The Innate and Adaptive Immune Response
to Measles Virus
A thesis submitted to Johns Hopkins University in conformity
with the requirements for the degree of Master of Science.
All Rights Reserved
Measles is one of the most important causes of childhood morbidity
and mortality worldwide. Although a vaccine is available, the high
transmission rate of measles virus requires population of 95% to interrupt its
transmission. The World Health Organization and the United Nations
Childrens Fund recommend that children that develop measles receive
vitamin A supplementation, as a safe, cheap, and efficacious way to reduce
the burden of disease. Due to differences between strains and confounding
data of measles stocks contaminated with defective interfering RNA
particles, the immune response to measles virus infection has not been well
defined. Furthermore, the mechanism by which vitamin A protects against
severe measles-induced disease is unknown.
In this thesis, I investigate the innate and adaptive immune response
to measles virus infection. Measles virus strains were purified of defective
interfering RNA particles and used for in vitro infections of monocyte derived
dendritic cells. Gene expression changes of interferon-stimulated genes and
viral stress-induced genes, IFIT1 and Mx1, were upregulated in response to
infection with the Edmonston measles virus vaccine strain, as well as the
wild-type strains of Bilthoven, IC-B, and C- and V-protein knockout strains,
as compared to mock infected cells. Unexpectedly, there were no differences
between transcript levels of these genes between C and V protein knockout
strains and the respective wild-type infection. Additionally, the absence of
type I interferon production supports the theory that measles virus induces
the transcription of these genes through the viral stress-induced pathway,
and not the interferon-stimulated pathway.
While a previous study had detected measles virus-specific IL-17-
producing T cells in measles virus-infected rhesus macaques, the Th17
response to measles virus has not been characterized. Th17 cell
differentiation was inhibited early after measles virus infection in vitro.
There was a significant decrease in IL-23A transcript ts and a significant
increase in IL-27 transcripts, both of which affect Th17 cell differentiation
negatively. However, in a rhesus macaque model of infection, a biphasic Th17
response was observed with peaks at days 18 and 56.
The effects of vitamin A supplementation following measles virus
infection on the immune response was explored in a rhesus macaque model
using supplemented and non-supplemented groups. While some data has yet
to be explored, major differences were not observed between the two groups
up to three months following infection, in regards to clearance of infectious
virus, immune cell composition, or immune cell function. Archived data will
elucidate the role of vitamin A in measles virus RNA persistence, and Th1
and T follicular helper cell responses. Data will continue to be analyzed out to
six months post infection. A larger cohort will be necessary to elucidate the
role of vitamin A in protection against severe disease and death due to
First and foremost, I would like thank my advisor, Dr. Diane E.
Griffin, for allowing me to do my masters research in her laboratory. Her
guidance and support was invaluable throughout my time here. I would like
to thank her for the opportunity to get involved in the dynamic, challenging,
and rewarding research that she had entrusted with me. Furthermore, a
huge thank you to Dr. Rupak Shivakoti for passing down his knowledge of
the basics of how to work with measles virus and acquainting me with Dr.
Griffins lab in general. Additional thanks go out to Rupak to teaching me
many, many techniques. Although he was available to ask questions while he
was here, it was helpful that he encouraged me to jump right in and
conduct my experiments independently early on. I would like to also thank
Rupak for being responsive to questions much after he had graduated from
the laboratory, which was especially helpful.
I would like to thank Dr. Wendy Lin, for providing me with her
knowledge of the logistics of working with measles virus in rhesus macaques.
Her ability to pass down her understandings and techniques was invaluable.
Furthermore, I would like to thank Wendy for taking time from her career at
Columbia University to come down to Baltimore to meet with us, as well as
making herself available to talk about techniques or data analysis.
Importantly, this project would not have run as smoothly as it did without
the help of Ashley Nelson, the PhD student with whom I shared
responsibility in this project. With Ashleys flexibility to work around my
schedule, we were able to make sure the assays for the monkey study could
be completed and analyzed in a timely manner so I could complete my thesis
work. I would also like to thank Ashley for her support and friendship
throughout my time here!
The vitamin A/monkey study was largely a success due to the time and
effort of Dr. Bob Adams and Dr. Tori Baxter. I would like to thank them
immensely for their time and expertise in handling the monkeys, obtaining
samples, and for being flexible with their schedules around the holidays,
while also granting this project many of their early mornings.
I would like to give a huge thanks to my roommate, Dr. Cailin Deal,
who was able to provide me her knowledge and skills in so many areas of
virology and immunology as a whole. Her expertise in writing in science was
crucial to the process of editing my thesis, as well as her general knowledge
of techniques and data analysis. Furthermore, I would like to thank Debbie
Hauer for her assistance in teaching me techniques and processing samples
that were essential for my projects and this thesis.
I would like to extend my warmest thanks to the rest of Dr. Griffins
laboratory for being so welcoming to me as a masters student, for passing
down their expertise and insight when I needed assistance, and for their
general support and friendship. Dr. Kim Shulz, Dr. Tori Baxter, Dr. Kirsten
Kulscar, Stephen Goldstein and Siva Manivannan, your help was
instrumental towards my experience as a student in this laboratory.
Additionally, I would like to acknowledge Gui Nilaratanakul, Rachy
Abraham, and Nina Martin for their presence and livelihood in the lab.
Finally, I would like to thank my mother, father, and brother Ryan, as
well as my extended family and friends for providing their unwavering
support. As a little girl, my parents told me I could do whatever I put my
mind to, and when I decided to pursue a science and research their
enthusiasm was there to match my own. This thesis is the product of the
hard work and support of many people, and I would again like to extend a
tremendous thanks to all of the people by my side!
Table of Contents
List of Tables...... ix
List of Figures. x
Chapter 1: Introduction to measles virus 1
Public health implications... 2
Measles virus pathogenesis. 2
Prevention of measles virus infection... 3
Measles virus virology.. 6
Measles virus infection. 7
Defective replication of measles virus genome 8
Innate immune response to viral infection.. 9
Cytoplasmic PRRs 10
Type I interferon.. 11
Interferon-inducible antiviral proteins... 12
Innate immune response to measles virus infection 12
Role of dendritic cells.. 14
Adaptive immune response to measles virus infection... 14
Antibody response 15
T lymphocyte response 16
Effector CD4+ T lymphocytes 16
Immunosuppression following measles virus infection.. 18
Chapter 2: Comparision of in vitro immune responses to wild-type
measles virus with C and V protein-knock out strains; wild-type and
vaccine strains of measles virus 24
Measles virus immune evasion. 25
Block of type I interferon production.. 25
Block of type I interferon signaling. 27
Interferon-stimulated genes (ISGs) and virus stress-induced
genes (VSIGs)... 28
Role of dendritic cells in measles virus infection.. 29
Defective interfering (DI) particles.. 29
Th17 response to viral infection... 30
Th17 response to measles virus infection.. 30
Materials and methods... 31
Chapter 3: Effects of vitamin A supplementation on the immune