Analysis of Melatonin's Role in Immunity and Disease: A Report

This report analyzes a study investigating the effects of melatonin on immunity and disease, specifically focusing on chronic Chagas' disease. The study examined melatonin's ability to modulate oxidative stress and the inflammatory response. The researchers used ELISA, flow cytometry, and TBARS to assess lipid peroxidation, cytokine levels, and immune cell activity. Key findings include melatonin's potential to reduce oxidative stress, modulate IL-17 production, and decrease MCP-1-producing T cells. The study highlights melatonin's potential protective role in down-modulating extreme inflammation in chronic Chagas’ illness, suggesting a novel therapeutic approach. The report discusses the study's methods, results, and conclusions, along with implications for future research, emphasizing the need for further investigation into melatonin's role in treating infectious diseases.

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IMMUNITY AND DISEASE
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Background and Introduction
Abstract
The research question or hypothesis being answered by this work is: Can the administration
of Melatonin for two months everyday modulate the oxidative stress alongside inflammatory
response in case of chronic infection? The researchers undertook for the treated group and
compared the results with the control group to answer this question. The investigators answered this
question by establishing that the administration of antioxidant compounds like is a novel and
auspicious intervention method for controlling oxidative stress resulting from the chronic Chagas’
disorder partly facilitated via the LPO abrogation alongside inflammatory response deterrence.
Introduction
The pleiotropic functions of melatonin comprise significant role of the primary pacemaker
of circadian and also acting as an extremely effective antioxidant. It has also been revealed that
Melatonin is playing a significant part in primary pacemaker thus suggesting melatonin as a target
for this study’s investigators. Literature has highlighted that melatonin acts as a highly efficient and
effective antioxidant and melatonin is an anti-oxidative enzyme gene expression regulator and
reduces oxidative stress in the entire body as well as triggering the augmented activity of various
antioxidant enzymes.
Gaps
The gap that this current study by Brazão et al. (2015) seeks to fill is the continual neglect
of Chagas’ illness with Trypanosoma cruzi (T. cruzi) as the causative agent despite being a tropical
disease. Brazão et al. (2015) hold that this area is ignored and hence needs an urgent attention.
Brazão et al. (2015) supports their call for urgent action into addressing the Chagas’ disease by
citing the study by showing that it has been discovered that the general human infection by T. cruzi
prevalence is approximated at 9.80M cases with an additional sixty million regard at risk of
contracting Chagas’ disease.

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Brazão et al. (2015) further support their call to action by highlighting how the increasing
migration of infected individuals from the endemic region to advanced economies like the US,
Spain, and Germany has broadened the Chagas’ disease scope and it threatens to exponentially
expand passed the conventionally infected regions. Brazão et al. (2015) study further shows that it
been established that thirteen million individuals from endemic Chagas’ illness nations are staying
in the US. Studies have also shown that between 80,000 and 120,000 amongst the thirteen people
living in the US from the endemic Chagas’ diseases economies have a long-lasting T. cruzi
infection (Brazão et al. 2015).
The current study thus highlight that premised on precarious lack of proof regarding the
actions of present obtainable drugs (nifurtimox and benznidazole) and lack of an operative
alongside possible Chagas’ disease treatment, a design of the rational therapeutic approach utilizing
melatonin as a novel intervention aiming to safeguard rats via LPO abrogation and inflammatory
response is effective. Thus, the investigators selected the administration of melatonin as a feasible
compound capable of modulating the immune response as well as inflammatory process to fill the
gap in the literature regarding the neglect of life-threatening Chagas’ disease.
Methods
ELISA
ELISA was used in this case to measure the F2- isoprostane in the serum to determine the
oxidative stress. In this case, it was used to measure the cytokines in the serum. It allowed the
investigators to measure multiple samples in one experiment. It was useful in detecting the secreted
product, cytokines
TBARS
The TBARS was undertaken to measure the oxidative stress as a marker of lipid
peroxidation. It measured the malondialdehyde concentration produced as a result of degradation of

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the unstable lipid peroxide. It helped determine the anti-oxidant enzyme activities. The TBARS
helped show the protracted T. cruzi infection through an upsurge in TBARS heights in infected
alongside not treated rats thereby allowing the investigators to compare the effect on melatonin
administration. Also, TBARS also help detect the melatonin antioxidant effects through protecting
lipids from the peroxidation based on the lowest and significant TBARS level.
12-H Light/12-H Dark and Sixty Days
This is required because light-dark cycles regulate the sleep-wake cycles significantly via
effects on synthesis of pineal glands and melatonin release. The pineal gland is synthesizing
melatonin and darkness promotes melatonin synthesis thereby triggering its release into the blood
plasma. Therefore, the concentration of melanin increase during the night. Both artificial and
natural light suppress melatonin synthesis and subsequent release, and hence its concentration drops
during daylight. The sixty-day period was required to help determine if administration of melatonin
was linked with alterations in IL-17 cytokine generation profile. This is infecting Wistar rats with
the T. cruzi would require this number of days to show whether the infection has taken place for
antibodies to release which would then help show an increase in the IL-17 levels for any affected
rat.
Results
Explaining and Discussing result in all figures
Figure 1 shows the effect of melatonin through lipid protection from peroxidation indicated
by lowermost alongside substantial levels of TBARS (p<0.0010).
Figure 2 has illustrated that NO reacts with O2- to yield ONOO which initiates LPO. The
investigators examined if administering melatonin orally affects NO generation and concluded that
concentration of nitrate on the 60th day hit the lowest significant values (p<05) for all animals
treated with melatonin as opposed to the infected and untreated group.
Figure 3 illustrates the determination if administering melatonin is linked to IL-17 cytokine
generation profile changes after the rats were infected with T. cruzi and left for 60 days and

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established that IL-17 significantly increased for all infected animals, unlike the uninfected ones. It
shows a rising pattern in IL-17A production for the infected as well as the melatonin-treated cohort
as opposed to infected and untreated ones.
Figure 4 (A and B) shows that MCP-1-generating CD4+ T alongside CD8+ T cells stood
significantly decreased in infected alongside treated animals’ spleen with (p<0.0010).
Figure 5 depicts the investigation of melatonin ability to modulate the CD11a LFA-1
expression in Chagas’ disease with a beta-2 integrin being incorporated in cells recruitments to
inflammation sites. The results showed that the proportion of CD8T cells the proportion of CD8T
cells that express CD11a or CD18 in the treated rats with melatonin stood declined as opposed to
control and not treated cohort. There were no observable differences in the proportion of CD4+ as
well as CD8+ T cells that expressed CD11a or CD18 in the rats infected, whether treated or not.
Figure 6 (A) showed the determination of actual melatonin contribution during infection
after examining if oral administration of melatonin affects the innate immune reaction. Analysis of
peritoneal macrophages showed a significant decline as opposed to infected and not treated rats.
Figure 6 (B) showed the results with spleen macrophages and the result was that melatonin
treatment never affected the proportion of such cells for any cohort, irrespective of the status of
infection.
Figure 7 showed the proportion of peritoneal macrophages that expressed RT1B in rates
infected and not treated staying higher as opposed to not affected rats. No difference was witnessed
in activation markets expression (RT1B) in peritoneal macrophages of the animals infected whether
treated or not.
Figure 8 (A) showed that infected and treated animals had a significant reduction in the cells
numbers as opposed to infected, yet not treated rats.
Figure 8 (B) demonstrated that melatonin treatment has no effect on spleen APC cells
proportions for any treated cohort, whether or not infected or not.
Unusual findings

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Administering chronic oral melatonin significantly abrogates greater circulating TBARS
levels.
Result Summary
Melatonin intervention demonstrated an increasing pattern in generating IL-17A
cytokines in T. cruzi-infected as well as treated rats with melatonin. Orally administering melatonin
significantly abrogates higher TBARS circulation level which indicates membrane LPO triggered
by ROS. A significant decline in MCP-1-generating CD4+ and CD8+ T cells percentages were
noted treated and infected rats. Melatonin administration during the long-lasting T. cruzi-infected
animals further showed significant shielding features, declining the spleen alongside peritoneal
macrophages alongside APC cells’ percentages. No significant variation was noted in the MHC
class II RT1B expressions in the infected and not treated rats.
Conclusions and Further Work
Major Novel Finding
The novel findings of this study are that there is a prospective protecting role of melatonin in
down-modulating extreme inflammation in case of chronic Chagas’ illness, partly mediated by LPO
abrogation.
Implications
The implication of this study is that there is a potential protective role of melatonin in
down-modulating extreme inflammation in the case of chronic Chagas’ illness, partly mediated by
LPO abrogation. This implies oxidative stress prevention in a long-lasting Chagas’ illness model via
the melatonin as an antioxidant compound becomes a novel and promising intervention for
additional probe on treatment trials for this infectious illness and others alike.
Research Avenues and Experimental Methods
There is a need for a study to probe the oxidative stress prevention in the enduring Chagas’
illness model within melatonin intervention using both ELISA and TBARS methods.

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References
Brazão, V., Colato, R.P., Santello, F.H., Filipin, M.D.V., Toldo, M.P.A., do Vale, G.T., Tirapelli,
C.R. and do Prado Júnior, J.C., 2015. Interleukin‐17, oxidative stress, and inflammation: role of
melatonin during Trypanosoma cruzi infection. Journal of pineal research, 59(4), pp.488-496.