen-presenting dendritic cells. J Immunol 1994;152:2613-2623.

    External Resources

  11. Cravens PD, Lipsky PE: Dendritic cells, chemokine receptors and autoimmune inflammatory diseases. Immunol Cell Biol 2002;80:497-505.
  12. Tschoep K, Manning TC, Harlin H, George C, Johnson M, Gajewski TF: Disparate functions of immature and mature human myeloid dendritic cells: implications for dendritic cell-based vaccines. J Leukoc Biol 2003;74:69-80.
  13. Gutzmer R, Langer K, Lisewski M, Mommert S, Rieckborn D, Kapp A, Werfel T: Expression and function of histamine receptors 1 and 2 on human monocyte-derived dendritic cells. J Allergy Clin Immunol 2002;109:524-531.
  14. van Rijn RM, van Marle A, Chazot PL, Langemeijer E, Qin Y, Shenton FC, Lim HD, Zuiderveld OP, Sansuk K, Dy M, Smit MJ, Tensen CP, Bakker RA, Leurs R: Cloning and characterization of dominant negative splice variants of the human histamine H4 receptor. Biochem J 2008;414:121-131.
  15. Sander K, Kottke T, Tanrikulu Y, Proschak E, Weizel L, Schneider EH, Seifert R, Schneider G, Stark H: 2,4-Diaminopyrimidines as histamine H4 receptor ligands - scaffold optimization and pharmacological characterization. Bioorg Med Chem 2009;17:7186-7196.
  16. Goordyal P, Isaacson PG: Immunocytochemical characterization of monocyte colonies of human bone marrow: a clue to the origin of Langerhans cells and interdigitating reticulum cells. J Pathol 1985;146:189-195.
  17. Bosma BM, Metselaar HJ, Tra WM, Mancham S, Kuipers EJ, Tilanus HW, Kwekkeboom J: Impairment of circulating myeloid dendritic cells in immunosuppressed liver transplant recipients. Clin Exp Immunol 2007;149:525-534.
  18. Idzko M, La Sala A, Ferrari D, Panther E, Herouy Y, Dichmann S, Mockenhaupt M, Di Virgilio F, Girolomoni G, Norgauer J: Expression and function of histamine receptors in human monocyte-derived dendritic cells. J Allergy Clin Immunol 2002;109:839-846.
  19. Amaral MM, Davio C, Ceballos A, Salamone G, Canones C, Geffner J, Vermeulen M: Histamine improves antigen uptake and cross-presentation by dendritic cells. J Immunol 2007;179:3425-3433.

    External Resources

  20. McIlroy A, Caron G, Blanchard S, Fremaux I, Duluc D, Delneste Y, Chevailler A, Jeannin P: Histamine and prostaglandin E up-regulate the production of Th2-attracting chemokines (CCL17 and CCl22) and down-regulate IFN-gamma-induced CXCL10 production by immature human dendritic cells. Immunology 2006;117:507-516.
  21. O'Doherty U, Steinman RM, Peng M, Cameron PU, Gezelter S, Kopeloff I, Swiggard WJ, Pope M, Bhardwaj N: Dendritic cells freshly isolated from human blood express CD4 and mature into typical immunostimulatory dendritic cells after culture in monocyte-conditioned medium. J Exp Med 1993;178:1067-1076.
  22. Sallusto F, Lanzavecchia A: Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha. J Exp Med 1994;179:1109-1118.
  23. Ferstl R, Akdis CA, O'Mahony L: Histamine regulation of innate and adaptive immunity. Front Biosci 2012;17:40-53.
  24. Mazzoni A, Siraganian RP, Leifer CA, Segal DM: Dendritic cell modulation by mast cells controls the Th1/Th2 balance in responding T cells. J Immunol 2006;177:3577-3581.

    External Resources

  25. Jutel M, Blaser K, Akdis CA: The role of histamine in regulation of immune responses. Chem Immunol Allergy 2006;91:174-187.
  26. Rissoan MC, Soumelis V, Kadowaki N, Grouard G, Briere F, de Waal Malefyt R, Liu YJ: Reciprocal control of T helper cell and dendritic cell differentiation. Science 1999;283:1183-1186.
  27. Langenkamp A, Messi M, Lanzavecchia A, Sallusto F: Kinetics of dendritic cell activation: impact on priming of Th1, Th2 and nonpolarized T cells. Nat Immunol 2000;1:311-316.
  28. Gschwandtner M, Mommert S, Kother B, Werfel T, Gutzmer R: The histamine H4 receptor is highly expressed on plasmacytoid dendritic cells in psoriasis and histamine regulates their cytokine production and migration. J Invest Dermatol 2011;131:1668-1676.
  29. Gschwandtner M, Bunk H, Kother B, Thurmond RL, Kietzmann M, Werfel T, Baumer W, Gutzmer R: Histamine down-regulates IL27 production in antigen-presenting cells. J Leukoc Biol 2012;92:21-29.
  30. van der Pouw Kraan TC, Snijders A, Boeije LC, de Groot ER, Alewijnse AE, Leurs R, Aarden LA: Histamine inhibits the production of interleukin-12 through interaction with H2 receptors. J Clin Invest 1998;102:1866-1873.
  31. Elenkov IJ, Webster E, Papanicolaou DA, Fleisher TA, Chrousos GP, Wilder RL: Histamine potently suppresses human IL-12 and stimulates IL-10 production via H2 receptors. J Immunol 1998;161:2586-2593.

    External Resources

  32. Caron G, Delneste Y, Roelandts E, Duez C, Bonnefoy JY, Pestel J, Jeannin P: Histamine polarizes human dendritic cells into Th2 cell-promoting effector dendritic cells. J Immunol 2001;167:3682-3686.

    External Resources

  33. Cassatella MA, Gasperini S, Calzetti F, Bertagnin A, Luster AD, McDonald PP: Regulated production of the interferon-gamma-inducible protein-10 (IP-10) chemokine by human neutrophils. Eur J Immunol 1997;27:111-115.
  34. Sallusto F, Lanzavecchia A: Understanding dendritic cell and T-lymphocyte traffic through the analysis of chemokine receptor expression. Immunol Rev 2000;177:134-140.
  35. Gasperini S, Marchi M, Calzetti F, Laudanna C, Vicentini L, Olsen H, Murphy M, Liao F, Farber J, Cassatella MA: Gene expression and production of the monokine induced by IFN-gamma (MIG), IFN-inducible T cell alpha chemoattractant (I-Tac), and IFN-gamma-inducible protein-10 (IP-10) chemokines by human neutrophils. J Immunol 1999;162:4928-4937.

    External Resources

  36. Flier J, Boorsma DM, Bruynzeel DP, Van Beek PJ, Stoof TJ, Scheper RJ, Willemze R, Tensen CP: The CXCR3 activating chemokines IP-10, MIG, and IP-9 are expressed in allergic but not in irritant patch test reactions. J Invest Dermatol 1999;113:574-578.
  37. Medoff BD, Sauty A, Tager AM, Maclean JA, Smith RN, Mathew A, Dufour JH, Luster AD: IFN-gamma-inducible protein 10 (CXCL10) contributes to airway hyperreactivity and airway inflammation in a mouse model of asthma. J Immunol 2002;168:5278-5286.

    External Resources

  38. Villagomez MT, Bae SJ, Ogawa I, Takenaka M, Katayama I: Tumour necrosis factor-alpha but not interferon-gamma is the main inducer of inducible protein-10 in skin fibroblasts from patients with atopic dermatitis. Br J Dermatol 2004;150:910-916.
  39. Giustizieri ML, Mascia F, Frezzolini A, De Pita O, Chinni LM, Giannetti A, Girolomoni G, Pastore S: Keratinocytes from patients with atopic dermatitis and psoriasis show a distinct chemokine production profile in response to T cell-derived cytokines. J Allergy Clin Immunol 2001;107:871-877.
  40. Fife BT, Kennedy KJ, Paniagua MC, Lukacs NW, Kunkel SL, Luster AD, Karpus WJ: CXCL10 (IFN-gamma-inducible protein-10) control of encephalitogenic CD4+ T cell accumulation in the central nervous system during experimental autoimmune encephalomyelitis. J Immunol 2001;166:7617-7624.

    External Resources

  41. Singh UP, Venkataraman C, Singh R, Lillard JW Jr: CXCR3 axis: role in inflammatory bowel disease and its therapeutic implication. Endocr Metab Immune Disord Drug Targets 2007;7:111-123.

 goto top of outline Author Contacts

Correspondence to: Dr. Franziska Glatzer
Division of Immunodermatology and Allergy Research
Department of Dermatology and Allergy, Hannover Medical School
Carl Neuberg Street 1, DE-30625 Hannover (Germany)
E-Mail Glatzer.Franziska@mh-hannover.de

 goto top of outline Article Information

Received: March 22, 2013
Accepted after revision: September 23, 2013
Published online: November 16, 2013
Number of Print Pages : 9
Number of Figures : 6, Number of Tables : 0, Number of References : 41

 goto top of outline Publication Details

International Archives of Allergy and Immunology

Vol. 163, No. 1, Year 2014 (Cover Date: January 2014)

Journal Editor: Valenta R. (Vienna), Bohle B. (Vienna)
ISSN: 1018-2438 (Print), eISSN: 1423-0097 (Online)

For additional information: http://www.karger.com/IAA

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