Th17-associated immune responses are required for resolution of Staphylococcus aureus nasal carriage Item Type dissertation Authors Archer, Nate Publication Date 2013 Abstract The anterior nares of humans are the major reservoir for Staphylococcus aureus colonization. Approximately 20% of the healthy human population is persistently and 80% intermittently colonized with S. aureus in the nasal cavity. Previous studies have ... Keywords IL-17; nasal carriage; Immune response; Interleukin-17; Th17 Cells; Staphylococcus aureus Download date 12/02/2023 01:11:38 Link to Item http://hdl.handle.net/10713/2971 Nathan K Archer, PhD Candidate School Address: Department of Microbial Pathogenesis School of Dentistry – 8th Floor South University of Maryland-Baltimore 650 W. Baltimore Street, Baltimore, Maryland, U.S.A. 21201 Tel. (410) 706-2263 Email: [email protected] EDUCATION Ph.D. Candidate in Microbiology and Immunology Expected graduation 2013 University of Maryland--Baltimore, Baltimore, MD, USA Advisor: Mark Shirtliff, Ph.D. August 2008 - B.S. University of Wisconsin - Madison, WI, USA Major: Biochemistry RESEARCH INTERESTS Nasal colonization, host-pathogen interactions, mucosal immunity, 2D electrophoresis, differential cytokine expression, biofilms, antimicrobial peptide production RELATED WORK EXPERIENCE Graduate Student and Research Assistant Baltimore, MD GPILS, Dept. of Microbiology and Immunology Sept 2008--Present Dental School, Dept. of Microbial Pathogenesis University of Maryland--Baltimore Advisor: Mark Shirtliff, PhD Independent Research Project Madison, WI University of Wisconsin - Madison Aug 2007-Aug 2008 Supervisor: Alan Rapraeger, PhD PROFESSIONAL ACTIVITIES Membership in Professional Organizations: 1. American Association for the Advancement of Science, 2009 – present. 2. American Society for Microbiology, 2009 – present. Posters 1. “Defining the Immune Response to Staphylococcus aureus Nasal Colonization”. 111th ASM General Meeting, New Orleans, LA, May 22, 2011. 2. “Defining the Immune Response to Staphylococcus aureus Nasal Colonization”. Maryland Branch ASM Dinner Meeting, Baltimore, MD, May 11, 2011. 3. “Effect of Th1 and Th2 mediated immune responses on Staphylococcus aureus biofilm infection”. International Conference on Gram-Positive Pathogens. Omaha, NE, October 10, 2010. 4. “A Th2-Mediated Immune Response Reduces Colonization of S. aureus on the Nasal Epithelium of Mice”. 110th ASM General Meeting. San Diego, CA, May 23, 2010. Presentations 1. “Clearance of Staphylococcus aureus Carriage is T-cell Mediated, B-cell Independent, and Relies on IL-17A”. 15th International Symposium on Staphylococci and Staphylococcal Infections, Lyon, France, August 27th, 2012. 2. “Defining the Immune Response to Staphylococcus aureus Nasal Colonization”. 34th Annual Graduate Research Conference, Baltimore, MD, April 5th, 2012. 3. “Staphylococcus aureus Nasal Colonization and the Host Immune Response”. 33rd Annual Graduate Research Conference, Baltimore, MD, April 7th, 2011. Local and National Service 1. Ad Hoc Reviewer – Antimicrobial Agents and Chemotherapy, 2013 – present. 2. Ad Hoc Reviewer – Biofouling, 2012 – present 3. Ad Hoc Reviewer – Annals of the New York Academy of Sciences , 2012 – present. SERVICE AND COMMITTEE ASSIGNMENTS 1. Campus Student Recruitment - Univ. of MD – Baltimore 2008-2012. 2. GSA Microbiology and Immunology Student Representative – Univ. of MD – Baltimore 2009-2010. 3. Speaker Invitation Committee Leader (Bacteriology) – Univ. of MD – Baltimore 2010-2012. 4. Microbial Pathogenesis Journal Club Organizer – Univ. of MD – Baltimore 2011- 2012. AWARDS 1. Best Oral Presentation Award. “Clearance of Staphylococcus aureus Carriage is T- cell Mediated, B-cell Independent, and Relies on IL-17A”. 15th International Symposium on Staphylococci and Staphylococcal Infections, Lyon, France, August 27th, 2012. PUBLICATIONS Peer-reviewed Journal Articles 1. Archer NK, Harro JM, Shirtliff ME. “Clearance of Staphylococcus aureus nasal carriage is T-cell dependent and mediated through IL-17A expression and neutrophil influx.” Infection and Immunity (2013). Print. 2. Archer NK, Mazaitis MJ, Costerton JW, Leid JG, Powers ME, Shirtliff ME. “Staphylococcus aureus biofilms: properties, regulation, and roles in human disease.” Virulence 2 (2011): 445-59. Print. 3. Harro, Janette M., Brian M. Peters, Graeme A. O'May, Nathan Archer, Patrick Kerns, Ranjani Prabhakara, and Mark E. Shirtliff. "Vaccine Development in Staphylococcus aureus: Taking the Biofilm Phenotype into Consideration." FEMS Immunology and Medical Microbiology 59 (2010): 306-23. Print. Book Chapters 1. Archer NK and Harro J, and Shirtliff ME. Utilizing the host immune response to battle biofilms. In Antibiofilm Agents: From Diagnosis to Treatment and Prevention. Ed. Kendra P. Rumbaugh and Iqbal Ahmad. Springer Series on Biofilms. 2013. - in preparation. 2. Archer NK, Leid JG, Costerton JW, and Shirtliff ME. Immunological methods for Staphylococcus aureus infection diagnosis and prevention. In Culture Negative Orthopedic Infections. Ed. DeMeo PJ, Costerton JW, Ehrlich GD, and Winkler H. Springer Series on Biofilms. Springer-Verlag Berlin Heidelberg. 2012. – in press. Published Abstracts and Symposia: 1. N. Archer and M.E. Shirtliff. “Clearance of Staphylococcus aureus Carriage is T- cell Mediated, B-cell Independent, and Relies on IL-17A”. 15th International Symposium on Staphylococci and Staphylococcal Infections, Lyon, France, August 27th, 2012. 2. N. Archer and M.E. Shirtliff. “Defining the Immune Response to Staphylococcus aureus Nasal Colonization”. 34th Annual Graduate Research Conference, Baltimore, MD, April 5th, 2012. 3. N. Archer and M.E. Shirtliff. “Defining the Immune Response to Staphylococcus aureus Nasal Colonization”. 111th ASM General Meeting, New Orleans, LA, May 22nd, 2011. 4. N. Archer and M.E. Shirtliff. “Staphylococcus aureus Nasal Colonization and the Host Immune Response”. 33rd Annual Graduate Research Conference, Baltimore, MD, April 7th, 2011. 5. Ranjani Prabhakara, Janette M. Harro, Megan L. Harris, Nate Archer, Jeff G. Leid, J. William Costerton, and Mark E. Shirtliff. “Effect of Th1 and Th2 mediated immune responses on Staphylococcus aureus biofilm infection”. International Conference on Gram-Positive Pathogens. Omaha, NE, October 10, 2010. 6. N. Archer and M.E. Shirtliff. “A Th2-Mediated Immune Response Reduces Colonization of S. aureus on the Nasal Epithelium of Mice”. 110th ASM General Meeting. San Diego, CA, May 23, 2010. FUNDING 1. T32 Training Program in Oral and Craniofacial Biology. May 1, 2011 – Present. Abstract Title of Dissertation: Th17-associated immune responses are required for resolution of Staphylococcus aureus nasal carriage Nathan Archer, Doctor of Philosophy, 2013 Dissertation Directed by: Mark E. Shirtliff, Ph.D., Professor, Department of Microbial Pathogenesis The anterior nares of humans are the major reservoir for Staphylococcus aureus colonization. Approximately 20% of the healthy human population is persistently and 80% intermittently colonized with S. aureus in the nasal cavity. Previous studies have shown a strong causal connection between S. aureus carriage and increased risk of nosocomial infection, as well as increased carriage due to immune dysfunction. However, the immune responses that permit persistence or mediate clearance are undefined. We developed a carriage model in C57BL/6J mice and showed that clearance begins 14 days post-inoculation. In contrast, SCID mice that have a deficient adaptive immune response are unable to eliminate S. aureus even after 28 days post-inoculation. Furthermore, decolonization was found to be T-cell mediated, but B-cell independent by evaluating carriage clearance in TCR-β/δ KO and IgH-µ KO mice, respectively. Up-regulation of IL-1β, KC, IL-17A and IL-17F occurred following inoculation with intra-nasal S. aureus. IL-17A production was crucial for clearance since IL-17A-deficient mice were unable to eliminate S. aureus carriage. In addition, inoculation of IL-17A/F KO mice displayed a significant inability to control S. aureus growth in the nares. Subsequently, cell differential counts were evaluated from nasal lavage fluid obtained from wild type and IL-17A-deficient colonized mice. These counts displayed IL-17A-dependent neutrophil migration. Antibody-mediated depletion of neutrophils in colonized mice caused reduced clearance compared to isotype treated controls. Th17-associated responses are reported to induce antimicrobial peptide production at epithelial surfaces. Therefore, we utilized RT- PCR to determine nasal tissue expression following inoculation with S. aureus of three antimicrobial peptides with anti-staphylococcal activity, mouse CRAMP, β-defensin-3 (mBD-3), and β-defensin-14 (mBD-14). We elucidated an IL-17A-dependent up- regulation of antimicrobial peptides post-S. aureus inoculation, and enhanced nasal tissue expression of mouse β-defensin-3 upon IL-17A stimulation. Additionally, we discovered that ex-vivo nasal tissue supernatants have anti-staphylococcal activity that is solute- dependent and heat-sensitive. Our data suggest that the Th17-associated immune response is required for nasal decolonization. This response is T-cell dependent, mediated via IL-17F and IL-17A production, neutrophil influx, and potentially antimicrobial peptide induction. Th17-associated immune responses may be targeted for strategies to mitigate distal infections originating from persistent S. aureus carriage in humans. Th17-associated immune responses are required for resolution of Staphylococcus aureus nasal carriage By Nathan Archer Dissertation submitted to the Faculty of the Graduate School of the University of Maryland, Baltimore in partial fulfillment of the requirements for the degree of Doctor of Philosophy 2013 ©Copyright 2010 by Nathan Archer All Rights Reserved Dedication This dissertation is dedicated to my family for teaching me the values of hard work, discipline, and pride that were necessary to achieve this level of education. Their focus to help me pursue higher education has forever changed the course of my life and I will be eternally grateful to them for it. This dissertation is also dedicated to my girlfriend, Migena, who has been a sounding board for new ideas, a shoulder to lean on during the tough times and there to celebrate with during the good times. I want to thank you all for your support, comfort, and love. iii