Transcript PPt for 520ev Feb 23 - Cal State LA

The Role of Exercise Stress and
Catecholamines in Immunity
Erika Vielmas
February 23, 2009
Biol 520
Background Information
Catecholamines (CAs)
Catecholamines
 Known as epinephrine and norepinephrine
(adrenaline/noradrenaline).
 Water soluble hormones that circulate in the
blood.
 Produced from chromaffin cells;
neuroendocrine cells found in the adrenal
medulla.
 β-adrenergic receptors: G-coupled protein
receptors that are targets for CAs.
Benschop et al 1994
http://en.wikipedia.org/wiki/Adrenergic_receptor
Catecholamine Synthesis
Tyrosine
hydroxylase
Aromatic amino
acid
decarboxylase
Tyrosine
+NH
3
CH2-CH-COO-
OH
OH
Dopamine
+NH
OH
3
CH2-CH2
Dopamine β-Hydroxylase
OH
Epinephrine
OH
Norepinephrine
+NH
H2N+-CH3
OH
CH-CH2
OH
OH
Phenylethanolamine
N-methyltransferase
3
CH-CH2
OH
Nelson & Cox 2005
The Hypothalamic–Pituitary–
Adrenal (HPA) Axis

PVN of the hypothalamus
releases Cortico releasing
hormone (CRH).

CRH stimulates the anterior
pituitary gland to release
adrenocorticotropin (ACTH)
hormone into circulation.

The ACTH induces the adrenal
glands to synthesize and secrete
glucocorticoids and
catecholamines.
PVN
A.P
www.ott.zynet.co.uk/.../images/encephf1.gif
Webster-Marketon et al 2008
Sympathetic Nervous System
The SNS is activated
by the secretion of
acetylcholine from
the pre ganglionic
sympathetic fibers
in the adrenal
medulla.
 SNS activation
induces secretion
of epinephrine into
blood circulation.
 Norepinephrine is
released from the
nerve terminals in
the vicinity of
immune cells.

Webster-Marketon et al 2008
users.rcn.com/.../BiologyPages/P/PNS.html
CAs can be Synthesized by
Immune Cells

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For many years, CAs were believed to be
synthesized only by neurons and endocrine
cells.
Recent studies suggest that lymphocytes and
mature CD3 T- cells can synthesize and secrete
CAs.
Cells need high levels of norepinephrine to
stimulate the production of more CAs.
Leposavic et al 2008
Lymphocyte Differentiation

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Mammalian stem cells differentiate into several
kinds of blood cells within the bone marrow by
a process called haematopoiesis.
All lymphocytes originate from a common
lymphoid progenitor before differentiating into
their distinct lymphocyte types.
The formation of lymphocytes is known as
lymphopoiesis.
http://en.wikipedia.org/wiki/Lymphocyte
www.textbookofbacteriology.net
Natural Killer Cells (NK)


Natural Killer Cells are large granular lymphoid-like cells
important in innate immunity.
NK cells are produced in the bone marrow.
These cells can detect virus-infected cells without having any
antigen specificity . (“natural killer”).
 To control their cytotoxic activity, NK cells possess two types of
surface receptors: activating receptors and inhibitory receptors.
 Inhibitory receptors recognize MHC I molecules and activating
receptors recognize interferons and cytokines (which may be
caused by stress); interferon γ (INF-γ) and interleukin-2 (IL-2) .
 80% of NK cells express CD8.

Calcagni et al 2006 Webster-Marketon et al 2008
http://jem.rupress.org/cgi/content/abstract/190/7/1005
Two Functionally Distinct Cells for NK
Cells : CD56dim & CD56bright

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CD56dim constitutes 90% of peripheral NK cells
and these are more cytotoxic than the
CD56bright . These cells express higher levels of
CD16 than their subset.
CD16 is a low-affinity immunoglobulin G (IgG)
Fc receptor that is expressed on natural killer
(NK) cells.
CD56bright cells have the capacity to produce
an abundant amount of cytokines.
Timmons et al 2006
T-lymphocytes
T lymphocytes
 Migrate to and mature in the thymus.
 Enter circulation and peripheral lymphoid
organs (spleen & lymph nodes)
 Patrol for invading pathogens and/or tumor
cells.
 Involved in adaptive immunity differentiate
further after exposure to an antigen.
http://en.wikipedia.org/wiki/Lymphocyte
T-lymphocytes

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
Have a special surface receptor that recognize
specific antigens. (Express CD3, and CD8).
CD8 cells differentiate into cytotoxic T cells or
can be expressed by NK cells.
CD3 complex is the complex of α:β or γ:δ T-cell
receptor chains.
Assist in the recognition and rejection of foreign
tissues.
CAs, can inhibit lymphocyte proliferation and
cytotoxicity.
http://en.wikipedia.org/wiki/Lymphocyte
Stress Induced by Exercise
Exhaustive Exercise
 Vigorous physical activity triggers
the response of CAs during exhaustive exercise.
 Know to induce a shift in peripheral blood NK
cells.
 Associated with significant shifts in peripheral
immune cell counts.
 Redistribution of lymphocytes among different
compartments of the immune system.
Imrich et al 2004, Zhang et al 2006
cache.boston.com/.../swim_06_30/swim13.jpg
The HPA axis and the
SNS are activated
which induces a
release of
glucocorticoids and
catecholamines
which can modulate
different aspects in
the immune system.
Webster-Marketon et al 2008
Hypothesis
Exercise-induced stress:
 Increases catecholamine levels
 Decreases natural killer cell cytotoxicity
 Decreases T-lymphocyte activity.
Overview
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We will be looking at Plasma concentrations of
CAs prior and after vigorous exercise.
NK cell and T-lymphocyte cell counts and
activity prior and after vigorous exercise.
Show the connection of CAs, stress (from
exercise), and immune cells.
Experimental Results
NK Cell Count Increase
Throughout Training
The training group consisted of 15 women in pre-season training for 1 month
training for 5h/day.
The control group consisted of 7 women instructed to avoid vigorous exercise.
Sample of blood from pre-training, during (10th day), end (1day prior to end
training, and post-training ( 1 week after).
Suzui et al 2004
Concentrations of the CAs and
Cytolytic Activity of the NKs
Catecholamine levels
were not affected.
There was no
significant difference
during the 1 month
training process.
Nk cell cytolytic activity
during and after training.
Suzui et al 2004
CAs levels and Cell Counts for
Immune Cells
The training group consisted of 10 healthy males.
6 weeks of endurance training: 1h run, 4 times a week
Followed by 5 days of bed rest.
Imrich et al 2004
Flow cytometry



Method for quantitating components or
structural features of cells & measures
fluorescence per cell or particle.
Scattered and emitted light from cells and
particles are converted to electrical pulses by
optical detectors. Parallel light waveforms is
picked up by confocal lenses focused at the
intersection point of cells and the light source.
After the different signals or pulses are
amplified they are processed by an Analog to
Digital Converter (ADC) which in turn allows for
events to be plotted on a graphical scale(One
Parameter, Two parameter Histograms).
http://biology.berkeley.edu/crl/flow_cytometry_basic.html
CD56dim & CD56bright Cell Distribution
Dot plot: CD56 and CD3
A total of 10,000 events were collected in the
lymphocyte gate based on forward- vs. side-scatter characteristics.
N=11 female adolescents age 14
Exercise: Vigorous Cycling
Rest: prior to exercise
Post: 60 min. after exercise
Recovery: 60 min. rest period after exercise.
Timmons et al 2006
NK Cell Count and CA
Concentrations
(pg.ml-1)
Epinephrine*
P< 0.001
Zhang et al 2006
Effects of Exercise on TLymphocytes
Subjects consisted of 7 males
Exercise: Cycling til’ exhaustion
Pre-exercise
60 min
Post-exercise
1h post-exercise
Trial 1: 6 day Intense Training Period
work out everyday.
Trial 2: 2 week training, 3-4 sessions per
week
Trial 3: Same as first trial
Lancaster et al 2004
NK cells in Lung Tissue
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NK cells have different activity in different
organs.
Kanemi and his team investigated the effect of
restraint stress on lung lymphocytes including
NK cells.
C57BL/6 female mice were exposed to 2 h
restraint stress.
Kanemi et al 2004
Plasma concentrations of
Epinephrine and
Norepinephrine
N=6
Samples measure by HPLC
After the mice were stressed, the
CAs levels immediately increased
but the value for Norepinephrine
was not shown to be significant
despite the increase.
Con= control
RS0= 0 hr
RS1= 1 hr
RS2= 2 hr
Kanemi et al 2004
Stress on pulmonary
NK cells
Y-axis is the number of cells.
Levels of lung NK cells
decreased significantly as well
as NK cells in the Plasma. Initial
decrease occurred during stress
and lasted throughout the
recovery phase.
Immunohistochemical analysis of
the lung tissue: a) control mice
b) stressed mice.
c) NK cells decreased
compared to control.
Kanemi et al 2004
Conclusion
Summary
Suzui et al 2004
Supports Hypothesis
Imrich et al 2004
Kanemi et al 2006
Partial support of
Hypothesis
Zhang et al 2006
Lancaster et al 2004
CA’s increased in girls,no change in boys;
Nk increased in girls, not in boys
Timmons et al 2006
Refutes Hypothesis
Take Home Message
Women
 Exercise decreased cytolytic activity despite an overall
increase in NK cells and T-cells. CAs are not affected.
Men
 Immune cells were not affected by exercise but CAs
levels increased significantly
Adolescents
 Increase in CAs for girls. Increase in cytolytic activity.
 Incease in CAs for boys. No significant change in
cytolytic activity
Mice
 Stress increased CAs.
 Decrease in NK cells.
Conclusion

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According to the data, my hypothesis was
partially supported.
CAs increase in certain instances as a result of
stress from exercise.
Cytotoxic activity is not directly mediated by
CAs increase.
Not enough studies done to show the
differences among men, women, adolescents,
and rodent models.
Future Experiments

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
Use proper controls.
Men: Trained vs. Untrained
Women: Trained vs. Untrained
Boys: Trained vs. Untrained
Girls: Trained vs. Untrained
Run a hormone analysis of testosterone, estrogen, and
CAs among adolescents and adults after exercise.
In vitro experiments to check cytotoxic activity of NK cells
and T-cells:
Using NK-sensitive and T-sensitive K562 target cell lines and
label with non-radioactive europium.
References
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Benschop, R.J., Nijkamp, F.P., Ballieux, R.E., Heijnen, C. (1994) The effects of B-adrenoreceptor Stimulation on Adhesion of
Human Natural Killer Cells to Cultured Endothelium. Br. Journal of Pharmacology 113, 1311-1316
Calcagni, E., Elenkov, I. (2006) Stress System Activity, Innate and T Helper Cytokines, and Susceptibility to Immune-Related
Diseases. Annual New York Academy of Sciences 1069, 62-76
Imrich, R., Tibenska, E., Koska, J., Ksinantova, L., Kvetnansky, R., Bergendiova-Sedlackova, K., Blazicek, P., Vigas, M. (2004)
Repeated Stress-Induced Stimulation of Catecholamines Response is not Followed by Altered Immune Cell Redistribution.
Anneals New York Academy of Sciences 1018, 266-272
Kanemi, O., Zhang, X., Sakamoto, Y., Ebina, M,. Nagatomi, R.(2005) Acute Stress Reduces Intraparenchymal Lung Natural
Killer Cells Via beta-adrenergic Stimulation. Clinical and Experimental Immunology, 139, 25–34
Lancaster G.I., Halson, S. L., Khan, Q., Drysdale, P., Wallace, F., Jeukendrup, A.E., Drayson, M.T., Gleeson, M. (2004) Effects of
Acute Exhaustive Exercise and Chronic Exercise Training on Type 1 and Type 2 T Lymphocytes. Exercise Immunology Rev 10,
91-106
Leposavic, G., Pilipovic, I., Radojevic, K., Pesic, V., Perisic, M., Kosec, D. (2008) Catecholamines as Immunomodulators: A
role for Adrenoceptor-mediated Mechanisms in Fine Tuning of T-cell Development. Autonomic Neuroscience:Basic and
Clinical 144, 1-12
Nelson, D.L., Cox, M.M. (2005)Lehninger Principles of Biochemistry. W.H. Freeman and Company.
Suzui, M., Kawai, T., Kimura, H., Takeda, K., Yagita, H., Okumura, K., Shek, P.N., Shephard, R.J. (2004) Natural Killer Cell Lytic
Activity and CD56dim and CD56bright Cell Distributions During and After Intensive Training. Journal of Applied Physiology
96: 2167–2173, 2004
Timmons, B.W., Tarnopolsky, M.A., Bar-Or, O.( 2006) Sex-based effects on the Distribution of NK Cell Subsets in Response to
Exercise and Carbohydrate Intake in Adolescents. Journal of Applied Physiology 100, 1513–1519
Webster-Marketon, J.I., Glaser, R. (2008) Stress Hormones and Immune Function.
Cellular Immunology 252, 16-26
Zhang, X., Matsuo, K., Farmawati, A., Higashi, K. O., Nagata, K., Nagatomi, R. (2006) Exhaustive Exercise Induces Differential
Changes in Serum Granulysin and Circulating Number of Natural Killer Cells. Journal of Experimental Medicine 210, 117-124
http://en.wikipedia.org/wiki/Adrenergic_receptor
http://en.wikipedia.org/wiki/Lymphocyte
http:// www.textbookofbacteriology.net
http://jem.rupress.org/cgi/content/abstract/190/7/1005
http://www.ott.zynet.co.uk/.../images/encephf1.gif
http://users.rcn.com/.../BiologyPages/P/PNS.html
http://cache.boston.com/.../swim_06_30/swim13.jpg
Questions/Comments??