HIV, ROS, immune recovery, antiretroviral therapy


Background: Reactive oxygen species (ROS) are generated at physiological levels as a result of cellular metabolism and contribute to cellular interaction and immune response. Elevated ROS may cause cell stress, damage, and apoptosis, and have been detected in different pathological states of infectious and non-infectious etiology.

Aim: To evaluate the association between intracellular ROS in T-cell subsets and HIV VL in chronic HIV infection.

Material and methods: Whole blood samples (Li-heparin, n=33) were analyzed during routine immune monitoring in two groups of HIV+ patients: A (n=21), on continuous cART for at least 2y, with sustained viral suppression (HIV VL<40 copies/ml) and group B (n=12) on cART for less than 2y, average HIV VL 92330 c/ml. Percentage and absolute counts (AC) of CD4+ and CD8+T cells were determined by flow cytometry (Multitest, BD Trucount™ tubes, FACS Canto II). Fluorometric ROS assay kit (Sigma-Aldrich) was adapted for flow cytometry analysis to detect intracellular ROS in CD4+ and CD8+ T-cells (FACSDiva 6.1.2).

Results: The average CD4AC did not differ significantly between group A and B (714 vs. 568, p>0.05), unlike the CD4/CD8 ratio (1.2 vs. 0.6, p<0.01). The mean fluorescence intensity (MFI) of CD4+T intracellular ROS was significantly lower in group A (mean MFI 1744 vs. 2492, p<0.05), unlike the CD8+T cell ROS content (1753 vs. 2129, p>0.05). Noteworthy, CD4+T intracellular ROS correlated positively with HIV VL (R=0.5, p<0.05), unlike CD8+T ROS. On the other hand, positive correlations between CD8+T ROS and cART duration, as well as age (R=0.5, p<0.05 for both) were observed in group A.

Conclusions: CD4+T ROS production may be an indicator of residual HIV activity in the settings of undetectable HIV VL. The combined effects of ageing and long-term cART affect mostly the CD8+T cell compartment.


Commoner B, Townsend, J, Pake G. Free Radicals in Biological Materials. Nature. 1954; 174: 689–691. doi: 10.1038/174689a0

Halliwell B, Gutteridge JMC. Oxigen - boon yet bane introducing oxygen toxicity and reactive species. In Free radicals in biology and medicine. Oxford University Press, 2015, 1-29. doi:10.1093/acprof: oso/9780198717478.001.0001

Ivanov AV, Valuev-Elliston VT, Ivanova ON, Kochetkov SN, Starodubova ES, Bartosch B, Isaguliants MG. Oxidative Stress during HIV Infection: Mechanisms and Consequences. Oxid Med Cell Longev. 2016; 8910396. doi: 10.1155/2016/8910396.

Ray PD, Huang BW, Tsuji Y. Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal. 2012; 24(5):981-90. doi: 10.1016/j.cellsig.2012.01.008.

Belikov AV, Schraven B, Simeoni L. T cells and reactive oxygen species. J Biomed Sci. 2015; 15-22:85. doi: 10.1186/s12929-015-0194-3.

Di Meo S, Reed TT, Venditti P, Victor VM. Role of ROS and RNS Sources in Physiological and Pathological Conditions. Oxid Med Cell Longev. 2016; 2016:1245049. doi: 10.1155/2016/1245049.

Couret J, Chang TL. Reactive Oxygen Species in HIV Infection. EC Microbiol. 2016; 3(6):597-604.

Anelli, T., Sannino, S. & Sitia, R. Proteostasis and “redoxtasis” in the secretory pathway: tales of tails from ERp44 and immunoglobulins. Free Radic. Biol. Med. 2015; 83, 323–330.

Garaude, J. Reprogramming of mitochondrial metabolism by innate immunity. Curr. Opin. Immunol. 2018; 56, 17–23.

Sies H, Jones DP, Reactive oxygen species (ROS) as pleiotropic physiological signalling agents. Nat Rev Mol Cell Biol. 2020; 21, 363–383

Trachootham D, Alexandre J, Huang P. Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat Rev Drug Discov. 2009; 8(7):579-91. doi: 10.1038/nrd2803.

Shukla Y, George J. Combinatorial strategies employing nutraceuticals for cancer development. Nutrition and Physical Activity in Aging, Obesity, and Cancer. 2011; 1229, 162-175.

Andersen JK. Oxidative stress in neurodegeneration: cause or consequence? Nat Med. 2004; 10 Suppl: S18-25. doi: 10.1038/nrn1434.

Szeto H.H. Mitochondria-targeted peptide antioxidants: novel neuroprotective agents. AAPS J. 2006; 8(3): E521–E531.

Paravicini TM, Touyz RM. Redox signaling in hypertension. Cardiovasc Res. 2006; 71(2):247-58. doi: 10.1016/j.cardiores.2006.05.001.

Giacco F, Brownlee M. Oxidative stress and diabetic complications. Circ Res. 2010; 107(9):1058-70. doi: 10.1161/CIRCRESAHA.110.223545.

Haigis MC, Yankner BA. The aging stress response. Mol Cell. 2010; 40(2):333-44. doi: 10.1016/j.molcel.2010.10.002.

Salmen S, Berrueta L: Immune modulators of HIV infection: The role of reactive oxygen species. J Clin Cell Immunol. 2012; 3:121. doi: 10.4172/2155-9899.1000121

Amarnath S, Dong L, Li J, Wu Y, Chen W. Endogenous TGF-beta activation by reactive oxygen species is key to Foxp3 induction in TCR-stimulated and HIV-1-infected human CD4+CD25- T cells. Retrovirology. 2007; 9; 4:57. doi: 10.1186/1742-4690-4-57.

Alimonti JB, Blake BT, Fowke KR. Mechanism of CD4+ T lymphocyte cell death in human immunodeficiency virus infection and AIDS. J Gen Virol. 2003; 7: 1649-1661.

Cummins NW, Badley AD. Mechanisms of HIV-associated lymphocyte apoptosis: 2010. Cell Death Dis. 2010; 1(11): e99. doi: 10.1038/cddis.2010.77.

Ibeh BO, Eze SE, Habu JB. Discordant Levels of Superoxide Dismutase and Catalase Observed in ART Naïve and Experienced HIV Patients in South Eastern Nigeria. J Infect Dis Ther. 2013; 1: 8-16.

Masiá M, Padilla S, Fernández M, Barber X, Moreno S, Iribarren JA, et al. Contribution of Oxidative Stress to Non-AIDS Events in HIV-Infected Patients. J Acquir Immune Defic Syndr. 2017; 75(2): e36–44

Vance DE. Aging with HIV: clinical considerations for an emerging population. The American Journal of Nursing. 2010; 110(3):42-7.

Eisele E, Siliciano RF. Redefining the viral reservoirs that prevent HIV-1 eradication. Immunity. 2012; 37(3): 377–388. doi: 10.1016/j.immuni.2012.08.010.

Vieillard V, Gharakhanian S, Lucar O, Katlama C, Launay O, Autran B, Ho Tsong Fang R, Crouzet J, Murphy RL, Debré P. Perspectives for immunotherapy: which applications might achieve an HIV functional cure? Oncotarget. 2016; 21;7(25):38946-38958. doi: 10.18632/oncotarget.7793.

Antela A, Rivero A, Llibre JM, Moreno S. Redefining therapeutic success in HIV patients: an expert view. Journal of Antimicrobial Chemotherapy. 2021; 76:10, 2501–2518,

Sun Y, Fu Y, Zhang Z, Tang T, Liu J, Ding H, Han X, Xu J, Chu Z, Shang H, Jiang Y. The investigation of CD4+T-cell functions in primary HIV infection with antiretroviral therapy. Medicine (Baltimore). 2017; 96(28): e7430. doi: 10.1097/MD.0000000000007430.

Serrano-Villar S, Sainz T, Lee SA, Hunt PW, Sinclair E, Shacklett BL, et al. HIV-infected individuals with low CD4/CD8 ratio despite effective antiretroviral therapy exhibit altered T cell subsets, heightened CD8+ T cell activation, and increased risk of non-AIDS morbidity and mortality. PLoS Pathog. 2014; 10(5): e1004078.

Yu F, Hao Y, Zhao H, Xiao J, Han N, Zhang Y, Dai G, Chong X, Zeng H, Zhang F. Distinct Mitochondrial Disturbance in CD4+T and CD8+T Cells from HIV-Infected Patients. J Acquir Immune Defic Syndr. 2017; 74(2):206-212. doi: 10.1097/QAI.0000000000001175.

Aquaro S, Scopelliti F, Pollicita M, Perno CF. Oxidative stress and HIV infection: Target pathways for novel therapies? Future HIV Therapy. 2008; 2(4):327-338.

Deguit CDT, Hough M, Hoh R, Krone M, Pilcher CD, Martin JN, Deeks SG, McCune JM, Hunt PW, Rutishauser RL. Some Aspects of CD8+ T-Cell Exhaustion Are Associated with Altered T-Cell Mitochondrial Features and ROS Content in HIV Infection. J Acquir Immune Defic Syndr. 2019; 82(2):211-219. doi: 10.1097/QAI.0000000000002121.




How to Cite

Emilova, R., Todorova, Y., Aleksova, M., Dimitrova, R., Alexiev, I., Grigorova, L., Yancheva, N., & Nikolova, M. (2022). DETERMINATION OF INTRACELLULAR REACTIVE OXYGEN SPECIES IN T-CELL SUBSETS OF HIV+ PATIENTS ON CONTINUOUS cART. PROBLEMS of Infectious and Parasitic Diseases, 50(1), 5–11.




Most read articles by the same author(s)