DOCSLIB.ORG

Cpg Oligodeoxynucleotide Inhibits Cockroach-Induced Asthma Via

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cpg Oligodeoxynucleotide Inhibits Cockroach-Induced Asthma Via Original Article Allergy Asthma Immunol Res. 2016 May;8(3):264-275. http://dx.doi.org/10.4168/aair.2016.8.3.264 pISSN 2092-7355 • eISSN 2092-7363 CpG Oligodeoxynucleotide Inhibits Cockroach-Induced Asthma via Induction of IFN-γ+ Th1 Cells or Foxp3+ Regulatory T Cells in the Lung Do-Hyun Kim,1,2 Jung-Ho Sohn,1,2,3 Hong-Jai Park,1,2 Jae-Hyun Lee,3 Jung-Won Park,3* Je-Min Choi1,2* 1Department of Life Science, College of Natural Sciences, Hanyang University, Seoul, Korea 2Research Institute for Natural Sciences, Hanyang University, Seoul, Korea 3Division of Allergy and Immunology, Department of Internal Medicine and Institute of Allergy, Yonsei University College of Medicine, Seoul, Korea This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Purpose: CpG oligodeoxynucleotide (CpG-ODN), a TLR9 agonist, activates innate immunity and induces Th1 response. Although the immune modu- latory effect of CpG-ODN has been extensively studied, its function in cockroach extract-induced allergic asthma has not been studied. Here, we in- vestigated the inhibitory function of CpG-ODN in cockroach extract-induced asthma in mice with different treatment schemes. Methods: Scheme 1: BALB/C mice were intra-nasally co-administered by cockroach extract and CpG-ODN twice a week for 3 weeks; Scheme 2: The mice were intra-na- sally pre-treated with CpG-ODN at day 0 and cockroach allergen challenge was performed from day 3 as in scheme 1. Scheme 3: Cockroach aller- gen challenge was performed as in scheme 1 and CpG-ODN was post-treated at day 21. Then, BAL cell count, flow cytometric analysis of alveolar macrophages, regulatory T cells, and lung tissue histology, Th1 and Th2 cytokines, serum IgE, cockroach specific IgE, IgG1/IgG2a ratio, and airway hyper-responsiveness were evaluated. Results: Mice with repeated intra-nasal exposure to CpG-ODN showed a dramatic decrease in eosinophilic inflammation, goblet cell hyperplasia, and airway hyper-responsiveness with reduction of IL-13, IL-5, and serum IgE, cockroach specific IgE and IgG1/IgG2a ratio. This inhibitory function might be related to the up-regulation of IL-10 and CD4+Foxp3+ regulatory T cells in the lung. Interestingly, one-time challenge of CpG-ODN either prior or posterior to cockroach extract exposure could modulate airway inflammation and hyper-responsive- ness via increase of Th1 response. Conclusions: Collectively, our data suggest that CpG-ODN treatment modulates Th2 inflammation in the lung by induction of regulatory T cells or Th1 response in a cockroach-induced asthma model. Key Words: CpG-ODN; cockroach allergen; Th2 inflammation; immune modulation INTRODUCTION immune responses similar to those in response to bacterial DNA.7,8 CpG-ODN treatment has been extensively studied to Asthma and allergies are the most common chronic inflam- stimulate both innate and adaptive immunity as it induces matory diseases associated with cockroach or house dust mite TNF-α expression in macrophages,9 augments IFN-γ and IL-12 (HDM) allergens characterized by airflow obstruction, airway expression and enhances the cytotoxic activity of NK cells10,11 eosinophilic inflammation, goblet cell metaplasia, airway re- and Th1 response, which can be used as immune adjuvants modeling, and airway hyper-responsiveness (AHR).1-3 Th2 cy- tokines such as IL-4, IL-5, IL-9, and IL-13 derived from helper T Correspondence to: Je-Min Choi, PhD, Department of Life Science, College cells and other cell types play a critical role in the pathophysiol- of Natural Sciences, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, ogy of asthma,4 and the balance between Th1 and Th2 cells is Seoul 04763, Korea. thought to be important in controlling disease symptoms.5 The Tel: +82-2-2220-4765; Fax: +82-2-2200-3495; E-mail: [email protected] suppression of Th2 responses by induction of Th1 response or Jung-Won Park, MD, PhD, Department of Internal Medicine, Institute of Allergy, regulatory T cells or by inhibiting Th2 inducing factors such as Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul IL-4, IL-5, and IL-13 have been extensively investigated for the 03722, Korea. Tel: +82-2-2228-1961; Fax: +82-2-362-8647; E-mail: [email protected] development of therapeutic drugs for asthma.6 Received: June 7, 2015; Revised: August 13, 2015; Accepted: August 28, 2015. CpG oligodeoxynucleotide (CpG-ODN) is a synthetic TLR9 li- •Do-Hyun Kim and Jung-Ho Sohn contributed equally to this work. gand, consisting of an unmethylated dinucleotide that activates •There are no financial or other issues that might lead to conflict of interest. 264 http://e-aair.org © Copyright The Korean Academy of Asthma, Allergy and Clinical Immunology • The Korean Academy of Pediatric Allergy and Respiratory Disease AAIR CpG Treatment Ameliorates Cockroach-Induced Asthma and to modulate Th2 responses.12,13 In addition, CpG-ODN has TCC ATG ACG TTC CTG ACG TT-3´). The control ODNs had been shown to enhance type I IFN production from plasmacy- equivalent lengths without functional CpG motifs (5´-TCG ATG toid dendritic cells (pDCs),14,15 and induce IL-10 producing reg- AGC TTC CTG AGT CT-3´). The ODNs were synthesized and ulatory T cells by indoleamine 2,3-dioxygenase (IDO) 1 pro- purified using a HPLC by Genotech, Inc. (Daejon, Korea). duction.16,17 Thus, CpG-ODN has been investigated as a potent preventive as well as a therapeutic immune modulator in aller- Cockroach-induced asthma model gic inflammatory diseases.18,19 For the co-treatment scheme, 6- to 8-week old BALB/C mice Cockroaches are known to be a significant environmental al- were intra-nasally co-sensitized by 120 μg cockroach extract lergen resulting in atopic asthma,20,21 and among the various and 3 μg of control ODN or CpG-ODN, twice a week for 3 environmental factors, cockroach allergen is recognized as a weeks. For the pre-treatment scheme, mice were administered major risk factor for pathogenesis of allergic asthma in hu- 3 μg of the control ODN or of CpG-ODN intra-nasally once on mans.22-24 Children who are exposed to high bedroom levels of day 0, and then 120 μg intra-nasal cockroach allergen challenge cockroach allergens showed an increased incidence of allergic was performed twice a week for 3 weeks. Four days after the last asthma, compared to other children.25,26 Also, among the many challenge, the mice were sacrificed and pathological changes of indoor allergens, only cockroaches were significantly related to the lungs were examined. For the post-treatment scheme, mice recurrent asthmatic wheezing.23 Previous preventive and thera- were sensitized by intra-nasal administration of 120 μg cock- peutic CpG-ODN treatment has been shown to suppress eosin- roach extract twice a week for 3 weeks. Two days after the last ophilic inflammation and hyper-responsiveness in the OVA/Al- challenge, 3 μg of control ODN or CpG-ODN was intra-nasally um-induced asthma model.27 More recently, CpG-ODN treat- administrated, and then the mice were sacrificed and analyzed ment also suppressed airway inflammation and hyper-respon- at 2 days later of CpG-ODN treatment. siveness in the HDM-induced asthma model.28 However, the immune modulatory effects of CpG-ODN in Measurements of airway hyper-responsiveness cockroach-induced asthma have not been investigated. The aim Four days after the last challenge, airway hyper-responsive- of the present study is to investigate the immune modulatory ef- ness (AHR) was measured with a flexiVent 5.1® small animal fect of intra-nasally administered CpG-ODN in a cockroach-in- ventilator (SCIREQ, PQ, Canada). In detail, mice were chal- duced asthma model with different treatment schemes. lenged with a saline control aerosol followed by increasing con- centrations of methacholine (MeCh) (Sigma-Aldrich, MO, USA; MATERIALS AND METHODS 3.1, 6.25, 12.5, 25, and 50 mg/mL). Aerosols were generated with an ultrasonic nebulizer and delivered to the inspiratory Animals line of the flexiVent using a bias flow of medical air. Each aero- Female BALB/C mice, 6 to 8 weeks old, were purchased from sol was delivered for 10 seconds during which time regular ven- Orient Bio (Daejeon, Korea). All mice were housed under spe- tilation was maintained. Two measurements were made at cific pathogen-free conditions, and maintained on a 12 hours 1-minute intervals following each aerosol.30 light-dark cycle with regular chow and autoclaved water ad libi- tum. All experiments described in this study were approved by Bronchoalveolar lavage fluid and lung homogenate the Animal Research Ethics Board of Yonsei University. To collect bronchoalveolar lavage (BAL) fluid, the lungs were lavaged with 1 mL of Hank’s balanced salt solution through an Cockroach allergen preparations intubation tube. Total cell numbers were counted with a hemo- Crude cockroach extract was prepared as previously de- cytometer. BAL fluid was centrifuged at 1,500 rpm for 3 minutes scribed.29 Briefly, 30 g of live or frozen Blattela germanica were at 4°C, and then smears of BAL cells were prepared by cytocen- pulverized in liquid nitrogen. The sample was then defatted in trifugation (Thermo, MA, USA) at 1,000 rpm for 3 minutes. All 200 mL of 1:1 ethyl ether/ethyl acetate and extracted overnight smears were stained with a Hemacolor staining kit (Merck, with slow stirring at 4°C in phosphate-buffered saline (pH 7.4) Darmstadt, Germany). Differential cell counts in BAL cells were containing 6 mM β-mercaptoethanol and 1 mg/mL 1-phenyl- done for at least 200 leukocytes, using standard hemocytologic 3-(2-thiazolyl)-2-thiourea to prevent melanization.
Load more

Recommended publications
  • United States Patent (10) Patent No.: US 7,935,351 B2 Klinman Et Al
    US007935351B2 (12) United States Patent (10) Patent No.: US 7,935,351 B2 Klinman et al. (45) Date of Patent: *May 3, 2011 (54) USE OF CPG OLIGODEOXYNUCLEOTIDES 35. A 3. 2. RS:udolph s et alal. TO INDUCE ANGOGENESIS 5,288,509 A 2f1994 Potman et al. 5.488,039 A 1, 1996 M tal. (75) Inventors: Dennis M. Klinman, Potomac, MD 5.492.899 A 2, 1996 NE al. (US); Mei Zheng, Augusta, GA (US); 5,585,479 A 12/1996 Hoke et al. Barry T. Rouse, Knoxville, TN (US) 5,602,1095,591,721 A 2/19971/1997 MasorAgrawal et etal. al. 5,612,060 A 3, 1997 A1 d (73) Assignees: The United States of America as 5,614,191 A 3, 1997 al represented by the Department of 5,650,156 A 7/1997 Grinstaffet al. Health and Human Services, 5,663,153 A 9, 1997 Hutcerson et al. Washington, DC (US); University of 3. A RE SEE, Tennessee Research Foundation, 5,700,590 A 12/1997 Masoretal Knoxville, TN (US) 5,712.256 A 1/1998 Kulkarni et al. 5,723,335 A 3, 1998 Hutcerson et al. (*) Notice: Subject to any disclaimer, the term of this 3. 2. A 38. El cal patent is extended or adjusted under 35 5.840,705 A 11/1998 Tsukuda U.S.C. 154(b) by 761 days. 5,849,719 A 12, 1998 Carson et al. This patent is Subject to a terminal dis- 33: A 23: SEE claimer. 5,922,766 A 7/1999 Acosta et al.
    [Show full text]
  • Human Naive B Cells Cells by Cpg Oligodeoxynucleotide-Primed T+
    Presentation of Soluble Antigens to CD8+ T Cells by CpG Oligodeoxynucleotide-Primed Human Naive B Cells This information is current as Wei Jiang, Michael M. Lederman, Clifford V. Harding and of September 27, 2021. Scott F. Sieg J Immunol 2011; 186:2080-2086; Prepublished online 14 January 2011; doi: 10.4049/jimmunol.1001869 http://www.jimmunol.org/content/186/4/2080 Downloaded from References This article cites 36 articles, 21 of which you can access for free at: http://www.jimmunol.org/content/186/4/2080.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 27, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2011 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Presentation of Soluble Antigens to CD8+ T Cells by CpG Oligodeoxynucleotide-Primed Human Naive B Cells Wei Jiang,*,† Michael M. Lederman,*,† Clifford V. Harding,†,‡ and Scott F. Sieg*,† Naive B lymphocytes are generally thought to be poor APCs, and there is limited knowledge of their role in activation of CD8+ T cells.
    [Show full text]
  • Toll-Like Receptor 9 and 21 Have Different Ligand Recognition Profiles
    Toll-like receptor 9 and 21 have different ligand recognition profiles and cooperatively mediate activity of CpG-oligodeoxynucleotides in zebrafish Da-Wei Yeha,b,1, Yi-Ling Liua,1, Yin-Chiu Loc, Chiou-Hwa Yuhd, Guann-Yi Yuc, Jeng-Fan Loe, Yunping Luof, Rong Xiangg, and Tsung-Hsien Chuanga,h,2 aImmunology Research Center, National Health Research Institutes, Miaoli 35053, Taiwan; bInstitute of Molecular Medicine, National Tsing-Hua University, HsinChu 30013, Taiwan; cNational Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Miaoli 35053, Taiwan; dInstitute of Molecular and Genomic Medicine, National Health Research Institutes, Miaoli 35053, Taiwan; eInstitute of Oral Biology, National Yang-Ming University, Taipei 11221, Taiwan; fDepartment of Immunology, School of Basic Medicine, Peking Union Medical College, Beijing 100005, People’s Republic of China; gSchool of Medicine, University of Nankai, Tianjin 300071, People’s Republic of China; and hProgram in Environmental and Occupational Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan Edited by Ken J. Ishii, National Institute of Biomedical Innovation, Ibaraki, Japan, and accepted by the Editorial Board October 29, 2013 (received for review March 21, 2013) CpG-oligodeoxynucleotides (CpG-ODNs) are potent immune stim- containing a GTCGTT motif. In contrast, the GTCGTT motif uli currently under investigation as antimicrobial agents for containing CpG-ODN generates stronger immune responses in different species. Toll-like receptor (TLR) 9 and TLR21 are the humans and various domestic animals (8, 9). cellular receptors of CpG-ODN in mammals and chickens, respec- Toll-like receptors (TLRs) are pattern recognition recep- tively. The avian genomes lack TLR9, whereas mammalian tors that play crucial roles in the initiation of host defense genomes lack TLR21.
    [Show full text]
  • Oligodeoxynucleotide in Murine Asthma Anti-Inflammatory Effects Of
    In Vivo Role of p38 Mitogen-Activated Protein Kinase in Mediating the Anti-inflammatory Effects of CpG Oligodeoxynucleotide in Murine Asthma This information is current as of September 23, 2021. Barun K. Choudhury, James S. Wild, Rafeul Alam, Dennis M. Klinman, Istvan Boldogh, Nilesh Dharajiya, William J. Mileski and Sanjiv Sur J Immunol 2002; 169:5955-5961; ; doi: 10.4049/jimmunol.169.10.5955 Downloaded from http://www.jimmunol.org/content/169/10/5955 References This article cites 37 articles, 21 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/169/10/5955.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 23, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology In Vivo Role of p38 Mitogen-Activated Protein Kinase in Mediating the Anti-inflammatory Effects of CpG Oligodeoxynucleotide in Murine Asthma1 Barun K.
    [Show full text]
  • P020210615609549638679.Pdf
    The Journal of Immunology SCARB2/LIMP-2 Regulates IFN Production of Plasmacytoid Dendritic Cells by Mediating Endosomal Translocation of TLR9 and Nuclear Translocation of IRF7 Hao Guo,*,† Jialong Zhang,* Xuyuan Zhang,*,† Yanbing Wang,* Haisheng Yu,*,† Xiangyun Yin,*,† Jingyun Li,*,† Peishuang Du,* Joel Plumas,‡ Laurence Chaperot,‡ ,x,{ ,‖,# , Jianzhu Chen,* Lishan Su,* Yongjun Liu,* ** and Liguo Zhang* Downloaded from Scavenger receptor class B, member 2 (SCARB2) is essential for endosome biogenesis and reorganization and serves as a receptor for both b-glucocerebrosidase and enterovirus 71. However, little is known about its function in innate immune cells. In this study, we show that, among human peripheral blood cells, SCARB2 is most highly expressed in plasmacytoid dendritic cells (pDCs), and its expression is further upregulated by CpG oligodeoxynucleotide stimulation. Knockdown of SCARB2 in pDC cell line GEN2.2 dramatically reduces CpG-induced type I IFN production. Detailed studies reveal that SCARB2 localizes in late endosome/ http://www.jimmunol.org/ lysosome of pDCs, and knockdown of SCARB2 does not affect CpG oligodeoxynucleotide uptake but results in the retention of TLR9 in the endoplasmic reticulum and an impaired nuclear translocation of IFN regulatory factor 7. The IFN-I production by TLR7 ligand stimulation is also impaired by SCARB2 knockdown. However, SCARB2 is not essential for influenza virus or HSV-induced IFN-I production. These findings suggest that SCARB2 regulates TLR9-dependent IFN-I production of pDCs by mediating endosomal translocation of TLR9 and nuclear translocation of IFN regulatory factor 7. The Journal of Immunology, 2015, 194: 4737–4749. ysosomes are ubiquitous acid membrane-bound organelles which also includes scavenger receptor class B, member 1 involved in the degradation of molecules, complexes, and (SCARB1), and CD36 (5).
    [Show full text]
  • Screening of Novel Immunostimulatory Cpg Odns and Their Anti-Leukemic Effects As Immunoadjuvants of Tumor Vaccines in Murine Acute Lymphoblastic Leukemia
    519-529.qxd 20/12/2010 01:05 ÌÌ ™ÂÏ›‰·519 ONCOLOGY REPORTS 25: 519-529, 2011 519 Screening of novel immunostimulatory CpG ODNs and their anti-leukemic effects as immunoadjuvants of tumor vaccines in murine acute lymphoblastic leukemia JIN WANG, WANGGANG ZHANG, AILI HE, WANHONG ZHAO and XINGMEI CAO Department of Hematology, Second Affiliated Hospital, Medical School of Xi'an Jiaotong University, Xi'an 710004, Shaanxi Province, P.R. China Received July 29, 2010; Accepted October 11, 2010 DOI: 10.3892/or.2010.1093 Abstract. Acute lymphoblastic leukemia (ALL) is a common Additional therapeutic strategies are required in order to malignant disease and a major cause of mortality due to further prolong remission duration and eradicate minimal recurrent disease. Immunotherapy is a promising strategy for residual disease, ideally with less toxicity than conventional eradicating minimal residual disease and thus preventing the chemotherapy. One of the these approaches is active immuno- relapse of leukemia. Apart from stem cell transplantation, therapy with novel vaccination regimens. We previously CpG oligodeoxynucleotides (ODNs) are excellent candidates launched a phase-I clinical trial and evaluated the efficacy for the immunotherapy of leukemia. However, the number of and toxicity of vaccination in patients with relapsed or usable CpG ODNs is limited. In this study, we tested a panel refractory acute leukemia, which proved to be a feasible, of CpG ODNs and obtained three CpG ODN sequences with safe, and capable way of eliciting anti-leukemic responses strong immunostimulatory activity by comparing their in vivo (1). capacity to activate lymphocytes. The data revealed that the As tumor cells are considered to be poorly immunogenic, flanking bases, the spacing of individual CpG motifs and poly- it is difficult to elicit an effective specific anti-tumor activity guanosine ends, contribute to the immunostimulatory activity from these cells alone.
    [Show full text]
  • Oligodeoxynucleotide Dendritic Cells to the TLR9 Agonist Cpg Distinct
    Distinct Responses of Lung and Spleen Dendritic Cells to the TLR9 Agonist CpG Oligodeoxynucleotide This information is current as Li Chen, Meenakshi Arora, Manohar Yarlagadda, Timothy of September 24, 2021. B. Oriss, Nandini Krishnamoorthy, Anuradha Ray and Prabir Ray J Immunol 2006; 177:2373-2383; ; doi: 10.4049/jimmunol.177.4.2373 http://www.jimmunol.org/content/177/4/2373 Downloaded from References This article cites 62 articles, 32 of which you can access for free at: http://www.jimmunol.org/content/177/4/2373.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 24, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2006 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Distinct Responses of Lung and Spleen Dendritic Cells to the TLR9 Agonist CpG Oligodeoxynucleotide1 Li Chen,* Meenakshi Arora,* Manohar Yarlagadda,* Timothy B. Oriss,* Nandini Krishnamoorthy,* Anuradha Ray,*† and Prabir Ray2*† Dendritic cells (DCs) sense various components of invading pathogens via pattern recognition receptors such as TLRs.
    [Show full text]
  • Oligonucleotide-Ficoll Conjugate Nanoparticle Adjuvant for Enhanced Immunogenicity of Anthrax Protective Antigen † † † † † † Bob Milley,*, Radwan Kiwan, Gary S
    This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. Article pubs.acs.org/bc Optimization, Production, and Characterization of a CpG- Oligonucleotide-Ficoll Conjugate Nanoparticle Adjuvant for Enhanced Immunogenicity of Anthrax Protective Antigen † † † † † † Bob Milley,*, Radwan Kiwan, Gary S. Ott, Carlo Calacsan, Melissa Kachura, John D. Campbell, ‡ † Holger Kanzler, and Robert L. Coffman † Dynavax Technologies Corporation, 2929 Seventh Street, Suite 100, Berkeley, California 94710, United States ‡ MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland 20878, United States ABSTRACT: We have synthesized and characterized a novel phosphorothioate CpG oligodeoxynucleotide (CpG ODN)-Ficoll conjugated nanoparticulate adjuvant, termed DV230-Ficoll. This adjuvant was constructed from an amine-functionalized-Ficoll, a heterobifunctional linker (succinimidyl-[(N-maleimidopropionamido)- hexaethylene glycol] ester) and the CpG-ODN DV230. Herein, we describe the evaluation of the purity and reactivity of linkers of different lengths for CpG-ODN-Ficoll conjugation, optimization of linker coupling, and conjugation of thiol-functionalized CpG to maleimide- functionalized Ficoll and process scale-up. Physicochemical character- ization of independently produced lots of DV230-Ficoll reveal a bioconjugate with a particle size of approximately 50 nm and covalent attachment of more than 100 molecules of CpG per Ficoll. Solutions of purified DV230-Ficoll were stable for at least 12 months at frozen and refrigerated temperatures and stability was further enhanced in lyophilized form. Compared to nonconjugated monomeric DV230, the DV230-Ficoll conjugate demonstrated improved in vitro potency for induction of IFN-α from human peripheral blood mononuclear cells and induced higher titer neutralizing antibody responses against coadministered anthrax recombinant protective antigen in mice.
    [Show full text]
  • Oligodeoxynucleotide Dendritic Cells to the TLR9 Agonist Cpg Distinct
    Distinct Responses of Lung and Spleen Dendritic Cells to the TLR9 Agonist CpG Oligodeoxynucleotide This information is current as Li Chen, Meenakshi Arora, Manohar Yarlagadda, Timothy of September 23, 2021. B. Oriss, Nandini Krishnamoorthy, Anuradha Ray and Prabir Ray J Immunol 2006; 177:2373-2383; ; doi: 10.4049/jimmunol.177.4.2373 http://www.jimmunol.org/content/177/4/2373 Downloaded from References This article cites 62 articles, 32 of which you can access for free at: http://www.jimmunol.org/content/177/4/2373.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 23, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2006 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Distinct Responses of Lung and Spleen Dendritic Cells to the TLR9 Agonist CpG Oligodeoxynucleotide1 Li Chen,* Meenakshi Arora,* Manohar Yarlagadda,* Timothy B. Oriss,* Nandini Krishnamoorthy,* Anuradha Ray,*† and Prabir Ray2*† Dendritic cells (DCs) sense various components of invading pathogens via pattern recognition receptors such as TLRs.
    [Show full text]
  • Toll-Like Receptors As Potential Therapeutic Targets for Multiple Diseases
    REVIEWS TOLL-LIKE RECEPTORS AS POTENTIAL THERAPEUTIC TARGETS FOR MULTIPLE DISEASES Claudia Zuany-Amorim, John Hastewell and Christoph Walker The family of Toll-like receptors (TLRs) is receiving considerable attention as potential regulators and controllers of the immune response through their ability to recognize pathogen-associated molecular patterns. The discovery that endogenous ligands, as well as microbial components, are recognized by TLRs, and that small-molecular-mass synthetic compounds activate TLRs, raised interest in these receptors as potential targets for the development of new therapies for multiple diseases. In this review, we discuss the current and future use of TLR agonists or antagonists in chronic inflammatory diseases and highlight potential problems that are associated with such approaches. 2 INNATE IMMUNITY The immune system has been divided traditionally into PAMPs . The first report of a mammalian TLR and its The early response of a host to an INNATE and ADAPTIVE component, each of which has involvement in host defence — TLR-4 as a receptor for infections by pathogens, such as different roles and functions in defending the organism LIPOPOLYSACCHARIDE (LPS)3 — was followed rapidly by the bacteria and viruses, before the against foreign agents, such as bacteria or viruses. The discovery that the human genome contains several antigen-specific, adaptive innate immune system has developed a series of con- TLRs — ten have been found so far, TLR-1–TLR-10. immune response is induced. served receptors, known as pattern-recognition recep- Members of the TLR family share characteristic extra- ADAPTIVE IMMUNITY tors (PRRs), that recognize specific pathogen-associated cellular and cytoplasmic domains.
    [Show full text]
  • Toll-Like Receptors and Their Potential Roles in Kidney Disease
    J Am Soc Nephrol 15: 854–867, 2004 Signaling Danger: Toll-Like Receptors and their Potential Roles in Kidney Disease HANS-JOACHIM ANDERS,* BERNHARD BANAS,† and DETLEF SCHLO¨ NDORFF* *Nephrological Center, Medizinische Poliklinik, Klinikum der Universität München-Junenstadt, Germany; and †Klinik u. Poliklinik f. Innere Medizin II, Universität Regensburg, Germany. Abstract. Toll-like receptors (TLR) are an emerging family of cells are among the non-immune cells that express TLR1, -2, receptors that recognize pathogen-associated molecular pat- -3, -4, and -6, suggesting that these TLR might contribute to terns and promote the activation of leukocytes and intrinsic the activation of immune responses in tubulointerstitial injury renal cells. Ligands of the TLR include exogenous microbial (e.g., bacterial pyelonephritis, sepsis, and transplant nephrop- components such as LPS (TLR4), lipoproteins and peptidogly- athy). In addition, TLR9 has been shown to be involved in cans (TLR1, -2, -6), viral RNA (TLR3), bacterial and viral antigen-induced immune complex glomerulonephritis and lu- unmethylated cytosin-guanosin dinucleotide (CpG)-DNA pus nephritis by regulating humoral and cellular immune re- (TLR9), and endogenous molecules including heat-shock pro- sponses. TLR are evolutionary conserved regulators of innnate teins and extracellular matrix molecules. Upon stimulation, and adaptive immune reponses. It is likely that TLR are in- TLR induce expression of inflammatory cytokines or costimu- volved in many if not all types of renal inflammation. Here the latory molecules via the MyD88-dependent and MyD88-inde- authors provide an overview on the biology of TLR, summa- pendent signaling pathways shared with the interleukin-1 re- rize the present data on their expression in the kidney, and ceptors.
    [Show full text]
  • Therapeutic Administration of a Synthetic Cpg Oligodeoxynucleotide Triggers Formation of Anti-Cpg Antibodies
    Published OnlineFirst June 27, 2012; DOI: 10.1158/0008-5472.CAN-12-0257 Cancer Priority Report Research Therapeutic Administration of a Synthetic CpG Oligodeoxynucleotide Triggers Formation of Anti-CpG Antibodies Julia Karbach1, Antje Neumann1, Claudia Wahle1, Kathrin Brand1, Sacha Gnjatic2, and Elke Jager€ 1 Abstract The synthetic oligodeoxynucleotide CpG 7909, which contains unmethylated cytosine/guanine (CpG) motifs, has potent immunostimulatory effects when coadministered with NY-ESO-1 peptides or recombinant NY-ESO-1 protein, resulting in an enhanced cellular and humoral immune response against the vaccine antigen. In this study, we report the development of anti-CpG-ODN antibodies in 21 of 37 patients who received CpG 7909 either alone or as a vaccine adjuvant. Specific anti-CpG immunoglobulin G (IgG) antibody titers ranged from 1:400 to 1:100,000. The anti-CpG antibodies cross-reacted with other synthetic CpG-ODNs but not with the DNA of mixed bacterial vaccine and were shown to be phosphorothioate backbone specific. Vaccine-related severe side effects observed in some patients were most likely not related to the development of anti-CpG antibodies. In addition, anti-CpG antibodies did not have negative effects on the vaccine immune response. These results show that anti- CpG antibodies develop in humans against short unmethylated CpG dinucleotide sequences after administration of CpG 7909. Our data therefore substantiate the potency of CpG 7909 to directly stimulate human B-cells and suggest that anti-CpG antibody monitoring should be a part of ongoing and planned clinical trials with CpG- ODNs. Cancer Res; 72(17); 4304–10. Ó2012 AACR. Introduction act on B-cell and monocyte activation.
    [Show full text]