Investigation of the Intrinsic Antibody Dependent Enhancement Hypothesis By Matthew H. Quinn ...

209 Pages·2011·8.52 MB·English

Investigation of the Intrinsic Antibody Dependent Enhancement
Hypothesis

By

Matthew H. Quinn

Submitted in Partial Fulfillment

of the

Requirements for the Degree

Doctor of Philosophy

Supervised by

Xia Jin MD, PhD

Department of Microbiology and Immunology

School of Medicine and Dentistry

University of Rochester

Rochester, New York

2011
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Dedication

I would like to dedicate this thesis to my parents, Hugh and Anna
Quinn, for always loving me and being there for me, and Kim Minchella, for
always supporting me and being there when I get home. Thank you.
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Curriculum Vitae

Matthew Quinn was born in Rome, New York on May 23, 1983 to Hugh
and Anna Quinn. He attended Camden High School, and graduated as
Valedictorian in 2001. He next moved to Rochester, NY, where he attended
Rochester Institute of Technology. There, he was a member of the RIT
Honors Program, the Research Scholars Program, and conducted
undergraduate research with Maureen C. Ferran for three years involving the
fate of the IκB protein during VSV infection. In 2005, he was inducted into the
βββ national biological honor society, and graduated RIT with High Honors,
obtaining a Bachelor of Science degree in Biotechnology with a minor in
Criminal Justice. That same year, he began graduate studies at the
University of Rochester School of Medicine and Dentistry in the Department
of Microbiology and Immunology in the Virology track. His interests remain in
the field of virology, and he began his professional training in the laboratory of
Xia Jin in 2006. In 2008, Matthew completed his Master of Science degree in
the study of dengue virus and began his PhD work investigating dengue viral
burst size and how it changes during infection. His work was supported in
part by a Public Health Service pre-doctoral training grant, the Ruth L.
Kirschstein National Research Service Award.
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Publications

1. Quinn M., Chen H., Kou Z., Martinez-Sobrido L., Schlesinger JJ., Jin X.
Increased viral burst size during antibody dependent enhancement of
dengue infection is not linked to increased virus uptake or interferon
suppression. (Virology, Submitted)

2. Arevalo MT., Simpson-Haidaris PJ., Chen H., Quinn MH., Jin X.
Syndecans mediate dengue viral infection of human vascular endothelial
cells. (Submitted)

3. Kou Z., Lim JY., Beltramello M., Quinn M., Chen H., Liu SN., Martinez-
Sobrido L., Diamond MS., Schlesinger JJ., de Silva A., Sallusto .F, Jin X.
Human antibodies against dengue enhance dengue viral infectivity
without suppressing type I interferon secretion in primary human
monocytes. Virology. 2011 Feb; 5;410(1):240-7.

4. Block OK., Rodrigo WW., Quinn M., Jin X., Rose RC., Schlesinger JJ. A
tetravalent recombinant dengue domain III protein vaccine stimulates
neutralizing and enhancing antibodies in mice. Vaccine. 2010. Nov;
29;28(51):8085-94.

5. Kou Z., Quinn M., Chen H., Rodrigo WW., Rose RC., Schlesinger JJ.,
Jin X. Monocytes, but not T or B cells, are the principal target cells for
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dengue virus (DV) infection among human peripheral blood mononuclear
cells. J. Med Virol. 2008. Jan;80(1):134-46.

6. Blackley S., Kou Z., Chen H., Quinn M., Rose RC., Schlesinger JJ.,
Coppage M., Jin X. Primary human splenic macrophages, but not T or B
cells, are the principal target cells for dengue virus infection in vitro. J
Virology. 2007. Dec;81(24):13325-34.
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Acknowledgements

I am incredibly grateful to my advisor Xia Jin for allowing me to begin
my professional training in his laboratory at the University of Rochester. He
has been one of the most influential people in shaping who I have become
and how I think. Xia has been a strong mentor, continually forthcoming with
advice on how to proceed in my research, how to focus my thinking to make
the most logical and direct decisions, both with respect to my research and in
the broader more general decisions in my life. I greatly appreciate the effort
he has put in to helping me throughout my graduate training. He has been
available and accessible morning through evening nearly every day, including
Saturdays, for advice and discussions. I very much appreciate all the hours
spent reading and rereading my documents and discussing results. The
approach Xia takes is a direct one. He is not afraid to tell you how he feels
and what he thinks, and that way of thinking, without room for excess niceties
or sugar coating, translates into research as precise, clearly thought out,
ordered experimental design and planning. This has helped me immensely
as I have prepared for talks, written and rewritten my paper, and planned out
experiments. For guidance and shaping at such a fundamental level, I will be
eternally grateful.
I am also very grateful to my committee members, Brian Ward, Jack
Schlesinger, Luis Martinez-Sobrido, and Toru Takimoto, for helping to guide
and shape my research project as much as they have been able to. I wish
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that we had been able to meet more frequently so that I could have had more
access to their criticisms, comments, and suggestions. Having done a
laboratory rotation in Brian’s lab, I knew that I needed him on my committee
because he is a very careful and thorough scientist who seems to always
asks probing questions that have helped me forge a deeper understanding of
the issues at hand. His inquisitive nature and attention to detail have proven
to be very important to me throughout the rest of my time at the university.
Jack has been an integral part of the unofficial dengue group since before I
began here. I have been very fortunate to interact with him from the
beginning of graduate school, during the dengue meetings that were held
when I rotated in Bob Rose’s lab. Jack has been a wealth of information,
support, and supplies, and has been immeasurable for my dengue research,
but more than that, he has always been encouraging and helpful, and this
work couldn’t have been done without him. I must thank Luis for joining my
committee after arriving here in the department. Luis has been helping me
since before he even arrived here, providing new tools, ideas, and support
every time we talk. He asks fundamental questions, and always seems to
know the most appropriate way to get at the desired answers. He has
brought a level of sophistication and excitement to virology, and even in the
relatively brief amount of time we have interacted, has set for me a goal that I
strive to achieve. Toru has been available and been helpful whenever I
needed it. He has provided clear comments and always been thorough and
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open with his comments and suggestions, which have continually served to
tighten and strengthen my project and focus me on the most important issues.
I would like to thank the current and past members of the Jin lab. At all
times Huiyuan has been helpful and efficient and the lab would not run
anywhere nearly as smoothly as it does without her keeping things in order.
Maria has been invaluable by sharing thoughts, protocols, reagents, and
study time as we prepared for the qualifying exam, talks, and made posters.
A large thank you to Kou for her efficiency, for teaching me Flow cytometry
and for all her help in planning and designing experiments. Theresa and
Shanley truly made the lab an enjoyable place to be in addition to helping me
whenever I needed it, and I do miss having a lab mate who appreciated the
temperature nice and low so I could work most efficiently. Dani was always
around when I entered Xia’s lab, is incredibly open, friendly, and easy to talk
to, and continues to help me even now. Many many thanks to Shanaka. No
one else has been as eager to help. Every time we talk, he is encouraging,
and is a fountain of help, knowledge, advice, and emotion. He is the gold
standard of productivity, and the place where I always set my initial sights.
Olivia, who was the first to help me during my first rotation. She is a true
friend and her initial help with primer design led to much more eventual help
with the many obstacles and checkpoints throughout graduate school, and
has continued to post-graduate and life decisions. I cannot express how
much you have all helped me.
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I would like to thank everyone else in the Microbiology and
Immunology department who has helped me, either directly with experiments,
or just in discussion of data, protocols, or other life issues. I would especially
like to thank Cristina for her help with fluorescent microscopy and Oksana for
help with real-time PCR. Kendra, Vinnie, Emily, and Ken have also been
helpful for additional troubleshooting and general discussions about graduate
school, science, life, and the future.
I would like to thank Ken, Vinnie, Vadim, Felix, Bess, Olivia, and Maria
for being good friends; for helping me when I needed it, and distracting me
when that was what was needed. Thank you to the members of the
departmental kickball, softball, and volleyball teams, as well as all of my
friends outside of work for providing me with support and friendship, and
helping to keep me balanced and sane during the more stressful times in the
past years.
I would like to thank everyone in the Infectious Disease Division for
housing me and helping things to run smoothly. I have basically lived in the
IDD these past years, and I have to thank Beverly, Wendy, Donna, and
Veronica for maintaining order in the IDD offices and helping me with faxing
and room reservations, and having a person to talk to when necessary.
Thank you to everyone who attended the IDD research conference talks, and
listened and commented on my talks. John, William, Carrie, Ed, and everyone
else were extremely helpful in bringing up new questions and continually
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helping to refine my approach and direction. I would like to especially thank
John for always being attentive and inquisitive and having a smile at the
ready and Carrie for having a kind word and additional comments and
suggestions.
Finally, I would like to thank all my friends who I have grown apart from
due to long hours and weekends in the lab, my family and Kim’s family for
always making me feel welcome and safe, and everyone else that I wasn’t
able to mention here. Thank you.