Stephanie B. H. Gumbs’s research while affiliated with University Medical Center Utrecht and other places

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Publications (5)


Fig. 1 Relationship of HIV-1 RNA levels (log 10cp/mL) measured in paired plasma and CSF samples in neuroasymptomatic and neurosymptomatic subjects. a Plots depicts the correlation between HIV RNA CSF and HIV RNA plasma for all subjects. Black dashed line represents line of identity. b, c, d Boxes depict median HIV RNA levels and IQR, measured in plasma (red) and CSF (blue) in all patients, neuroasymptomatic and neurosymptomatic subjects. e, f Boxes depict CSF (blue) and plasma (red) median HIV RNA levels and IQR between neuroasymptomatic and neurosymptomatic subjects. * = Statistically significant (p < 0.05)
Characterization of HIV variants from paired Cerebrospinal fluid and Plasma samples in primary microglia and CD4+ T-cells
  • Article
  • Full-text available

May 2024

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48 Reads

Journal of NeuroVirology

Stephanie B H Gumbs

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Tania Mudrikova

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Despite antiretroviral therapy (ART), HIV persistence in the central nervous system (CNS) continues to cause a range of cognitive impairments in people living with HIV (PLWH). Upon disease progression, transmigrating CCR5-using T-cell tropic viruses are hypothesized to evolve into macrophage-tropic viruses in the CNS that can efficiently infect low CD4-expressing cells, such as microglia. We examined HIV-1 RNA concentration, co-receptor usage, and CSF compartmentalization in paired CSF and blood samples from 19 adults not on treatment. Full-length envelope CSF- and plasma-derived reporter viruses were generated from 3 subjects and phenotypically characterized in human primary CD4 ⁺ T-cells and primary microglia. Median HIV RNA levels were higher in plasma than in CSF (5.01 vs. 4.12 log10 cp/mL; p = 0.004), and coreceptor usage was mostly concordant for CCR5 across the paired samples (n = 17). Genetically compartmentalized CSF viral populations were detected in 2 subjects, one with and one without neurological symptoms. All viral clones could replicate in T-cells (R5 T cell-tropic). In addition, 3 CSF and 1 plasma patient-derived viral clones also had the capacity to replicate in microglia/macrophages and, therefore have an intermediate macrophage tropic phenotype. Overall, with this study, we demonstrate that in a subset of PLWH, plasma-derived viruses undergo genetic and phenotypic evolution within the CNS, indicating viral infection and replication in CNS cells. It remains to be studied whether the intermediate macrophage-tropic phenotype observed in primary microglia represents a midpoint in the evolution towards a macrophage-tropic phenotype that can efficiently replicate in microglial cells and propagate viral infection in the CNS.

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Clinical and virological data of IciS-05 (a) and IciS-11 (b) pre- and post-allo-HSCT. HIV-1 proviral DNA expressed as LTR DNA copies/million PBMC, and HIV-1 RNA as copies/mL for plasma and chimerism. Open symbols represent values under the level of quantification. Abbreviations: allo-HSCT, allogeneic hematopoietic stem cell transplantation. PBMCs, peripheral blood mononuclear cells.
Results of HIV-1-DNA quantification and characterization in patient IciS-05.
Results of HIV-1-DNA quantification and characterization in patient IciS-11.
Autopsy Study Defines Composition and Dynamics of the HIV-1 Reservoir after Allogeneic Hematopoietic Stem Cell Transplantation with CCR5Δ32/Δ32 Donor Cells

September 2022

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122 Reads

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9 Citations

Allo-HSCT with CCR5Δ32/Δ32 donor cells is the only curative HIV-1 intervention. We investigated the impact of allo-HSCT on the viral reservoir in PBMCs and post-mortem tissue in two patients. IciS-05 and IciS-11 both received a CCR5Δ32/Δ32 allo-HSCT. Before allo-HSCT, ultrasensitive HIV-1 RNA quantification; HIV-1-DNA quantification; co-receptor tropism analysis; deep-sequencing and viral characterization in PBMCs and bone marrow; and post-allo-HSCT, ultrasensitive RNA and HIV-1-DNA quantification were performed. Proviral quantification, deep sequencing, and viral characterization were done in post-mortem tissue samples. Both patients harbored subtype B CCR5-tropic HIV-1 as determined genotypically and functionally by virus culture. Pre-allo-HSCT, HIV-1-DNA could be detected in both patients in bone marrow, PBMCs, and T-cell subsets. Chimerism correlated with detectable HIV-1-DNA LTR copies in cells and tissues. Post-mortem analysis of IciS-05 revealed proviral DNA in all tissue biopsies, but not in PBMCs. In patient IciS-11, who was transplanted twice, no HIV-1-DNA could be detected in PBMCs at the time of death, whereas HIV-1-DNA was detectable in the lymph node. In conclusion, shortly after CCR5Δ32/Δ32, allo-HSCT HIV-1-DNA became undetectable in PBMCs. However, HIV-1-DNA variants identical to those present before transplantation persisted in post-mortem-obtained tissues, indicating that these tissues play an important role as viral reservoirs.


Table 1 ).
Shock and kill within the CNS: A promising HIV eradication approach?

September 2022

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91 Reads

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14 Citations

Journal of Leukocyte Biology

The most studied HIV eradication approach is the “shock and kill” strategy, which aims to reactivate the latent reservoir by latency reversing agents (LRAs) and allowing elimination of these cells by immune‐mediated clearance or viral cytopathic effects. The CNS is an anatomic compartment in which (persistent) HIV plays an important role in HIV‐associated neurocognitive disorder. Restriction of the CNS by the blood–brain barrier is important for maintenance of homeostasis of the CNS microenvironment, which includes CNS‐specific cell types, expression of transcription factors, and altered immune surveillance. Within the CNS predominantly myeloid cells such as microglia and perivascular macrophages are thought to be a reservoir of persistent HIV infection. Nevertheless, infection of T cells and astrocytes might also impact HIV infection in the CNS. Genetic adaptation to this microenvironment results in genetically distinct, compartmentalized viral populations with differences in transcription profiles. Because of these differences in transcription profiles, LRAs might have different effects within the CNS as compared with the periphery. Moreover, reactivation of HIV in the brain and elimination of cells within the CNS might be complex and could have detrimental consequences. Finally, independent of activity on latent HIV, LRAs themselves can have adverse neurologic effects. We provide an extensive overview of the current knowledge on compartmentalized (persistent) HIV infection in the CNS and on the “shock and kill” strategy. Subsequently, we reflect on the impact and promise of the “shock and kill” strategy on the elimination of persistent HIV in the CNS. Review on the obstacles and limitations of using the shock and kill strategy to eradicate the HIV CNS reservoir.


Characterization of HIV-1 Infection in Microglia-Containing Human Cerebral Organoids

April 2022

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169 Reads

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42 Citations

Abstract: The achievement of an HIV cure is dependent on the eradication or permanent silencing of HIV-latent viral reservoirs, including the understudied central nervous system (CNS) reservoir. This requires a deep understanding of the molecular mechanisms of HIV’s entry into the CNS, latency establishment, persistence, and reversal. Therefore, representative CNS culture models that reflect the intercellular dynamics and pathophysiology of the human brain are urgently needed in order to study the CNS viral reservoir and HIV-induced neuropathogenesis. In this study, we characterized a human cerebral organoid model in which microglia grow intrinsically as a CNS culture model to study HIV infection in the CNS. We demonstrated that both cerebral organoids and isolated organoid-derived microglia (oMG), infected with replication-competent HIVbal reporter viruses, support productive HIV infection via the CCR5 co-receptor. Productive HIV infection was only observed in microglial cells. Fluorescence analysis revealed microglia as the only HIV target cell. Susceptibility to HIV infection was dependent on the co-expression of microglia-specific markers and the CD4 and CCR5 HIV receptors. Altogether, this model will be a valuable tool within the HIV research community to study HIV–CNS interactions, the underlying mechanisms of HIV-associated neurological disorders (HAND), and the efficacy of new therapeutic and curative strategies on the CNS viral reservoir.


Human microglial models to study HIV infection and neuropathogenesis: a literature overview and comparative analyses

February 2022

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98 Reads

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20 Citations

Journal of NeuroVirology

HIV persistence in the CNS despite antiretroviral therapy may cause neurological disorders and poses a critical challenge for HIV cure. Understanding the pathobiology of HIV-infected microglia, the main viral CNS reservoir, is imperative. Here, we provide a comprehensive comparison of human microglial culture models: cultured primary microglia (pMG), microglial cell lines, monocyte-derived microglia (MDMi), stem cell–derived microglia (iPSC-MG), and microglia grown in 3D cerebral organoids (oMG) as potential model systems to advance HIV research on microglia. Functional characterization revealed phagocytic capabilities and responsiveness to LPS across all models. Microglial transcriptome profiles of uncultured pMG showed the highest similarity to cultured pMG and oMG, followed by iPSC-MG and then MDMi. Direct comparison of HIV infection showed a striking difference, with high levels of viral replication in cultured pMG and MDMi and relatively low levels in oMG resembling HIV infection observed in post-mortem biopsies, while the SV40 and HMC3 cell lines did not support HIV infection. Altogether, based on transcriptional similarities to uncultured pMG and susceptibility to HIV infection, MDMi may serve as a first screening tool, whereas oMG, cultured pMG, and iPSC-MG provide more representative microglial culture models for HIV research. The use of current human microglial cell lines (SV40, HMC3) is not recommended.

Citations (4)


... The intact proviral DNA assay (IPDA) 23 detected potentially intact proviruses in two samples that had been obtained during ART-suppressed viremia 17 and 32 months before allo-HSCT in the context of his participation in the Swiss HIV cohort study (Fig. 2c). By contrast, potentially intact proviruses were never detected following people with HIV who did not achieve full donor chimerism 12,17 or in tissue sanctuaries analyzed in necropsy studies 18 . Moreover, during the weeks following allo-HSCT, a window of vulnerability occurs when highly activated CD4 + T cells from both donor and recipient coexist 19 , thereby increasing the risk of reservoir reseeding if infection of donor cells is not prevented by pharmacological or genetic and host barriers. ...

Reference:

Sustained HIV remission after allogeneic hematopoietic stem cell transplantation with wild-type CCR5 donor cells
Autopsy Study Defines Composition and Dynamics of the HIV-1 Reservoir after Allogeneic Hematopoietic Stem Cell Transplantation with CCR5Δ32/Δ32 Donor Cells

... We have discussed a few approaches to achieve reduced viral replication in the CNS. One approach discussed by Nühn et al. in their comprehensive 2022 review is that of "shock and kill" [167]. This approach aims to reactivate the latent reservoir with latency reversal agents, including epigenetic modifiers, intracellular signalling modulators, cytokine or immune receptor agonists and transcription elongation factors, which will then be eliminated by the immune system or by viral cytolysis, achieving viral cure. ...

Shock and kill within the CNS: A promising HIV eradication approach?

Journal of Leukocyte Biology

... In the context of immune cell-associated genes, there were also increases in the genes that encode T cell CD8α and CD8β chains in 5XFAD/MR1 KO mice, whereas CD4 gene expression was decreased in these mice. The expression changes are across all cell populations; therefore, it is important to note that CD4 can be upregulated on microglia during an human immunodeficiency virus infection 22 and CD8 has been found on microglia/macrophages after a stroke. 23 How this might contribute to the reduced pathology in the MR1-deficient 5XFAD mice remains to be determined. ...

Characterization of HIV-1 Infection in Microglia-Containing Human Cerebral Organoids

... Recent studies have begun to use organoids to model the impact of HIV and antiretroviral drugs on the central nervous system (Donadoni et al., 2024;Premeaux et al., 2021). Many of these studies have focused on generating organoids with integrated microglia to allow for the investigation of productive infection of HIV and its effect on neural cell types (Gumbs et al., 2022;Kong et al., 2024). Only a few have focused on the direct impact of antiretroviral drugs, including DTG, and shown its impact on morphogenesis, organoid structure, and expression of the folate receptor alpha (Caiaffa et al., 2024;Kirkwood-Johnson et al., 2021). ...

Human microglial models to study HIV infection and neuropathogenesis: a literature overview and comparative analyses

Journal of NeuroVirology