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Microbes & Immunity Oxidative toxicity and folate in HIV on DTG-ART

Taken together, accumulating evidence underscores 2.2.2. Eligibility criteria
that, while DTG remains a highly efficacious antiretroviral Inclusion criteria were as follows:
agent with substantial public health benefits, its long- (i) Aged 18–55 years
term safety profile requires continued vigilance. Concerns (ii) Confirmed HIV serostatus (positive or negative)
spanning neuropsychiatric events, folate metabolism, (iii) Written informed consent
oxidative stress, and metabolic outcomes highlight the need (iv) For the DTG-experienced group: ≥24 weeks of
for integrated pharmacovigilance, mechanistic research, continuous treatment with DTG-based ART.
and context-specific clinical guidance, particularly in
regions with limited dietary folate intake and rising ART Exclusion criteria included:
scale-up. • Pregnancy
• Concurrent opportunistic infections (e.g.,
2. Materials and methods tuberculosis)
2.1. Materials • Known metabolic, hematological, renal, or hepatic
disorders
2.1.1. Study design and setting • Use of antioxidant supplements, cytotoxic drugs, or
This was a quasi-experimental, prospective cohort study medications known to interfere with oxidative stress
conducted at the University of Nigeria Teaching Hospital or folate metabolism.
(UNTH), Ituku-Ozalla, Enugu State, Nigeria. The study 2.2.3. Ethical considerations
was designed to evaluate the effects of DTG-based ART
on hematological indices, micronutrient status, oxidative Ethical approval for the study was obtained from the
stress markers, and toxicity biomarkers among people Health Research Ethics Committee of the UNTH
living with HIV (PLWH). (NHREC/05/01/2008B-FWA00002458-1RB00002323).
All procedures were conducted in accordance with the
2.1.2. Study population and group distribution principles of the Declaration of Helsinki. Written informed
A total of 120 participants were recruited through consent was obtained from all participants before
purposive sampling and categorized into three groups of enrolment. Confidentiality and data protection were
equal size (n = 40 per group): maintained through data anonymization and restricted
(i) Group 1 - Treatment-naïve HIV-positive group: Newly access to personal identifiers.
diagnosed, ART-naïve HIV-positive individuals
(ii) Group 2 - DTG-experienced HIV-positive group: 2.2.4. Sample collection and processing
HIV-positive individuals who had received DTG- Venous blood (5 mL) was collected aseptically from each
based ART for a minimum of 24 weeks participant into two tubes:
(iii) Group 3 - HIV-negative control group: Age- and sex- (i) Ethylenediaminetetraacetic acid tubes: For
matched HIV-negative individuals without chronic hematological profiling and plasma separation
illness or known hematological disorders. (ii) Plain vacutainer tubes: For serum separation.

Recruitment was conducted at the UNTH HIV clinic Samples in plain tubes were allowed to clot, then
and the general outpatient department. HIV diagnosis centrifuged at 3,000 rpm for 10 min, and serum aliquots
was confirmed using the national HIV testing algorithm, were stored at −20°C until biochemical and immunological
while HIV-negative status was verified through serological analyses were performed. Plasma and serum samples were
screening. thawed only once to minimize degradation of analytes.
2.2. Methods 2.2.5. Laboratory procedures
2.2.1. Sample size determination a. Determination of plasma folate levels
The minimum sample size was estimated using Fisher’s Plasma folate concentrations were quantified using
formula for cross-sectional studies, based on the national the Maglumi 600 fully automated chemiluminescence
HIV prevalence rate in Nigeria of 2.1%. A minimum of immunoassay analyzer (Snibe Co., Ltd., China). The
20
102 participants was required to achieve adequate statistical assay was based on a competitive chemiluminescent
power at a 95% confidence level and a 5% margin of error. immunoassay in which folic acid was labeled with N-(4-
To compensate for possible attrition, 120 participants were aminobutyl)-N-ethylisoluminol and immobilized onto
enrolled, distributed equally across the three study groups magnetic microbeads through folate-binding protein
(40 per group). antibodies.

Volume 2 Issue 4 (2025) 124 doi: 10.36922/MI025310074