Snímek 1

Snímek 1

About OMICS Group OMICS Group International is an amalgamation of Open Access publications and worldwide international science conferences and events. Established in the year 2007 with the sole aim of making the information on Sciences and technology Open Access, OMICS Group publishes 400 online open access scholarly journals in all aspects of Science, Engineering, Management and Technology journals. OMICS Group has been instrumental in taking the knowledge on Science & technology to the doorsteps of ordinary men and women. Research Scholars, Students, Libraries, Educational Institutions, Research centers and the industry are main stakeholders that benefitted greatly from this knowledge dissemination. OMICS Group also organizes 300 International conferences annually across the globe, where knowledge transfer takes place through debates, round table discussions, poster presentations, workshops, symposia and exhibitions. About OMICS Group Conferences OMICS Group International is a pioneer and leading science event organizer, which publishes around 400 open access journals and conducts over 300 Medical, Clinical, Engineering, Life Sciences, Phrama scientific conferences all over the globe annually with the support of more than 1000 scientific associations and 30,000 editorial board members and 3.5 million followers to its credit. OMICS Group has organized 500 conferences, workshops and national symposiums across the major cities including San Francisco, Las Vegas, San Antonio, Omaha, Orlando, Raleigh, Santa Clara, Chicago, Philadelphia, Baltimore, United Kingdom, Valencia, Dubai, Beijing, Hyderabad, Bengaluru and Mumbai. The cellular basis of protective immunity against experimental infection caused by Francisella tularensis Kubelkova K., Orlikova A., Krocova Z., Pejchal J., Macela A., Stulik J. Faculty of Military Health Sciences, University of Defense, 500 01 Hradec Kralove, Czech Republic 3 Francisella tularensis Francisellae facultative intracellular bacterial pathogens Small, nonmotile, obligate anaerobe One of the most infectious bacterial agens (10 CFU) Francisella proliferates inside macrophages, neutrophils, dendritic cells and hepatocytes

Geographic distribution of four existing Francisella tularensis subtypes (holarctica Type B, tularensis Type A1 and A2, mediasiatica, novicida) Tularemia Zoonotic infection Vectors mainly ticks and mosquitoes Broad spectrum of clinical manifestations with dominant symptoms granulomas and secondary atypical pneumonia Treatment ATB (Gen, Tet) The cellular basis of protective immunity against experimental infection caused by Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic Francisella tularensis 4 Kubelkova K. and Macela A.: Putting Jigsaw Together - A Brief Insight Into the Tularemia, Open Life Sciences, 2015, Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic Ready to publish. 5 B cell involvement B cells and antibodies are necessary for mice to develop their natural resistance to primary and secondary LVS infections .The role of antibodies in the protection against intracellular pathogen F. tularensis still remains poorly understood ! Extracellular phase in the host, which makes it accessible to humoral immune responses Ab responses containing both Th-1 and Th-2 antibody isotypes are detectable as early as 3 days following i.d. infection Confer early as well as long term immunity Immune response against LPS (as a major protective antigen) No naturally B cell-deficient murine strain has been identified yet Serum Ab against bacterial proteins FopA, OmpA, DsbA, GroEL, KatG etc. Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 6 The role of antibodies in protective immune response

Traditional view Antibodies have little (if any) protective role against tularemia Late 1970ies Antibodies can confer protection against attenuated Francisella tularensis strains and can confer some degree of protection against virulent strains of holarctica subtype /Macela A.: Thesis, 1980. Late 1990ies B-cells but not circulating antibodies are indispensable in protective immunity against tularemia /Elkins K.et al: Infect Immun.,1999. New millennium: Passive transfer of immunity protects against the same subtype Passive transfer of immunity against tularemia is possible Antibody-dependent cell-mediated cytotoxicity (ADCC) / Fulop M. et al: Vaccine, 2001. , Stenmark S. et al: Microb Pathog., 2003., Sanapala et al. 2012, Kubelkova Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic K. et al. Microbial Pathogen, 2012 7 The role of antibodies in protective immune response Balb/c mice Cobalt 60

Dose of 4 Gy Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 8 Fenotypization of spleen cells of immunosuprimmised Balb/c mice CD3+ CD8+ T cells 104 R7 104 R8 PE-Cy7 Log Comp PE-Cy7 Log Comp 103 103 102 102 101 100 100 R9 R8 R9 R10 101 R10

101 102 103 PE Log Comp R7 104 Obr..3: CD3+CD8+T lymfocyty 100 100 101 102 103 PE Log Comp 104 Passive transfer of antibodies protects irradiated mice against F. tularensis holarctica LVS infection Co60 (4 Gy) Immune or normal sera (0.2 ml serum, i.p.) 48 hrs Francisella tularensis LVS (1.3 x 102 live microbes, s.c.) 2 hrs 21 days Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 10 Passive transfer of immunity protects irradiated mice against primary as well as secondary F. tularensis infection Co60 (4 Gy) Immune or normal sera (0.2 ml serum, i.p.)

3 days Francisella tularensis LVS (1.3 x 102 live microbes, s.c.) 2 hrs 22 days 21 days Francisella tularensis SchuS4 (1.2 x 102 live microbes, s.c.) Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 11 Live microbes induce protective Abs in immunosuprised individuals Irradiation 1KL Number of mice 10 2KL 10 - 3KL 10 - 4KL 10 -

5KL 10 - 6KL 10 - 7KL 10 - Interval of immunization 2 hod. before infection 2 hod. before infection 2 hod. before infection 2 hod. before infection 2 hod. before infection 2 hod. before infection 2 hod. before - Route immun. i.p. Immunization [200ul] Ab 4Gy+LVS i.p.

F. tularensis LVS Dose LVS [200ul] 106 bb/mouse Route infect. s.c. Ab 4Gy+LVS F. tularensis LVS 107 bb/mouse s.c. i.p. Ab 4Gy+LVS F. tularensis LVS 108 bb/mouse s.c. i.p. Ab 4Gy+LVS F. tularensis LVS 109 bb/mouse s.c. i.p. PBS F. tularensis LVS 107 bb/mouse

s.c. i.p. PBS F. tularensis LVS 108 bb/mouse s.c. i.p. PBS F. tularensis LVS 109 bb/mouse s.c. infection Percent survival 100 Infection 10 6 LVS, Ab 2h 10 7 LVS, Ab 2h 10 8 LVS, Ab 2h 10 9 LVS, Ab 2h 80 60 40 10 7 LVS, PBS 10 8 LVS, PBS 10 9 LVS, PBS 20 0

0 3 6 9 12 Time interval 15 18 21 Live microbes induce protective Abs in immunosuprised individuals Number of mice Irradiation Immunization [200ul] Interval of immunization Route/ immun. Infection Dose LVS [200ul] Route/infect. 1B 10 4 Gy

Ab 4Gy+LVS 2 hod. before infection i.p. F. tularensis LVS 102 CFU/mouse s.c. 2B 10 4 Gy Ab 4Gy+LVS 2 hod. before infection i.p. F. tularensis LVS 3B 10 4 Gy Ab 4Gy+LVS 2 hod. before infection i.p. F. tularensis LVS 104CFU/mouse s.c.

4B 10 4 Gy PBS 2 hod. before infection i.p. F. tularensis LVS 102 CFU/mouse s.c. 5B 10 4 Gy PBS 2 hod. before infection i.p. F. tularensis LVS 102 CFU/mouse Percent survival 100 10 2 LVS, Ab 2h 10 3 LVS, Ab 2h 10 4 LVS, Ab 2h 80

60 10 2 LVS, PBS 10 3 LVS, PBS 40 20 0 0 3 6 9 12 Time interval 15 18 21 103CFU/mouse s.c. s.c. More efficient protection - using Ab 4Gy+LVS in irradiated animals Live microbes induce protective Abs in immunosuprised individuals Number of mice Gy Immunization

Time Route/ immun. Infection Route/inf. Dose LVS [200ul] 1A 5 - Ab 4Gy+LVS 2 hod. before infection i.p. F. tularensis LVS s.c. 108 CFU/mouse 2A 5 - Ab 4Gy+LVS 2 hod. before infection i.p. F. tularensis

LVS s.c. 3A 5 4 Gy Ab 4Gy+LVS 2 hod. before infection i.p. F. tularensis LVS s.c. 4A 5 4 Gy Ab 4Gy+LVS 2 hod. before infection i.p. F. tularensis LVS s.c. 5A 5 4 Gy

PBS 2 hod. before infection i.p. F. tularensis LVS s.c. 109CFU/mouse 103 CFU/mouse 104 CFU/mouse 101 CFU/mouse Secondary infection with hypervirulent F. tularensis SchuS4 strain Number of mice Infection Route of infection Dose SchuS4 [200ul] Protection 1A 5 F. tularensis SchuS4 s.c. 102 CFU/my 100% 2A 5 F. tularensis SchuS4

s.c. 102 CFU/my 100% 3A 5 F. tularensis SchuS4 s.c. 10 CFU/my 100 % 4A 5 F. tularensis SchuS4 s.c. 102 CFU/my 100 % 5A 5 F. tularensis SchuS4 s.c. 102 CFU/my 0% 2 Cytokine profile of immunocompromised mice

The description of cytokine changes as a factor of importance during Francisella infection in nave and immunocompromised mice ELISA kits (Invitrogen) IL-1b, IL-4, IL-6, TNF-a, IFN-g Passive transfer of immunity rather normalize the cytokine levels Dominant disproportion exists in the levels of IFN-g in blood and tissue homogenates, which suggests the high consumption of this cytokine in the sites of production Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 15 Identification of immunoreactive Francisella proteins GPS Explorer Software v. 3.6 (Applied Biosystems), which integrates the Mascot search algorithm against F. tularensis LVS genome databases http:// www.ncbi.nlm.nih.gov/COG/old/xognitor.ht ml http://www.cbs.dtu.dk/services/SignalP http://www.psort.org/psortb/ Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 16 Identification of immunoreactive Francisella proteins Signal trans duction mechanisms TCell divis ion and chromos ome partitioning D- Trans cription K-

Secondary metabolites biosynthes is, transport and catabolis m Q- DNA replication, recombination and repair Ltrans lation, ribosomal s tructure and biogenes is J- General function prediction only R- aminoacid transport and metabolism Ecoenzyme metabolism H- No related COG cell envelope biogenes is , outer membrane M- function unknown S- Nucleotide transport and metabolis m F- pos ttranslation modification, protein turnover, chaperones O- inorganic ion trans port and metabolism Plipid metabolism I- energy production and conversion C- carbohydrate transport and metabolis m G- http://www.ncbi.nlm.nih.gov/COG/old/xognitor.html Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic

17 Conserved hypothetical proteins: FTT0086 FTT0848 FTT0655 The result applications Changing the vaccine strategy Important role of circulating antibodies during the interaction of F. tularensis with host immunoregulatory system Passively protected mice were also able to survive primary LVS and the subsequent challenge with the hypervirulent strain F. tularensis SchuS4 Functional passive immunization protocol for nave, as well as immunocompromised animals Cytokine production of immunocompromised mice has been characterized as a part the host response to Francisella infection Combination of passive transfer of antibodies and subsequent active immunization represents the safety way to protective immunity against tularemia New immunoreactive proteins monoclonal Abs Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 19 With thanks to colleagues and collaborators

UNIVERSITY OF DEFENCE Macela Ales Stulik Jiri Krocova Zuzana Safarova Maria Zakova Jitka FDA, CBER Elkins Karen DePascalis Roberto Kurtz Sherry Kubelkova K., Krocova Z., Balonova L., Stulik J., Macela A.: Specific antibodies protect gamma-irradiated mice against Francisella tularensis strain 15 and live vaccine strain (LVS) infection. Microbial Pathogenesis, 2012, 53, 259-268. Kubelkova K., Krocova Z., Plzakova L., Macela A.: The Role of B cells in Intracellular Bacterial Pathogen Infection, B cells: Molecular Biology, Developmental Origin and Impact on the Immune Systm. 2013, ISBN: 978-1-62808-541-9, p.1-44. Plzakova L., Kubelkova K., Krocova Z., Zarybnicka L., Sinkorova Z., Macela A.: B cell subsets are activated and produce cytokines in the course of early phases of Francisella tularensis LVS infection. Microbial Pathogenesis, 2014, http://dx.doi.org/10.1016/j.micpath.2014.08.009. Kubelkova K. and Macela A.: Putting Jigsaw Together - A Brief Insight Into the Tularemia, Open Life Sciences, 2015, Ready to publish. This work was supported by the Grant Long-term organization plan 1011 received from the Czech Ministry of Education, Youth and Sports and by Grant No. P302/11/1631 from the Czech Science Foundation. Centre of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic 20 Let Us Meet Again We welcome you all to our future conferences of OMICS Group International Please Visit: www.omicsgroup.com www.conferenceseries.com

Recently Viewed Presentations

  • Literacy Irrational Rational Denominator Factors Research Why are

    Literacy Irrational Rational Denominator Factors Research Why are

    3. Rationalise the Denominator. a) b) c) Irrational. Rational. Denominator. Factors. Rationalise the denominator - no surds. Why are irrational numbers called surds? A Sequence of numbers is:-What are the next 3 terms? Does this triangle have a right angle
  • Trusts V Foundations the Cyprus International Trust Option

    Trusts V Foundations the Cyprus International Trust Option

    THE CYPRUS INTERNATIONAL TRUST OPTION BY PHANI SCHIZA ANTONIOU Tel Aviv Conference 24/2/2011 SUCCESSFUL BUSINESS PEOPLE AND WEALTHY FAMILIES MAY POSSESS VALUABLE ASSETS WORLDWIDE NEED FOR EFFICIENT STRUCTURE TO HOLD, PROTECT AND MANAGE THEM TO RESPOND TO THE DIFFERENT SYSTEMS...
  • Presentation Title Goes Here - Our Mission

    Presentation Title Goes Here - Our Mission

    Cheminformatics for Computational Chemistry and Computer-Aided Molecular Discovery R. S. Pearlman1, Y. Wu1, K. M. Smith2, and B. B. Masek2 1 Laboratory for the Development of CADD Software, College of Pharmacy, University of Texas, Austin TX
  • Nessun titolo diapositiva - AISV

    Nessun titolo diapositiva - AISV

    Nelle parole, significativa e stabile frequenza delle fricative e sono ben rappresentate le occlusive e le laterali. Nell'italiano infantile prevalgono le nasali e le liquide; discreta anche la presenza di affricate. Nelle prime parole dei bambini americani c'è una prevalenza...
  • Invertebrate Zoology Lecture 17: Phylum Arthropoda, Part 1

    Invertebrate Zoology Lecture 17: Phylum Arthropoda, Part 1

    Intermediate phyla Why considered intermediate? Phylum Onychophora Phylum Tardigrada Bauplan basics Other features Tagmatization Paired jointed appendages "Primitive condition": appendages associated with each segment, Lack motile cilia, except for some sperm Why? Which other group has non-motile cilia?
  • Regulation of Import Competition and Unfair Trade

    Regulation of Import Competition and Unfair Trade

    2000 Congress granted permanent normal trade status to China effective on China's admission to WTO. China admitted to WTO - Dec. 2001. Subject to political events and tensions . Concern over Taiwan - admitted to WTO as part of "Separate...
  • STAGES IN COST REDUCTION Dennis Geyer Geyer Management

    STAGES IN COST REDUCTION Dennis Geyer Geyer Management

    It seeks to improve the relationship among supply chain members to enhance competitiveness. Costs are organized by links in the value chain. Reductions achieved by consolidation of links, sharing information, better coordination, and exploiting synergies among supply chain members.
  • TDW Positive Material Identification Kenny Greene August 20,

    TDW Positive Material Identification Kenny Greene August 20,

    AUT B-scanner (C-Scan display) Automated Ball Indention (ABI) Optical Emissions Spectrometry (OES) Magnetic Particle Testing (MT) These five NDE methods make up the PMI process. During this process we use UTT to verify Actual Wall Thickness (AWT).