Anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitors (TKIs) show potent efficacy in several ALK-driven tumors, but the development of resistance limits their long-term clinical impact. Although resistance mechanisms have been studied extensively in ALK-driven non-small cell lung cancer, they are poorly understood in ALK-driven anaplastic large cell lymphoma (ALCL). Here, we identify a survival pathway supported by the tumor microenvironment that activates phosphatidylinositol 3-kinase γ (PI3K-γ) signaling through the C-C motif chemokine receptor 7 (CCR7). We found increased PI3K signaling in patients and ALCL cell lines resistant to ALK TKIs. PI3Kγ expression was predictive of a lack of response to ALK TKI in patients with ALCL. Expression of CCR7, PI3Kγ, and PI3Kδ were up-regulated during ALK or STAT3 inhibition or degradation and a constitutively active PI3Kγ isoform cooperated with oncogenic ALK to accelerate lymphomagenesis in mice. In a three-dimensional microfluidic chip, endothelial cells that produce the CCR7 ligands CCL19/CCL21 protected ALCL cells from apoptosis induced by crizotinib. The PI3Kγ/δ inhibitor duvelisib potentiated crizotinib activity against ALCL lines and patient-derived xenografts. Furthermore, genetic deletion of CCR7 blocked the central nervous system dissemination and perivascular growth of ALCL in mice treated with crizotinib. Thus, blockade of PI3Kγ or CCR7 signaling together with ALK TKI treatment reduces primary resistance and the survival of persister lymphoma cells in ALCL.

• MeSH
• anaplastická lymfomová kináza MeSH
• anaplastický velkobuněčný lymfom * farmakoterapie   genetika   patologie   MeSH
• endoteliální buňky metabolismus   MeSH
• fosfatidylinositol-3-kinasy MeSH
• inhibitory proteinkinas farmakologie   terapeutické užití   MeSH
• inhibitory tyrosinkinasy MeSH
• krizotinib farmakologie   terapeutické užití   MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí MeSH
• nádory plic * farmakoterapie   MeSH
• nemalobuněčný karcinom plic * farmakoterapie   MeSH
• receptory CCR7 genetika   MeSH
• tyrosinkinasové receptory metabolismus   MeSH
• tyrosinkinasy MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Increasing evidence points to host genetics as a factor in COVID-19 prevalence and outcome. CCR5 is a receptor for proinflammatory chemokines that are involved in host responses, especially to viruses. The CCR5-delta32 minor allele is an interesting variant, given the role of CCR5 in some viral infections, particularly HIV-1. Recent studies of the impact of CCR5-delta32 on COVID-19 risk and severity have yielded contradictory results. This ecologic study shows that the CCR5-delta32 allelic frequency in a European population was significantly negatively correlated with the number of COVID-19 cases (p=0.035) and deaths (p=0.006) during the second pandemic wave. These results suggest that CCR5-delta32 may be protective against SARS-CoV-2 infection, as it is against HIV infection, and could be predictive of COVID-19 risk and severity. Further studies based on samples from populations of different genetic backgrounds are needed to validate these statistically obtained findings.

• MeSH
• COVID-19 genetika   imunologie   mortalita   virologie   MeSH
• fenotyp MeSH
• frekvence genu MeSH
• genetická predispozice k nemoci MeSH
• interakce hostitele a patogenu MeSH
• lidé MeSH
• mutace * MeSH
• ochranné faktory MeSH
• prevalence MeSH
• receptory CCR5 genetika   MeSH
• rizikové faktory MeSH
• SARS-CoV-2 imunologie   patogenita   MeSH
• stupeň závažnosti nemoci MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

Infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes coronavirus disease (COVID-19), has spread widely around the globe. Significant inter-individual differences have been observed during the course of the infection, which suggests that genetic susceptibility may be a contributing factor. CC chemokine receptor 5 (CCR5), which acts as a co-receptor for the entry of HIV-1 into cells, is promising candidate whose can have an influence on SARS-CoV-2 infection. A genetic mutation known as CCR5Delta32, consisting of a 32-nucleotide deletion, encodes a truncated protein that protects homozygous carriers of the deletion from HIV-1 infection. Similarly, inhibition of CCR5 seems to be protective against COVID-19. In our study, we successfully genotyped 416 first-wave SARS-CoV-2-positive infection survivors (164 asymptomatic and 252 symptomatic) for CCR5?32, comparing them with a population based sample of 2,404 subjects. We found the highest number (P=0.03) of CCR5Delta32 carriers in SARS-CoV-2-positive/COVID-19-asympto-matic subjects (23.8 %) and the lowest number in SARS-CoV-2-positive/COVID-19-symptomatic patients (16.7 %), with frequency in the control population in the middle (21.0 %). We conclude that the CCR5?32 I/D polymorphism may have the potential to predict the severity of SARS-CoV-2 infection.

• MeSH
• COVID-19 diagnóza   genetika   virologie   MeSH
• fenotyp MeSH
• frekvence genu MeSH
• genetická predispozice k nemoci MeSH
• genetické asociační studie MeSH
• hodnocení rizik MeSH
• lidé MeSH
• ochranné faktory MeSH
• receptory CCR5 genetika   MeSH
• rizikové faktory MeSH
• sekvenční delece * MeSH
• studie případů a kontrol MeSH
• stupeň závažnosti nemoci MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Following nerve injury, disintegrated axonal mitochondria distal to the injury site release mitochondrial formylated peptides and DNA that can induce activation and inflammatory profiling of Schwann cells via formyl peptide receptor 2 (Fpr2) and toll-like receptor 9 (TLR9), respectively. We studied RT4 schwannoma cells to investigate the regulation of Fpr2 and TLR9 after stimulation with fMLF as a prototypical formylated peptide. RT4 cells were treated with fMLF at various concentrations and times with and without pretreatment with inhibitors (chloroquine for activated TLR9, PBP10 for Fpr2). Western blots of Fpr2, TLR9, p-p38, p-NFκB, and IL-6 were compared in relation to inflammatory profiling of RT4 cells and chemokine receptors (CCR2, CXCR4) as potential co-receptors of Fpr2. fMLF stimulation upregulated Fpr2 in RT4 cells at low concentrations (10 nM and 100 nM) but higher concentrations were required (10 µM and 50 µM) when the cells were pretreated with an activated TLR9 inhibitor. Moreover, the higher concentrations of fMLF could modulate TLR9 and inflammatory markers. Upregulation of Fpr2 triggered by 10 nM and 100 nM fMLF coincided with higher levels of chemokine receptors (CCR2, CXCR4) and PKCβ. Treating RT4 cells with fMLF, as an in vitro model of Schwann cells, uncovered Schwann cells' complex responses to molecular patterns of release from injured axonal mitochondria.

• MeSH
• chlorochin farmakologie   MeSH
• krysa rodu rattus MeSH
• N-formylmethionin-leucyl-fenylalanin farmakologie   MeSH
• nádorové buněčné linie MeSH
• neurilemom metabolismus   patologie   MeSH
• receptory CCR2 genetika   metabolismus   MeSH
• receptory CXCR4 genetika   metabolismus   MeSH
• receptory pro formylované peptidy antagonisté a inhibitory   genetika   metabolismus   MeSH
• Schwannovy buňky cytologie   účinky léků   metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• toll-like receptor 9 antagonisté a inhibitory   genetika   metabolismus   MeSH
• upregulace účinky léků   MeSH
• zánět metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus 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

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

• MeSH
• biologická terapie etika   metody   MeSH
• cílená genová oprava etika   metody   MeSH
• CRISPR-Cas systémy genetika   MeSH
• editace genu * etika   metody   MeSH
• fertilizace in vitro metody   MeSH
• genetická terapie * etika   metody   MeSH
• genetické inženýrství etika   metody   MeSH
• HIV genetika   MeSH
• lidé MeSH
• receptory CCR5 genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• novinové články MeSH

Úvod: Vstup viru lidské imunitní nedostatečnosti typu 1 (HIV-1) do cílových buněk je umožněn CD4 receptorem a jedním ze dvou koreceptorů CXCR4 nebo CCR5. Delece úseku 32 nukleotidů v genu pro CCR5 (CCR5Δ32) v obou alelách poskytuje silnou, avšak ne absolutní odolnost proti infekci HIV-1 a delece v jedné alele zpomaluje postup nemoci směřující k rozvinutí AIDS. Zde jsme analyzovali prevalenci a roli heterozygotního výskytu CCR5Δ32 na postup onemocnění u HIV pozitivních jedinců v České republice. Metody: Celkem 92 HIV-1 séropozitivních osob včetně 80 Čechů z AIDS centra v Nemocnici Na Bulovce v Praze bylo zařazeno do genotypizace CCR5, která byla součástí studie role HIV fitness na průběh onemocnění. Z periferních mononukleárních buněk pacienta byla získána DNA, která byla použita k amplifikaci pomocí PCR v reálném čase použitím specifických sond, které detekovaly divokou variantu CCR5 a variantu s 32 nt delecí. Podmnožina 74 pacientů bez antiretrovirové léčby, kteří byli sledováni déle než jeden rok, byla použita k určení role heterozygotního fenotypu CCR5 na průběh nemoci. Výsledky: Heterozygotní CCR5Δ32 varianta byla nalezena u 23,8 % z 80 českých HIV-1 séropozitivních osob, což je velmi podobné jako publikovaná 21 % a 24 % prevalence u HIV negativní české populace. Homozygotní mutovaná varianta nebyla nalezena. U skupiny osob s heterozygotním CCR5 fenotypem jsme pozorovali slabě zvýšený průměrný počet CD4-pozitivních T-lymfocytů a nižší průměrné hodnoty virémie v plazmě. Závěr: Celkově, naše studie neukázala žádný zřejmý užitek přítomnosti heterozygotní CCR5Δ32 varianty na přenos HIV a pouze malý užitek na průběh nemoci u české HIV-1 pozitivní kohorty.

Background: Entry of human immunodeficiency virus type 1 (HIV-1) in target cells is enabled by CD4 receptor and one of two co-receptors, CXCR4 or CCR5. Deletion of 32 bp in CCR5 gene (CCR5Δ32) in both alleles provides strong but not absolute resi-stance to HIV-1 infection and deletion in one allele slows disease progression to AIDS. Here, we analyzed the prevalence and the role of CCR5Δ32 heterozygosity on the disease progression in HIV positive patients in the Czech Republic. Patients and methods: A total of 92 HIV-1 seropositive subjects that included 80 Czech individuals from the AIDS center in the Hospital Na Bulovce in Prague were enrolled in CCR5 genoty-ping as a part of a study of the role of HIV fitness on disease progression. DNA was extracted from patient’s peripheral blood mononuclear cells and subjected to real-time PCR with specific probes detecting wild-type and 32 bp-deleted CCR5 variants. A subgroup of 74 antiretroviral therapy-naive patients with more than one year of follow-up was used to determine the role of the CCR5Δ32 heterozygous phenotype in disease progression. Results: CCR5Δ32 was found heterozygous in 23.8% of 80 Czech HIV-1 seropositive individuals which is very similar to 21% and 24% prevalence reported in HIV negative Czech population. Homozygous mutant variant was not detected. In CCR5Δ32 he-terozygous group we observed slightly higher mean CD4+ T-cell count and lower mean plasma viremia levels.Conclusions: Overall, our study indicates no obvious benefit of CCR5Δ32 heterozygosity on HIV transmission and only small benefit on disease progression in the Czech HIV-1 cohort.

• MeSH
• genotypizační techniky MeSH
• HIV-1 * genetika   izolace a purifikace   MeSH
• kohortové studie MeSH
• lidé MeSH
• polymorfismus genetický MeSH
• progrese nemoci MeSH
• receptory CCR5 genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

OBJECTIVE: Cardiac surgery is known to trigger a systemic inflammatory response. While the use of conventional cardiopulmonary bypass (CPB) results in profound inflammation, modified mini-CPB is considered less harmful. We evaluated the impact of cardiac surgery on the expression of CD162, CD166, CD195 molecules and their association with the type of CPB used. METHODS AND RESULTS: Twenty-four patients were enrolled in our study. Twelve of them were operated using conventional CPB while the other twelve patients underwent surgery with mini-CPB. Blood samples were analysed by flow cytometry. We observed a significant increase in median fluorescence intensity of CD162 and CD195 that peaked instantly after surgery and normalized to the baseline value on the 1st day post surgery, whereas CD166 was initially down-regulated and its median fluorescence intensity (MFI) value increased to the baseline in the next few days. CONCLUSION: We observed immediate changes in the expression of CD162, CD166, and CD195 molecules on the neutrophils after surgery in both study groups of patients. The intensity of the observed changes was significantly greater in the group of patients who underwent conventional CPB compared to patients who underwent mini-CPB cardiac surgery.

• MeSH
• CD antigeny analýza   imunologie   MeSH
• fetální proteiny analýza   imunologie   MeSH
• kardiopulmonální bypass škodlivé účinky   přístrojové vybavení   metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové glykoproteiny analýza   imunologie   MeSH
• miniinvazivní chirurgické výkony škodlivé účinky   přístrojové vybavení   metody   MeSH
• molekuly buněčné adheze neuronové analýza   imunologie   MeSH
• neutrofily imunologie   MeSH
• receptory CCR5 analýza   imunologie   MeSH
• senioři MeSH
• zánět etiologie   imunologie   prevence a kontrola   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

OBJECTIVE: Inclusion body myositis (IBM) is characterized by a combination of inflammatory and degenerative changes affecting muscle. While the primary cause of IBM is unknown, genetic factors may influence disease susceptibility. To determine genetic factors contributing to the etiology of IBM, we conducted the largest genetic association study of the disease to date, investigating immune-related genes using the Immunochip. METHODS: A total of 252 Caucasian patients with IBM were recruited from 11 countries through the Myositis Genetics Consortium and compared with 1,008 ethnically matched controls. Classic HLA alleles and amino acids were imputed using SNP2HLA. RESULTS: The HLA region was confirmed as the most strongly associated region in IBM (P = 3.58 × 10(-33) ). HLA imputation identified 3 independent associations (with HLA-DRB1*03:01, DRB1*01:01, and DRB1*13:01), although the strongest association was with amino acid positions 26 and 11 of the HLA-DRB1 molecule. No association with anti-cytosolic 5'-nucleotidase 1A-positive status was found independent of HLA-DRB1*03:01. There was no association of HLA genotypes with age at onset of IBM. Three non-HLA regions reached suggestive significance, including the chromosome 3 p21.31 region, an established risk locus for autoimmune disease, where a frameshift mutation in CCR5 is thought to be the causal variant. CONCLUSION: This is the largest, most comprehensive genetic association study to date in IBM. The data confirm that HLA is the most strongly associated region and identifies novel amino acid associations that may explain the risk in this locus. These amino acid associations differentiate IBM from polymyositis and dermatomyositis and may determine properties of the peptide-binding groove, allowing it to preferentially bind autoantigenic peptides. A novel suggestive association within the chromosome 3 p21.31 region suggests a role for CCR5.

• MeSH
• autoprotilátky imunologie   MeSH
• běloši genetika   MeSH
• genetická predispozice k nemoci MeSH
• genetické asociační studie MeSH
• HLA-DRB1 řetězec genetika   MeSH
• lidé MeSH
• lidské chromozomy, pár 3 genetika   MeSH
• myozitida s inkluzními tělísky genetika   imunologie   MeSH
• receptory CCR5 genetika   MeSH
• věk při počátku nemoci MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH