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RESULTS
Kenya Joseph,1 Joseph S. Marino2, and Jeanette M. Bennett3
1Biology, 2Kinesiology, 3Psychology, The University of North Carolina at Charlotte
Blood Lymphocytes Cytokine Gene Expression from Glucocorticoid &
Lipopolysaccharide Exposure in Healthy Smokers & Non-Smokers
• We also conducted a glucocorticoid & nicotine receptor
function assay
• Saliva, serum, plasma and Isolated lymphocytes
(control & LPS stimulated) stored at -80 C
• The ability of white blood cells to effectively signal each
other and trigger various actions is a key component in
immune response to pathogens.
• Cigarette smoking negatively impacts the immune system
leading to longer and more frequent illness in smokers
and chronic inflammation.
• This elevated inflammation often leads to development of
chronic disorders such as asthma, chronic bronchitis,
COPD and autoimmune disorders.
• First line treatment for these inflammation-related chronic
disease is steroid or glucocorticoid medications;
unfortunately patients who smoke do not respond as well
as non-smokers.
• Glucocorticoids are hormones that can aid in the
reduction of inflammation by inhibiting immune cells;
however, reduced responsiveness to steroid medications
could indicate that smokers may have glucocorticoid
resistance when the chronic disease develops.
• The difference in the LPS and LPS+DEX observed was
due to reduced activation from LPS in smokers.
• The results indicates that immune system dysregulation
may occur long before any chronic disease or systemic
inflammation presents clinically.
• Future work includes examining the potential mechanism
for the loss of LPS activation in healthy smokers’
lymphocytes.
REFERENCES
1. Garlichs, C. D., Cicha, I., Raaz, D., Meyer, L., Stumpf, C., Klinghammer, L., … Daniel, W. G.
(2009). CD40/CD154 system and pro-inflammatory cytokines in young healthy male
smokers without additional risk factors for atherosclerosis. Inflammation Research,
58(6), 306–311. http://doi.org/10.1007/s00011-008-8084-8
2. Gellman, M. D., & Turner, J. R. (Eds.). (2013). Encyclopedia of Behavioral Medicine. New
York, NY: Springer New York. Retrieved from http://link.springer.com/10.1007/978-1-
4419-1005-9
3. Schlotz, W., Yim, I. S., Zoccola, P. M., Jansen, L., & Schulz, P. (2011). The perceived stress
reactivity scale: Measurement invariance, stability, and validity in three countries.
Psychological Assessment, 23(1), 80–94. http://doi.org/10.1037/a0021148
4. Zijlstra, F. J. (1998). Smoking and nicotine in inflammatory bowel disease: good or bad
for cytokines? Mediators of Inflammation, 7(3), 153–155.
INTRODUCTION BACKGROUND METHODS – CONT’D
DISCUSSION
REFERENCES
• To examine differences in cytokine gene expression in
peripheral blood lymphocytes to an immune trigger or
lipopolysacchride (LPS) and a immune suppressor or
dexamethasone (DEX) between healthy individuals who
never smoked and healthy smokers who have not yet
exhibited clinical disease.
OBJECTIVE
This research was funded via internal funds from UNC Charlotte. Thank you to all the undergraduate and
graduate research assistants in the StressWAVES Biobehavioral Research Lab.
ACKNOWLEDGEMENTS
• 48 Healthy adults were brought into the lab between 7:30 & 8:30a
• Whole Blood was drawn, donors were in a fasted state -
1 serum tube, 1 EDTA tube and 3 sodium heparin treated tubes
• Glucocorticoid Receptor (GCR) Expression Assay – whole blood
was diluted and divided into 4 treatments: none, Dex, LPS, & Dex
+ LPS
• Samples were plated and incubated for 2 hrs at 38 C and 5% CO2
• Next, lymphocytes were washed, RNA isolated, and RT-PCR
conducted.
Dexamethasone
METHODS
CONCLUSION
• DEX significantly reduced the lymphocytes’ IL-6, TNF-α
and IFN-γ mRNA expression response to LPS.
• The immunosuppressive effect of Dex was greater for
never smokers vs. smokers for TNF-α and IFN-γ.
RESULTS

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UNCCURC Poster 2016 FINAL

  • 1. RESULTS Kenya Joseph,1 Joseph S. Marino2, and Jeanette M. Bennett3 1Biology, 2Kinesiology, 3Psychology, The University of North Carolina at Charlotte Blood Lymphocytes Cytokine Gene Expression from Glucocorticoid & Lipopolysaccharide Exposure in Healthy Smokers & Non-Smokers • We also conducted a glucocorticoid & nicotine receptor function assay • Saliva, serum, plasma and Isolated lymphocytes (control & LPS stimulated) stored at -80 C • The ability of white blood cells to effectively signal each other and trigger various actions is a key component in immune response to pathogens. • Cigarette smoking negatively impacts the immune system leading to longer and more frequent illness in smokers and chronic inflammation. • This elevated inflammation often leads to development of chronic disorders such as asthma, chronic bronchitis, COPD and autoimmune disorders. • First line treatment for these inflammation-related chronic disease is steroid or glucocorticoid medications; unfortunately patients who smoke do not respond as well as non-smokers. • Glucocorticoids are hormones that can aid in the reduction of inflammation by inhibiting immune cells; however, reduced responsiveness to steroid medications could indicate that smokers may have glucocorticoid resistance when the chronic disease develops. • The difference in the LPS and LPS+DEX observed was due to reduced activation from LPS in smokers. • The results indicates that immune system dysregulation may occur long before any chronic disease or systemic inflammation presents clinically. • Future work includes examining the potential mechanism for the loss of LPS activation in healthy smokers’ lymphocytes. REFERENCES 1. Garlichs, C. D., Cicha, I., Raaz, D., Meyer, L., Stumpf, C., Klinghammer, L., … Daniel, W. G. (2009). CD40/CD154 system and pro-inflammatory cytokines in young healthy male smokers without additional risk factors for atherosclerosis. Inflammation Research, 58(6), 306–311. http://doi.org/10.1007/s00011-008-8084-8 2. Gellman, M. D., & Turner, J. R. (Eds.). (2013). Encyclopedia of Behavioral Medicine. New York, NY: Springer New York. Retrieved from http://link.springer.com/10.1007/978-1- 4419-1005-9 3. Schlotz, W., Yim, I. S., Zoccola, P. M., Jansen, L., & Schulz, P. (2011). The perceived stress reactivity scale: Measurement invariance, stability, and validity in three countries. Psychological Assessment, 23(1), 80–94. http://doi.org/10.1037/a0021148 4. Zijlstra, F. J. (1998). Smoking and nicotine in inflammatory bowel disease: good or bad for cytokines? Mediators of Inflammation, 7(3), 153–155. INTRODUCTION BACKGROUND METHODS – CONT’D DISCUSSION REFERENCES • To examine differences in cytokine gene expression in peripheral blood lymphocytes to an immune trigger or lipopolysacchride (LPS) and a immune suppressor or dexamethasone (DEX) between healthy individuals who never smoked and healthy smokers who have not yet exhibited clinical disease. OBJECTIVE This research was funded via internal funds from UNC Charlotte. Thank you to all the undergraduate and graduate research assistants in the StressWAVES Biobehavioral Research Lab. ACKNOWLEDGEMENTS • 48 Healthy adults were brought into the lab between 7:30 & 8:30a • Whole Blood was drawn, donors were in a fasted state - 1 serum tube, 1 EDTA tube and 3 sodium heparin treated tubes • Glucocorticoid Receptor (GCR) Expression Assay – whole blood was diluted and divided into 4 treatments: none, Dex, LPS, & Dex + LPS • Samples were plated and incubated for 2 hrs at 38 C and 5% CO2 • Next, lymphocytes were washed, RNA isolated, and RT-PCR conducted. Dexamethasone METHODS CONCLUSION • DEX significantly reduced the lymphocytes’ IL-6, TNF-α and IFN-γ mRNA expression response to LPS. • The immunosuppressive effect of Dex was greater for never smokers vs. smokers for TNF-α and IFN-γ. RESULTS