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. 2024 Feb 20:15:1334236.
doi: 10.3389/fimmu.2024.1334236. eCollection 2024.

Low unspliced cell-associated HIV RNA in early treated adolescents living with HIV on long suppressive ART

Kathleen Gärtner  1 , Sara Domínguez-Rodríguez  2   3 , Judith Heaney  4 , Triantafylia Gkouleli  1   5 , Paul Grant  4 , Karim Dorgham  6 , Delphine Sauce  6 , Cathia Soulie  7 , Eloise J Busby  8 , Denise M O'Sullivan  8   9 , Moira Spyer  1 , Johannes C Botha  1   4 , Maria Angeles Muñoz-Fernandez  10 , Alfredo Tagarro  2   3   11 , Nicola Cotugno  9   12   13 , Jim F Huggett  8   14 , Nigel Klein  1 , Paolo Palma  9   13 , Pablo Rojo Conejo  2 , Caroline Foster  15 , Carlo Giaquinto  16 , Paolo Rossi  12 , Deborah Persaud  17 , Anita De Rossi  16 , Anne-Geneviève Marcelin  7 , Eleni Nastouli  1   4
Affiliations

Low unspliced cell-associated HIV RNA in early treated adolescents living with HIV on long suppressive ART

Kathleen Gärtner et al. Front Immunol. .

Abstract

Introduction: Initiation of antiretroviral treatment (ART) in patients early after HIV-infection and long-term suppression leads to low or undetectable levels of HIV RNA and cell-associated (CA) HIV DNA and RNA. Both CA-DNA and CA-RNA, overestimate the size of the HIV reservoir but CA-RNA as well as p24/cell-free viral RNA can be indicators of residual viral replication. This study describes HIV RNA amounts and levels of cytokines/soluble markers in 40 well-suppressed adolescents who initiated ART early in life and investigated which viral markers may be informative as endpoints in cure clinical trials within this population.

Methods: Forty adolescents perinatally infected with HIV on suppressive ART for >5 years were enrolled in the CARMA study. HIV DNA and total or unspliced CA-RNA in PBMCs were analyzed by qPCR/RT-qPCR and dPCR/RT-dPCR. Cell-free HIV was determined using an ultrasensitive viral load (US-VL) assay. Plasma markers and p24 were analyzed by digital ELISA and correlations between total and unspliced HIV RNA and clinical markers, including age at ART, Western Blot score, levels of cytokines/inflammation markers or HIV CA-DNA, were tested.

Results: CA-RNA was detected in two thirds of the participants and was comparable in RT-qPCR and RT-dPCR. Adolescents with undetectable CA-RNA showed significantly lower HIV DNA compared to individuals with detectable CA-RNA. Undetectable unspliced CA-RNA was positively associated with age at ART initiation and Western Blot score. We found that a higher concentration of TNF-α was predictive of higher CA-DNA and CA-RNA. Other clinical characteristics like US-VL, time to suppression, or percent CD4+ T-lymphocytes were not predictive of the CA-RNA in this cross-sectional study.

Conclusions: Low CA-DNA after long-term suppressive ART is associated with lower CA-RNA, in concordance with other reports. Patients with low CA-RNA levels in combination with low CA-DNA and low Western Blot scores should be further investigated to characterize candidates for treatment interruption trials. Unspliced CA-RNA warrants further investigation as a marker that can be prioritized in paediatric clinical trials where the sample volume can be a significant limitation.

Keywords: HIV-1; adolescents; cell-associated DNA; cell-associated RNA; cytokines; early treated; suppressed.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.

Figures

Figure 1
Figure 1
Analysis of HIV-1 cell-associated nucleic acids in children on ART; (A) LTR- and pol-primer/probe binding sites in the HIV-1 genome; (B) Detection of HIV-1 DNA, total and unspliced CA-RNA in PBMCs; (C) Level of CA-DNA in children with undetectable (brown column) and detectable (green columns) total and unspliced CA-RNA; Wilcoxon matched-pairs signed rank test *p<0.05, ***p<0.01.
Figure 2
Figure 2
Detection of ultra-sensitive VL in CARMA participants. Samples with detectable CA-RNA plus CA-DNA are black, samples with negative CA-RNA are depicted with a green circle and samples with negative CA-DNA have a red center. Double-negative samples are red with a green circle. Shown is the mean of all samples with 95% confidence interval.
Figure 3
Figure 3
Clinical markers in groups of participants with detectable and undetectable CA-RNA; (A) Age at ART, (B) Baseline VL, (C) Time to suppression, (D) %CD4 T-lymphocytes, (E) %CD8 T-lymphocytes, (F) Western Blot Score. Shown are medians with 95% confidence interval; Wilcoxon matched-pairs signed rank test *p<0.05, ***p<0.01.
Figure 4
Figure 4
Importance of different cytokines and inflammation markers for prediction of increasing (A) HIV CA-DNA; (B) total CA-RNA; (C) unspliced CA-RNA.
See this image and copyright information in PMC

References

    1. Consolidated Guidelines on HIV Prevention, Diagnosis, Treatment and Care for Key Populations - 2016 Update. Geneva: WHO; (2016). Guidelines Approved by the Guidelines Review Committee. - PubMed
    1. Violari A, Cotton MF, Gibb DM, Babiker AG, Steyn J, Madhi SA, et al. Early antiretroviral therapy and mortality among HIV-infected infants. N Engl J Med (2008) 359(21):2233–44. doi: 10.1056/NEJMoa0800971 - DOI - PMC - PubMed
    1. Foster C, Pace M, Kaye S, Hopkins E, Jones M, Robinson N, et al. Early antiretroviral therapy reduces HIV DNA following perinatal HIV infection. AIDS (2017) 31(13):1847–51. doi: 10.1097/QAD.0000000000001565 - DOI - PubMed
    1. Tagarro A, Chan M, Zangari P, Ferns B, Foster C, De Rossi A, et al. Early and highly suppressive antiretroviral therapy are main factors associated with low viral reservoir in european perinatally HIV-infected children. J Acquir Immune Defic Syndr (2018) 79(2):269–76. doi: 10.1097/QAI.0000000000001789 - DOI - PMC - PubMed
    1. Li JZ, Etemad B, Ahmed H, Aga E, Bosch RJ, Mellors JW, et al. The size of the expressed HIV reservoir predicts timing of viral rebound after treatment interruption. AIDS (2016) 30(3):343–53. doi: 10.1097/QAD.0000000000000953 - DOI - PMC - PubMed

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