CXCL12 Chemokine Research Articles

CXCL8 and CXCL11 chemokine secretion in dermal fibroblasts is differentially modulated by vanadium pentoxide.

An increase in skin rashes or atopic dermatitis has been observed in individuals working with vanadium. However, to the best of our knowledge no invivo or invitro studies have evaluated the effect of exposure to vanadium in dermal fibroblasts. Cells viability and proliferation were assessed by WST‑1 assay, cells were treated with increasing concentrations of V2O5 (1, 10 and 100nM). CXCL8 and CXCL11 concentrations were measured in the supernatants using an ELISA assay. V2O5 was not observed as having a significant effect on dermal fibroblast's viability and proliferation. However, it was revealed that V2O5 was able to induce the secretion of CXCL8 and CXCL11 chemokines into dermal fibroblasts. V2O5 synergistically increased the effect of interferon (IFN)γ on CXCL11 secretion. In addition, V2O5 synergistically increased the effect of the tumor necrosis factor α on CXCL8 secretion and abolished the inhibitory effect of IFNγ. V2O5 induction of CXCL8 and CXCL11 chemokines may lead to the appearance and perpetuation of an inflammatory reaction into the dermal tissue. Further studies are required to evaluate dermal integrity and manifestations in subjects occupationally exposed, or living in polluted areas.

Requirement of CXCL11 chemokine production for induction of protection against pulmonary cryptococcosis

Abstract Cryptococcus neoformans is an opportunistic fungal pathogen that causes cryptococcal pneumonia in immunocompromised and immunocompetent individuals. We previously have shown that mice given an experimental pulmonary infection with a genetically modified strain of H99 that produces murine IFN-γ, H99γ, are able to completely resolve the pulmonary infection via the upregulation of the IFN-γ/STAT1 signaling pathway. Activation of STAT1 signaling leads to the production of CXCL9, CXCL10 and CXCL11 chemokines. These chemokines each serve as ligands for the CXCR3 receptor that is present on host immune cells including T cells, NK cells, dendritic cells (DCs) and plasmacytoid DCs and facilitate their chemotaxis to the site of infection. C57BL/6 (B6) mice produce a non-functional CXCL11 protein. Our studies show that B6 mice infected with C. neoformans strain H99γ have a 40% survival, compared to 100% survival in BALB/c mice and an 80% survival of F1 generation mice (first generation of BALB/c × B6) at day 60-post inoculation. After 60 days post-inoculation F1 mice infected with H99γ had a significant decrease in fungal burden compared to B6 mice. We also demonstrated that B6 mice do not produce CXCL11, as expected, at day 7 and 14 post-inoculation whereas F1 mice show a significant increase in the production of CXCL11 compared to B6 mice. Lastly, treatment of B6 mice given a pulmonary C. neoformans infection with recombinant CXCL11 resulted in a significant decrease in pulmonary fungal burden compared to untreated B6 mice infected with C. neoformans. These data suggest that activation of the STAT1-mediated signaling pathway and CXCL11 chemokine production is necessary for optimal protection against pulmonary C. neoformans infection.

Urine levels of CCL2 and CXCL10 chemokines as potential biomarkers of kidney allograft function: a pilot study.

Abstract Introduction Early prognostic markers that identify high-risk patients could lead to increased surveillance, personalized immunosuppression, and improved outcomes. The vigilance and rapid detection of allograft injury is what would result in the identification of patients who will require more intensive monitoring to ensure that their new allografts function well for the longest possible time. The aim of the study was to assess whether urine levels of CCL2 and CXCL10 can be valuable, noninvasive source of information about inflammation, tubular injury, progression of fibrosis and other damage features in the kidney allograft. Materials and methods In this prospective study 40 patients who underwent a protocolar biopsy within 1 year post kidney transplant were included. The levels of chemokines CCL2 and CXCL10 with reference to creatinine in urine were assayed in all patients. Based on biopsy results research group (N=25) with diagnosis of BKV nephropathy, IFTA II–III, peritubular capillaritisis, C4d or rejection was selected. Patients with normal biopsy results were included as control group (N=15). Results Median urinary CCL2-to-creatinine (Cr) ratio [ng/mmol] was significantly elevated in research group compared to control group (21.76 ± 18.57 vs 8.99 ±5.12, p=0.003). There was no statistical difference between groups regarding CXCL10/Cr ratio (7.96 ± 9.08 vs 5.58 ±5.7 p=0.32). Patients with BKV nephropathy (N=3) had much higher level of CCL2 [ng/l] than control group but the difference did not reach statistical significance (429.69 ± 166.49 vs 78.04 ±53.61, p=0.06). Conclusion CCL2/Cr may be used as a non-ivasive tool useful for monitoring graft condition and predicting recipients with high risk of transplant kidney loss.

Topically applied azaphenothiazines inhibit experimental psoriasis in mice

The therapeutic efficacy of topically applied azaphenothiazine derivatives: 9-chloro-6-acetylaminobutylquinobenzo[3,2-b][1,4]thiazine (compound 4) and 6-chloroethylureidoethyldiquino[3,2-b;2′;3′-e][1,4]thiazine (compound 5) in the amelioration of inflammatory symptoms of imiquimod-induced psoriasis in mice was investigated. Clobederm®, containing clobetasol propioniate, served as a reference drug. The application of the compounds led to thinning of the epidermis and reduction of the cell layers. The suppressive actions of the compounds were even stronger with regard to pathological changes of the dermis. The compounds also exerted generalized, anti-inflammatory effects by decreasing the number of circulating leukocytes, lowering subiliac lymph node weight and partially normalizing an altered blood cell composition. The changes in the composition of main cell types in the epidermis and dermis were less affected by the compounds. In addition, both compounds inhibited to a similar degree production of tumor necrosis factor α (TNF α) in human whole blood cell culture. Whereas compound 5 strongly inhibited IL-8 and CXCL10 chemokines in human keratinocytes – KERTr cell line, transfected with poly(I:C), the suppressive action of compound 4 in this model was weak. In addition, compound 5, but not compound 4, exhibited at low doses proapoptotic properties with regard to colonic cell lines. In summary, we demonstrated the therapeutic potential of two selected azaphenotiazines in the amelioration of the skin pathology elicited in a mouse experimental model of psoriasis.

LyeTxI-b, a Synthetic Peptide Derived From Lycosa erythrognatha Spider Venom, Shows Potent Antibiotic Activity in Vitro and in Vivo.

The antimicrobial peptide LyeTxI isolated from the venom of the spider Lycosa erythrognatha is a potential model to develop new antibiotics against bacteria and fungi. In this work, we studied a peptide derived from LyeTxI, named LyeTxI-b, and characterized its structural profile and its in vitro and in vivo antimicrobial activities. Compared to LyeTxI, LyeTxI-b has an acetylated N-terminal and a deletion of a His residue, as structural modifications. The secondary structure of LyeTxI-b is a well-defined helical segment, from the second amino acid to the amidated C-terminal, with no clear partition between hydrophobic and hydrophilic faces. Moreover, LyeTxI-b shows a potent antimicrobial activity against Gram-positive and Gram-negative planktonic bacteria, being 10-fold more active than the native peptide against Escherichia coli. LyeTxI-b was also active in an in vivo model of septic arthritis, reducing the number of bacteria load, the migration of immune cells, the level of IL-1β cytokine and CXCL1 chemokine, as well as preventing cartilage damage. Our results show that LyeTxI-b is a potential therapeutic model for the development of new antibiotics against Gram-positive and Gram-negative bacteria.

Transcriptomics profiling of breast cancer cells, MDA‐MB‐231, exposed to TNF‐α: focus on pro‐survival and pro‐inflammatory regulated pathways.

Triple‐negative breast cancer (TNBC) is characterized by the absence of estrogen, progesterone and HER2 receptors. TNBC is considered to have a poor prognosis due to the limited treatment options with the exclusion of hormone receptor‐based chemotherapies. Aggressive TNBC growth is accompanied by infiltration of tumor‐associated macrophages which can establish a perpetual state of inflammation rendering sustained release of proinflammatory cytokines such as tumor necrosis factor (TNF)‐α, which can lead to immunological evasion. In this work, we evaluated the effects of TNF‐α on the changes to the whole transcriptome: including mRNAs, long intergenic non‐coding RNA transcripts (lincRNA) in MDA‐MB‐231 cells. The data show enormous upregulation (60‐fold change FC, p<.0001) in CXCL8 chemokine (C‐X‐C motif) ligand 8, with a statistically significant elevation of other transcripts including IL‐6 (+10.47), TNFRSF9 (9.58), TNFAIP3 (+7.52) and CCL2 (+3.5). On the other side, there was a reduction in numerous non‐coding microRNA including MIR29A (−8.8), MIRLET7A2 (−8.7), MIR222 (−4.91), MIR100 (−4.91), MIR27B (−4.68) and MIR181B1 (−7.95) in addition to Glutathione S – transferase (GSTT2) (−5.59). Pathway analysis shows that the most relevant effects of TNF‐α are on driving NF‐kB related survival, cytokines, and inflammatory pathway elements and TNF‐α related pathways.Support or Funding InformationSupported by grants from the National Institute on Minority Health and Health Disparities RCMI grant G12MD007582 and COE grant P20 MD006738.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

FRI-510 - Serum CXCL10, CXCL11, CXCL12 and CXCL14 chemokine patterns in patients with acute liver injury

FRI-510 - Serum CXCL10, CXCL11, CXCL12 and CXCL14 chemokine patterns in patients with acute liver injury

Production of Novel Camelid Anti-CXCL10 Specific Polyclonal Antibodies and Evaluation of Their Bioreactivity

CXCL10 chemokine is a member of CXC chemokine family. It is secreted from a variety of cells in response to IFN-γ and stimulates a range of inflammatory responses via binding to its receptor, CXCR3. CXCL10 has a pivotal role in the pathogenesis of various infectious diseases, cancers, and inflammatory and autoimmune diseases. It has been put forward as a potential biomarker and therapeutic target in diagnosis and treatment of these diseases. In the present study, production of camel heavy chain antibodies (HCAbs) specific for the CXCL10 is reported. In this regard, recombinant CXCL10 was used for immunization of camel and subsequently the CXCL10 HCAbs were obtained. Afterwards, three subclasses of IgG were separated using protein A and protein G affinity columns, characterized with SDS-PAGE and confirmed for specific binding to the CXCL10 using ELISA. These IgG subclasses successfully recognized CXCL10 and a strong and specific reactivity towards CXCL10 were observed. Therefore, the selected HCAbs and their corresponding expression library could be used to develop a recombinant variable domain of these HCAbs (nanobody or VHH) as a new possible strategy for treatment of multiple sclerosis and other autoimmune diseases.

Differential modulation by vanadium pentoxide of the secretion of CXCL8 and CXCL11 chemokines in thyroid cells

Recently it has been hypothesized that vanadium serves a carcinogenic role in the thyroid. However, to date, no invivo or invitro studies have evaluated thyroid disruption in humans and/or animals following exposure to vanadium. The present study evaluated the effect of vanadium pentoxide (V2O5) on cell viability and proliferation, and chemokine (C‑X‑C motif) ligand (CXCL)8 and CXCL11 secretion in normal thyrocytes. The results demonstrated that V2O5 had no effect on thyroid follicular cell viability and proliferation. However, V2O5 was able to induce the secretion of CXCL8 and CXCL11 chemokines from thyrocytes. Notably, V2O5 synergistically increased the effect of the interferon (IFN)‑γ on CXCL11 secretion. In addition, V2O5 synergistically increased the effect of tumor necrosis factor‑α on CXCL8 secretion, and abolished the inhibitory effect of IFN‑γ. Overall this induction of CXCL8 and CXCL11 secretion may lead to the induction and perpetuation of an inflammatory reaction in the thyroid. Further studies are now required to evaluate thyroid function and nodule development in subjects who are occupationally exposed, or living in polluted areas.

MiR200-regulated CXCL12\u03b2 promotes fibroblast heterogeneity and immunosuppression in ovarian cancers

High-grade serous ovarian cancers (HGSOC) have been subdivided into molecular subtypes. The mesenchymal HGSOC subgroup, defined by stromal-related gene signatures, is invariably associated with poor patient survival. We demonstrate that stroma exerts a key function in mesenchymal HGSOC. We highlight stromal heterogeneity in HGSOC by identifying four subsets of carcinoma-associated fibroblasts (CAF-S1-4). Mesenchymal HGSOC show high content in CAF-S1 fibroblasts, which exhibit immunosuppressive functions by increasing attraction, survival, and differentiation of CD25+FOXP3+ T lymphocytes. The beta isoform of the CXCL12 chemokine (CXCL12β) specifically accumulates in the immunosuppressive CAF-S1 subset through a miR-141/200a dependent-mechanism. Moreover, CXCL12β expression in CAF-S1 cells plays a crucial role in CAF-S1 immunosuppressive activity and is a reliable prognosis factor in HGSOC, in contrast to CXCL12α. Thus, our data highlight the differential regulation of the CXCL12α and CXCL12β isoforms in HGSOC, and reveal a CXCL12β-associated stromal heterogeneity and immunosuppressive environment in mesenchymal HGSOC.

Vanadium pentoxide induces the secretion of CXCL9 and CXCL10 chemokines in thyroid cells

Vanadium is a grey metal, existing in different states of oxidation, whose most common form in commercial products is vanadium pentoxide(V2O5). All vanadium compounds have been considered toxic. A carcinogenic role of vanadium on the thyroid has recently been proposed. However no invivo or invitro studies have evaluated thyroid disruption in humans and/or animals after exposure to vanadium. In the present study we evaluate the effect of V2O5 on proliferation, and chemokine secretion in normal thyrocytes. Our study demonstrated that V2O5 has no effect on thyroid follicular cell viability or proliferation, but it is able to induce the secretion of T-helper(Th)1 chemokines into the thyroid, synergistically increasing the effect of important Th1 cytokines such as interferon (IFN)γ and tumor necrosis factor (TNF)α. Through this process, V2O5 promotes the induction and perpetuation of an inflammatory reaction in the thyroid. Further studies are necessary to evaluate thyroid function, and nodules, in subjects occupationally exposed, or living in polluted areas.

Neuroinflammation after Traumatic Brain Injury Is Enhanced in Activating Transcription Factor 3 Mutant Mice

Traumatic brain injury (TBI) induces a neuroinflammatory response resulting in astrocyte and microglia activation at the lesion site. This involves upregulation of neuroinflammatory genes, including chemokines and interleukins. However, so far, there is lack of knowledge on transcription factors (TFs) modulating this TBI-associated gene expression response. Herein, we analyzed activating transcription factor 3 (ATF3), a TF encoding a regeneration-associated gene (RAG) predominantly studied in peripheral nervous system (PNS) injury. ATF3 contributes to PNS axon regeneration and was shown before to regulate inflammatory processes in other injury models. In contrast to PNS injury, data on ATF3 in central nervous system (CNS) injury are sparse. We used Atf3 mouse mutants and a closed-head weight-drop-based TBI model in adult mice to target the rostrolateral cortex resulting in moderate injury severity. Post-TBI, ATF3 was upregulated already at early time points (i.e,. 1-4 h) post-injury in the brain. Mortality and weight loss upon TBI were slightly elevated in Atf3 mutants. ATF3 deficiency enhanced TBI-induced paresis and hematoma formation, suggesting that ATF3 limits these injury outcomes in wild-type mice. Next, we analyzed TBI-associated RAG and inflammatory gene expression in the cortical impact area. In contrast to the PNS, only some RAGs (Atf3, Timp1, and Sprr1a) were induced by TBI, and, surprisingly, some RAG encoding neuropeptides were downregulated. Notably, we identified ATF3 as TF-regulating proneuroinflammatory gene expression, including CCL and CXCL chemokines (Ccl2, Ccl3, Ccl4, and Cxcl1) and lipocalin. In Atf3 mutant mice, mRNA abundance was further enhanced upon TBI compared to wild-type mice, suggesting immune gene repression by wild-type ATF3. In accord, more immune cells were present in the lesion area of ATF3-deficient mice. Overall, we identified ATF3 as a new TF-mediating TBI-associated CNS inflammatory responses.

Plasma CXCL1 levels and TRAF3IP2 variants in patients with myocardial infarction.

IL-17A plays an important role in inflammatory responses in myocardial infarction (MI). IL-17A signals through its receptor, for which, Act1 (TRAF3IP2) functions as a key upstream adaptor in the pathway. To compare frequencies of functional polymorphisms of TRAF3IP2 (rs13210247, rs33980500) between patients with MI and healthy controls. The selected SNPs were studied in 201 Iranian MI patients and 201 healthy blood donors from Fars Province by PCR-RFLP in association with clinicopathologic criteria of patients. CXCL1 plasma levels in 126 MI patients and 50 normal subjects were measured by ELISA. A significant increase in the mutant (T) allele of TRAF3IP2 rs33980500 in patients with diastolic dysfunction of the heart (P=.01) was observed. The highest correlation, however, was observed between the TRAF3IP2 rs33980500 TT genotype and T allele with left main coronary artery stenosis (P=.01, P<.001; OR=31.03). T allele of TRAF3IP2 rs33980500 was also associated with female gender, family history of cardiovascular disease, and mechanical complications of heart (P=.04, P=.02, and P=.01, respectively). Moreover, TRAF3IP2 rs13210247 (G) correlated with mechanical complications of the heart (P=.01). A significant increase in the plasma levels of CXCL1 chemokine in patients (P=.0006) associated with TT genotype of TRAF3IP2 (rs33980500) was observed (P=.04). The gene variants of Act1 adaptor are associated with correlates of poor outcome in patients with MI and plasma CXCL1 levels.

CXCR4 receptor blockage reduces the contribution of tumor and stromal cells to the metastatic growth in the liver.

The liver is a common site for the metastatic spread of primary malignancies including colorectal cancer, and liver metastasis is a main cause of death in cancer patients. This is due to the complexity of the interactions taking place in the liver between tumor and stromal cells. In fact, cancer‑associated fibroblasts (CAFs) have been shown to support tumor growth through the CXCL12/CXCR4 axis. However, along with cancer cells, myeloid‑derived suppressor cells (MDSCs), immature dendritic cells with immunosuppressive potential, also express CXCR4. It has recently been demonstrated that reducing CXCL12 availability in the tumor microenvironment decreases liver metastasis. Therefore, blocking CXCL12 chemokine receptor CXCR4 may be a successful approach to diminish the metastatic spread of colorectal cancer to the liver. However, the subjacent mechanisms by which this chemokine influences the tumor are not fully understood. Thus, in order to uncover the role of CXCR4 during tumor cell/liver fibroblast crosstalk driving liver metastasis, the CXCR4 antagonist AMD3100 was used for invitro studies and in an invivo approach using an orthotopic model of liver metastasis in immune competent mice through intrasplenic injection of grafted C26 cells. Invitro blockage of CXCR4 led to an impaired migratory potential of tumor and hepatic stellate cells (HSCs) and a reduced tumor response to CXCL12. Invivo administration of AMD3100 to tumor‑bearing mice resulted in attenuated metastatic development in the liver, which was accompanied by an impaired infiltration of αSMA‑expressing cells within the tumors. In addition, a reduced CD11+Ly6G+ cell count in the liver was directly correlated with a reduction in MDSC numbers in the blood of AMD3100‑treated mice compared to the vehicle‑treated mice. Therefore, disruption of the CXCR4/CXCL12 axis by CXCR4 antagonist AMD3100 blocked the contribution of both cancer and stromal cells to the metastatic cascade in the liver.

Molecular characterization and functional activity of CXCL8_L3 in large yellow croaker Larimichthys crocea

CXCL8, also called interleukin-8, is a typical CXC chemokine that plays a key role in promoting inflammation. Phylogenetically, fish CXCL8 chemokines can be divided into three subgroups, CXCL8_L1, CXCL8_L2, and CXCL8_L3, of which CXCL8_L3 is a new subgroup. The CXCL8_L3 gene sequences have been reported in many fish species, but their function remains unknown. Here, a CXCL8_L3 (LycCXCL8_L3) gene was cloned from large yellow croaker Larimichthys crocea. Its open reading frame (ORF) was 309 nucleotides long, encoding a peptide of 102 amino acids. The deduced LycCXCL8_L3 protein contains an 18-aa signal peptide and an 84-aa mature polypeptide, which has four conserved cysteine residues (C30, C32, C57, and C73) as found in other known CXCL8 chemokines. Phylogenetic analysis showed LycCXCL8_L3 formed a major clade with CXCL8_L3 sequences from other fish species. The LycCXCL8_L3 transcript was constitutively expressed in all examined tissues and significantly up-regulated in the spleen and head kidney tissues by inactivated trivalent bacterial vaccine. The LycCXCL8_L3 transcript was also detected in peripheral blood leukocytes (PBLs), primary head kidney macrophages (PKM), and large yellow croaker head kidney cell line (LYCK), with the highest levels in PKM. Recombinant LycCXCL8_L3 (rLycCXCL8_L3) protein could not only chemotactically attract lymphocytes and eosinophils in PBLs, but also enhance the respiratory burst activity of PKM. These results indicate that LycCXCL8_L3 may play an important role in the inflammatory response of large yellow croaker. To our knowledge, this is the first report on functional study of the CXCL8_L3 in fish.

Contribution of Regulatory T Cells in Nucleotide-Binding Oligomerization Domain 2 Response to Influenza Virus Infection.

Influenza A virus (IAV) is recognized to cause severe pulmonary illnesses in humans, particularly in elderly and children. One of the features associated with IAV infection is an excessive lung inflammation due to an uncontrolled immune response. The nucleotide-binding oligomerization domain 2 (NOD2) receptor is known to recognize ssRNA viruses such as IAV, but its role in the inflammatory process during viral infections remains to be clarified. In a previous report, we have shown that activation of NOD2 with muramyl dipeptide (MDP) significantly reduces both viral loads and lung inflammation and also improves pulmonary function during IAV infection. These findings prompted us to further investigate whether NOD2 receptor may contribute to regulate inflammation during viral infection. In the present study, we show that administration of MDP to mice infected with IAV stimulates the migration of regulatory T (Treg) cells to the lungs. Such a presence of Treg cells was also accompanied with a reduction of neutrophils in the lungs during IAV infection, which correlated, with a significant decrease of Th17 cells. In our model, Treg cell recruitment is dependent of CXCL12 and CCL5 chemokines. Moreover, we show that the presence of Ly6Clow patrolling monocytes is required for Treg cells mobilization to the lung of mice treated with MDP. In fact, following monocyte depletion by administration of clodronate liposome, mobilization of Treg cells to the lungs of treated mice was found to occur when circulating Ly6Clow monocytes begin to reemerge. In addition, we also detected an increased production of TGF-β, a cytokine contributing to Treg activity when blood Ly6Clow monocytes are restored. Together, our results demonstrate that MDP treatment can promote an anti-inflammatory environment through the mobilization of Treg cells to the lung, a mechanism that requires the presence of Ly6Clow monocytes during IAV infection. Overall, our results suggest that activation of NOD2 receptor could be an appealing approach to control pulmonary inflammation in patients infected with IAV.

Modulation of HIV replication in monocyte derived macrophages (MDM) by steroid hormones.

Significant sex specific differences in the progression of HIV/AIDS have been reported. Several studies have implicated steroid hormones in regulating host factor expression and modulating HIV transmission and replication. However, the exact mechanism exerted by steroid hormones estrogen and progesterone in the regulation of HIV-1 replication is still unclear. Results from the current study indicated a dose dependent down regulation of HIV-1 replication in monocyte derived macrophages pre-treated with high concentrations of estrogen or progesterone. To elucidate the molecular mechanisms associated with the down regulation of HIV-1 replication by estrogen and progesterone we used PCR arrays to analyze the expression profile of host genes involved in antiviral responses. Several chemokines, cytokines, transcription factors, interferon stimulated genes and genes involved in type-1 interferon signaling were down regulated in cells infected with HIV-1 pre-treated with high concentrations of estrogen or progesterone compared to untreated HIV-1 infected cells or HIV-1 infected cells treated with low concentrations of estrogen or progesterone. The down regulation of CXCL9, CXCL10 and CXCL11 chemokines and IL-1β, IL-6 cytokines in response to high concentrations of estrogen and progesterone pre-treatment in HIV-1 infected cells was confirmed at the protein level by quantitating chemokine and cytokine concentrations in the culture supernatant. These results demonstrate that a potent anti-inflammatory response is mediated by pre-treatment with high concentrations of estrogen and progesterone. Thus, our study suggests a strong correlation between the down-modulation of anti-viral and pro-inflammatory responses mediated by estrogen and progesterone pre-treatment and the down regulation of HIV-1 replication. These findings may be relevant to clinical observations of sex specific differences in patient populations and point to the need for further investigation.

339: Maternal and fetal umbilical cord levels of chemokines in normal and PreE pregnancies - A prospective trial

Preeclampsia (PreE) is the de novo development of hypertension and proteinuria after 20 weeks of gestation affecting 3-8% of pregnancies, and has a link to alterations of feto-placental stress that pass to the offspring causing detrimental effects. We aimed at comparing the clinical outcomes and chemokines from the maternal and cord blood between patients with and without PreE. We recruited 20 normal pregnant (NP) and PreE patients in an IRB approved prospective cohort at our institution. We evaluated maternal age, body mass index (BMI), blood pressure (BP), and gestational age (GA) at delivery along with measured serum levels of six chemokines in maternal-fetal paired samples using ELISA test. The markers are; Human CCL4/MIP-1beta, Human CXCL10/IP-10, Eotaxin/CCL11, RANTES (CCL5), Human MCP1, Human M-CSF. Maternal: The systolic blood pressure (SBP) and diastolic blood pressure (DBP) were higher in PreE (SBP 162 ± 17; DBP 97 ± 12) compared to NP (SBP 121 ± 3; DBP 75 ± 9; p <0.05). GA at delivery in weeks was lower in PreE (35.1 ± 3.8) compared to NP (39.2 ± 0.3; p <0.05), as well as birth weight in PrE (2556 ± 977) compared to NP (3385 ± 328; p <0.011) indicating a link to intrauterine growth restriction (IUGR). We did not find a difference in maternal BMI between the two groups. Biomarkers: For all six chemokines CCL4, CXCL10, CCL11, CCL5, MCP1, and M-CSF, maternal and fetal cord serum chemokines levels differ between subjects with and without PreE (p < 0.001). There is also a difference between the maternal and fetal levels for the six chemokines (<0.01). The fetal levels were compared to associated maternal levels for each of the six chemokines using t test and Mann-Whitney U test. Women with PreE had higher BP, preterm deliveries, and fetal IUGR. However, in this cohort the women were of similar age, size, and parity prior to the onset of pregnancy. The six chemokines in the samples from maternal and fetal circulation are greater in the PreE group, and each of the six chemokines samples from the fetuses had values greater than maternal samples which could implicate placental cells as the source. PreE alters the intrauterine environment and activates the detrimental signaling that is transferred to the fetus resulting in the aforementioned complications.

Vascular smooth muscle cell response to cyclic mechanical stretch and aortic dissection

Acute aortic dissection is the most common life-threatening vascular disease, with sudden onset of severe pain and a high fatality rate. The pulsatile nature of blood flow exposes vascular smooth muscle cells (VSMCs) in the vessel wall to cyclic mechanical stretch (CMS), which evokes VSMC death, phenotypic switching, and migration, leading to aortic dissection. We have revealed that CMS of rat aortic smooth muscle cells (RASMCs) caused JNK- and p38-dependent cell death and that a calcium channel blocker, azelnidipine and an angiotensin II receptor antagonist, olmesartan decreased the phosphorylation of JNK and p38 and, subsequently, decreased cell death by CMS. JNK and p38 inhibitors also inhibited CMS-induced cell death. In addition, we showed that the expression of Cxcl1 and Cx3cl1 chemokines was induced by CMS in a JNK-dependent manner. Expression of Cxcl1 was also induced in VSMCs by hypertension produced by abdominal aortic constriction in mouse. In addition, antagonists against the receptors for CXCL1 and CX3CL1 increased cell death, indicating that CXCL1 and CX3CL1 protect RASMCs from CMS-induced cell death. We also revealed that STAT1 is activated in RASMCs subjected to CMS. Taken together, these results indicate that CMS of VSMCs induces inflammation-related gene expression, including that of CXCL1 and CX3CL1, and activates JNK and p38 MAP kinases, which may play important roles in the stress response against CMS caused by acute rise in blood pressure.

NLRP3 inflammasome driven liver injury and fibrosis: Roles of IL-17 and TNF in mice.

The NLRP3 inflammasome, a caspase-1 activation platform, plays a key role in the modulation of liver inflammation and fibrosis. Here, we tested the hypothesis that interleukin 17 (IL-17) and tumor necrosis factor (TNF) are key cytokines involved in amplifying and perpetuating the liver damage and fibrosis resulting from NLRP3 activation. To address this hypothesis, gain-of-function Nlrp3A350V knock-in mice were bred onto il17a and Tnf knockout backgrounds allowing for constitutive Nlrp3 activation in myeloid derived cells in mice deficient in IL-17 or TNF. Livers of Nlrp3A350V knock-in mice exhibited severe liver inflammatory changes characterized by infiltration with neutrophils, increased expression of chemokine (C-X-C motif) ligand (CXCL) 1 and CXCL2 chemokines, activated inflammatory macrophages, and elevated levels of IL-17 and TNF. Mutants with ablation of il17a signal showed fewer neutrophils when compared to intact Nlrp3A350V mutants, but still significant inflammatory changes when compared to the nonmutant il17a knockout littermates. The severe inflammatory changes associated with mutant Nlrp3 were almost completely rescued by Tnf knockout in association with a marked decrease in circulating IL-1β levels. Intact Nlrp3A350V mutants showed changes in liver fibrosis, as evidenced by morphometric quantitation of Sirius Red staining and increased mRNA levels of profibrotic genes, including connective tissue growth factor and tissue inhibitor of matrix metalloproteinase 1. Il17a lacking mutants exhibited amelioration of the aforementioned fibrosis, whereas Tnf-deficient mutants showed no signs of fibrosis when compared to littermate controls. Conclusion: Our study uncovers key roles for TNF and, to a lesser extent, IL-17 as mediators of liver inflammation and fibrosis induced by constitutive NLRP3 inflammasome activation in myeloid-derived cells. These findings may lead to therapeutic strategies aimed at halting the progression of liver injury and fibrogenesis in various liver pathogeneses driven by NLRP3 activation. (Hepatology 2018;67:736-749).