Monique Nijhuis | University Medical Center Utrecht ,The Netherlands (original) (raw)

Papers by Monique Nijhuis

Clinical Infectious Diseases, Apr 23, 2014

The "Berlin patient" is the first patient cured of HIV-1 infection after allogeneic transplantati... more The "Berlin patient" is the first patient cured of HIV-1 infection after allogeneic transplantation with nonfunctional CCR5 coreceptor stem cells. We demonstrate that CXCR4predicted minority viruses present prior to transplantation were unable to rebound after transplantation due to their dependence on CCR5 for replication and high genetic barrier toward CXCR4 usage.

Retrovirology, Aug 6, 2012

Human Immunodeficiency Virus (HIV) maturation plays an essential role in the viral life cycle by ... more Human Immunodeficiency Virus (HIV) maturation plays an essential role in the viral life cycle by enabling the generation of mature infectious virus particles through proteolytic processing of the viral Gag and GagPol precursor proteins. An impaired polyprotein processing results in the production of non-infectious virus particles. Consequently, particle maturation is an excellent drug target as exemplified by inhibitors specifically targeting the viral protease (protease inhibitors; PIs) and the experimental class of maturation inhibitors that target the precursor Gag and GagPol polyproteins. Considering the different target sites of the two drug classes, direct cross-resistance may seem unlikely. However, coevolution of protease and its substrate Gag during PI exposure has been observed both in vivo and in vitro. This review addresses in detail all mutations in Gag that are selected under PI pressure. We evaluate how polymorphisms and mutations in Gag affect PI therapy, an aspect of PI resistance that is currently not included in standard genotypic PI resistance testing. In addition, we consider the consequences of Gag mutations for the development and positioning of future maturation inhibitors.

Science Translational Medicine, May 6, 2020

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the only medical intervention w... more Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the only medical intervention which has led to an HIV cure. Whereas the size of the HIV reservoir sharply decreases after allo-HSCT, the dynamics of the T-cell reconstitution has not been comprehensively described. We analyzed the activation and differentiation of CD4 + and CD8 + T-cells, and the breadth and quality of HIV-and CMV-specific CD8 + T-cell responses in 16 patients with HIV who underwent allo-HSCT (including 5 individuals who received cells from CCR532/32 donors) to treat their underlying hematological malignancy and who remained on antiretroviral therapy (ART). We found that reconstitution of the T-cell compartment was slow and heterogeneous with an initial expansion of activated CD4 + T-cells that preceded the expansion of CD8 + T-cells; full immune reconstitution was not achieved after allo-HSCT. Although HIV-specific CD8 + T-cells disappeared immediately after allo-HSCT, weak ex vivo HIV-specific CD8 + T-cell responses were detectable several weeks after transplant, and could still be detected at the time of full T-cell chimerism, indicating that de novo priming, and hence antigen exposure, occurred during the time of T-cell expansion. These HIV-specific T-cells had limited functionality compared to CMVspecific CD8 + T-cells, and persisted years after allo-HSCT. In conclusion, immune reconstitution was slow, heterogeneous and incomplete and coincided with de novo detection of weak HIVspecific T-cell responses. The initial short phase of high T-cell activation, in which HIV antigens were present, may constitute a window of vulnerability for the reseeding of viral reservoirs, emphasizing the importance of maintaining ART directly after allo-HSCT.

PLOS ONE, Jan 9, 2013

Background: Optimal regimen choice of antiretroviral therapy is essential to achieve long-term cl... more Background: Optimal regimen choice of antiretroviral therapy is essential to achieve long-term clinical success. Integrase inhibitors have swiftly been adopted as part of current antiretroviral regimens. The purpose of this study was to review the evidence for integrase inhibitor use in clinical settings. Methods: MEDLINE and Web-of-Science were screened from April 2006 until November 2012, as were hand-searched scientific meeting proceedings. Multiple reviewers independently screened 1323 citations in duplicate to identify randomized controlled trials, nonrandomized controlled trials and cohort studies on integrase inhibitor use in clinical practice. Independent, duplicate data extraction and quality assessment were conducted. Results: 48 unique studies were included on the use of integrase inhibitors in antiretroviral therapy-naive patients and treatment-experienced patients with either virological failure or switching to integrase inhibitors while virologically suppressed. On the selected studies with comparable outcome measures and indication (n = 16), a meta-analysis was performed based on modified intention-to-treat (mITT), on-treatment (OT) and as-treated (AT) virological outcome data. In therapy-naive patients, favorable odds ratios (OR) for integrase inhibitor-based regimens were observed, (mITT OR 0.71, 95% CI 0.59-0.86). However, integrase inhibitors combined with protease inhibitors only did not result in a significant better virological outcome. Evidence further supported integrase inhibitor use following virological failure (mITT OR 0.27; 95% CI 0.11-0.66), but switching to integrase inhibitors from a high genetic barrier drug during successful treatment was not supported (mITT OR 1.43; 95% CI 0.89-2.31). Integrase inhibitor-based regimens result in similar immunological responses compared to other regimens. A low genetic barrier to drug-resistance development was observed for raltegravir and elvitegravir, but not for dolutegravir. Conclusion: In first-line therapy, integrase inhibitors are superior to other regimens. Integrase inhibitor use after virological failure is supported as well by the meta-analysis. Careful use is however warranted when replacing a high genetic barrier drug in treatment-experienced patients switching successful treatment.

Antiviral Therapy, 2009

Objective: Bevirimat is the first drug of a new class of antivirals that hamper the maturation of... more Objective: Bevirimat is the first drug of a new class of antivirals that hamper the maturation of HIV. The objective of this study was to evaluate the sequence variability of the gag region targeted by bevirimat in HIV subtype-B isolates. Methods: Of 484 HIV subtype-B isolates, the gag region comprising amino acids 357-382 was sequenced. Of the patients included, 270 were treatment naive and 214 were treatment experienced. In the latter group, 48 HIV isolates harboured mutations associated with reverse transcriptase inhibitor resistance only, and 166 HIV isolates carried mutations associated with protease inhibitor resistance. Results: In the treatment-naive patient population, approximately 30% harboured an HIV isolate with at least one mutation associated with a reduced susceptibility to bevirimat (H358Y, L363M, Q369H, V370A/M/del and T371del). In HIV isolates with protease inhibitor resistance, the prevalence of bevirimat resistance mutations increased to 45%. Accumulation of mutations at four positions in the bevirimat target region, S368C, Q369H, V370A and S373P, was significantly observed. Mutations associated with bevirimat resistance were detected more frequently in HIV isolates with three or more protease inhibitor resistance mutations than in those with less than three protease inhibitor mutations. Conclusion: Reduced bevirimat activity can be expected in one-third of treatment-naive HIV subtype-B isolates and significantly more in protease inhibitor-resistant HIV. These data indicate that screening for bevirimat resistance mutations before administration of the drug is essential.

Journal of the International AIDS Society, 2012

Journal of the International AIDS Society, Jul 1, 2018

Journal of Clinical Virology, Sep 1, 2016

Clinical outcome of HCV genotype-‐1 therapy combina8on of pegylated-‐interferon-‐alpha (IFN), ... more Clinical outcome of HCV genotype-‐1 therapy combina8on of pegylated-‐interferon-‐alpha (IFN), ribavirin (RBV) and protease-‐inhibitors (PIs) depends on host and viral factors. This non-‐interven8onal study collects data from PI-‐resistance-‐associated-‐muta8ons within the NS3 gene, viral quasispecies distribu8on and host factors, in order to predict clinical outcome using the geno2pheno[HCV]-‐tool. METHODS NS5B+NS3 sequences from plasma samples were obtained. Subtyping (including GT-‐1a samples clade classifica8on) and resistance against Boceprevir (BOC) and Telaprevir (TPV) was determined with geno2pheno[HCV] (hSp://hcv.bioinf.mpi-‐inf.mpg.de/). If possible, host IL28B-‐polymorphism was tested. RESULTS 130 GT1-‐infected pa8ents are up-‐to-‐date enrolled. IL28B polymorphism was available for 121 pa8ents: 29xCC, 75xCT, 17xTT. 74/130 samples were 1a and 56/130 1b. Baseline BOC/TPV-‐resistance-‐associated-‐muta8ons were detected in 22/130 (16.9%) baseline-‐samples: 9x132V, 4x117H, 1x132V+174F, 1x168G, 1x36L, 1x36A, 1x36M+80K+155K, 2x54S, 1x54S+80R, 1x54S+155K. 73 GT 1a-‐samples could be further classified: 44/73 clade-‐I and 29/73 clade-‐II. The muta8on Q80K, conferring treatment problems with simeprevir, was observed in 20/35 clade-‐I baseline samples. CONCLUSION Analysis of NS5B+NS3 with geno2pheno[HCV] interpreta8on allows subtyping, clade classifica8on, and predic8on of PIs suscep8bility. PI resistance-‐muta8ons exist at baseline in higher extent as reported in previous studies. Incorpora8ng addi8onal viral and clinical data will con8nuously improve geno2pheno[HCV] for beSer therapy response predic8on. Fig. 2. Genotyping/ subtyping success rate for cohort 1. Percentages are calculated considering prior subtyping based on 5 ́-‐UTR (the amount of samples). Fig. 3. Genotype/ subtype distribuJon within the cohort 1. Fig. 4. NS3 amplificaJon and sequencing success rate. Amplifica8on (blue) and sequencing (green) rates are decipted as bars (below percentage is the amount of samples). Fig. 5. Baseline genotype/ subtype distribuJon within the cohort 2. Fig. 6. Geno-‐ /subtyping and NS3 sequencing success rate in week 0. Geno-‐ /subtyping (red) and sequencing (green) rates are decipted as bars (below percentage is the amount of samples). Fig. 7. Genotype/ subtype distribuJon within the cohort 2, week 4. Fig. 8. Geno-‐ /subtyping and NS3 sequencing success rate in week 4. Geno-‐ /subtyping (red) and sequencing (green) rates are decipted as bars (below percentage is the amount of samples). SUMMARY & CONCLUSION

Journal of the International AIDS Society, Nov 1, 2014

Clinical Infectious Diseases, Mar 9, 2017

Background. In Western countries emergence of human immunodeficiency virus (HIV) drug resistance ... more Background. In Western countries emergence of human immunodeficiency virus (HIV) drug resistance has tremendously decreased, and transmission of drug resistance has merely stabilized in recent years. However, in many endemic settings with limited resources rates of emerging and transmitted drug resistance are not regularly assessed. Methods. We performed a survey including all HIV-infected individuals who received resistance testing in 2010-2015 in Aruba, a highly endemic HIV area in the Caribbean. Transmitted HIV drug resistance was determined using World Health Organization (WHO) criteria. Transmission dynamics were investigated using phylogenetic analyses. In a subset, baseline samples were re-analyzed using next generation sequencing (NGS). Results. Baseline resistance testing was performed in 104 newly diagnosed untreated individuals (54% of all newly diagnosed individuals in 2010-2015): 86% were men, 39% were foreign-born, and 22% had AIDS at diagnosis. And 33% (95% CI: 24-42%) was infected with a drug-resistant HIV variant. The prevalence of resistance to non-nucleoside reverse transcriptase inhibitors (NNRTIs) reached 45% (95% CI: 27-64%) in 2015, all based on the prevalence of mutation K103N. NGS did not demonstrate additional minority K103N-variants compared to routine resistance testing. K103N-harboring strains were introduced into the therapy-unexposed population via at least 6 independent transmissions epidemiologically linked to the surrounding countries. Virological failure of the WHO-recommended first-line NNRTI-based regimen was higher in the presence of K103N. Conclusions. The prevalence of resistant HIV in Aruba has increased to alarming levels, compromising the WHO-recommended first-line regimen. As adequate surveillance as advocated by the WHO is limited, the Caribbean region could face an unidentified rise of NNRTI-resistant HIV.

Retrovirology, Dec 1, 2014

Background: Different patterns of drug resistance are observed in treated and therapy naïve HIV-1... more Background: Different patterns of drug resistance are observed in treated and therapy naïve HIV-1 infected populations. Especially the NRTI-related M184I/V variants, which are among the most frequently encountered mutations in treated patients, are underrepresented in the antiretroviral naïve population. M184I/V mutations are known to have a profound effect on viral replication and tend to revert over time in the new host. However it is debated whether a diminished transmission efficacy of HIV variants with a reduced replication capacity can also contribute to the observed discrepancy in genotypic patterns. As dendritic cells (DCs) play a pivotal role in HIV-1 transmission, we used a model containing primary human Langerhans cells (LCs) and DCs to compare the transmission efficacy M184 variants (HIV-M184V/I/T) to HIV wild type (HIV-WT). As control, we used HIV harboring the NNRTI mutation K103N (HIV-K103N) which has a minor effect on replication and is found at a similar prevalence in treated and untreated individuals. Results: In comparison to HIV-WT, the HIV-M184 variants were less efficiently transmitted to CCR5 + Jurkat T cells by both LCs and DCs. The transmission rate of HIV-K103N was slightly reduced to HIV-WT in LCs and even higher than HIV-WT in DCs. Replication experiments in CCR5 + Jurkat T cells revealed no apparent differences in replication capacity between the mutant viruses and HIV-WT. However, viral replication in LCs and DCs was in concordance with the transmission results; replication by the HIV-M184 variants was lower than replication by HIV-WT, and the level of replication of HIV-K103N was intermediate for LCs and higher than HIV-WT for DCs. Conclusions: Our data demonstrate that drug resistant M184-variants display a reduced replication capacity in LCs and DCs which directly impairs their transmission efficacy. As such, diminished transmission efficacy may contribute to the lower prevalence of drug resistant variants in therapy naive individuals.

Frontiers in Immunology

Cellular immune responses are of pivotal importance to understand SARS-CoV-2 pathogenicity. Using... more Cellular immune responses are of pivotal importance to understand SARS-CoV-2 pathogenicity. Using an enzyme-linked immunosorbent spot (ELISpot) interferon-γ release assay with wild-type spike, membrane and nucleocapsid peptide pools, we longitudinally characterized functional SARS-CoV-2 specific T-cell responses in a cohort of patients with mild, moderate and severe COVID-19. All patients were included before emergence of the Omicron (B.1.1.529) variant. Our most important finding was an impaired development of early IFN-γ-secreting virus-specific T-cells in severe patients compared to patients with moderate disease, indicating that absence of virus-specific cellular responses in the acute phase may act as a prognostic factor for severe disease. Remarkably, in addition to reactivity against the spike protein, a substantial proportion of the SARS-CoV-2 specific T-cell response was directed against the conserved membrane protein. This may be relevant for diagnostics and vaccine design...

Nature Medicine

Despite scientific evidence originating from two patients published to date that CCR5Δ32/Δ32 hema... more Despite scientific evidence originating from two patients published to date that CCR5Δ32/Δ32 hematopoietic stem cell transplantation (HSCT) can cure human immunodeficiency virus type 1 (HIV-1), the knowledge of immunological and virological correlates of cure is limited. Here we characterize a case of long-term HIV-1 remission of a 53-year-old male who was carefully monitored for more than 9 years after allogeneic CCR5Δ32/Δ32 HSCT performed for acute myeloid leukemia. Despite sporadic traces of HIV-1 DNA detected by droplet digital PCR and in situ hybridization assays in peripheral T cell subsets and tissue-derived samples, repeated ex vivo quantitative and in vivo outgrowth assays in humanized mice did not reveal replication-competent virus. Low levels of immune activation and waning HIV-1-specific humoral and cellular immune responses indicated a lack of ongoing antigen production. Four years after analytical treatment interruption, the absence of a viral rebound and the lack of i...

PLOS ONE

The rapid emergence and spread of numerous severe acute respiratory syndrome coronavirus 2 (SARS-... more The rapid emergence and spread of numerous severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants across the globe underscores the crucial need for continuous SARS-CoV-2 surveillance to ensure that potentially more pathogenic variants are detected early and contained. Whole genome sequencing (WGS) is currently the gold standard for COVID-19 surveillance; however, it remains cost-prohibitive and requires specialized technical skills. To increase surveillance capacity, especially in resource-scarce settings, supplementary methods that are cost- and time-effective are needed. Real-time multiplex PCR genotyping assays offer an economical and fast solution for screening circulating and emerging variants while simultaneously complementing existing WGS approaches. In this study we evaluated the AllplexTM SARS-CoV-2 Variants II multiplex real-time PCR genotyping assay, Seegene (South Korea), and implemented it in retrospectively characterizing circulating SARS-CoV-2 variants ...

South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde, 2022

We describe a case of prolonged SARS-CoV-2 RNA shedding in an HIV-negative 21-year-old man recove... more We describe a case of prolonged SARS-CoV-2 RNA shedding in an HIV-negative 21-year-old man recovering from abdominal and thoracic trauma. Nasopharyngeal (NP) swabs collected at 12 time points over a 95-day span all tested positive for SARS-CoV-2 by reverse transcription polymerase chain reaction (RT-PCR). Genotyping revealed canonical beta-variant E484K and N501Y mutations at earlier time points. Human rhinovirus, coronavirus NL63 and respiratory syncytial virus B were detected at different time points by RT-PCR. Full blood analysis at time point 9 (day 82) showed leukopenia with lymphocytosis. The patient's NP swab tested negative for SARS-CoV-2 by RT-PCR 101 days after the first positive test. The prolonged duration of SARS-CoV-2 RNA shedding in the context of trauma presented here is unique and has important implications for COVID-19 diagnosis, management and policy guidelines.

Nature Medicine, 2021

,16,17 ✉ and The International AIDS Society (IAS) Global Scientific Strategy working group* Despi... more ,16,17 ✉ and The International AIDS Society (IAS) Global Scientific Strategy working group* Despite the success of antiretroviral therapy (ART) for people living with HIV, lifelong treatment is required and there is no cure. HIV can integrate in the host genome and persist for the life span of the infected cell. These latently infected cells are not recognized as foreign because they are largely transcriptionally silent, but contain replication-competent virus that drives resurgence of the infection once ART is stopped. With a combination of immune activators, neutralizing antibodies, and therapeutic vaccines, some nonhuman primate models have been cured, providing optimism for these approaches now being evaluated in human clinical trials. In vivo delivery of gene-editing tools to either target the virus, boost immunity or protect cells from infection, also holds promise for future HIV cure strategies. In this Review, we discuss advances related to HIV cure in the last 5 years, highlight remaining knowledge gaps and identify priority areas for research for the next 5 years.

Open Forum Infectious Diseases, 2021

Severe acute respiratory syndrome coronavirus 2 infection after coronavirus disease 2019 vaccinat... more Severe acute respiratory syndrome coronavirus 2 infection after coronavirus disease 2019 vaccination raises concerns about the emergence of vaccine escape variants. Here we characterize 14 breakthrough infections among 5860 fully vaccinated Dutch health care workers ≥14 days after the final dose of vaccination with either BNT162b2, mRNA-1273, or Ad26.COV2.S. These breakthrough infections presented with regular B.1.1.7 (Alpha) and B.1.617.2 (Delta) variants and high viral loads, despite normal vaccine-induced B- and T-cell immune responses detected by live virus neutralization assays and ELISpot. High-risk exposure settings, such as in households, indicate a potential risk of viral transmission despite full vaccination.

Journal of Virus Eradication, 2017

Background: Allogeneic stem cell transplantation (allo-SCT) in HIV-infected subjects with severe ... more Background: Allogeneic stem cell transplantation (allo-SCT) in HIV-infected subjects with severe hematological malignancies is the only described strategy capable to dramatically reduce HIV latent reservoir. Whether this putative eradication strategy is associated with seroreversion has not been established yet. Within the IciStem consortium, we explored the longitudinal serostatus of HIV+ individuals after allo SCT. Methods: Longitudinal plasma samples from 13 HIV+ allotransplanted patients under cART were analyzed. HIV-1 serostatus was tested in a qualitative western blot assay (New Lav Blot I, Biorad). For 7 subjects with longer follow-up (>2years) additional analysis was done using the standard and low-sensitive (LS) versions of the VITROS anti-HIV-1 assay (Ortho-Clinical Diagnostics) and the LAg avidity assay. Results: Evolution of the HIV-specific antibodies in plasma was studied for 13 allo-SCT patients, all of them under cART. We observed that p24 and/or p31 disappeared in 9/13 patients, sometimes only three months after allo-SCT. gp140, gp160, and gp120 bands persisted in most individuals. Surprisingly, in two cases we found an undetermined (Pt#19 and Pt#28) western blot. LAg avidity assay was negative in 6/7 individuals with longer follow. LS-VITROS detuned assay showed that transplanted patients presented lower antibody levels than viremic and successfully suppressed HIV+ controls. These levels started to decrease directly after allo-SCT. Remarkably Pt#19 and Pt#28 presented antibody levels close to HIV negative donors. Conclusions: We conclude that allo-SCT not only remarkably decreased the HIV latent reservoir but also reduced the level of HIV antibodies in presence of cART. We have observed evidence of seroreversion a few years after allo-SCT. Future cART discontinuation will unravel the role of the antibodies dynamics in the HIV cure.