Molecular Biology is the study of the tiny building blocks of life, like DNA and proteins, that make up cells. By studying molecular biology, we can understand how cells work, and even find ways to cure diseases!

Graves# Disease is a form of hyperthyroidism often accompanied by Thyroid Associated Ophthalmopathy (TAO), an autoimmune-mediated inflammation of the extraocular connective tissue. Despite numerous studies to locate the specific factors that regulate the swelling and produce an effective cure, neither of these goals has been achieved. Recently, scientists discovered that Prostaglandin E(2) (PGE(2)) is present in significantly increased levels in the orbital tissues of patients with TAO, and must be an important factor in the inflammatory response around the eyes. The primary goal of this research project was to determine the mechanisms that lie behind the up-regulation of PGE(2) in patients with TAO and establish correlations among Th1 and Th2 cytokines, growth factors, and PGE(2).

What drives the painful eye swelling in Graves' disease? Patients with Thyroid Associated Ophthalmopathy (TAO) have high levels of Prostaglandin E2 (PGE2) in their eye tissues. PGE2 is a molecule that promotes inflammation.

You culture three types of cells in petri dishes:
- Graves' orbital fibroblasts
- normal orbital fibroblasts
- dermal fibroblasts (control)

After two weeks you treat each cell type with different cytokines (immune signaling molecules). Then you measure PGE2 levels using a western blot and ELISA protocol.

The Th2 cytokine IL-4 significantly increases PGE2 levels. TGF-B actually inhibits PGE2, which was unexpected. Both IL-4 and IL-1B share a sixteen-hour optimal response time.


Manipulation of PGE(2) Levels with Various Cytokines of Thyroid Associated Ophthalmopathy

Objectives/Goals:

Graves# Disease is a form of hyperthyroidism often accompanied by Thyroid
Associated Ophthalmopathy
(TAO), an autoimmune-mediated inflammation of the extraocular connective tissue.
Despite numerous
studies to locate the specific factors that regulate the swelling and produce an
effective cure, neither of
these goals has been achieved. Recently, scientists discovered that
Prostaglandin E(2) (PGE(2)) is present
in significantly increased levels in the orbital tissues of patients with TAO,
and must be an important
factor in the inflammatory response around the eyes. The primary goal of this
research project was to
determine the mechanisms that lie behind the up-regulation of PGE(2) in patients
with TAO and establish
correlations among Th1 and Th2 cytokines, growth factors, and PGE(2).

Methods/Materials:

I experimented with three types of cells: Graves# orbital fibroblasts, normal
orbital fibroblasts, and dermal
fibroblasts (control). I cultured each type of cell in petri dishes for two
weeks. Then I treated these cells
with each cytokine at different time intervals. I used the cytokine IL-1B,
already proven to increase
PGE(2) levels, as my positive control treatment. After treating the cells, I
harvested my cultures, which
included lysing and scraping the cells. I was able to solubilize the proteins
and extract the supernatan to
conduct a protein assay. Then I performed a western blot, using a PGE(2) Elisa
Kit Protocol to test for
PGE(2) levels.

Results:

This research experiment led to significant insights concerning the mechanisms
that lie behind TAO.
Contrary to expectations, TGF-B was found to exhibit the potential to inhibit
PGE(2). On the other hand,
a significant relationship between the Th2 cytokine IL-4 and the up-regulation
of PGE(2) was established.
Both IL-4 and IL-1B shared many important similarities, including a sixteen-hour
optimal time interval.

Conclusions/Discussion:

The potential implications of these findings are considerable. Not only have we
disproved a query that
only Th1 cells are responsible for significant up-regulation of PGE2, but we
have now identified another
cytokine important for the progression of TAO. Although IL-4 has remained a
rather obscure cytokine,
these findings have revealed its important involvement with TAO. There must be
some common link
between IL-4 and IL-1B. Further studies concerning the mechanisms associated
with IL-1B and IL-4 will
indisputably be followed.

Summary Statement:

The primary goal of this research project was to determine the mechanisms that
lie behind the
up-regulation of PGE(2) in patients with TAO and establish correlations among
Th1 and Th2 cytokines,
growth factors, and PGE(2).


Cytokines and PGE2 in Thyroid Eye Disease

Manipulation of PGE(2) Levels with Various Cytokines of Thyroid Associated Ophthalmopathy

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Biochemistry is the study of the chemistry of living things. Biochemists study how different molecules like DNA and proteins work together to sustain life. The groundbreaking discoveries in biochemistry continue to lead to the discovery of new medicines and cutting-edge treatments for diseases.

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Cytokines and PGE2 in Thyroid Eye Disease

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