DOCSLIB.ORG

Galectin-3 Inhibits Paracoccidioides Brasiliensis Growth and Impacts

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Galectin-3 Inhibits Paracoccidioides Brasiliensis Growth and Impacts bioRxiv preprint doi: https://doi.org/10.1101/557918; this version posted February 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Galectin-3 inhibits Paracoccidioides brasiliensis growth and 2 impacts paracoccidioidomycosis through multiple 3 mechanisms 4 5 Otavio Hatanaka1, Caroline Patini Rezende1, Pedro Moreno1, Fabrício Freitas 6 Fernandes2, Patrícia Kellen Martins Oliveira Brito2, Roberto Martinez3, 7 Carolina Coelho4,5, Maria Cristina Roque-Barreira2, Arturo Casadevall6, Fausto 8 Almeida1* 9 10 1Department of Biochemistry and Immunology, Ribeirao Preto Medical School, 11 University of Sao Paulo, Ribeirao Preto, SP, 14049-900, Brazil 12 2Department of Cellular and Molecular Biology, Ribeirao Preto Medical School, 13 University of Sao Paulo, Ribeirao Preto, SP, 14049-900, Brazil 14 3Department of Internal Medicine, Ribeirao Preto Medical School, University of Sao 15 Paulo, Ribeirao Preto, SP, 14048-900, Brazil 16 4Department of Biosciences, College of Life and Environmental Sciences, University 17 of Exeter, Exeter, UK 18 5Medical Research Council Centre for Medical Mycology, University of Aberdeen, 19 Aberdeen, UK 20 6Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg 21 School of Public Health, Baltimore, MD, 21205, USA 22 23 * Correspondence: 24 Fausto Almeida 25 [email protected] bioRxiv preprint doi: https://doi.org/10.1101/557918; this version posted February 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 26 ABSTRACT 27 28 The thermodimorphic pathogenic fungi Paracoccidioides brasiliensis and 29 Paracoccioidioides lutzii are the etiologic causes of paracoccidioidomycosis (PCM), 30 the most prevalent systemic mycosis in Latin America. Galectin-3 (Gal-3), an animal 31 β-galactoside-binding protein, modulates important roles during microbial infections, 32 such as triggering a Th2-polarized immune response in PCM. Herein, we demonstrate 33 that Gal-3 also plays other important roles in P. brasiliensis infection. We verified 34 Gal-3 levels are upregulated in human and mice infections and establish that Gal-3 35 inhibits P. brasiliensis growth by inhibiting budding. Furthermore, Gal-3 affects 36 disruption and internalization of extracellular vesicles (EV) from P. brasiliensis by 37 macrophages. Our results suggest important roles for Gal-3 in P. brasiliensis infection, 38 indicating that increased Gal-3 production during P. brasiliensis infection may 39 account for affecting the fungal growth and EV stability, promoting a benefic course 40 of experimental PCM. 41 42 Keywords: Galectin-3, Paracoccidioides brasiliensis, Extracellular vesicles, fungal 43 infection. 44 45 46 bioRxiv preprint doi: https://doi.org/10.1101/557918; this version posted February 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 47 IMPORTANCE 48 49 Paracoccidiodomycosis (PCM) is the most prevalent systemic mycosis in Latin 50 America. Although the immune mechanisms to control PCM are still not fully 51 understood, several events of the host innate and adaptive immunity are crucial to 52 determine the progress of the infection. Mammalian β-galactoside-binding protein 53 Galectin-3 (Gal-3) plays significant roles during microbial infections, and has been 54 studied for its immunomodulatory roles but it can also have direct antimicrobial 55 effects. We asked whether this protein plays a role in P. brasiliensis. We report herein 56 that Gal-3 indeed has direct effects on fungal pathogen, inhibiting fungal growth and 57 reducing extracellular vesicles stability. Our results suggest a direct role for Gal-3 58 in P. brasiliensis infection, with beneficial effects for the mammalian host. 59 60 61 INTRODUCTION 62 63 Paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin 64 America, is caused by the thermodimorphic human pathogens Paracoccidioides 65 brasiliensis and Paracoccioidioides lutzii. After inhalation of airborne propagules 66 from fungal mycelia phase, in the lungs the fungi convert into the infectious form - 67 yeast phase (1-3). The yeast can spread to several organs causing systemic disease (4). 68 Human defense against PCM depends on satisfactory cellular immune response and 69 cytokine production (5, 6). Immune mechanisms that prevented cell division and 70 budding of the fungal cells could aid in the control the PCM. bioRxiv preprint doi: https://doi.org/10.1101/557918; this version posted February 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 71 Extracellular vesicles (EVs) are produced by all living cells, and actively 72 participate as key regulators of physiopathological mechanisms during fungal 73 infections (7, 8). Fungal EVs carry several virulence factors and other important 74 molecules, contributing to fungal pathogenicity and host immunomodulation (9-14). 75 Since EVs plays significant roles in the host-pathogen relationship, the vesicular 76 stability is important to ensure suitable delivery of their cargo into host cells (13, 15). 77 Bacterial and eukaryotic pathogens present surface glycans that may be 78 recognized by host carbohydrate-binding proteins. These interactions commonly 79 affect the microorganism pathogenesis, the host immune response or the success of 80 intracellular parasitism (16-19). Recently, we have reported that Galectin-3 (Gal-3), a 81 β-galactoside-binding animal lectin, plays significant roles in cryptococcal infection 82 (13). Gal-3 interferes the C. neoformans infection, inhibiting C. neoformans growth 83 and promoting vesicle disruption (13). Also, Gal-3 has been reported to influence the 84 outcome of other mycoses, such as Candida albicans (20) and Histoplasma 85 capsulatum (21). In Paracoccidioides brasiliensis, Gal-3 was reported to play an 86 immunomodulatory role in the host response (22). Since Gal-3 can influence host 87 response against PCM, as well as several other microbial infections, and regulates 88 different functions in the physiopathology of infections, we explored whether Gal-3 89 influence P. brasiliensis growth and vesicle stability. 90 In this work, we assessed the Gal-3 levels in humans and mice with PCM. Also, 91 we demonstrated the influence of Gal-3 in the P. brasiliensis growth and stability of 92 EVs. Our results demonstrate that Gal-3 inhibits growth and budding of P. 93 brasiliensis yeast cells, and promotes vesicle disruption. Our results suggest that Gal- 94 3 can impacts the interaction of P. brasiliensis with host cells. 95 96 bioRxiv preprint doi: https://doi.org/10.1101/557918; this version posted February 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 97 RESULTS 98 Gal-3 is up-regulated during PCM 99 Since increased Gal-3 expression was previously described during human and 100 experimental inflammatory diseases (23, 24), and recently in C. neoformans infection 101 (13), we measured Gal-3 levels in serum samples from individuals suffering of PCM, 102 either acute or chronic PCM form. Compared to the healthy individuals, the acute and 103 chronic form patients showed higher Gal-3 serum levels, as shown previously for 104 other infections (Figure 1). There was no significant difference (P value: 0.4204, 105 unpaired t-test) between the Gal-3 levels in sera of acute and chronic patients with 106 PCM. 107 Subsequently, we measured Gal-3 levels in tissues and serum of C57BL/6 mice 108 on days 30 and 60 post-infection with P. brasiliensis (Figure 2). In comparison with 109 control animals (PBS), infected mice had higher Gal-3 levels in all examined tissues 110 (lungs and spleen, Figure 2B and 2C, respectively) and serum samples (Figure 2A). 111 As previously reported for C. neoformans infection, there is a correlation between P. 112 brasiliensis infection and increased levels of serum Gal-3, which could reflect the 113 inflammatory conditions caused by these infectious diseases. 114 Gal-3 inhibits P. brasiliensis growth 115 Since P. brasiliensis cell division and budding is crucial to successful PCM, 116 and Gal-3 inhibits C. neoformans growth (13), we evaluated whether Gal-3 could 117 affect the growth and budding of P. brasiliensis. P. brasiliensis growth in culture, 118 measured by MTT assay, was compared between Gal-3 treated, PBS- and denatured 119 Gal-3-treated yeasts. Gal-3 inhibited P. brasiliensis growth by approximately 50% bioRxiv preprint doi: https://doi.org/10.1101/557918; this version posted February 23, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 120 after 72 h compared with the controls (denatured Gal-3 treated or PBS) (Figure 3). To 121 verify whether Gal-3 treatment of fungal yeast induces yeast death, we performed 122 viability assays using fluorescein diacetate/ethidium bromide staining. Gal-3 treated 123 and control cultures contained similar proportions of viable cells up until 72 h after 124 Gal-3 treatment, and all cultures were >80% viable (data not shown). To further 125 characterize Gal-3 effects in the growth of P.
Load more

Recommended publications
  • Lectin Free Diet
    THERMO-TECH INC. Lectins Free Diet Lectins are proteins in plants that have been associated with both positive and negative health effects. Some plant-based foods, such as beans and legumes, whole grains, nuts, seeds and certain vegetables contain a high amount of lectins. Lectins are a type of protein that, in humans, may help cells interact with one another. Some scientists also believe that lectins provide a form of defense in plants to keep insects away. These proteins also contain nitrogen, which is needed for plants to grow. While many parts of plants contain lectins, the seed is the part that people eat most often. Lectins may impact health in multiple ways, ranging from digestive disturbances (a dysbiotic condition of the gut) to chronic disease risk (inflammation and autoimmune disease including: celiac disease, psoriasis, diabetes, rheumatoid arthritis, MS and Parkinson's Disease to name a few). They have also been shown to cause red blood cells to cluster together thus not carrying the nutrients and oxygen to our cells this can cause malnutrition although our diet is varied and plentiful. Lectins are categorized as anti-nutrients since they block the absorption of some nutrients. Lectins may cause an upset stomach when plant foods are eaten uncooked. They are also the reason why it can be dangerous to eat undercooked legumes. The lectins in red kidney beans is called phytohaemagglutinin is responsible for red kidney bean poisoning, which results from eating raw or undercooked kidney beans. According to the United States Food and Drug Administration (FDA), consuming just four raw kidney beans could cause symptoms including severe nausea, vomiting, and diarrhea.
    [Show full text]
  • Binding and the Effect of the Red Kidney Bean Lectin, Phytohaemagglutinin, In
    Downloaded from https://www.cambridge.org/core British Journal of Nutrition (2006), 95, 105–115 DOI: 10.1079/BJN20051612 q The Authors 2006 Binding and the effect of the red kidney bean lectin, phytohaemagglutinin, in . IP address: the gastrointestinal tract of suckling rats 170.106.202.58 Ann Linderoth1*, Olena Prykhod’ko1, Bo Ahre´n2, Frida Fa˚k1, Stefan G. Pierzynowski1 and Bjo¨rn R. Westro¨m1 1Department of Cell and Organism Biology, Lund University, Helgonava¨gen 3B, SE-223 62 Lund, Sweden , on 2Department of Medicine, University Hospital, Lund University, Lund, Sweden 29 Sep 2021 at 02:15:37 (Received 7 April 2005 – Revised 8 July 2005 – Accepted 17 July 2005) Enteral exposure of suckling rats to phytohaemagglutinin (PHA) has been shown to induce growth and precocious functional maturation of the gastrointestinal tract. The aim of the present study was to explore the mechanism of this action. Suckling rats, 14 d old, were fed a single dose , subject to the Cambridge Core terms of use, available at of PHA (0·05 mg/g body weight) or saline. The binding of PHA to the gut epithelium and its effect on the morphology and functional properties of the gut and pancreas were studied up to 3 d after treatment. Initially, at 1–24 h, the PHA bound along the gut mucosal lining, resulting in dis- turbed gut morphology with villi shortening and rapid decreases in disaccharidase activities and macromolecular absorption capacity. During a later phase, between 1 and 3 d, the PHA binding had declined, and an uptake by enterocytes was observed.
    [Show full text]
  • Part I: Abstracts of Members' Proffered
    ABSTRACTS OF MEMBERS PAPERS 401 PART I: ABSTRACTS OF MEMBERS' PROFFERED PAPERS ENHANCEMENT OF CHEMICALLY- Pregnant rats were injected with ethyl- INDUCED NEOPLASIA BY PROXI- nitrosourea to induce cerebral gliomas in a MAL ENTERECTOMY. R. C. N. WIL- high proportion of the offspring. With a dose LIAMSON, F. L. R. BAUER and R. A. MALT, of 40-50 mg/kg body weight the average United Bristol Hospitals and MlVassachusetts latent period for these brain tumours is 246 General Hospital, Boston, U.S.A. (Introduced days. Cultures have been prepared at by M. 0. Symes). different times after transplacental exposure Azoxymethane-induced tumours in the but before a tumour is visible. It has been rat, morphologically indistinguishable from reported previously (Roscoe and Claisse those arising in man, affect the duodenum, (1976) Nature, Lond., 262, 314) that there is jejunum and colon, but usually spare the a marked difference in the behaviour of ileum (Ward, J. M. (1975) Vet. Pathol., 12, cultures prepared 138-145 days p.i. and 2 165). Proximal small-bowel resection (PSBR) days p.i. These experiments have been causes prompt ileal hyperplasia, with a lesser extended and are reported here. Cultures response in the colon. To test the possible prepared 111-112 days p.i. were similar to adjuvant effect of postresectional hyper- those derived 138-145 days p.i. They con- plasia on intestinal carcinogenesis, rats tained cells like those found in tumour (n= 76) were submitted to 5000 PSBR 10 cultures which can predominate in culture days after the last of 16 weekly injections of and are tumourigenic.
    [Show full text]
  • Lymphocyte Responses to DR1/4 Restricted Ann Rheum Dis: First Published As 10.1136/Ard.53.3.171 on 1 March 1994
    Annals of the Rheumatic Diseases 1994; 53: 171-177 171 Lymphocyte responses to DR1/4 restricted Ann Rheum Dis: first published as 10.1136/ard.53.3.171 on 1 March 1994. Downloaded from peptides in rheumatoid arthritis Margot A Skinner, Lisa Watson, Arie Geursen, Paul L J Tan Abstract examination of the synovium has shown a Objective-To determine whether analog prominent infiltrate of lymphocytes, the and unrelated DR1/4 binding peptides majority of which are activated T cells.2 In alter DR1/4 restricted responses of addition, several experimental treatment peripheral blood lymphocytes (PBL) from strategies which act primarily through their patients with rheumatoid arthritis (RA). effect on T cells are reported to be effective in Methods-PBL from 25 patients with RA the treatment of RA.' 3 and 12 healthy controls were cultured with The association of RA with major histo- DR1/4 restricted peptides of the influenza compatibility genes is well established4 and the haemagglutinin, amino acids 307-319 HILA DR4 (DRB1*04) and DR1 (DRBl*01) (HA) and matrix proteins, amino acids genes involved is well documented.5 Disease 17-29 (IM). Responses were determined susceptibility has been localised to the third by 3H-thymidine uptake proliferation hypervariable region of the i chain of DR assays and limiting dilution analysis. and in white groups is most commonly Competitor peptides were analogs HA- associated with Dw4 (DRB1*0401), R312 and HA-K313 differing from HA by one containing the amino acid sequence amino acid at the 312 or 313 position LLEQKRAA at positions 67-74 and VG at respectively or unrelated peptides which residues 85-86.5 This disease-associated bind to DR1I/4.
    [Show full text]
  • Dry Kidney Beans Need to Be Boiled
    THE OHIO STATE UNIVERSITY OHIO STATE UNIVERSITY EXTENSION OHIO AGRICULTURAL RESEARCH AND DEVELOPMENT CENTER April 12, 2013 Dry kidney beans By Martha Filipic 614-292-9833 need to be boiled [email protected] Editor: A few weeks ago, I contacting their doctor. Other types of beans also This column was reviewed by Linnette Goard, field specialist soaked some dry kidney contain PHA, but it’s much more in food safety, selection and beans to prepare them concentrated in red kidney beans. For management for Ohio State for some chili. My sister example, the unit of measurement University Extension, the told me that before for the toxin is called “hau,” for outreach arm of the College “hemagglutinating unit.” Raw red of Food, Agricultural, and adding them to the kidney beans have anywhere from Environmental Sciences. chili, I should boil them 20,000 to 70,000 hau, but that drops to 200 to 400 hau when the beans to make sure the beans Communications and are fully cooked — not enough to be wouldn’t make us sick. I Technology a problem. White kidney beans, or Strategic Communications did, but was that really cannellini beans, contain only about 2021 Coffey Road necessary? one-third of the toxin as red kidney Columbus, OH 43210-1044 614-292-2011 You’ve got a knowledgeable sister. beans. Broad beans, or fava beans, Many people don’t know the risk contain just 5 to 10 percent of what’s 208 Research Services posed by dry red kidney beans when in red kidney beans. Building they’re not cooked properly.
    [Show full text]
  • Phytohaemagglutinin-M (PHA-M) Liquid
    Ref : FT.L3010an Page : 1/2 Technical data sheet Version date : 13/02/2014 Phytohaemagglutinin-M (PHA-M) liquid CAT N° : L3010 Storage conditions : Store frozen medium at -20°C After thawing, the PHA-M is stable for at least 1 month at +2/+8°C. The PHA-M may appear cloudy at +2/+8°C, but this turbidity has no effect on the activity of the product. Shelf life : 36 months Composition : After thawing, each ml of the solution contains 5 to 10 mg of protein. Recommended use : - Respect storage conditions of the product - Do not use the product after its expiry date - Store the product in a dry area - Wear clothes adapted to the manipulation of the product to avoid contamination (e.g. : gloves, mask, hygiene cap, overall…) - Protect the product from any form of humidity - Use, in one time, after opening, the entire quantity of product of the container, without making a concentrated solution (to avoid the formation of precipitates). If it is not possible, close the container immediately after sampling the quantity of powder required. - Supplements can be added prior to sterile filtration of the medium or aseptically introduced to sterile medium (respect the final concentration of the media). The nature of the supplements may affect storage conditions and shelf life of the medium. The product is intended to be used in vitro, in laboratory only. Do not use it in therapy, human or veterinary applications. Application : Phytohaemagglutinin is a lectin extracted from red kidney beans (Phaseolus vulgaris). The protein consists of two molecular species : a leucoagglutinin (PHA-L)and an erythroagglutinin (PHA-E).
    [Show full text]
  • Effect of the 13-Adrenoceptor Agonist Clenbuterol and Phytohaemagglutinin on Growth, Protein Synthesis and Polyamine Metabolism of Tissues of the Rat 'S
    Br. J. Pharmacol. (1992), 106, 476-482 '." Macmillan Press Ltd, 1992 Effect of the 13-adrenoceptor agonist clenbuterol and phytohaemagglutinin on growth, protein synthesis and polyamine metabolism of tissues of the rat 'S. Bardocz, D.S. Brown, G. Grant, A. Pusztai, J.C. Stewart & R.M. Palmer Rowett Research Institute, Bucksburn, Aberdeen AB2 9SB 1 The kidney bean lectin, phytohaemagglutinin (PHA), induced a marked atrophy of skeletal muscle which was evident from the changes in tissue composition (protein, RNA, DNA and polyamine content) and from the reduction in weight and protein synthesis of hind leg muscles of rats fed on kidney bean-diets for four days. The P-adrenoceptor agonist, clenbuterol, induced skeletal muscle hypertrophy by transiently stimulating protein synthesis. As a consequence, the muscle loss caused by a short exposure to PHA was, in part, ameliorated by clenbuterol treatment. 2 Cardiac muscle was affected to a lesser extent than skeletal muscle by both clenbuterol and the lectin. However, there was evidence that protein synthesis in heart was reduced by PHA. 3 PHA had opposite effects on the gut, the lectin-induced hyperplasia of the jejunum was accompanied by a large increase in protein synthesis. Clenbuterol alone had no effect on the jejunum whereas a combination of PHA and clenbuterol appeared to exacerbate the effect of the lectin on gut. 4 Both the lectin-induced gut growth and the hypertrophy of skeletal muscle caused by clenbuterol were preceded by the accumulation of polyamines in the respective tissues. Of particular note was the observation that a significant increase in the proportion of the intraperitoneally injected '4C-labelled spermidine or putrescine taken up by the growing tissues could be detected by the second day.
    [Show full text]
  • Binding and the Effect of the Red Kidney Bean Lectin, Phytohaemagglutinin, in the Gastrointestinal Tract of Suckling Rats
    Downloaded from https://www.cambridge.org/core British Journal of Nutrition (2006), 95, 105–115 DOI: 10.1079/BJN20051612 q The Authors 2006 Binding and the effect of the red kidney bean lectin, phytohaemagglutinin, in . IP address: the gastrointestinal tract of suckling rats 170.106.33.22 Ann Linderoth1*, Olena Prykhod’ko1, Bo Ahre´n2, Frida Fa˚k1, Stefan G. Pierzynowski1 and Bjo¨rn R. Westro¨m1 1Department of Cell and Organism Biology, Lund University, Helgonava¨gen 3B, SE-223 62 Lund, Sweden , on 2Department of Medicine, University Hospital, Lund University, Lund, Sweden 26 Sep 2021 at 12:43:14 (Received 7 April 2005 – Revised 8 July 2005 – Accepted 17 July 2005) Enteral exposure of suckling rats to phytohaemagglutinin (PHA) has been shown to induce growth and precocious functional maturation of the , subject to the Cambridge Core terms of use, available at gastrointestinal tract. The aim of the present study was to explore the mechanism of this action. Suckling rats, 14 d old, were fed a single dose of PHA (0·05 mg/g body weight) or saline. The binding of PHA to the gut epithelium and its effect on the morphology and functional properties of the gut and pancreas were studied up to 3 d after treatment. Initially, at 1–24 h, the PHA bound along the gut mucosal lining, resulting in dis- turbed gut morphology with villi shortening and rapid decreases in disaccharidase activities and macromolecular absorption capacity. During a later phase, between 1 and 3 d, the PHA binding had declined, and an uptake by enterocytes was observed.
    [Show full text]
  • Porifera Lectins: Diversity, Physiological Roles and Biotechnological Potential
    Mar. Drugs 2015, 13, 5059-5101; doi:10.3390/md13085059 OPEN ACCESS marine drugs ISSN 1660-3397 www.mdpi.com/journal/marinedrugs Review Porifera Lectins: Diversity, Physiological Roles and Biotechnological Potential Johan Gardères 1,2,3, Marie-Lise Bourguet-Kondracki 1, Bojan Hamer 2, Renato Batel 2, Heinz C. Schröder 3 and Werner E. G. Müller 3, * 1 Unité Molécules de Communication et Adaptation des Microorganismes, UMR 7245 CNRS, Muséum National d’Histoire Naturelle, CP 54, 57 rue Cuvier, Paris 75005, France; E-Mails: [email protected] (J.G.); [email protected] (M.-L.B.-K.) 2 Laboratory for Marine Molecular Biology, Center for Marine Research, Ruđer Bošković Institute, G. Paliaga 5, 52210 Rovinj, Croatia; E-Mails: [email protected] (B.H.); [email protected] (R.B.) 3 ERC Advanced Investigator Grant Research Group at Institute for Physiological Chemistry, University Medical Center of Johannes Gutenberg University Mainz, Duesbergweg 6, Mainz D-55128, Germany; E-Mail: [email protected] * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +49-6131-39-25910; Fax: +49-6131-39-25243. Academic Editor: Anake Kijjoa Received: 30 April 2015 / Accepted: 27 July 2015 / Published: 7 August 2015 Abstract: An overview on the diversity of 39 lectins from the phylum Porifera is presented, including 38 lectins, which were identified from the class of demosponges, and one lectin from the class of hexactinellida. Their purification from crude extracts was mainly performed by using affinity chromatography and gel filtration techniques. Other protocols were also developed in order to collect and study sponge lectins, including screening of sponge genomes and expression in heterologous bacterial systems.
    [Show full text]
  • In Vivo Fluorescence Imaging of E-Selectin: Quantitative Detection of Endothelial Activation in Arthritis
    In Vivo Fluorescence Imaging of E-Selectin: Quantitative Detection of Endothelial Activation in Arthritis A thesis submitted for the degree of PhD by Luke Gompels August 2010 Kennedy Institute of Rheumatology Faculty of Medicine Imperial College SUPERVISORS Dr Ewa Paleolog Prof. Dorian Haskard 1 Abstract Rheumatoid arthritis (RA) is a chronic progressive systemic inflammatory disease, characterized by synovial inflammation and localized destruction of cartilage and bone. Heterogeneity in the clinical presentation of RA and uncertainty about which patients will respond to treatment makes diagnosis and management challenging. Fluorescent imaging in the near infrared (NIR) spectrum significantly decreases tissue autofluorescence offering unique potential to detect specific molecular targets in vivo. E-selectin or endothelial adhesion molecule-1 (ELAM-1), a 115kDa glycoprotein induced on endothelial cells in response to pro-inflammatory cytokines involved in RA, such as interleukin (IL)-1 and tumour necrosis factor (TNF. E-selectin has been well validated as a potential biomarker of disease activity. My study aimed to investigate whether E-selectin targeted optical imaging in vivo could be developed as a sensitive, specific and quantifiable preclinical molecular imaging technique, and also whether this approach could be used to delineate the molecular effects of novel therapies. I utilised anti-E-selectin antibody labelled with NIR fluorophore in a mouse model of paw swelling induced by intra-plantar injection of TNFand in acute collagen-induced arthritis (CIA) in DBA/1 mice, a widely used model of RA. E-selectin generated signal, localised to points of maximal clinical inflammation in the inflamed mouse paw in both models with significant differences to control antibody.
    [Show full text]
  • Normal Human Alveolar Macrophages Obtained by Bronchoalveolar Lavage Have a Limited Capacity to Release Interleukin-1
    Normal human alveolar macrophages obtained by bronchoalveolar lavage have a limited capacity to release interleukin-1. M D Wewers, … , P B Bitterman, R G Crystal J Clin Invest. 1984;74(6):2208-2218. https://doi.org/10.1172/JCI111647. Research Article Interleukin-1 (IL-1) is a mediator released by stimulated mononuclear phagocytes that is thought to play an important role in modulating T and B lymphocyte activation as well as in contributing to the febrile response and other inflammatory processes. Circulating mononuclear phagocytes, blood monocytes, readily release IL-1 when stimulated. However, the ability of lung mononuclear phagocytes, alveolar macrophages, to dispose of the large daily burden of inhaled antigens without stimulating an inflammatory response suggests that the release of IL-1 by alveolar macrophages may differ significantly from that of blood monocytes. To evaluate this hypothesis, normal autologous alveolar macrophages, obtained by bronchoalveolar lavage, were compared with blood monocytes for their ability to release IL-1 in response to a standard stimulus, lipopolysaccharide (LPS). Alveolar macrophages were found to be at least 1,000 times less sensitive to LPS than blood monocytes. Furthermore, alveolar macrophages released significantly less IL-1 than blood monocytes (26 +/- 11 vs. 128 +/- 21 U/10(6) cells X 24 h, respectively, after stimulation with 10 micrograms/ml of LPS, P less than 0.001). This difference was not due to the release of substances by macrophages, which inhibited lymphocyte proliferation in response to IL-1, or to degradation of IL-1 by macrophages. Culturing macrophages in the presence of indomethacin and dialysis of macrophage supernatants did not affect the difference, and culturing macrophages with monocytes […] Find the latest version: https://jci.me/111647/pdf Normal Human Alveolar Macrophages Obtained by Bronchoalveolar Lavage Have a Limited Capacity to Release Interleukin-1 Mark D.
    [Show full text]
  • Galectin-3: Is This Member of a Large Family of Multifunctional Lectins (Already) a Therapeutic Target?
    Expert Opinion on Therapeutic Targets ISSN: 1472-8222 (Print) 1744-7631 (Online) Journal homepage: https://www.tandfonline.com/loi/iett20 Galectin-3: is this member of a large family of multifunctional lectins (already) a therapeutic target? Antonio Romero & Hans-Joachim Gabius To cite this article: Antonio Romero & Hans-Joachim Gabius (2019): Galectin-3: is this member of a large family of multifunctional lectins (already) a therapeutic target?, Expert Opinion on Therapeutic Targets, DOI: 10.1080/14728222.2019.1675638 To link to this article: https://doi.org/10.1080/14728222.2019.1675638 Accepted author version posted online: 01 Oct 2019. Submit your article to this journal View related articles View Crossmark data Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=iett20 Publisher: Taylor & Francis & Informa UK Limited, trading as Taylor & Francis Group Journal: Expert Opinion on Therapeutic Targets DOI: 10.1080/14728222.2019.1675638 Galectin-3: is this member of a large family of multifunctional lectins (already) a therapeutic target? Antonio Romero1* and Hans-Joachim Gabius2* 1Structural and Chemistry Department, Centro de Investigaciones Biológicas (CIB), CSIC, Ramiro de Maeztu 9, 28040 Madrid, Spain 2Institute of Physiological Chemistry, Faculty of Veterinary Medicine, Ludwig-Maximilians- University Munich, 80539 Munich, Germany Corresponding author: Dr. A. Romero (Research Professor of the Spanish Research Council, CSIC): Phone: +34 91 1098044, Fax: +34 91 5360432, e-mail: [email protected] Prof. Dr. Dr. h.c. H.-J. Gabius: Tel.+49-89-2180 2290, Fax: +49-89-2180 992290, e-mail: [email protected] or [email protected] Accepted Manuscript Information Classification: General ABSTRACT Introduction: The discoveries that sugars are a highly versatile platform to generate biochemical messages and that glycan-specific receptors (lectins) are a link between these signals and their bioactivity explain the interest in endogenous lectins such as galectins.
    [Show full text]