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Gene & Protein in Disease Interleukins-1β, interleukin-8, and the SARS-CoV-2

for the SARS that caused the coronavirus disease 2019 hyperinflammation and patient death. This study aimed
(COVID-19) pandemic, which was decreed on March 11, to evaluate the mRNA expression of important genes
2020, by the World Health Organization. 2,3 associated with cytokine storm, such as IL-1β, IL-8,
The clinical manifestations of individuals infected and IL-1RN, in patients with and without SARS-CoV-2
with SARS-CoV-2 range from acute and mild respiratory infection. At the time of this study, some patients had
conditions, such as influenza syndrome, to SARS. In been vaccinated, whereas others were still unvaccinated.
4,5
general, the severity of COVID-19 is affected by the Moreover, the frequency of clinical manifestations was
dominance of the viral pathogenicity or inflammatory evaluated in patients with SARS-CoV-2 infection who
response of the host, with the latter having the probability were either vaccinated or unvaccinated.
of causing systemic hyperinflammation. 6 The primary outcome measures evaluated in
Studies have shown that patients who died or our study were based on the elevated levels of pro-
required intensive care units (ICUs) with SARS due to inflammatory mediators in patients with COVID-19.
COVID-19 had increased pro-inflammatory cytokine These primary outcome measures could justify the
levels. 6-11 Some diseases, such as rheumatic, tumoral, systemic hyperinflammation (cytokine storm) associated
with worse clinical symptoms and signs. Our secondary
and infectious diseases, can result in an exacerbated and outcome measure compared the levels of pro-inflammatory
uncontrolled release of pro-inflammatory cytokines, mediators in patients who tested positive for COVID-19
11
such as interleukins (ILs) and tumor necrosis factor-alpha and had different or similar severity. We compared patients
(TNF-α), leading to a cytokine storm, in which systemic who had been vaccinated with the CoronaVac/Sinovac
inflammatory processes and multiple organ failure can
occur. 6,10-12 vaccine, which was available at the time of this study, with
those who had not been vaccinated.
Pro-inflammatory cytokines correspond to extracellular
glycoproteins that perform important functions in the 2. Methods
immune response. Their intensified synthesis can result in 2.1. Materials
systemic, metabolic, and hemodynamic instability, which
can worsen several pathologies. 13,14 ILs consist of cytokines The following materials were used in this study: QIAmp
produced primarily by leukocytes and can be considered Viral RNA/QIAGEN kit (catalog number 52906, QIAGEN,
pro-inflammatory when they possess the ability to maximize Hilden, Germany); BIOGENE kit RNA/viral DNA
different stages of the inflammatory process in the face of extraction/Bioclin (reference K204, Biogene Shirley, NY,
antigens that may mediate an immune response. 15 USA); Nanodrop 2000 spectrophotometer, automatic
thermal cycler GeneAmp polymerase chain reaction
IL-1β is a pro-inflammatory cytokine that is primarily (PCR) System 9700, and ABI Prism 7500 Fast Sequence
responsible for fever. It is mainly produced by macrophages Detection System Equipment (Thermo Scientific, Waltham,
and monocytes, with higher levels of production during MA, USA); and High-Capacity Complementary DNA
viral infections. 15,16 IL-1g/AR (receptor antagonist) acts (cDNA) Reverse Transcription Kit (Applied Biosystems ,
™
as an endogenous autoregulator competing with IL-1 USA). Quantitative PCR (qPCR) was performed using
receptors (α and β); hence, it can be used to prevent the TaqMan assays (Applied Biosystems) to evaluate IL-1β
negative effects from the exacerbated levels of IL-1. 13,14 (Hs 01555410_m1), IL-8 (Hs9999034_m1), and IL-1RN
IL-8 is a pro-inflammatory cytokine belonging to the (Hs00893626_m1). We also used UBC (Hs00824723_m1)
CXC chemokine subfamily. It is primarily produced by and GPDH (02758991_m1).
macrophages and can increase the levels of chemotactic
factors by stimulating innate immunity and cell-mediated 2.2. Genetic material extraction
immunity. 15,17 Viral and human RNA extraction for detecting SARS-
COVID-19 vaccines are currently approved for use CoV-2 was performed using the protocol described in
in adults, adolescents, and children and other vaccines the QIAmp Viral RNA/QIAGEN kit (catalog number
are still being tested. Nevertheless, there is no scientific 52906) to detect SARS-CoV-2 by reverse transcription
18
evidence for specific effective treatments that can cure PCR (RT-PCR). For gene amplification, viral/human
COVID-19. Hence, it is essential to understand the RNA extraction was performed using the BIOGENE kit
6,19
inflammatory response and factors, including pro- RNA/viral DNA extraction/Bioclin (reference K204). The
inflammatory cytokines that can influence the physiological Nanodrop 2000 spectrophotometer (Thermo Scientific,
response, for constructing therapeutic alternatives to treat Waltham, MA, USA) was used to determine and adjust
individuals with COVID-19, thereby preventing systemic RNA concentrations.

Volume 3 Issue 4 (2024) 2 doi: 10.36922/gpd.4076