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Features of pro- and anti-inflammatory cytokines in the
blood of HIV-positive children
Doniyor KARIMOV
Botir ERMATOV
Nigora SUNNATOVA
Tashkent State Dental Institute
Institute of Human Immunology and Genomics of the Academy of Sciences of the Republic of Uzbekistan
Tashkent Medical Academy
ARTICLE INFO
ABSTRACT
Article history:
Received February 2024
Received in revised form
15 February 2024
Accepted 15 March 2024
Available online
25 April 2024
This article describes the results of the study and the
characteristics of the cytokines IL-4, IL-18, IFN-
γ
and TNF-
α
in the
blood plasma of HIV-positive and HIV-negative children. The study
design was a cross-sectional study in which blood samples were
collected from study subjects during ART. Cytokines IL-4, IL-18, TNF-
α
and INF-
γ
were determined using an ELISA kit (Vector Best,
Russian Federation). The results showed the concentration of IFN-
gamma, a regulator of the humoral immune response, in the group of
patients with HIV infection was statistically significantly higher than
in the control group (p=0.00001). On the contrary, the concentration
of another regulator of the humoral immune response, IL-4, differs
slightly in the main and control groups (p=0.1425). The content of
the pro-inflammatory cytokine TNF-alpha in the HIV-positive group
was significantly higher than in the HIV-negative group (p=0.00001).
Also, the concentration of the pro-inflammatory cytokine IL-18 was
significantly higher in the main group than in the control group
(p=0.0002). Although cytokine profiles during antiretroviral therapy
among people living with HIV (PLHIV) have been documented, no
clearly defined pattern of effects of antiretroviral therapy on cytokine
profiles has been identified.
2181-
1415/©
2024 in Science LLC.
https://doi.org/10.47689/2181-1415-vol5-iss2-pp151-156
This is an open access article under the Attribution 4.0 International
(CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/deed.ru)
Keywords:
interleukins,
HIV,
CD4 cells.
1
Assistant, Central Asian University, Tashkent State Dental Institute. E-mail: d.karimov@centralasian.uz
2
DSc, Senior Researcher, Institute of Human Immunology and Genomics of the Academy of Sciences of the Republic
of Uzbekistan. E-mail: doc.zulfiya@bk.ru
3
PhD, Associate Professor, Tashkent Medical Academy. E-mail: mrashidova7@gmail.com
4
Assistant, Tashkent State Dental Institute. E-mail: bakhtiyorbegmatov.office@gmail.com
5
Assistant, Tashkent Medical Academy. E-mail: dani_karimov@list.ru
6
PhD, Assistant, Tashkent State Dental Institute. E-mail: mrashidova7@gmail.com
7
Assistant, Central Asian University. E-mail: n.sunnatova@centralasian.uz
8
DSc, Associate Professor, Tashkent State Dental Institute. E-mail: aselmaxatova1998@gmail.com
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Oiv-
musbat bolalar qonida yallig‘lanish va yallig‘lanishga
qarshi sitokinlarning xususiyatlari
ANNOTATSIYA
Kalit so‘zlar
:
interleykinlar,
OIV,
CD4 hujayralari.
Ushbu maqolada tadqiqot natijalari va OIV-musbat va
OIV-manfiy bolalarning qon plazmasidagi IL-4, IL-18,
IFN-gamma
va
TNF-alfa
sitokinlarining
xususiyatlari
tasvirlangan. Tadqiqot dizayni kesma tadqiqot bo‘lib, unda qon
namunalari ART paytida o‘rganilayotgan subyektlardan olindi.
Особенности
про
-
и
противовоспалительных
цитокинов в крови у ВИЧ
-
позитивных детей
АННОТАЦИЯ
Ключевые слова:
интерлейкины,
ВИЧ,
CD4 клетки
.
В данной статье описаны результаты исследования и
особенности цитокинов IL
-4, IL-18, IFN-
γ и TNF
-
α в плазме
крови у ВИЧ
-
положительных и ВИЧ
-
негативных детей.
Дизайн исследования представлял собой перекрестное
исследование, в ходе которого у субъектов исследования
вовремя
АРТ собирались образцы крови. Цитокины IL
-4, IL-
18, TNF-
α и INF
-
γ определяли с помощью набора ИФА
(«Вектор Бест», Российская Федерация). Результаты показали
концентрация ИФН
-
гамма
–
ррегулятора гуморального
иммунного ответа в группе больных с ВИЧ
-
инфекцией
статистически значимо выше,
чем в контрольной группе
(p=0.00001). Напротив, концентрация другого регулятора
гуморального иммунного ответа –
ИЛ
-
4 в основной и
контрольной группах различается незначительно (p=0.1425).
Содержание провоспалительного цитокина ФНО
-
альфа в
ВИЧ
-
положительной группе оказалось значительно выше,
чем в ВИЧ
-
отрицательной группе (p=0.00001). Также,
концентрация
провоспалительного
цитокина
ИЛ
-18
оказалась в основной группе значительно выше, чем в
контрольной (p=0.0002). Хотя профили цитокинов во время
антиретровирусной терапии среди людей, живущих
с ВИЧ (ЛЖВ), документированы, четко определенной
закономерности влияния антиретровирусной терапии на
профили цитокинов выявлено не было.
INTRODUCTION
Cytokines are a broad group of soluble proteins and peptides that contribute
significantly to the immune response to intracellular and extracellular pathogens. They
are mainly classified into anti-inflammatory cytokines and pro-inflammatory cytokines
(Tudela et al., 2014), and a physiological balance between these two groups is required
for homeostasis. Cytokines produced by CD4+ T cells play an important role in the fight
against intracellular infections such as HIV.
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One of the known factors required to direct CD4+ T helper 1 cells to an appropriate
immune response are cytokines produced by cells of the innate immune system (Tudela
et al., 2014). Antigen-presenting cells such as dendritic cells release interleukin 12, which
induces naïve T cells into CD4+ T helper 1 (TH1) cells. In fact, TH1 cells produce
interleukin 2 and interferon-gamma, which are pro-inflammatory cytokines responsible
for the intracellular clearance of pathogens.
Prescribing antiretroviral therapy for the treatment of HIV infection has become a
very effective tool for controlling HIV infection among patients. According to da Cunha
and Maselli (da Cunha et al., 2015), currently highly active antiretroviral therapy
(HAART) is the most effective and safe option for combating HIV-1 infection. It improves
immune system recovery while reducing the risk of mortality. It is important to note that
although ART aims to reduce viral load, the impact on the cytokine profile must be taken
into account in drug production, as dysregulated cytokine profile is a hallmark of marked
progression to more severe disease (AIDS).
PURPOSE OF THE STUDY
The purpose of the research was to study the characteristics of cytokines IL-4,
IL-18, IFN-
γ
and TNF-
α
in the blood plasma of HIV-positive and HIV-negative children.
MATERIALS AND METHODS
Study population.
A total of 77 participants, including 69 HIV-infected individuals and 8 HIV-negative
controls, were randomly selected for this study. Blood sampling and research were
carried out at the Republican AIDS Center of Uzbekistan and the Institute of Immunology
and Genomics of the Academy of Sciences of Uzbekistan, Tashkent. Blood samples were
collected at one point during antiretroviral therapy.
A. HIV positive patients
HIV-infected individuals (n=69) were aged 9
–
19 (m=16.33) years and their blood
samples were collected during ART. The number of male and female participants was
42 and 27, respectively.
B. Participants for control
Children in the control group (n=8) were HIV-seronegative and aged from 8 to
14 years (m=12.0). Among them are 5 boys and 3 women.
Inclusion and exclusion criteria
Individuals younger than 19 years of age who initiated ART were eligible to
participate. Individuals with active opportunistic infections, inflammatory conditions,
and diarrhea were excluded. Control subjects were apparently healthy HIV-seronegative
individuals younger than 19 years of age and eligible for the study.
Study design
The study design was a cross-sectional study in which blood samples were
collected from study subjects during ART.
Sample collection
From all study participants, 5 milliliters of fasting blood was collected by
venipuncture. Blood samples were appropriately aliquoted into serum separation tubes
(SST) to obtain pure serum for cytokine determination (IL-4, IL-18, TNF-
α
, and INF-
γ
).
Sera were separated and stored at ≤ −20°C until analysis.
Cytokine measurement
Cytokines IL-4, IL-18, TNF-
α and INF
-
γ were determined using an ELISA kit (Vector
Best, Russian Federation). The quantitative sandwich enzyme-linked immunosorbent
assay method was used.
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Statistical analysis
Results were presented as mean ± standard deviation. Differences between the
results of the control group and HIV-positive subjects receiving ART were analyzed using
Student's t-test. The statistical package MS Excel version 2010 was used to analyze the
data. Significant levels were considered at P < 0.05.
RESULTS
The average value of IL-4 in the blood of participants in the main and control
groups was 1.68 pg/ml and 1.16 pg/ml, respectively. There was no statistically
significant difference between the IL-4 values in the groups (p=0.1425). The average
concentration of IL-18 in the groups of participants with HIV and without HIV was
301.20 pg/ml and 165.25 pg/ml, respectively. The level of IL-18 in the blood plasma of
patients with HIV was significantly higher than that of participants with negative HIV
status (p=0.0002). In the blood serum of participants in the main and control groups,
the average IFN-gamma content was 54.72 pg/ml and 12.07 pg/ml, respectively. It was
found that the concentration of IFN-gamma in the main group was statistically
significantly higher than in the control group (p=0.00001). The results of this study
showed that the concentration of TNF-alpha in the blood plasma of patients in the main
group (M=19.45 pg/ml) was significantly higher than that in the group of HIV-negative
participants (M=9.99 pg/ml) (p=0.00001).
DISCUSSION
Studies have shown (Tudela et al., 2014) (Osuji et al., 2018) that pro-inflammatory
cytokines are released in significant quantities at various stages of HIV infection, but as
the viral load decreases, the number of CD4+ T helper cells in the blood increases, which
leads to increase the release of pro-inflammatory cytokines. The results of our study
were similar to the results of recent studies (Tudela et al., 2014) (Osuji et al., 2018). Thus,
in our study, the content of the pro-inflammatory cytokine TNF-alpha in the HIV-positive
group was significantly higher than in the HIV-negative group (p = 0.00001). Also, the
concentration of the pro-inflammatory cytokine IL-18 was significantly higher in the
main group than in the control group (p=0.0002). However, these results contrast with a
study by Fantauzzi, Floridia (Fantauzzi et al., 2015) and Musve, Oktedalen (Amoani,
Sakyi, Barnie, et al., 2021), who found relatively low levels of TNF-
α
among HIV+ patients.
According to a study (JR & NG, 2016), ART significantly reduces the incidence and
severity of opportunistic diseases. It gives the div the opportunity to restore its
immune function. During antiretroviral therapy, there is a decrease in plasma IFN-
γ
levels, which helps suppress inflammation and activate the immune system. This
decrease in the level of the above-mentioned cytokine may be due to the restoration of
the immune system after suppression of the virus. On the contrary, according to the
results of our study, the concentration of IFN-gamma, a regulator of the humoral immune
response, in the group of patients with HIV infection was statistically significantly higher
than in the control group (p = 0.00001).
CONCLUSION
The concentration of IFN-gamma, a regulator of the humoral immune response, in
the group of patients with HIV infection was statistically significantly higher than in the
control group (p=0.00001). On the contrary, the concentration of another regulator of the
humoral immune response, IL-4, differs slightly in the main and control groups
(p=0.1425). The content of the pro-inflammatory cytokine TNF-alpha in the HIV-positive
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group was significantly higher than in the HIV-negative group (p=0.00001).
The concentration of the pro-inflammatory cytokine IL-18 was significantly Also higher
in the main group than in the control group (p=0.0002).
The dynamics of rise and fall of cytokine levels during ART makes cytokines a good
prognostic tool for monitoring their effectiveness. Although cytokine profiles during
antiretroviral therapy among people living with HIV (PLHIV) have been documented, no
clearly defined pattern of effects of antiretroviral therapy on cytokine profiles has been
identified. While some studies report a pro-inflammatory effect of antiretroviral therapy,
others illustrate the opposite.
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