Increased levels of the chemokine CCL2 in cancer patients are associated with poor prognosis. Experimental evidence suggests that CCL2 correlates with inflammatory monocyte recruitment and induction of vascular activation, but the functionality remains open. Here, we show that endothelial Ccr2 facilitates pulmonary metastasis using an endothelial-specific Ccr2-deficient mouse model (Ccr2ecKO). Similar levels of circulating monocytes and equal leukocyte recruitment to metastatic lesions of Ccr2ecKO and Ccr2fl/fl littermates were observed. The absence of endothelial Ccr2 strongly reduced pulmonary metastasis, while the primary tumor growth was unaffected. Despite a comparable cytokine milieu in Ccr2ecKO and Ccr2fl/fl littermates the absence of vascular permeability induction was observed only in Ccr2ecKO mice. CCL2 stimulation of pulmonary endothelial cells resulted in increased phosphorylation of MLC2, endothelial cell retraction, and vascular leakiness that was blocked by an addition of a CCR2 inhibitor. These data demonstrate that endothelial CCR2 expression is required for tumor cell extravasation and pulmonary metastasis. IMPLICATIONS: The findings provide mechanistic insight into how CCL2-CCR2 signaling in endothelial cells promotes their activation through myosin light chain phosphorylation, resulting in endothelial retraction and enhanced tumor cell migration and metastasis.

• MeSH
• chemokin CCL2 metabolismus   MeSH
• endoteliální buňky metabolismus   patologie   MeSH
• kapilární permeabilita MeSH
• karcinom plic Lewisové krevní zásobení   metabolismus   patologie   sekundární   MeSH
• lehké řetězce myosinu metabolismus   MeSH
• metastázy nádorů MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• nádory plic krevní zásobení   metabolismus   patologie   sekundární   MeSH
• pohyb buněk fyziologie   MeSH
• receptory CCR2 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Glial cells activated by peripheral nerve injury contribute to the induction and maintenance of neuropathic pain by releasing neuromodulating cytokines and chemokines. We investigated the activation of microglia and astrocytes as well as the cellular distribution of the chemokine CCL2 and its receptor CCR2 in the trigeminal subnucleus caudalis (TSC) ipsilateral and contralateral to infraorbital nerve ligature (IONL). The left infraorbital nerve was ligated under aseptic conditions, and sham controls were operated without nerve ligature. Tactile hypersensitivity was significantly increased bilaterally in vibrissal pads of both sham- and IONL-operated animals from day 1 to 7 and tended to normalize in sham controls surviving for 14 days. Activated microglial cells significantly increased bilaterally in the TSC of both sham- and IONL-operated animals with a marked but gradual increase in the ipsilateral TSC from 1 to 7 days followed by a decrease by day 14. In contrast, robust activation of astrocytes was found bilaterally in the TSC of IONL-operated rats from 3 to 14 days with a transient activation in the ipsilateral TSC of sham-operated animals. Cellular distribution of CCL2 varied with survival time. CCL2 immunofluorescence was detected in neurons within 3 days and in astrocytes at later time points. In contrast, CCR2 was found only in astrocytes at all time points with CCR2 intensity being dominant in the ipsilateral TSC. In summary, our results reveal bilateral activation of microglial cells and astrocytes as well as changes in the cellular distribution of CCL2 and its receptor CCR2 in the TSC during the development and maintenance of orofacial neuropathic pain.

• MeSH
• chemokin CCL2 metabolismus   MeSH
• krysa rodu rattus MeSH
• modely nemocí na zvířatech MeSH
• neuralgie metabolismus   MeSH
• neuroglie metabolismus   MeSH
• potkani Wistar MeSH
• receptory CCR2 metabolismus   MeSH
• substantia gelatinosa metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Neuropathic pain after spinal cord injury lacks any effective treatments, often leading to chronic pain. This study tested whether the daily administration of fully characterized polyphenolic extracts from grape stalks and coffee could prevent both reflexive and non-reflexive chronic neuropathic pain in spinal cord-injured mice by modulating the neuroimmune axis. Female CD1 mice underwent mild spinal cord contusion and received intraperitoneal extracts in weeks one, three, and six post-surgery. Reflexive pain responses were assessed weekly for up to 10 weeks, and non-reflexive pain was evaluated at the study's end. Neuroimmune crosstalk was investigated, focusing on glial activation and the expression of CCL2/CCR2 and CX3CL1/CX3CR1 in supraspinal pain-related areas, including the periaqueductal gray, rostral ventromedial medulla, anterior cingulate cortex, and amygdala. Repeated treatments prevented mechanical allodynia and thermal hyperalgesia, and also modulated non-reflexive pain. Moreover, they reduced supraspinal gliosis and regulated CCL2/CCR2 and CX3CL1/CX3CR1 signaling. Overall, the combination of polyphenols in these extracts may offer a promising pharmacological strategy to prevent chronic reflexive and non-reflexive pain responses by modifying central sensitization markers, not only at the contusion site but also in key supraspinal regions implicated in neuropathic pain. Overall, these data highlight the potential of polyphenolic extracts for spinal cord injury-induced chronic neuropathic pain.

• MeSH
• chemokin CCL2 metabolismus   MeSH
• chemokin CX3CL1 metabolismus   MeSH
• CX3C chemokinový receptor 1 metabolismus   MeSH
• glióza * farmakoterapie   metabolismus   MeSH
• hyperalgezie farmakoterapie   MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• neuralgie * farmakoterapie   metabolismus   etiologie   prevence a kontrola   MeSH
• polyfenoly * farmakologie   aplikace a dávkování   MeSH
• poranění míchy * komplikace   farmakoterapie   metabolismus   MeSH
• receptory CCR2 metabolismus   MeSH
• rostlinné extrakty * farmakologie   aplikace a dávkování   MeSH
• signální transdukce * účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Poznatky získané v posledních letech defi nují aterosklerózu jako chronické systémové zánětlivé onemocnění, které je charakterizováno degenerativními změnami a extracelulární akumulací lipidů. Zánět je důležitým patogenetickým mechanismem aterosklerózy, jejíž klinické projevy zahrnují onemocnění různé lokalizace a závažnosti. V procesu aterogeneze hrají klíčovou úlohu chemotaktické cytokiny a jejich receptory. Expresi a funkci těchto molekul ovlivňuje variabilita (polymorfi smy) genů, které je kódují. Snahou badatelů v translačním výzkumu je na základě nových znalostí patobiologie kardiovaskulárních onemocnění rozšířit současné spektrum vyšetřovaných parametrů o nové ukazatele, které by pomohly upřesnit míru rizika vzniku a progrese onemocnění. Jednou z cest k vytčenému cíli je studium kandidátních molekul uplatňujících se v patogenezi aterosklerózy, diskutovanou kandidátní molekulou je také mononukleární chemoatraktant MCP-1 (CCL2) a jeho receptor CCR2.

Atherosclerosis has been defi ned as systemic, chronic infl ammatory disease characterized by degenerative processes and extracellular accumulation of lipids. Infl ammation is important pathogenetic mechanism of atherosclerosis whose clinical manifestations comprise diseases of distinct location. Chemotactic cytokines and their receptors play key role in atherogenesis. Expression and function of these molecules is infl uenced by variability (polymorphisms) in their encoding genes. Translational research in pathobiology of cardiovascular disorders aims at extending the current spectrum of laboratory parameters with new markers, which could help us to defi ne the patients at risk more precisely and to determine disease course more accurately. Investigation of atherosclerosis candidate molecules is one of the possible routes to this aim. Mononuclear chemoattractant MCP-1 (CCL2) and its receptor CCR2 represent such molecules.

• Klíčová slova
• mononukleární chemoatraktant MCP-1 (CCL2), receptor CCR2,
• MeSH
• angina pectoris MeSH
• ateroskleróza genetika   imunologie   MeSH
• chemokin CCL2 genetika   krev   MeSH
• infarkt myokardu MeSH
• klinické zkoušky jako téma MeSH
• lidé MeSH
• mediátory zánětu MeSH
• polymorfismus genetický MeSH
• receptory CCR2 MeSH
• zánět MeSH
• Check Tag
• lidé MeSH

BACKGROUND: Chemokines and chemokine receptors are major mediators of leukocyte trafficking into the sites of the immune response. They participate in defence against microbial infection, in Th1/Th2 polarization of the immune response, allograft rejection and angiogenesis/angiostasis as well as in tumorigenesis and metastasis. To date, several functional polymorphisms of chemokine and chemokine receptor genes have been discovered that are able to deregulate chemokine system and, therefore, they may interfere with the pathogenesis of a large number of inflammatory and other diseases. In this review we focus on the known polymorphisms of two chemokines: CCL2, CCL5 and their corresponding receptors (CCR2, CCR5) and we also discuss their associations with susceptibility and progression to selected immune-mediated diseases. METHODS AND RESULTS: Based on relevant literature this article gives a short overview of case-control and family studies regarding effect of the genetic factors on diseases such as coronary artery disease, systemic lupus erythematosus, diabetes mellitus, lung diseases and others. CONCLUSION: Recent advance in the identification of chemokine genetic background of the diseases could provide opportunity for pharmacological treatment. However, we need more information about posttranscriptional events to understand functional relevance of polymorphisms and to discovery new avenues to blocking disease development.

• MeSH
• chemokin CCL2 genetika   MeSH
• chemokin CCL5 genetika   imunologie   fyziologie   MeSH
• lidé MeSH
• polymorfismus genetický MeSH
• receptory CCR2 genetika   imunologie   fyziologie   MeSH
• receptory CCR5 genetika   imunologie   fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

• MeSH
• albuminurie * farmakoterapie   MeSH
• chemokin CCL2 * terapeutické užití   MeSH
• diabetické nefropatie patofyziologie   MeSH
• lidé MeSH
• randomizované kontrolované studie jako téma MeSH
• receptory CCR2 antagonisté a inhibitory   MeSH
• Check Tag
• lidé MeSH

Several lines of evidence suggest that chemokines play an important role in asthma and allergy. We analysed polymorphisms at -2518A/G and -2076A/T of MCP-1 and V64I of CCR2 gene in healthy subjects (n = 306) and allergic patients (n = 332). Allele and genotype frequencies did not differ significantly between groups. Nevertheless, MCP-1 variants were associated with allergen sensitization. The results suggest that MCP-1, but not CCR2 gene variants, may participate in the pathogenesis of allergic phenotypes at least in the Caucasian population.

• MeSH
• alely MeSH
• alergie epidemiologie   genetika   MeSH
• chemokin CCL2 genetika   MeSH
• dospělí MeSH
• financování organizované MeSH
• frekvence genu MeSH
• genotyp MeSH
• lidé středního věku MeSH
• lidé MeSH
• polymorfismus genetický MeSH
• receptory CCR2 genetika   MeSH
• studie případů a kontrol MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Geografické názvy
• Česká republika MeSH

Peripheral nerve injuries (PNIs) may result in cellular and molecular changes in supraspinal structures possibly involved in neuropathic pain (NPP) maintenance. Activated glial cells in specific supraspinal subregions may affect the facilitatory role of descending pathways. Sterile chronic compression injury (sCCI) and complete sciatic nerve transection (CSNT) in rats were used as NPP models to study the activation of glial cells in the subregions of periaqueductal gray (PAG) and rostral ventromedial medulla (RVM). Molecular markers for activated astrocytes (glial fibrillary acidic protein, GFAP) and microglial cells (OX42) were assessed by quantitative immunohistochemistry and western blotting. The cellular distribution of CCL2/CCR2 was monitored using immunofluorescence. sCCI induced both mechanical and thermal hypersensitivity from day 1 up to 3 weeks post-injury. Unilateral sCCI or CSNT for 3 weeks induced significant activation of astrocytes bilaterally in both dorsolateral (dlPAG) and ventrolateral PAG (vlPAG) compared to naïve or sham-operated rats. More extensive astrocyte activation by CSNT compared to sCCI was induced bilaterally in dlPAG and ipsilaterally in vlPAG. Significantly more extensive activation of astrocytes was also found in RVM after CSNT than sCCI. The CD11b immunopositive region, indicating activated microglial cells, was remarkably larger in dlPAG and vlPAG of both sides from sCCI- and CSNT-operated rats compared to naïve or sham-operated controls. No significant differences in microglial activation were detected in dlPAG or vlPAG after CSNT compared to sCCI. Both nerve injury models induced no significant differences in microglial activation in the RVM. Neurons and activated GFAP+ astrocytes displayed CCL2-immunoreaction, while activated OX42+ microglial cells were CCR2-immunopositive in both PAG and RVM after sCCI and CSNT. Overall, while CSNT induced robust astrogliosis in both PAG and RVM, microglial cell activation was similar in the supraspinal structures in both injury nerve models. Activated astrocytes in PAG and RVM may sustain facilitation of the descending system maintaining NPP, while microglial activation may be associated with a reaction to long-lasting peripheral injury. Microglial activation via CCR2 may be due to neuronal and astrocytal release of CCL2 in PAG and RVM following injury.

• Publikační typ
• časopisecké články MeSH

Blood vessel functionality is crucial for efficient tumor-targeted drug delivery. Heterogeneous distribution and perfusion of angiogenic blood vessels contribute to suboptimal accumulation of (nano-) therapeutics in tumors and metastases. To attenuate pathological angiogenesis, an L-RNA aptamer inhibiting the CC motif chemokine ligand 2 (CCL2) was administered to mice bearing orthotopic 4T1 triple-negative breast cancer tumors. The effect of CCL2 inhibition on tumor blood vessel functionality and tumor-targeted drug delivery was evaluated via multimodal and multiscale optical imaging, employing fluorophore-labeled polymeric (10 nm) and liposomal (100 nm) nanocarriers. Anti-CCL2 treatment induced a dose-dependent anti-angiogenic effect, reflected by a decreased relative blood volume, increased blood vessel maturity and functionality, and reduced macrophage infiltration, accompanied by a shift in the polarization of tumor-associated macrophages (TAM) towards a less M2-like and more M1-like phenotype. In line with this, CCL2 inhibitor treatment improved the delivery of polymers and liposomes to tumors, and enhanced the antitumor efficacy of free and liposomal doxorubicin. Together, these findings demonstrate that blocking the CCL2-CCR2 axis modulates TAM infiltration and polarization, resulting in vascular normalization and improved tumor-targeted drug delivery.

• MeSH
• chemokin CCL2 * farmakologie   MeSH
• ligandy MeSH
• makrofágy MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory * patologie   MeSH
• nanomedicína MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: The choroid plexus is located in the cerebral ventricles. It consists of a stromal core and a single layer of cuboidal epithelial cells that forms the blood-cerebrospinal barrier. The main function of the choroid plexus is to produce cerebrospinal fluid. Subarachnoid hemorrhage due to aneurysm rupture is a devastating type of hemorrhagic stroke. Following subarachnoid hemorrhage, blood and the blood degradation products that disperse into the cerebrospinal fluid come in direct contact with choroid plexus epithelial cells. The aim of the current study was to elucidate the pathophysiological cascades responsible for the inflammatory reaction that is seen in the choroid plexus following subarachnoid hemorrhage. METHODS: Subarachnoid hemorrhage was induced in rats by injecting non-heparinized autologous blood to the cisterna magna. Increased intracranial pressure following subarachnoid hemorrhage was modeled by using artificial cerebrospinal fluid instead of blood. Subarachnoid hemorrhage and artificial cerebrospinal fluid animals were left to survive for 1, 3, 7 and 14 days. Immunohistochemical staining of TLR4, TLR9, FPR2, CCL2, TNFα, IL-1β, CCR2 and CX3CR1 was performed on the cryostat sections of choroid plexus tissue. The level of TLR4, TLR9, FPR2, CCL2, TNFα, IL-1β was detected by measuring immunofluorescence intensity in randomly selected epithelial cells. The number of CCR2 and CX3CR1 positive cells per choroid plexus area was manually counted. Immunohistochemical changes were confirmed by Western blot analyses. RESULTS: Immunohistochemical methods and Western blot showed increased levels of TLR9 and a slight increase in TLR4 and FRP2 following both subarachnoid hemorrhage as well as the application of artificial cerebrospinal fluid over time, although the individual periods were different. The levels of TNFα and IL-1β increased, while CCL2 level decreased slightly. Accumulation of macrophages positive for CCR2 and CX3CR1 was found in all periods after subarachnoid hemorrhage as well as after the application of artificial cerebrospinal fluid. DISCUSSION: Our results suggest that the inflammation develops in the choroid plexus and blood-cerebrospinal fluid barrier in response to blood components as well as acutely increased intracranial pressure following subarachnoid hemorrhage. These pro-inflammatory changes include accumulation in the choroid plexus of pro-inflammatory cytokines, innate immune receptors, and monocyte-derived macrophages.

• Publikační typ
• časopisecké články MeSH