scholarly journals Examination of Early Interactions betweenHaemophilus ducreyi and Host Cells by Using Cocultured HaCaT Keratinocytes and Foreskin Fibroblasts

1999 ◽  
Vol 67 (10) ◽  
pp. 5352-5360 ◽  
Author(s):  
Franca R. Zaretzky ◽  
Thomas H. Kawula
Keyword(s):  
Hacat Cell ◽  
Inflammatory Cells ◽  
Etiologic Agent ◽  
Host Cells ◽  
Hacat Cells ◽  
Tissue Destruction ◽  
Necrosis Factor Alpha ◽  
Potent Inducer ◽  
Ulcer Disease

ABSTRACT Haemophilus ducreyi is the etiologic agent of chancroid, a sexually transmitted genital ulcer disease. Keratinocytes are likely the first cell type encountered by H. ducreyiupon infection of human skin; thus, the interaction between H. ducreyi and keratinocytes is probably important for the ability of H. ducreyi to establish infection. We have used the HaCaT keratinocyte cell line grown in monolayers and in cocultures with HS27 fibroblasts to investigate H. ducreyi interactions with keratinocytes and the host-cell response to H. ducreyiinfection. Using quantitative adherence and gentamicin protection assays, we determined that approximately 13% of H. ducreyiadhered to HaCaT cell monolayers, while only a small proportion (0.0052%) was intracellular. By transmission electron microscopy, we observed numerous H. ducreyi organisms adherent to but rarely within HaCaT cells cocultured with fibroblasts. Both liveH. ducreyi and purified H. ducreyilipooligosaccharide (LOS) induced significant interleukin 8 (IL-8) expression from HaCaT cell-HS27 cell cocultures. However, the level of IL-8 expression in response to LOS alone was not as pronounced.H. ducreyi LOS was a more potent inducer of IL-8 from cocultures than Escherichia coli lipopolysaccharide (LPS) at the same concentration, suggesting a unique effect of H. ducreyi LOS on cocultures. Neither live H. ducreyinor purified H. ducreyi LOS or E. coli LPS induced tumor necrosis factor alpha expression from cocultures.H. ducreyi induced drastically different cytokine profiles from cocultures than from HS27 or HaCaT cells cultured separately. IL-8 expression by skin cells in response to H. ducreyiinfection in vivo may be responsible for the massive influx of polymorphonuclear leukocytes and other inflammatory cells to the site of infection. This influx of inflammatory cells may be partly responsible for the tissue destruction characteristic of chancroid.

Cigarette smoking, emphysema, and damage to alpha 1-proteinase inhibitor

1994 ◽  
Vol 266 (6) ◽  
pp. L593-L611 ◽  
Author(s):  
M. D. Evans ◽  
W. A. Pryor
Keyword(s):  
Cigarette Smoke ◽  
Proteinase Inhibitor ◽  
Inflammatory Cells ◽  
Lavage Fluid ◽  
Lung Lavage ◽  
Tissue Destruction ◽  
Phagocytic Cells ◽  
Lung Fluid

The proteinase-antiproteinase theory for the pathogenesis of emphysema proposes that the connective tissue destruction associated with emphysema arises from excessive proteinase activity in the lower respiratory tract. For this reason, the relative activities of neutrophil elastase and alpha 1-proteinase inhibitor (alpha 1-PI) are considered important. Most emphysema is observed in smokers; therefore, alpha 1-PI has been studied as a target for smoke-induced damage. Damage to alpha 1-PI in lung fluid could occur by several mechanisms involving species delivered to the lung by cigarette smoke and/or stimulated inflammatory cells. Oxidative damage to alpha 1-PI has received particular attention, since both cigarette smoke and inflammatory cells are rich sources of oxidants. In this article we review almost two decades of research on mechanistic studies of damage to alpha 1-PI by cigarette smoke and phagocytic cells in vitro, studies emphasizing the importance of elastinolytic activity in the pathogenesis of emphysema in vivo and studies of human lung lavage fluid to detect defects in alpha 1-PI at the molecular and functional levels.

scholarly journals In Vivo Vascularization of Anisotropic Channeled Porous Polylactide-Based Capsules for Islet Transplantation: The Effects of Scaffold Architecture and Implantation Site

2015 ◽  
pp. S75-S84 ◽  
Author(s):  
N. KASOJU ◽  
D. KUBIES ◽  
E. FÁBRYOVÁ ◽  
J. KŘÍŽ ◽  
M. M. KUMOREK ◽  
...  
Keyword(s):  
Islet Transplantation ◽  
Porous Scaffold ◽  
Oxygen Demand ◽  
Inflammatory Cells ◽  
Host Cells ◽  
Cell Infiltration ◽  
Greater Omentum ◽  
Brown Norway ◽  
Porous Scaffolds

The replacement of pancreatic islets for the possible treatment of type 1 diabetes is limited by the extremely high oxygen demand of the islets. To this end, here we hypothesize to create a novel extra-hepatic highly-vascularized bioartificial cavity using a porous scaffold as a template and using the host body as a living bioreactor for subsequent islet transplantation. Polylactide-based capsular-shaped anisotropic channeled porous scaffolds were prepared by following the unidirectional thermally-induced phase separation technique, and were implanted under the skin and in the greater omentum of Brown Norway rats. Polyamide mesh-based isotropic regular porous capsules were used as the controls. After 4weeks, the implants were excised and analyzed by histology. The hematoxylin and eosin, as well as Masson's trichrome staining, revealed a) low or no infiltration of giant inflammatory cells in the implant, b) minor but insignificant fibrosis around the implant, c) guided infiltration of host cells in the test capsule in contrast to random cell infiltration in the control capsule, and d) relatively superior cell infiltration in the capsules implanted in the greater omentum than in the capsules implanted under the skin. Furthermore, the anti-CD31 immunohistochemistry staining revealed numerous vessels at the implant site, but mostly on the external surface of the capsules. Taken together, the current study, the first of its kind, is a significant step-forward towards engineering a bioartificial microenvironment for the transplantation of islets.

scholarly journals Relative Importance of NF-κB p50 in Mycobacterial Infection

2001 ◽  
Vol 69 (11) ◽  
pp. 7100-7105 ◽  
Author(s):  
Hiroyuki Yamada ◽  
Satoru Mizuno ◽  
Mohammad Reza-Gholizadeh ◽  
Isamu Sugawara
Keyword(s):  
Transforming Growth Factor ◽  
Interleukin 2 ◽  
Transforming Growth Factor Β ◽  
Mycobacterial Infection ◽  
Host Cells ◽  
Mrna Levels ◽  
Necrosis Factor Alpha ◽  
Airborne Infection ◽  
Normal Ranges

ABSTRACT To understand the role of NF-κB in the development of murine tuberculosis in vivo, NF-κB p50 knockout mice were infected withMycobacterium tuberculosis by placing them in the exposure chamber of an airborne-infection apparatus. These mice developed multifocal necrotic pulmonary lesions or lobar pneumonia. Compared with the levels in wild-type mice, pulmonary inducible nitric oxide synthase, interleukin-2 (IL-2), gamma interferon, and tumor necrosis factor alpha mRNA levels were significantly low but expression of IL-10 and transforming growth factor β mRNAs were within the normal ranges. The pulmonary IL-6 mRNA expression level was higher. Therefore, NF-κB and its interaction with host cells play an important role in the pathogenesis of tuberculosis.

scholarly journals Molecular Docking Studies of Curcumin Analogues against SARS-CoV-2 Spike Protein

2021 ◽  
Author(s):  
Jéssica Nogueira ◽  
Flávia Verza ◽  
Felipe Nishimura ◽  
Umashankar Das ◽  
Ícaro Caruso ◽  
...  
Keyword(s):  
Molecular Docking ◽  
Virus Disease ◽  
Etiologic Agent ◽  
Host Cells ◽  
Spike Protein ◽  
Therapeutic Effects ◽  
Host Cell Membrane ◽  
Curcumin Analogues

Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) is the etiologic agent of the current pandemic of corona virus disease 2019 (COVID-19) that has inflicted the loss of thousands of lives worldwide. The coronavirus surface spike (S) glycoprotein is a class I fusion with a S1 domain which is attached to the human angiotensin converting enzyme 2 (ACE2) receptor, and a S2 domain which enables fusion with the host cell membrane and internalization of the virus. Curcumin has been suggested as a potential drug to control inflammation and as a potential inhibitor of S protein, but its therapeutic effects are hampered by poor bioavailability. We performed a molecular docking and dynamic study using 94 curcumin analogues designed to have improved metabolic stability against the SARS-CoV-2 spike protein and compared their affinity with curcumin and other potential inhibitors. The docking analysis suggested that the S2 domain is the main target of these compounds and compound 2606 displayed a higher binding affinity (-9.6 kcal mol-1) than curcumin (-6.8 kcal mol-1) and the Food and Drug Administration (FDA) approved drug hydroxychloroquine (-6.3 kcal mol-1). Further additional validation in vitro and in vivo of these compounds against SARS-CoV-2 may provide insights into the development of a drug that prevents virus entry into host cells.

Novel Calcium Aminoclay (CaAC)-Vitamin C Hybrid: In-Vitro Cytotoxicity Assessment in HaCaT Cells and In-Vivo Embryotoxicity Assay in Zebrafish

2021 ◽  
Vol 21 (7) ◽  
pp. 3667-3672
Author(s):  
Vinh Van Tran ◽  
Vu Khac Bui Hoang ◽  
Hang-Suk Chun ◽  
Ju-Young Moon ◽  
Young-Chul Lee
Keyword(s):  
Vitamin C ◽  
Cell Viability ◽  
Hacat Cell ◽  
In Vitro Cytotoxicity ◽  
Hatching Rate ◽  
Hacat Cells ◽  
Promising Candidate ◽  
Cytotoxicity Assessment

Vitamin C (VC) is well-known as a hydrophilic antioxidant commonly used in cosmeceutical formulations due to its protection and maintenance of youthful skin. Aminoclay (AC), a synthetic organic-nanoclay, has shown great potential for delivery of VC. However, the practical cosmeceutical applications of aminoclay for delivery of VC are severely limited due to the paucity of reported research on its cytotoxicity to human skin. Therefore, in the present study, we evaluated the biosafety of a calcium aminoclay-vitamin C (CaAC-VC) hybrid through an In-Vitro cytotoxicity assessment in HaCaT cells and an In-Vivo embryotoxicity assay in zebrafish. HaCaT cell viability and changes in the morphology and hatching rate of the zebrafish were investigated. The results indicated that the CaAC-VC hybrid showed a lower cytotoxicity relative to pure VC and that as such, it should be considered to be a promising candidate for VC-delivery applications.

scholarly journals The role of Leishmania GP63 in the modulation of innate inflammatory response to Leishmania major infection

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0262158
Author(s):  
Aretha Chan ◽  
Jose-Mauricio Ayala ◽  
Fernando Alvarez ◽  
Ciriaco Piccirillo ◽  
George Dong ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Leishmania Major ◽  
Critical Role ◽  
Protozoan Parasite ◽  
Inflammatory Cells ◽  
Host Cells ◽  
Cell Infection ◽  
Zinc Metalloprotease

Leishmaniasis is a disease caused by the protozoan parasite Leishmania and is known to affect millions of individuals worldwide. In recent years, we have established the critical role played by Leishmania zinc-metalloprotease GP63 in the modulation of host macrophage signalling and functions, favouring its survival and progression within its host. Leishmania major lacking GP63 was reported to cause limited infection in mice, however, it is still unclear how GP63 may influence the innate inflammatory response and parasite survival in an in vivo context. Therefore, we were interested in analyzing the early innate inflammatory events upon Leishmania inoculation within mice and establish whether Leishmania GP63 influences this initial inflammatory response. Experimentally, L. major WT (L. majorWT), L. major GP63 knockout (L. majorKO), or L. major GP63 rescue (L. majorR) were intraperitoneally inoculated in mice and the inflammatory cells recruited were characterized microscopically and by flow cytometry (number and cell type), and their infection determined. Pro-inflammatory markers such as cytokines, chemokines, and extracellular vesicles (EVs, e.g. exosomes) were monitored and proteomic analysis was performed on exosome contents. Data obtained from this study suggest that Leishmania GP63 does not significantly influence the pathogen-induced inflammatory cell recruitment, but rather their activation status and effector function. Concordantly, internalization of promastigotes during early infection could be influenced by GP63 as fewer L. majorKO amastigotes were found within host cells and appear to maintain in host cells over time. Collectively this study provides a clear analysis of innate inflammatory events occurring during L. major infection and further establish the prominent role of the virulence factor GP63 to provide favourable conditions for host cell infection.

scholarly journals A potential role for Galectin-3 inhibitors in the treatment of COVID-19

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9392 ◽  
Author(s):  
John L. Caniglia ◽  
Maheedhara R. Guda ◽  
Swapna Asuthkar ◽  
Andrew J. Tsung ◽  
Kiran K. Velpula
Keyword(s):  
Disease Process ◽  
Standard Of Care ◽  
Host Cells ◽  
World Health ◽  
Necrosis Factor Alpha ◽  
Viral Attachment ◽  
Tnf Α ◽  
Galectin 3

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV2), the causative agent of coronavirus disease 2019 (COVID-19), has been declared a global pandemic by the World Health Organization. With no standard of care for the treatment of COVID-19, there is an urgent need to identify therapies that may be effective in treatment. Recent evidence has implicated the development of cytokine release syndrome as the major cause of fatality in COVID-19 patients, with elevated levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) observed in patients. Galectin-3 (Gal-3) is an animal lectin that has been implicated in the disease process of a variety of inflammatory conditions. Inhibitors of the small molecule Gal-3 have been shown to reduce the levels of both IL-6 and TNF-α in vitro and have shown anti-inflammatory effects in vivo. Additionally, a key domain in the spike protein of β-coronaviridae, a genus which includes SARS-CoV2, is nearly identical in morphology to human Gal-3. These spike proteins are critical for the virus’ entry into host cells. Here we provide a systematic review of the available literature and an impetus for further research on the use of Gal-3 inhibitors in the treatment of COVID-19. Further, we propose a dual mechanism by which Gal-3 inhibition may be beneficial in the treatment of COVID-19, both suppressing the host inflammatory response and impeding viral attachment to host cells.

scholarly journals Unconventional Maturation of Dendritic Cells Induced by Particles from the Laminated Layer of Larval Echinococcus granulosus

2014 ◽  
Vol 82 (8) ◽  
pp. 3164-3176 ◽  
Author(s):  
Cecilia Casaravilla ◽  
Álvaro Pittini ◽  
Dominik Rückerl ◽  
Paula I. Seoane ◽  
Stephen J. Jenkins ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Echinococcus Granulosus ◽  
Thick Layer ◽  
Host Cells ◽  
Interleukin 12 ◽  
Necrosis Factor Alpha ◽  
The Impact ◽  
Laminated Layer

ABSTRACTThe larval stage of the cestode parasiteEchinococcus granulosuscauses hydatid disease in humans and livestock. This infection is characterized by the growth in internal organ parenchymae of fluid-filled structures (hydatids) that elicit surprisingly little inflammation in spite of their massive size and persistence. Hydatids are protected by a millimeter-thick layer of mucin-based extracellular matrix, termed the laminated layer (LL), which is thought to be a major factor determining the host response to the infection. Host cells can interact both with the LL surface and with materials that are shed from it to allow parasite growth. In this work, we analyzed the response of dendritic cells (DCs) to microscopic pieces of the native mucin-based gel of the LL (pLL).In vitro, this material induced an unusual activation state characterized by upregulation of CD86 without concomitant upregulation of CD40 or secretion of cytokines (interleukin 12 [IL-12], IL-10, tumor necrosis factor alpha [TNF-α], and IL-6). When added to Toll-like receptor (TLR) agonists, pLL-potentiated CD86 upregulation and IL-10 secretion while inhibiting CD40 upregulation and IL-12 secretion.In vivo, pLL also caused upregulation of CD86 and inhibited CD40 upregulation in DCs. Contrary to expectations, oxidation of the mucin glycans in pLL with periodate did not abrogate the effects on cells. Reduction of disulfide bonds, which are known to be important for LL structure, strongly diminished the impact of pLL on DCs without altering the particulate nature of the material. In summary, DCs respond to the LL mucin meshwork with a “semimature” activation phenotype, bothin vitroandin vivo.

scholarly journals Dietary Glycine Prevents Peptidoglycan Polysaccharide-Induced Reactive Arthritis in the Rat: Role for Glycine-Gated Chloride Channel

2001 ◽  
Vol 69 (9) ◽  
pp. 5883-5891 ◽  
Author(s):  
Xiangli Li ◽  
Blair U. Bradford ◽  
Michael D. Wheeler ◽  
Stephen A. Stimpson ◽  
Heather M. Pink ◽  
...  
Keyword(s):  
Chloride Channel ◽  
Reactive Arthritis ◽  
Control Diet ◽  
Glycine Receptor ◽  
Inflammatory Cells ◽  
Necrosis Factor Alpha ◽  
Splenic Macrophages ◽  
Tnf Α ◽  
Chloride Influx

ABSTRACT Peptidoglycan polysaccharide (PG-PS) is a primary structural component of bacterial cell walls and causes rheumatoid-like arthritis in rats. Recently, glycine has been shown to be a potential immunomodulator; therefore, the purpose of this study was to determine if glycine would be protective in a PG-PS model of arthritis in vivo. In rats injected with PG-PS intra-articularly, ankle swelling increased 21% in 24 to 48 h and recovered in about 2 weeks. Three days prior to reactivation with PG-PS given intravenously (i.v.), rats were divided into two groups and fed a glycine-containing or nitrogen-balanced control diet. After i.v. PG-PS treatment joint swelling increased 2.1 ± 0.3 mm in controls but only 1.0 ± 0.2 mm in rats fed glycine. Infiltration of inflammatory cells, edema, and synovial hyperplasia in the joint were significantly attenuated by dietary glycine. Tumor necrosis factor alpha (TNF-α) mRNA was detected in ankle homogenates from rats fed the control diet but not in ankles from rats fed glycine. Moreover, intracellular calcium was increased significantly in splenic macrophages treated with PG-PS; however, glycine blunted the increase about 50%. The inhibitory effect of glycine was reversed by low concentrations of strychnine or chloride-free buffer, and it increased radiolabeled chloride influx nearly fourfold, an effect also inhibited by strychnine. In isolated splenic macrophages, glycine blunted translocation of the p65 subunit of NF-κB into the nucleus, superoxide generation, and TNF-α production caused by PG-PS. Further, mRNA for the beta subunit of the glycine receptor was detected in splenic macrophages. This work supports the hypothesis that glycine prevents reactive arthritis by blunting cytokine release from macrophages by increasing chloride influx via a glycine-gated chloride channel.

scholarly journals Biological Characterization of Lipopolysaccharide from Treponema pectinovorum

2002 ◽  
Vol 70 (1) ◽  
pp. 211-217 ◽  
Author(s):  
Lakshmyya Kesavalu ◽  
Clinton W. Falk ◽  
Kenneth J. Davis ◽  
Michelle J. Steffen ◽  
Xiaoping Xu ◽  
...  
Keyword(s):  
Lethal Dose ◽  
Biological Properties ◽  
Host Cells ◽  
Kinetic Assay ◽  
Necrosis Factor Alpha ◽  
Monocyte Chemoattractant Protein 1 ◽  
Heat Labile ◽  
Tnf Α ◽  
Macrophage Colony

ABSTRACT This study investigated the endotoxic and biological properties of purified lipopolysaccharide (LPS) isolated from an oral spirochete, Treponema pectinovorum. Endotoxicity, measured by Limulus amoebocyte lysate kinetic assay, showed that the LPS contained 1.28 endotoxin units per μg of purified LPS, which was approximately 4,000 times less than Escherichia coli O55:B5 LPS. To determine in vivo endotoxicity, LPS responder mice were administered LPS following galactosamine (GalN) sensitization. The LPS induced neither endotoxic symptoms nor lethality for 96 h, suggesting negligible or very low endotoxicity. In contrast, infection with live T. pectinovorum induced 100% lethality within 12 h in GalN-sensitized LPS responder mice, indicating an endotoxin-like property of this treponeme. Heat-killed microorganisms exhibited no lethality in GalN-sensitized mice, suggesting that the endotoxicity was associated with heat-labile components. To determine cytokine and chemokine induction by LPS, human gingival fibroblasts were stimulated and secretion of interleukin 1β (IL-1β), granulocyte-macrophage colony-stimulating factor, gamma interferon, IL-6, IL-8, and monocyte chemoattractant protein 1 (MCP-1) was assessed. The purified LPS induced significant amounts of only IL-6, IL-8, and MCP-1, although they were substantially lower than levels after challenge with live T. pectinovorum. After injection of LPS or live or heat-killed T. pectinovorum, serum was collected from mice and analyzed for proinflammatory cytokines IL-1β, tumor necrosis factor alpha (TNF-α), and IL-6. LPS induced only IL-6 consistently. Both live and heat-killed T. pectinovorum induced serum IL-6, which was higher than the level detected following LPS administration. Importantly, live bacteria elicited systemic TNF-α and IL-1β levels similar to those induced by a lethal dose of live E. coli O111. The results indicated that T. pectinovorum LPS has very low or no endotoxicity, although it can elicit low levels of cytokines from host cells. In contrast to the LPS, live T. pectinovorum demonstrated in vivo toxicity, which was associated with serum IL-1β, TNF-α, and IL-6, suggesting an endotoxin-like property of a heat-labile molecule(s) of the spirochete.

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