he Most Interesting Viru In the World
Dec 30, 2015
The Most Interesting VirusIn the World
Presented By: Kunqian Li
Mentor: Dr. Xiaoyong Bao Ph.D.
UTMB Galveston
Pediatrics
Independent Study Mentorship: HST – Fall 2014
ISM Teacher: Mrs. Click
DISCLAIMER: SOME INFORMATION IN THIS PRESENTATION COMES FROM OUTSIDE SOURCES
The Scientific Research of
Human Metapneumovirus
What is Independent Study Mentorship (ISM)?
This course is a project-based learning experience at the professional level.
Provides opportunities for an in depth study in a particular curricular area.
Students must:Document 3-5 hours of work each weekSubmit weekly logs of documentationMeet with mentor on a regular basisPresent a final project
A little bit about my mentor
Dr. Xiaoyong Bao Ph.D
-Assistant Professor of Pediatrics in the
Division of Clinical and Experimental
Immunology and Infectious Disease
-Ph.D. in Cellular Physiology and
Molecular Biophysics from UTMB
-Research focuses on host-virus
interaction, with the goal to generate
suitable vaccine candidates to prevent
paramyxovirus infection
"UTMB Department of Pediatrics." Welcome. N.p., n.d. Web. 12 Oct. 2014.
The University of Texas Medical Branch
• Established in 1891
• More than 70 major buildings, more than 2,500 students and more than
1,000 faculty.
• UTMB’s mission is to improve health for the people of Texas and around
the world.
"The University of Texas Medical Branch at Galveston." Welcome to UTMB Health. N.p., n.d. Web. 12 Oct. 2014.
What is Human Metapneumovirus (hMPV)?
Identified in 2001 in the Netherlands
Can cause upper and lower respiratory tract infections in people of all ages
Most people with hMPV have mild symptoms but some can have severe illness including pneumonia and a flare-up of asthma
More serious in children under the age of 1 and in the elderly"Human Metapneumovirus - American Lung Association."
American Lung Association. N.p., n.d. Web. 12 Oct. 2014.
"American Society for MicrobiologyClinical Microbiology Reviews." Human Metapneumovirus: Lessons
Learned over the First Decade. N.p., n.d. Web. 12 Oct. 2014.
Purpose of This ProjectResearch hMPV and the effects it can have
Educate audience on respiratory viruses
Discover unknown information that will lead to vaccines
Pictures taken from Microsoft Office Clipart
RED
Current Research on hMPVhMPV is a single negative-stranded RNA
enveloped virus
hMPV is second to RSV as the most common cause of respiratory tract infection
hMPV lacking the M2-2 Protein made it a promising vaccine candidate
hMPV is the first identified human pathogen belonging to the genus “Metapneumovirus in the Pneumovirinae subfamily of the Paramyxoviridae family”
"Human Metapneumovirus." - MicrobeWiki. N.p., n.d. Web. 07 Dec. 2014. <https://microbewiki.kenyon.edu/index.php/Human_Metapneumovirus>.
Current Research on hMPV microRNA and small-interfering RNA both suppress gene
expression by complementary binding to target mRNAs
Northern blots and real-time polymerase chain reactions techniques are used to investigate hMPV
hMPV leads to significant changes in miRNAs in human airway epithelial cells
hMPV infection does not induce tRFs
In a CDC test, hMPV was detected in 6% of children
“From 2003-2009 the average annual rate of hMPV hospitalization was 1/1000 children less than five years old”
"Human Metapneumovirus." - MicrobeWiki. N.p., n.d. Web. 07 Dec. 2014. <https://microbewiki.kenyon.edu/index.php/Human_Metapneumovirus>.
Survey To gather more information on the subject at
hand, a survey was conducted through Survey Monkey
Here are the questions to my survey:
Are you male or female?
What is your age?
Have you taken healthcare related courses?
Have you heard of Human Metapneumovirus or Respiratory Syncytial Virus?
Which of the symptoms have you suffered from in the past 12 weeks?
High fever
Severe cough
Difficulty breathing
Rapid breathing
Wheezing
Vomiting
Diarrhea
None
How many times do you wash your hands per day?
How long do you wash your hands for?
What do you sneeze into?
Have you shared food and/or eating utensils in the past 2 weeks?
How often do you touch your eyes, mouth, or nose?
Results of the SurveyI had hypothesized that:
Health care professionals are more aware of hMPV because they are more exposed to viruses
40% of all participants exhibit at least two risk factors or severe symptoms of hMPV
Women are taking better precautionary measures in preventing hMPV infection than men are due to their natural maternal instinct
Results ContinuedMost people are unaware of the virus regardless
of education background
Has taken healthcare related courses
Hasn’t taken healthcare related courses
Results ContinuedOn average, females wash their hands longer
than men
FemalesMales
Results ContinuedMales tend to touch their face more
Females Males
Online Weeblyhttp://kunqianli.weebly.com
Laboratory Procedures Dr. Bao observing the
growth of cells in minimum essential medium
45 wells
3 time groups (6,15, 24 hours)
5 different hMPV variables with a mock (control)
3 wells for each variable
Cell Infection
Inoculate Cells
Dispose of serum
Introduce serum mixed with hMPV variables
Observe Cells under microscope after appropriate time
A few of the laboratory equipment necessary in cell infection
96 Well PlateElectronic Pipette
Fixing 96 – Well Plate1. Wash with 150 microliters of (DPBSx1)
for 3 times
2. Fix solution:
(10ml of Methanol +667microliters of Hydrogen Peroxide 30wt), mix well.
3. Add 100microliters of fixing solution to each well, then incubate for 30 minutes
4. Wash after the 30 min incubation for 3 more times
5. Add 100microliters of (DPBSx1) to each well
6. Store in 4°C
A549 Cells Cells located in the basal layer of the epidermis
The primary targeting cells of hMPV are airway epithelial cells
A549 Cells can serve as an airway epithelial cell model for hMPV infection.
Evidence that A549 Cells can be used successfully in this study
X, Bao, Liu T, Spetch L, Kolli D, Garofalo R.P, and Casola A. "Images in This Article." NIH Public Access. U.S. National Library of Medicine, 25 July 2007. Web. 12 Oct. 2014.
Sandwich ELISAIn order to identify the plaques growing on the
cells, the Sandwich ELISA method was employed
Pros to this method:High specificity, since two antibodies are used the
antigen/analyte is specifically captured and detected
Suitable for complex samples, since the antigen does not require purification prior to measurement
Flexibility and sensitivity, since both direct and indirect detection methods can be used
"Sandwich ELISA, Highly Sensitive." Sandwich ELISA. N.p., n.d. Web. 07 Dec. 2014. <http://www.elisa-antibody.com/ELISA-Introduction/ELISA-types/sandwich-elisa>.
Data Gathered at the Lab
Final Product
• Poster board
displaying data
gathered over a
specific time
period
• Focus on RANTES
and IL-8
RANTES A cytokine that is a member of the interleukin-8 superfamily of
cytokines. RANTES is a protein. It is a selective attractant for memory
T lymphocytes and monocytes. It binds to CCR5, a coreceptor of HIV.
RANTES is an acronym for Regulated on Activation, Normal T
Expressed and Secreted. It is also known as CCL5.
"RANTES (Regulated on Activation, Normal T Cell Expressed and Secreted), Inflammation, Obesity, and the Metabolic Syndrome." RANTES (Regulated on Activation, Normal T Cell Expressed and Secreted), Inflammation, Obesity, and the Metabolic Syndrome. N.p., n.d. Web. 07 Dec. 2014. <http://circ.ahajournals.org/content/115/8/946.full>.
Interleukin-8 Interleukin 8 (IL-8) or CXCL8 is a chemokine produced by
macrophages and other cell types such a epithelial cells.
Through a chain of biochemical reactions, IL-8 is secreted and is an important mediator of the immune reaction in the innate immune system response.
IL-8, also known as neutrophil chemotactic factor, has two primary functions: It induces chemotaxis in target cells causing them to migrate
towards the site of infection. It induces phagocytosis once those cells arrive.
"Result Filters." National Center for Biotechnology Information. U.S. National Library of Medicine, n.d. Web. 07 Dec. 2014. <http://www.ncbi.nlm.nih.gov/pubmed/7657403>.
Significance of this Data
WT 15 Hrs Mult 1 15 Hrs Mult 2 15 Hrs0
1000
2000
3000
4000
5000
6000
3119.8
4816.94
3901.413666.86
4761.534308.8
3397.14
4928.735293.72
IL-8 (54)Obs Conc
Trial 1Trial 2Trial 3
WT 15 Hrs Mult 1 15 Hrs Mult 2 15 Hrs0
50
100
150
200
250
33.08
138.86
194.26
51.95
135.25
178.1
59.8
135.04
216.46
RANTES (37)Obs Conc
Trial 1Trial 2Trial 3
ConclusionThis semester has been one of the most
influential semesters I’ve had in high school
Much has been learned through hands-on experiences in a professional field
My knowledge on hMPV has increased tremendously
Picture from Microsoft Office Clipart
QuestionsHelpful Sources
kunqianli.weebly.comArticle published by mentor
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2386556/
Contact [email protected]
Thank You
Special thanks to:My outstanding mentor:
Dr. Xiaoyong Bao
And of CourseMy wonderful ISM teacher:
Mrs. ClickMy Parents:
George Li and Congying WangMy generous and terrific evaluators:
Mr. Thomas and Mr. MayberryAnd all those who supported me this
semester
Works Cited "American Society for MicrobiologyClinical Microbiology Reviews." Human Metapneumovirus:
Lessons Learned over the First Decade. N.p., n.d. Web. 12 Oct. 2014.
"Human Metapneumovirus - American Lung Association." American Lung Association. N.p., n.d. Web. 12 Oct. 2014.
"Human Metapneumovirus." - MicrobeWiki. N.p., n.d. Web. 07 Dec. 2014. <https://microbewiki.kenyon.edu/ index.php/Human_Metapneumovirus>.
"RANTES (Regulated on Activation, Normal T Cell Expressed and Secreted), Inflammation, Obesity, and the Metabolic Syndrome." RANTES (Regulated on Activation, Normal T Cell Expressed and Secreted), Inflammation, Obesity, and the Metabolic Syndrome. N.p., n.d. Web. 07 Dec. 2014. <http:// circ.ahajournals.org/content/115/8/946.full>.
"Respiratory Syncytial Virus Infection of Human Lung Endothelial Cells Enhances Selectively Intercellular Adhesion Molecule-1 Expression." Respiratory Syncytial Virus Infection of Human Lung Endothelial Cells Enhances Selectively Intercellular Adhesion Molecule-1 Expression. N.p., n.d. Web. 07 Dec. 2014. <http://www.jimmunol.org/content/174/11/7359.full>.
"Result Filters." National Center for Biotechnology Information. U.S. National Library of Medicine, n.d. Web. 07 Dec. 2014. <http://www.ncbi.nlm.nih.gov/pubmed/7657403>.
"Sandwich ELISA, Highly Sensitive." Sandwich ELISA. N.p., n.d. Web. 07 Dec. 2014. <http://www.elisa- antibody.com/ELISA-Introduction/ELISA-types/sandwich-elisa>.
"The University of Texas Medical Branch at Galveston." Welcome to UTMB Health. N.p., n.d. Web. 12 Oct. 2014.
"UTMB Department of Pediatrics." Welcome. N.p., n.d. Web. 12 Oct. 2014.
X, Bao, Liu T, Spetch L, Kolli D, Garofalo R.P, and Casola A. "Images in This Article." NIH Public Access. U.S. National Library of Medicine, 25 July 2007. Web. 12 Oct. 2014.