Blocking ATP-releasing channels prevents high extracellular ATP levels and airway hyperreactivity in an asthmatic mouse model.

Allergic asthma is a chronic airway inflammatory response to different triggers like inhaled allergens. Excessive ATP in fluids from patients with asthma is considered an inflammatory signal and an important autocrine/paracrine modulator of airway physiology. Here, we investigated the deleterious effect of increased extracellular ATP (eATP) concentration on the mucociliary clearance (MCC) effectiveness and determined the role of ATP releasing channels during airway inflammation in an ovalbumin (OVA)-sensitized mouse model. Our allergic mouse model exhibited high levels of eATP measured in the tracheal fluid with a luciferin-luciferase assay and reduced MCC velocity determined by microspheres tracking in the trachea ex vivo. Addition of ATP had a dual effect on MCC, where lower ATP concentration (µM) increased microspheres velocity, whereas higher concentration (mM) transiently stopped microspheres movement. Also, an augmented ethidium bromide uptake by the allergic tracheal airway epithelium suggests an increase in ATP release channel functionality during inflammatory conditions. The use of carbenoxolone, a nonspecific inhibitor of connexin and pannexin1 channels reduced the eATP concentration in the allergic mouse tracheal fluid and dye uptake by the airway epithelium, providing evidence that these ATP release channels are facilitating the net flux of ATP to the lumen during airway inflammation. However, only the specific inhibition of pannexin1 with 10Panx peptide significantly reduced eATP in bronchoalveolar lavage and decreased airway hyperresponsiveness in OVA-allergic mouse model. These data provide evidence that blocking eATP may be a pharmacological alternative to be explored in rescue therapy during episodes of airflow restriction in patients with asthma.

Harnessing the therapeutic potential of antibodies targeting connexin hemichannels.

BACKGROUND: Connexin hemichannels have been implicated in pathology-promoting conditions, including inflammation, numerous widespread human diseases, including cancer and diabetes, and several rare diseases linked to pathological point mutations. METHODS: We analysed the literature focusing on antibodies capable of modulating hemichannel function, highlighting generation methods, applications to basic biomedical research and translational potential. RESULTS: Anti-hemichannel antibodies generated over the past 3 decades targeted mostly connexin 43, with a focus on cancer treatment. A slow transition from relatively unselective polyclonal antibodies to more selective monoclonal antibodies resulted in few products with interesting characteristics that are under evaluation for clinical trials. Selection of antibodies from combinatorial phage-display libraries, has permitted to engineer a monoclonal antibody that binds to and blocks pathological hemichannels formed by connexin 26, 30 and 32. CONCLUSIONS: All known antibodies that modulate connexin hemichannels target the two small extracellular loops of the connexin proteins. The extracellular region of different connexins is highly conserved, and few residues of each connexins are exposed. The search for new antibodies may develop an unprecedented potential for therapeutic applications, as it may benefit tremendously from novel whole-cell screening platforms that permit in situ selection of antibodies against membrane proteins in native state. The demonstrated efficacy of mAbs in reaching and modulating hemichannels in vivo, together with their relative specificity for connexins overlapping epitopes, should hopefully stimulate an interest for widening the scope of anti-hemichannel antibodies. There is no shortage of currently incurable diseases for which therapeutic intervention may benefit from anti-hemichannel antibodies capable of modulating hemichannel function selectively and specifically.

Characterization of three connexin32 genes and their role in inflammation-induced ATP release in the Japanese flounder Paralichthys olivaceus.

Extracellular ATP (eATP) is a potent singling molecule in activation of fish innate immunity while the molecular determinants for eATP release in fish were not completely understood. Connexin32 (Cx32) is a member of gap junction protein family that plays important immunological functions in mammals. However, the immune relevance of Cx32 and its role in ATP release in fish has not been investigated. Here, we identified, characterized three Cx32 isoform genes (Cx32.2, Cx32.2x and Cx32.7) from the Japanese flounder Paralichthys olivaceus, and investigated their role in inflammation-induced ATP release in fish. Expression analysis revealed that even though all the three Cx32 genes are constitutively expressed in all examined Japanese flounder tissues, Cx32.2 and Cx32.2x are dominantly expressed in liver, and Cx32.7 is highly expressed in intestine and head kidney macrophages. In addition, we showed that gene expression of all the three Cx32 isoforms was modulated by cAMP stimulation and inflammatory challenges. Furthermore, we revealed that Cx32 expression was upregulated in TNF-alpha overexpressed Japanese flounder FG-9307 cells. Moreover, overexpression of the three Cx32 isoforms significantly reduced the gene expression level of LPS-induced pro-inflammatory cytokine IL-8 and TNF-alpha, indicating that Cx32 is involved in modulating inflammatory response in fish. Finally, we showed that inflammation-induced ATP release was significantly increased in Cx32-overexpressed Japanese flounder FG-9307 cells, and this increased ATP release could be attenuated by pre-incubation with gap junction protein blocker carbenoxolone. Taken together, we for the first time reported the involvement of Cx32 in fish immunity. Our findings suggested that in addition to Cx43 and pannexin1 channels, Cx32 also plays a role in inflammation-induced ATP release in fish.

A potent antagonist antibody targeting connexin hemichannels alleviates Clouston syndrome symptoms in mutant mice.

BACKGROUND: Numerous currently incurable human diseases have been causally linked to mutations in connexin (Cx) genes. In several instances, pathological mutations generate abnormally active Cx hemichannels, referred to also as "leaky" hemichannels. The goal of this study was to assay the in vivo efficacy of a potent antagonist antibody targeting Cx hemichannels. METHODS: We employed the antibody to treat Cx30A88V/A88V adult mutant mice, the only available animal model of Clouston syndrome, a rare orphan disease caused by Cx30 p.A88V leaky hemichannels. To gain mechanistic insight into antibody action, we also performed patch clamp recordings, Ca2+ imaging and ATP release assay in vitro. FINDINGS: Two weeks of antibody treatment sufficed to repress cell hyperproliferation in skin and reduce hypertrophic sebaceous glands (SGs) to wild type (wt) levels. These effects were obtained whether mutant mice were treated topically, by application of an antibody cream formulation, or systemically, by intraperitoneal antibody injection. Experiments with mouse primary keratinocytes and HaCaT cells revealed the antibody blocked Ca2+ influx and diminished ATP release through leaky Cx30 p.A88V hemichannels. INTERPRETATION: Our results show anti-Cx antibody treatment was effective in vivo and sufficient to counteract the effects of pathological connexin expression in Cx30A88V/A88V mice. In vitro experiments suggest antibodies gained control over leaky hemichannels and contributed to restoring epidermal homeostasis. Therefore, regulating cell physiology by antibodies targeting the extracellular domain of Cxs may enforce an entirely new therapeutic strategy. These findings support the further development of antibodies as drugs to address unmet medical needs for Cx-related diseases. FUND: Fondazione Telethon, GGP19148; University of Padova, SID/BIRD187130; Consiglio Nazionale delle Ricerche, DSB.AD008.370.003\TERABIO-IBCN; National Science Foundation of China, 31770776; Science and Technology Commission of Shanghai Municipality, 16DZ1910200.

Gap Junctions and Breast Cancer Dormancy.

Breast cancer (BC) relapse, despite clinical advancement, remains one of the biggest issues in the field. Intercellular communication, specifically via connexin (Cx)-mediated gap junctions (GJs), play a key role in the long-term survival of these, treatment-resistant breast cancer stem cells (CSCs), allowing for relapse. Both basic and clinical evidence reveal dual roles for GJs, in tumor suppression, generally referred to as dormancy, and progression and metastasis. GJ intercellular communication (GJIC) can be mediated by multiple types of Cxs, depending on the organ to which the BC cells metastasize. This review expands on the differential expression of Cx-mediated GJIC between CSCs and niche cells within a given microenvironment.

Expression analysis of Pannexin1 channel gene in response to immune challenges and its role in infection-induced ATP release in tilapia (Oreochromis niloticus).

ATP released from immune cells plays an important role in activation of host innate immunity. However, the molecular mechanisms for pathogen infection-induced ATP release in fish remains unclear. Pannexin1 (Panx1) is a recently identified ATP release channel important for controlling immune responses. The immune relevance of Panx1 in fish, however, is still poorly understood. In this study, we characterized a Panx1 gene homologue (termed tPanx1) from Nile tilapia (Oreochromis niloticus) and analyzed its expression in response to different immune challenges. We also investigated the role of tPanx1 channel in bacterial infection-induced ATP release. Real-time quantitative PCR analysis revealed that tPanx1 gene is expressed in all tested tissues with predominant expression in intestine. Immune challenges with lipopolysaccharide, polyinosinic-polycytidylic acid and zymosan led to increased gene expression of tPanx1 in tilapia head kidney cells and peripheral blood leucocytes. In addition, tPanx1 gene was up-regulated in hepatopancreas, muscle, spleen, gill, head kidney and blood after Aeromonas hydrophila infection. Furthermore, pharmacological inhibition of tPanx1 channel activity with Panx1 channel inhibitor, carbenoxolone, significantly attenuated A. hydrophila infection-induced ATP release in tilapia head kidney cells. Taken together, our findings suggested that tPanx1 is an important immune response gene involved in bacterial infection-induced ATP release in tilapia O. niloticus.

Pathways of airway oxidant formation by house dust mite allergens and viral RNA converge through myosin motors, pannexons and Toll-like receptor 4.

INTRODUCTION: Intracellular reactive oxidant species (ROS) are generated in human airway epithelial cells by the prothrombinase action of Group 1 house dust mite (HDM) allergens and by ligation of viral RNA sensor Toll-like receptors (TLRs). We explored signaling convergence between HDM allergens and TLRs in ROS generation because epithelial cells form the primary barrier against inhaled substances and dictate host responses to allergens and viruses. METHODS: ROS formation by Calu-3 human airway cells was studied by measuring dihydrorhodamine 123 oxidation after activation by polyinosinic:polycytidylic acid (to activate TLR3), CL097 (to activate TLR7), a natural mixture of HDM allergens, or BzATP. RESULTS: TLR4 activation was identified as an indispensable response element for all stimuli, operating downstream from myosin motor activation, pannexon gating for ATP release and the endogenous activation of prothrombin. Exogenous prothrombin activation by HDM allergens was prevented by SGUL 1733, a novel inhibitor of the proteolytic activity of Group 1 HDM allergens, which thus prevented TLR4 from being activated at source. CONCLUSIONS: Our data identify for the first time that endogenously-generated prothrombin and TLR4 form a shared effector mechanism essential to intracellular ROS generation activated by a group 1 HDM allergen (itself a prothrombinase) or by ligation of viral RNA-sensing TLRs. These stimuli operate a confluent signaling pathway in which myosin motors, gating of pannexons, and ADAM 10 lead to prothrombin-dependent activation of TLR4 with a recycling activation of pannexons.

The Potential for Connexin Hemichannels to Drive Breast Cancer Progression through Regulation of the Inflammatory Response.

Over the past few decades, connexin hemichannels have become recognized as major players in modulating the inflammatory response. Chronic inflammation is documented to promote tumorigenesis and is a critical component of tumor progression. Furthermore, inflammation is strongly linked to angiogenesis, immunotolerance, invasiveness, metastasis, and resistance in breast cancers. In this review, the literature on the role of connexin hemichannels in inflammation is summarized, and the potential role for hemichannel-mediated inflammation in driving breast cancer progression is discussed. Lastly, the potential for connexin-based therapeutics to modulate the inflammatory component of the tumor microenvironment as an avenue for the treatment of breast cancer is also discussed.

Pannexin1 Channels Are Required for Chemokine-Mediated Migration of CD4+ T Lymphocytes: Role in Inflammation and Experimental Autoimmune Encephalomyelitis.

Pannexin1 (Panx1) channels are large high conductance channels found in all vertebrates that can be activated under several physiological and pathological conditions. Our published data indicate that HIV infection results in the extended opening of Panx1 channels (5-60 min), allowing for the secretion of ATP through the channel pore with subsequent activation of purinergic receptors, which facilitates HIV entry and replication. In this article, we demonstrate that chemokines, which bind CCR5 and CXCR4, especially SDF-1α/CXCL12, result in a transient opening (peak at 5 min) of Panx1 channels found on CD4(+) T lymphocytes, which induces ATP secretion, focal adhesion kinase phosphorylation, cell polarization, and subsequent migration. Increased migration of immune cells is key for the pathogenesis of several inflammatory diseases including multiple sclerosis (MS). In this study, we show that genetic deletion of Panx1 reduces the number of the CD4(+) T lymphocytes migrating into the spinal cord of mice subjected to experimental autoimmune encephalomyelitis, an animal model of MS. Our results indicate that opening of Panx1 channels in response to chemokines is required for CD4(+) T lymphocyte migration, and we propose that targeting Panx1 channels could provide new potential therapeutic approaches to decrease the devastating effects of MS and other inflammatory diseases.

[An increase in the ratio of peripheral blood CD4+/CD8+ T lymphocytes and the levels of connexin 40 and inflammatory factors in spontaneously hypertensive rats].

OBJECTIVE: To analyze the relationships of connexin 40 (Cx40) with peripheral blood CD4(+) and CD8(+) T lymphocyte subsets and inflammatory factors of spontaneously hypertensive (SH) rats. METHODS: Flow cytometry was used to detect CD4(+), CD8(+) lymphocytes and Cx40 expression on the cells in the peripheral blood of Wistar-Kyoto (WKy) rats and SH rats. ELISA was performed to test the levels of interleukin 2 (IL-2), interferon γ (IFN-γ), IL-4 and IL-6. RESULTS: Compared with WKy rats, the systolic blood pressure, the percentage of CD4(+) T lymphocytes, the expression of Cx40 on the surface of CD4(+) and CD8(+) T lymphocytes and the ratio of CD4(+)/CD8(+) in the peripheral blood of SH rats were significantly higher, with the exception of the percentage of CD8(+) lymphocytes which was lower. Also, we found that the serum levels of IL-2, IL-4 and IL-6 in the SH rats were significantly higher than those of WKy rats. However, there was no significant difference in the IFN-γ level between SH and WKy rats. CONCLUSION: The ratio of peripheral blood CD4(+)/CD8(+) T lymphocytes and the levels of Cx40, IL-2, IL-4 and IL-6 are significantly elevated in SH rats.

Conflicting effects by antibodies against connexin36 during the action of intracellular Cyclic-AMP onto electrical synapses of retinal ganglion cells.

Alpha-type retinal ganglion cells (alpha cells) of the same class in mammalian retina are connected by gap junctions. Electrical synapses between alpha cells were examined using combined techniques of dual patch-clamp recordings, intracellular labeling and electron microscopy in the albino rat retina. In simultaneous dual whole-cell recordings from pairs of neighboring alpha cells, bidirectional electrical synapses with symmetrical junction conductance were observed in pairs with cells of the same morphological type. Regulatory domains of gap junction protein subunit connexins in electrical synapses between alpha cells by extracellular and intracellular ligands investigated by dual whole-patch clamp recordings. I examined how passage currents through electrical synapses between alpha cells are modulated by specific antibodies against connexin36 proteins, and extracellular or intracellular application of ligands. Control conditions led us to observe large passage currents between connected cells and adequate transjunctional conductance (Gj) (1.35[Formula: see text][Formula: see text][Formula: see text]0.51[Formula: see text]nS). Experimental results show that high level of intracellular cyclic AMP within examined cells suppress electrical synapses between the neighboring cells. Gj between examined cells reduced to 0.15[Formula: see text][Formula: see text][Formula: see text]0.04[Formula: see text]nS. Under application of dopamine (1.25[Formula: see text][Formula: see text][Formula: see text]0.06[Formula: see text]nS) or intracellular cyclic GMP (0.98[Formula: see text][Formula: see text][Formula: see text]0.23[Formula: see text]nS), however, Gj also remains as in the control level. Intracellular application of an antibody against the cytoplasmic loop of connexin36 reduced Gj (0.98[Formula: see text][Formula: see text][Formula: see text]0.23[Formula: see text]nS). Cocktail of the antibody against cytoplasmic connexin36 and intracellular cyclic AMP leaves Gj as in the level by single involvement of the cytoplasmic antibody. The elimination of Gj by the cytoplasmic antibody was in a dose-dependent manner. These results suggest that binding domains against cyclic AMP may be present in the cytoplasmic sites of connexin proteins to regulate channel opening of gap junctions between mammalian retinal alpha ganglion cells.

Connexins: sensors of epidermal integrity that are therapeutic targets.

Gap junction proteins (connexins) are differentially expressed throughout the multiple layers of the epidermis. A variety of skin conditions arise with aberrant connexin expression or function and suggest that maintaining the epidermal gap junction network has many important roles in preserving epidermal integrity and homeostasis. Mutations in a number of connexins lead to epidermal dysplasias giving rise to a range of dermatological disorders of differing severity. 'Gain of function' mutations reveal connexin-mediated roles in calcium signalling within the epidermis. Connexins are involved in epidermal innate immunity, inflammation control and in wound repair. The therapeutic potential of targeting connexins to improve wound healing responses is now clear. This review discusses the role of connexins in epidermal integrity, and examines the emerging evidence that connexins act as epidermal sensors to a variety of mechanical, temperature, pathogen-induced and chemical stimuli. Connexins thus act as an integral component of the skin's protective barrier.

The role of pannexin1 in the induction and resolution of inflammation.

Extracellular ATP is an important signaling molecule throughout the inflammatory cascade, serving as a danger signal that causes activation of the inflammasome, enhancement of immune cell infiltration, and fine-tuning of several signaling cascades including those important for the resolution of inflammation. Recent studies demonstrated that ATP can be released from cells in a controlled manner through pannexin (Panx) channels. Panx1-mediated ATP release is involved in inflammasome activation and neutrophil/macrophage chemotaxis, activation of T cells, and a role for Panx1 in inducing and propagating inflammation has been demonstrated in various organs, including lung and the central and peripheral nervous system. The recognition and clearance of dying cells and debris from focal points of inflammation is critical in the resolution of inflammation, and Panx1-mediated ATP release from dying cells has been shown to recruit phagocytes. Moreover, extracellular ATP can be broken down by ectonucleotidases into ADP, AMP, and adenosine, which is critical in the resolution of inflammation. Together, Panx1, ATP, purinergic receptors, and ectonucleotidases contribute to important feedback loops during the inflammatory response, and thus represent promising candidates for new therapies.

Cogan's syndrome--clinical guidelines and novel therapeutic approaches.

Cogan's syndrome (CS) is a rare chronic inflammatory disorder, classically characterized by interstitial keratitis and sensorineural hearing loss. Recurrent episodes of inner ear disease might result in deafness. In some patients, it may also be accompanied by systemic vasculitis. Diagnosis of CS is often missed or delayed due to its rarity, the nonspecific clinical signs at onset, and the lack of a confirmatory diagnostic test. The mechanisms responsible for CS are unknown; however, in the last decade, the pathogenesis has been somewhat elucidated, suggesting that the disease is a result of inner ear autoimmunity. The autoimmune hypothesis postulates the triggering of the disease by a viral infection via a number of mechanisms, which are mainly as follows: antigenic mimicry, self-perpetuating inflammation by cytokine release, and unveiling hidden epitopes. Aside from its clinical resemblance to other autoimmune disorders, some autoantigen has apparently been identified, namely, CD148 and connexine 26. Treatment should begin as early as possible. While treatment is based primarily on glucocorticoids, there is no standard alternative for patients who respond poorly. Failure of conventional treatment could lead to profound sensorineural hearing loss. From the limited data we have, infliximab seems to be the most promising biological remedy, enabling steroid tapering and leading to improvement in auditory/ocular disease, with better results when administered in early stages. Proposed guidelines for the use of infliximab in CS are found in the last table of the review, in an attempt to define the proper timing for initiating infliximab treatment in order to avoid permanent disability.

P2X7 receptor-Pannexin1 interaction mediates stress-induced interleukin-1 beta expression in human periodontal ligament cells.

BACKGROUND AND OBJECTIVE: Pannexin 1 (Panx1) has been found to form nonjunctional hemichannels. It is also proposed to combine with the P2X7 receptor, forming a complex involved in adenosine triphosphate (ATP)-induced interleukin-1beta (IL-1ß) release in macrophages. Previously, we reported that mechanical stress induced IL-1ß expression via the ATP/P2X7 receptor-dependent pathway in human periodontal ligament (HPDL) cells and that ATP was released through the connexin 43 (Cx43) hemichannel. In the present work, we examined the role of Panx1 in stress-induced IL-1ß induction in HPDL cells. MATERIAL AND METHODS: Cultured HPDL cells were treated with compressive loading or ATP to stimulate IL-1ß expression. Inhibitors, antagonists and the small interfering RNA technique were used to investigate the involvement of Panx1 in IL-1ß induction. Co-immunoprecipitation (Co-IP) and immunostaining were used to determine the association of Panx1 with the P2X7 receptor. The IL-1ß release mechanism was analyzed using inhibitors. RESULTS: Blocking Panx1 significantly decreased ATP release, as well as IL-1ß up-regulation, upon stimulation with stress or ATP. Co-IP revealed the association of Panx1 and the P2X7 receptor in HPDL cells, which was increased in response to mechanical loading. Pretreatment with vesicular trafficking inhibitors significantly reduced the amount of IL-1ß released from stimulated cells, suggesting that IL-1ß might be released through vesicles. CONCLUSION: We clearly illustrated the contribution of Panx1 in ATP release, as well as in IL-1ß induction in HPDL cells. The association of Panx1 and the P2X7 receptor might be required for IL-1ß induction, and their possible novel role in IL-1ß vesicular release was indicated.

Antibodies targeting extracellular domain of connexins for studies of hemichannels.

Hemichannels are transmembrane channels composed of either a connexin or pannexin hexamer. The docking of the extracellular domains of connexin hemichannels contributed by neighboring cells forms a gap junction channel that joins the cytoplasm of adjacent cells. Connexins are expressed ubiquitously in different organs, but some subtypes are expressed exclusively in certain tissues and tumors. Both gap junction channels and hemichannels participate in diverse physiological and pathological responses. However, the lack of specific reagents that inhibit only gap junction channels or hemichannels is a challenge that makes it different to discern the specific roles of either channel. Fortunately, the available information regarding the connexin sequence, secondary and tertiary structure, and their biochemical and physiological properties permits the development of strategies to block exclusively the hemichannel activity exclusively, with no effect on gap junction activity. This task is accomplished through the use of specifics antibodies that target the extracellular sites of desired connexin subtype. However, the underlying mechanism of how antibodies targeting extracellular connexin epitopes actually inhibit hemichannels remains unknown. Although these antibodies are being used for detecting and blocking of hemichannels in normal and tumor cells, they can also be potentially used for tissue-specific treatment and drug delivery in clinical applications. In this article, we will first review the literature concerning the structure of connexins and the unique properties of extracellular loop domains of the connexins. Furthermore, we will discuss briefly the development of connexin (Cx) 43(E2) antibody, a specific antibody which detects the second extracellular loop of Cx43 and specifically prevents the opening of Cx43 hemichannels. We will then summarize the reported studies of specific reagents used for the inhibition of connexin hemichannels including antibodies developed against extracellular loop domains. This article is part of the Special Issue Section entitled 'Current Pharmacology of Gap Junction Channels and Hemichannels'.

Investigation of 20 non-HLA (human leucocyte antigen) psoriasis susceptibility loci in Chinese patients with psoriatic arthritis and psoriasis vulgaris.

BACKGROUND: Recently, a number of non-HLA (human leucocyte antigen) psoriasis genetic susceptibility loci have been identified through genome-wide association studies, but data on their association with psoriatic arthritis (PsA) are lacking. OBJECTIVES: To investigate recently identified psoriasis susceptibility loci in a cohort of Chinese patients with PsA, psoriasis vulgaris (PsV) and healthy controls. METHODS: Twenty single-nucleotide polymorphisms (SNPs) from 20 loci were selected for genotyping in 379 patients with PsA, 595 patients with PsV and 1181 healthy controls using the MassARRAY platform (Sequenom, San Diego, CA, U.S.A.). Data handling, quality control and association were performed using PLINK software, v. 1.07. The Cochran-Armitage trend test was used to test the genotype-phenotype association. RESULTS: PsA showed a significant association with markers at TNIP1 (rs17728338, P = 2.20 × 10(-8)), IL28RA (rs4649203, P = 5.04 × 10(-6)), IL12B (rs2082412, P = 3.82 × 10(-5)), ERAP1 (rs27524, P = 1.25 × 10(-3)), PTTG1 (rs2431697, P = 1.22 × 10(-3)) and GJB2 (rs3751385, P = 1.48 × 10(-3)) when compared with the control group. In PsV a significant association was found for IL28RA (rs4649203, P = 9.53 × 10(-7)), TNIP1 (rs17728338, P = 1.21 × 10(-4)) and ERAP1 (rs27524, P = 1.17 × 10(-3)). The allele frequencies were not statistically different between PsA and PsV except for SNPs at IL12B and ZNF816A with a nominal P-value of 0.04 and 0·01, respectively. CONCLUSIONS: This study provides evidence for the involvement of ERAP1, IL28RA, GJB2 and PTTG1 loci in PsA susceptibility and confirmed the previously reported association with PsA and PsV. These results support the hypothesis that genetic aetiology of psoriasis is the same in both PsA and PsV and also support the higher genetic component of PsA than PsV.

Aberrant connexin 43 and 26 expression in cervical dysplasia.

OBJECTIVE: To determine whether connexin (Cx) expression is altered in cervical dysplasia. STUDY DESIGN: Cx proteins form gap junctions and are expressed in squamous epithelia including ectocervix. We used multispectral imaging to perform a quantitative immunohistochemical survey of Cx43 Cx26 in 37 archival human cervical specimens. RESULTS: Cx43 expression was very low in normal cervix (100%), but was increased in low-grade squamous intraepithelial lesions (LSILs) (64%), primarily in a parabasal distribution. High-grade squamous intraepithelial lesions (HSILs) showed weak full-thickness Cx43 staining (53%) or lacked Cx43 (47%). An aberrant increase in Cx43 expression was often (62%) present in histologically normal areas of specimens that elsewhere harbored dysplasia. Cx26 was highly expressed in the basal layer of normal ectocervix (100%). In LSIL, 57% showed a decrease in Cx26 and the rest showed no change relative to the normal pattern. In HSIL, Cx26 was expressed in the full thickness of the epithelium, at a high level in 80% of cases and a low level in the rest. CONCLUSION: Cx alteration is moderately consistent in cervical dysplasia, and for Cx43 can precede histologic changes. The resulting changes in Cx signaling may be important in the pathogenesis of cervical intraepithelial neoplasia.

Suppression of connexin 26 is related to protease-activated receptor 2-mediated pathway in patients with allergic rhinitis.

BACKGROUND: Connexin (Cx) 26 plays a key role in maintaining the integrity of tight junctions. However, the expression and modulation of Cx26 in allergic rhinitis (AR) has not been well understood. METHODS: We detected the expression of Cx26 in house-dust mite (HDM)-sensitized AR patients and investigated the Cx26 production and modulation in primary human nasal epithelial cells (HNECs) and BEAS-2B cells after treatment with the allergen Der p 1 from Dermatophagoides pteronyssinus. RESULTS: We found that the mRNA and protein levels of Cx26 were significantly down-regulated in AR patients compared with the control. Der p 1 was found to induce protease-activated receptor 2 (PAR2) expression and suppress Cx26 production significantly in vitro. PAR2 siRNA was shown to prevent the suppression of Cx26 induced by Der p 1 in BEAS-2B cells. CONCLUSION: The suppression of Cx26 in HDM-sensitized AR patients is related to a PAR2-mediated pathway and might serve during the initiation and maintenance of AR. Targeting the PAR2-mediated Cx26 suppression may be a potential means of preventing allergic sensitization.