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. 2018 Dec 17;15(1):191.
doi: 10.1186/s12985-018-1101-9.

Monitoring HIV DNA and cellular activation markers in HIV-infected humanized mice under cART

Mary-Aude Rochat  1 , Erika Schlaepfer  1 , Stefan P Kuster  1 , Duo Li  1 , Annette Audige  1 , Sandra Ivic  1 , Audrey Fahrny  1 , Roberto F Speck  2
Affiliations

Monitoring HIV DNA and cellular activation markers in HIV-infected humanized mice under cART

Mary-Aude Rochat et al. Virol J. .

Abstract

Background: The major obstacle to cure of HIV type-1 infection is the presence of the HIV reservoir, hidden from the immune system and insensitive to combined antiretroviral therapy (cART). Eradication approaches have been hindered by the difficulty for accurately monitoring its size in vivo, especially in the lymphoid organs. Humanized mouse models are a valuable tool for systematically assess the efficacy of therapeutic interventions in reducing the HIV reservoir. Nonetheless, persistence of the HIV reservoir over time, in the presence of cART, has yet to be analyzed in this in vivo model.

Findings: We found that the proviral DNA as well as the total DNA were very stable in the spleen and mesenteric lymph node irrespective of the length of cART. Notably, the amount of proviral DNA was very similar in the spleen and lymph node. Furthermore, we observed a correlation between the percentage of splenic human CD4+ T-cells with total HIV DNA, between the number of human CD38 + CD8+ T-cells in the spleen with the amount of integrated HIV DNA, and eventually between the hCD4/hCD8 ratio in the spleen with integrated as well as total HIV DNA implying that the CD8+ T cells influence the size of the HIV reservoir.

Conclusions: Here, we demonstrated the stability of this reservoir in humanized mice irrespective of the length of cART, confirming the relevancy of this model for HIV latency eradication investigations. Notably, we also found correlates between the frequency of CD4+ T-cells, their activation status and viral parameters, which were analogous to the ones in HIV-infected patients. Thus, hu-mice represent a very valuable HIV latency model.

Keywords: Alu-PCR; HIV reservoir size; HIV-1; Humanized mice; cART.

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Conflict of interest statement

Ethics approval and consent to participate

All humanized mouse experiments as well as the procurement of human cord blood were approved by ethical committees of the University of Zurich and the Federal Veterinary Departments. The experiments were following the local guidelines (TschV, Zurich) and the Swiss animal protection law (TschG). Human cord blood was obtained with informed written consent of the parents and processed anonymously.

Consent for publication

All authors consent for publication.

Competing interests

The authors declare that they have no competing interests.

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Figures

Fig. 1
Fig. 1
Stability of the HIV reservoir in humanized mice in the spleen and lymph node. Infected NSG mice (n = 36) were euthanized prior to cART (n = 3) and after documented viral suppression at 5 (n = 2), 6 (n = 10), 9 (n = 9), 13 (n = 7) and 17 (n = 5) weeks of treatment. a Viral load in hu-mice over time before and after treatment with cART. Spleen and lymph nodes were removed at euthanasia and HIV provirus (b) and total HIV DNA/millions of hCD45+ cells (c) quantified over time shown in blue for the lymph node and in red for the spleen (mean ± SEM). d Correlation between proviruses/millions of hCD45+ cells between the spleen and the lymph node from the same animal or pool of animals. e Correlation between total HIV DNA between the lymph node and the spleen from the same animal or pool of animals, (P = 0.2089, r = 0.311). f and g Correlation between proviruses and total HIV DNA in the spleen or the lymph node, respectively. Because of the low yield of lymphatic tissues in some mice, we were forced to pool the lymphatic tissues of some mice, which had a similar peak viral load. In fact in some mice we were not able to retrieve lymph node tissue. Thus, the data presented for the lymph nodes are based on 26 mice euthanized and eventually 14 (b) and 16 (c) data points, and for the spleen on 36 mice with 24 data points
Fig. 2
Fig. 2
Association between the reservoir size, viral load, hCD4+ T cells and immune activation. a, b Correlation between the viral load and the percentage of HLADR+CD38 + hCD4+ (a) and hCD8+ T-cells (b) in the blood at 4 weeks p.i.. c Correlation between number of proviruses/millions of hCD45+ cells in the spleen with the baseline viral load (P = 0.4626, r = 0.157). d Correlation between total HIV DNA/millions of hCD45+ cells in the spleen with the blood frequency of CD4+ T cells at 4 weeks p.i.. e and f Correlation between proviruses and total HIV DNA with the CD4+/CD8+ T-cell ratio in the spleen. g Correlation between the ratio proviruses/total HIV DNA with the frequency of CD4+ T cells in the spleen. h Correlation between the number of proviruses/millions of hCD45+ cells with the percentage of CD38+ hCD8+ T cells in the spleen. i Correlation between the total HIV DNA/millions of hCD45+ cells with the frequency of HLADR+CD38+ hCD8+
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