interleukin-8 and Lyme-Disease

We estimated serum concentrations of cytokines IL-1, IL-6, IL-8, TNF-alfa and IL-6R of patients with diagnosed Lyme disease treated with beta-lactam antibiotics. Detection of proinflammatory cytokines was performed in ELISA tests. The examination was performed before and after treatment. Comparison with control group stated statistically significant higher concentration of IL-1 and IL-6 before and after treatment. There were no differences in concentration of TNF-alfa, IL-8 and IL-6R. Comparing concentrations of cytokines before and after treatment there was no differences either. Lack of changes in concentration of proinflammatory cytokines during beta-lactam therapy could be explained by too short period of therapy or immunologic background of inflammatory process in Lyme disease which was only initiated by spirochete Borrelia burgdorferi.

Topics: Adult; Cytokines; Female; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Lyme Disease; Male; Middle Aged; Receptors, Interleukin-6; Tumor Necrosis Factor-alpha

In Lyme borreliosis, the skin constitutes a major interface for the host, the bacteria and the tick. Skin immunity is provided by specialized immune cells but also by the resident cells: the keratinocytes and the fibroblasts. Discoveries on the role of the microbiome in the modulation of skin inflammation and immunity have reinforced the potential importance of the skin in vector-borne diseases. In this study, we analyzed in vitro the interaction of human primary keratinocytes and fibroblasts with Borrelia burgdorferi sensu stricto N40 in presence or absence of bacterial commensal supernatants. We aimed to highlight the role of resident skin cells and skin microbiome on the inflammation induced by B. burgdorferi s.s.. The secretomes of Staphylococcus epidermidis, Corynebacterium striatum and Cutibacterium acnes showed an overall increase in the expression of IL-8, CXCL1, MCP-1 and SOD-2 by fibroblasts, and of IL-8, CXCL1, MCP-1 and hBD-2 in the undifferentiated keratinocytes. Commensal bacteria showed a repressive effect on the expression of IL-8, CXCL1 and MCP-1 by differentiated keratinocytes. Besides the inflammatory effect observed in the presence of Borrelia on all cell types, the cutaneous microbiome appears to promote a rapid innate response of resident skin cells during the onset of Borrelia infection.

Topics: Animals; Borrelia burgdorferi; Humans; Immunity, Innate; Inflammation; Interleukin-8; Ixodes; Lyme Disease; Secretome

Even after appropriate treatment, a proportion of Lyme disease patients suffer from a constellation of symptoms, collectively called Post-Treatment Lyme Disease Syndrome (PTLDS). Brain PET scan of patients with PTLDS have demonstrated likely glial activation indicating persistent neuroinflammatory processes. It is possible that unresolved bacterial remnants can continue to cause neuroinflammation. In previous studies, we have shown that non-viable Borrelia burgdorferi can induce neuroinflammation and apoptosis in an oligodendrocyte cell line. In this follow-up study, we analyze the effect of sonicated remnants of B. burgdorferi on primary rhesus frontal cortex (FC) and dorsal root ganglion (DRG) explants. Five FC and three DRG tissue fragments from rhesus macaques were exposed to sonicated B. burgdorferi and analyzed for 26 inflammatory mediators. Live bacteria and medium alone served as positive and negative control, respectively. Tissues were also analyzed for cell types mediating inflammation and overall apoptotic changes. Non-viable B. burgdorferi induced significant levels of several inflammatory mediators in both FC and DRG, similar to live bacteria. However, the levels induced by non-viable B. burgdorferi was often (several fold) higher than those induced by live ones, especially for IL-6, CXCL8 and CCL2. This effect was also more profound in the FC than in the DRG. Although the levels often differed, both live and dead fragments induced the same mediators, with significant overlap between FC and DRG. In the FC, immunohistochemical staining for several inflammatory mediators showed the presence of multiple mediators in astrocytes, followed by microglia and oligodendrocytes, in response to bacterial remnants. Staining was also seen in endothelial cells. In the DRG, chemokine/cytokine staining was predominantly seen in S100 positive (glial) cells. B. burgdorferi remnants also induced significant levels of apoptosis in both the FC and DRG. Apoptosis was confined to S100 + cells in the DRG while distinct neuronal apoptosis was also detected in most FC tissues in response to sonicated bacteria. Non-viable B. burgdorferi can continue to be neuropathogenic to both CNS and PNS tissues with effects likely more profound in the former. Persistence of remnant-induced neuroinflammatory processes can lead to long term health consequences.

Topics: Animals; Apoptosis; Borrelia burgdorferi; Chemokine CCL2; Female; Frontal Lobe; Ganglia, Spinal; In Vitro Techniques; Inflammation Mediators; Interleukin-6; Interleukin-8; Lyme Disease; Macaca mulatta; Male; Neuroinflammatory Diseases

Our aim was to identify the mechanisms of immune and cytokine regulation in different clinical forms of Ixodes tick-borne borreliosis.. The clinical observations performed on 581 patients with erythemic (113 patients), non-eritemic (242 patients) forms of Ixodes tick- borne borreliosis and Borrelia co-infection tick-borne encephalitis (226 patients) in the manifestation of the disease. The examination included the determination of the levels of cytokines (interleukin 1β, 4, 8, and tumor necrosis factor α), indicators of cell immunity (CD3+, CD4+, CD8+) and phagocytosis (phagocytic index, the number of phagocytic neutrophils) during the height of the disease and during convalescence.. We established the correlations of cytokine production, lymphocyte subpopulations and phagocytosis. Revealed divergent immune-mediated mechanisms of pathogenesis, depending on the clinical forms of Ixodes tick-borne borreliosis and period of the disease, indicating the complexity and diversity of the immune system rearrangements. Cytokine regulation of immune response, mainly consists in the synthesis of the chemoattractant interleukin 8 production which is inversely correlated with the number of phagocytic neutrophils (r = -0.440, p < 0.001) with erythemic form of the disease, is directly correlated with the level of interleukin 4 (r = 0.313, p < 0.001) at non-eritemic form borreliosis, with the production of interleukin 1β (r = 0.367, p < 0.001) and interleukin 4 (r = 0.348, p < 0.001) in Borrelia co-infection tick-borne encephalitis.. In the manifestation of acute Borrelia infection, regardless of the clinicalform was typical Th1/Th2 type of immune response and then switches to the period of convalescence for Th1 (cellular) immune response in the form of erythemic Ixodes tick-borne borreliosis and Borrelia co-encephalitic tick-borne infection, and Th2 (humoral) immune response when non-eritemic form of the disease.

Topics: Adult; Animals; Case-Control Studies; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cytokines; Encephalitis, Tick-Borne; Female; Humans; Interleukin-8; Ixodes; Lyme Disease; Middle Aged; Neutrophils; Phagocytosis; Th1 Cells; Tumor Necrosis Factor-alpha

Lyme neuroborreliosis (LNB), caused by the spirochete Borrelia burgdorferi, affects both the peripheral and the central nervous systems. Radiculitis or nerve root inflammation, which can cause pain, sensory loss, and weakness, is the most common manifestation of peripheral LNB in humans. We previously reported that rhesus monkeys infected with B. burgdorferi develop radiculitis as well as inflammation in the dorsal root ganglia (DRG), with elevated levels of neuronal and satellite glial cell apoptosis in the DRG. We hypothesized that B. burgdorferi induces inflammatory mediators in glial and neuronal cells and that this inflammatory milieu precipitates glial and neuronal apoptosis.. To model peripheral neuropathy in LNB we incubated normal rhesus DRG tissue explants with live B. burgdorferi ex vivo and identified immune mediators, producer cells, and verified the presence of B. burgdorferi in tissue sections by immunofluorescence staining and confocal microscopy. We also set up primary cultures of DRG cells from normal adult rhesus macaques and incubated the cultures with live B. burgdorferi. Culture supernatants were subjected to multiplex ELISA to detect immune mediators, while the cells were evaluated for apoptosis by the in situ TUNEL assay. A role for inflammation in mediating apoptosis was assessed by evaluating the above phenomena in the presence and absence of various concentrations of the anti-inflammatory drug dexamethasone. As Schwann cells ensheath the dorsal roots of the DRG, we evaluated the potential of live B. burgdorferi to induce inflammatory mediators in human Schwann cell (HSC) cultures.. Rhesus DRG tissue explants exposed to live B. burgdorferi showed localization of CCL2 and IL-6 in sensory neurons, satellite glial cells and Schwann cells while IL-8 was seen in satellite glial cells and Schwann cells. Live B. burgdorferi induced elevated levels of IL-6, IL-8 and CCL2 in HSC and DRG cultures and apoptosis of sensory neurons. Dexamethasone reduced the levels of immune mediators and neuronal apoptosis in a dose dependent manner.. In this model, B. burgdorferi induced an inflammatory response and neuronal apoptosis of DRG. These pathophysiological processes could contribute to peripheral neuropathy in LNB.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Borrelia burgdorferi; Chemokine CCL2; Culture Media; Cytoplasm; Dexamethasone; Fluorescent Antibody Technique; Ganglia, Spinal; Humans; In Situ Nick-End Labeling; Inflammation; Inflammation Mediators; Interleukin-6; Interleukin-8; Lyme Disease; Macaca mulatta; Microscopy, Confocal; Neurons; Satellite Cells, Perineuronal; Schwann Cells

In Lyme borreliosis, the skin is the key site of bacterial inoculation by the infected tick, and of cutaneous manifestations, erythema migrans and acrodermatitis chronica atrophicans. We explored the role of fibroblasts, the resident cells of the dermis, in the development of the disease. Using microarray experiments, we compared the inflammation of fibroblasts induced by three strains of Borrelia burgdorferi sensu stricto isolated from different environments and stages of Lyme disease: N40 (tick), Pbre (erythema migrans) and 1408 (acrodermatitis chronica atrophicans). The three strains exhibited a similar profile of inflammation with strong induction of chemokines (CXCL1 and IL-8) and IL-6 cytokine mainly involved in the chemoattraction of immune cells. Molecules such as TNF-alpha and NF-κB factors, metalloproteinases (MMP-1, -3 and -12) and superoxide dismutase (SOD2), also described in inflammatory and cellular events, were up-regulated. In addition, we showed that tick salivary gland extracts induce a cytotoxic effect on fibroblasts and that OspC, essential in the transmission of Borrelia to the vertebrate host, was not responsible for the secretion of inflammatory molecules by fibroblasts. Tick saliva components could facilitate the early transmission of the disease to the site of injury creating a feeding pit. Later in the development of the disease, Borrelia would intensively multiply in the skin and further disseminate to distant organs.

Topics: Animals; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Borrelia burgdorferi; Dermis; Extracellular Matrix; Fibroblasts; Gene Expression Profiling; Humans; Inflammation; Interleukin-8; Lyme Disease; Oligonucleotide Array Sequence Analysis; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Ticks; Transcription, Genetic; Up-Regulation

Topics: Animals; Antibodies, Bacterial; Antimicrobial Cationic Peptides; beta-Defensins; Borrelia burgdorferi; Cathelicidins; Cells, Cultured; Defensins; Humans; Immunity, Innate; Interleukin-8; Keratinocytes; Lyme Disease; Salivary Glands; Skin; Tissue Extracts

Lyme borreliosis is a spirochetal infection caused by the Borrelia burgdorferi sensu lato complex that can proceed towards an inflammatory joint manifestation known as Lyme arthritis. Production of chemokines orchestrating neutrophil infiltration is supposed to be key to early arthritic pathogenesis. Using PMA-differentiated macrophage-like THP-1 (mTHP-1) cells we identified by antibody array methodology or mRNA analysis IL-8, GRO-alpha, NAP-2, and SDF-1alpha as being among those chemokines that are upregulated by bacterial lysates obtained from B. burgdorferi. Based on these observations, we set out to characterize in detail mechanisms mediating IL-8 release in this cellular model. TLR2 blocking antibodies, analysis of p65 translocation, and electromobility-shift analysis revealed activation of the TLR2/NF-kappaB axis by B. burgdorferi. The functional importance of this pathway was substantiated by suppression of IL-8 after inhibition of IkappaB kinase. Notably, MAP kinases, specifically the MEK1/2-ERK1/2 pathway, were essential for IL-8 secretion. Those data were confirmed by using freshly isolated adherent peripheral blood mononuclear cells. On the contrary, B. burgdorferi-induced IL-8 in mTHP-1 was unlikely related to flagellin, alpha3beta1-integrin signaling, lipopolysaccharide, bacterial DNA, NOD1/NOD2 agonists, or to intermediate production of IL-1beta and TNF-alpha. Induction of IL-8 by B. burgdorferi was not due to amplification of constitutive AP-1 DNA-binding activity detectable in mTHP-1 cells. Data presented herein validate that TLR2, particularly on mTHP-1 cells, holds a central position in mediating IL-8 secretion associated with extracellular B. burgdorferi and beyond that suggest inhibition of IkappaB kinase and MEK1/2 kinases as promising pharmacological strategies aiming at IL-8 in early Lyme arthritis.

Topics: Animals; Borrelia burgdorferi; Cell Line; Chemokines; Enzyme Inhibitors; Humans; Interleukin-8; Lyme Disease; MAP Kinase Kinase 1; MAP Kinase Kinase 2; Microarray Analysis; Mitogen-Activated Protein Kinases; Monocytes; NF-kappa B; Signal Transduction; Tissue Extracts; Toll-Like Receptor 2; Tumor Necrosis Factor-alpha

Components of the spirochete Borrelia burgdorferi sensu lato ( B. burgdorferi s.l.) do not have chemotactic activity. However, B. burgdorferi s.l. causes a chemotactic response, probably by stimulating synthesis of cytokines of the chemokine family by host cells. Our aim was to confirm that the synthesis of chemokines is increased in Lyme borreliosis and that they may account for leukocyte migration, thus being involved in inflammatory response.. We measured concentrations of chemokines: interleukin 8 (Il-8) and macrophage inflammatory protein 1alpha and 1beta (MIP-1alpha, -1beta) in serum of 20 patients with erythema migrans (early localized infection, group I), of 19 patients with Lyme arthritis (chronic infection, group II), and in serum and cerebrospinal fluid (CSF) of 20 patients with neuroborreliosis (early disseminated infection, group III), before and after 2 weeks of antibiotic therapy (examinations 1 and 2), as well as in the sera of 12 healthy blood donors and CSF of ten patients in whom Lyme borreliosis and meningitis were excluded (control group). Interleukin 1beta (Il-1beta) level in serum and CSF and pleocytosis of CSF were assessed simultaneously.. The mean concentrations of all studied chemokines in serum were significantly elevated in all study groups in examination 1 and decreased in examination 2. The concentration of Il-8 in serum was higher in group I and the concentration of MIP-1alpha in group III was higher in comparison with group II. Serum concentrations of all chemokines in group I and III correlated with the concentration of Il-1beta, while in group II this correlation appeared only for Il-8 in examination 2. Concentrations of all chemokines in CSF were significantly increased, but as for MIP-1alpha and 1beta they remained lower than in serum. The concentration of Il-8 in CSF was variable and reached values several fold higher than in the serum in some patients. There was no correlation between chemokine concentrations and CSF pleocytosis.. The synthesis of chemokines (Il-8, MIP-1alpha and 1beta) is increased in Lyme borreliosis and, at least in the early stages of the disease, is related to the synthesis of Il-1beta. Chemokine concentrations depend on the clinical form of Lyme borreliosis, with a tendency for higher values in early infection (erythema migrans and neuroborreliosis). Of the chemokines studied, Il-8 created a chemotactic gradient towards the inflammation site, and thus might be responsible for leukocyte migration.

Topics: Adult; Aged; Blood Donors; Case-Control Studies; Cell Movement; Chemokine CCL3; Chemokine CCL4; Chemotaxis; Disease Progression; Female; Humans; Infant, Newborn; Inflammation; Interleukin-8; Leukocytes; Lyme Disease; Macrophage Inflammatory Proteins; Male; Middle Aged

Reverse transcription-polymerase chain reaction (RT-PCR) with interleukin-1alpha (IL-1alpha)-specific primers using total RNA from lipopolysaccharide (LPS)-stimulated lung macrophages resulted in the amplification of two distinct cDNA fragments. Cloning and sequencing of the canine and feline fragments revealed that, except for the absence of a specific 174 nucleotide sequence, the short and the long transcripts were identical. The in-frame 174 nucleotide deletion corresponds to exon 5 of the human and murine IL-1alpha gene, which encodes the cleavage site for calpain, a protein necessary for the processing of the IL-1alpha precursor into mature IL-1alpha. The two transcripts were found in the dog, cat and pig; analysis by RT-PCR, Southern and Northern blot hybridization showed no expression of the shorter IL-1alpha mRNA in equine or bovine macrophages. Expression of the two canine IL-1alpha transcripts was also detected in synovial membranes and was coordinately up-regulated in response to Borrelia burgdorferi infection under both in-vitro and in-vivo conditions.

Topics: Alternative Splicing; Animals; Base Sequence; Blotting, Northern; Blotting, Southern; Cats; Cattle; Dogs; Horses; Interleukin-1; Interleukin-8; Lyme Disease; Macrophages; Molecular Sequence Data; Reverse Transcriptase Polymerase Chain Reaction; Sequence Homology, Nucleic Acid; Swine; Synovial Membrane; Up-Regulation

Canine synovial membrane explants were exposed to high- or low-passage Borrelia burgdorferi for 3, 6, 12, and 24 h. Spirochetes received no treatment, were UV light irradiated for 16 h, or were sonicated prior to addition to synovial explant cultures. In explant tissues, mRNA levels for the proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha), interleukin-1alpha (IL-1alpha), IL-1beta, and IL-8 were surveyed semiquantitatively by reverse transcription-PCR. Culture supernatants were examined for numbers of total and motile (i.e., viable) spirochetes, TNF-like and IL-1-like activities, polymorphonuclear neutrophil (PMN) chemotaxis-inducing activities, and IL-8. During exposure to synovial explant tissues, the total number of spirochetes in the supernatants decreased gradually by approximately 30%, and the viability also declined. mRNAs for TNF-alpha, IL-1alpha, IL-1beta, and IL-8 were up-regulated in synovial explant tissues within 3 h after infection with untreated or UV light-irradiated B. burgdorferi, and mRNA levels corresponded to the results obtained with bioassays. During 24 h of coincubation, cultures challenged with untreated or UV light-irradiated spirochetes produced similar levels of TNF-like and IL-1-like activities. In contrast, explant tissues exposed to untreated B. burgdorferi generated significantly higher levels of chemotactic factors after 24 h of incubation than did explant tissues exposed to UV light-treated spirochetes. In identical samples, a specific signal for IL-8 was identified by Western blot analysis. High- and low-passage borreliae did not differ in their abilities to induce proinflammatory cytokines. No difference in cytokine induction between untreated and sonicated high-passage spirochetes was observed, suggesting that fractions of the organism can trigger the production and release of inflammatory mediators. The titration of spirochetes revealed a dose-independent cytokine response, where 10(3) to 10(7) B. burgdorferi organisms induced similar TNF-like activities but only 10(7) spirochetes induced measurable IL-1-like activities. The release of chemotactic factors was dose dependent and was initiated when tissues were infected with at least 10(5) organisms. We conclude that intact B. burgdorferi or fractions of the bacterium can induce the local up-regulation of TNF-alpha, IL-1alpha, and IL-1beta in the synovium but that the interaction of viable spirochetes with synovial cells leads to the release of IL-8, whi

Topics: Animals; Antibodies, Blocking; Borrelia burgdorferi Group; Chemotactic Factors; Chemotaxis, Leukocyte; Colony Count, Microbial; Culture Media, Conditioned; Cytotoxicity Tests, Immunologic; Dogs; In Vitro Techniques; Interleukin-1; Interleukin-8; Lyme Disease; Neutrophils; Polymerase Chain Reaction; Recombinant Proteins; RNA, Messenger; Sonication; Synovial Membrane; Tumor Necrosis Factor-alpha; Ultraviolet Rays; Up-Regulation

Previous studies have shown that Borrelia burgdorferi, the spirochetal agent of Lyme disease, promotes inflammation by stimulating endothelial cells to upregulate adhesion molecules for leukocytes and to produce a soluble agent that is chemotactic for neutrophils. We determined that interleukin-8 (IL-8) was the chemotactic agent for neutrophils present in conditioned media from cultured human umbilical vein endothelial cells stimulated with B. burgdorferi. As few as one spirochete per endothelial cell stimulated production of IL-8 within 8 h of coincubation. When 10 spirochetes per endothelial cell were added, IL-8 was detected after 4 h of coculture. Production of IL-8 continued in a linear fashion for at least 24 h. Neutralizing antibodies against IL-8 reduced migration of neutrophils across spirochete-stimulated endothelial monolayers by 93%. In contrast, pretreatment of neutrophils with antagonists of platelet-activating factor did not inhibit migration. Increases in production of IL-8 and expression of the adhesion molecule E-selectin by endothelial cells in response to B. burgdorferi were not inhibited by IL-1 receptor antagonist or a neutralizing monoclonal antibody directed against tumor necrosis factor alpha, used either alone or in combination. These results suggest that activation of endothelium by B. burgdorferi is not mediated through the autocrine action of secreted IL-1 or tumor necrosis factor alpha. Rather, it appears that B. burgdorferi must stimulate endothelium either by a direct signaling mechanism or by induction of a novel host-derived proinflammatory cytokine.

Topics: Borrelia burgdorferi Group; Cell Movement; Cells, Cultured; Endothelium, Vascular; Humans; Interleukin-1; Interleukin-8; Lyme Disease; Neutrophils; Tumor Necrosis Factor-alpha

Twenty 6-week-old specific-pathogen-free beagles were infected with Borrelia burgdorferi by tick challenge, and five uninfected dogs served as controls. During the study, all dogs were monitored for infection, clinical signs, and antibody response against B. burgdorferi. During episodes of lameness or postmortem, synovial fluids from each dog were examined for volume, cell number, polymorphonuclear leukocyte (PMN) content, cell viability, and chemotactic activity. Twenty-five tissues collected postmortem from each dog were tested for interleukin-8 (IL-8) mRNA, tumor necrosis factor alpha (TNF-alpha) mRNA, presence of live spirochetes, and histopathological changes. Thirteen infected dogs (group A), which seroconverted rapidly (maximum titers within 50 to 90 days), developed acute and severe mono- or oligoarthritis almost exclusively in the limb closest to the tick bite (median incubation period, 66 days). Synovial fluids of the arthritic joints collected during episodes of lameness had significantly elevated volume, cell count, PMN proportion, cell viability, and chemotactic activity for PMNs. The remaining joints of the same animals contained synovial fluids with elevated chemotactic activity and cell viability. Twelve dogs tested positive for IL-8 mRNA in multiple tissues (synovia, pericardium, and peritoneum), and 10 dogs expressed TNF-alpha mRNA, but only in the tributary lymph nodes of the inflamed joints. Histological examinations revealed severe poly- or oligoarthritis and moderate to severe cortical hyperplasia in draining lymph nodes of the inflamed joints in all 13 dogs. Seven infected dogs with mild or no clinical signs (group B) seroconverted slowly (peak titers after 90 days), and only some joint fluids showed chemotactic activity, which on average was lower than that in inflamed and noninflamed joints from dogs in group A. Four dogs expressed IL-8 mRNA (in the synovia and pericardium), and three dogs had TNF-alpha mRNA in tributary lymph nodes. Histologically, nonsuppurative arthritis was found in multiple joints, and mild to moderate cortical hyperplasia was found in draining lymph nodes. Five uninfected dogs without lameness (group C) had normal synovial fluids and tissues. In all infected dogs, live spirochetes were demonstrated more frequently in tissues of the somatic quadrant closest to the tick bite than in tissues further from the site of infection, suggesting that dissemination of B. burgdorferi occurs more by migration than by bloo

Topics: Animals; Borrelia burgdorferi Group; Dogs; Interleukin-8; Joints; Lyme Disease; Synovial Membrane; Up-Regulation