Lentiviral vector-mediated gene transfer in T cells from Wiskott-Aldrich syndrome patients leads to functional correction

Wiskott–Aldrich syndrome (WAS) is an X-linked primary immunodeficiency with a median survival below the age of 20 due to infections, severe hemorrhage, and lymphomas. Transplantation of hematopoietic stem cells from HLA-identical sibling donors is a resolutive treatment, but is available for a minority of patients. Transplantation of genetically corrected autologous hematopoietic stem cells or T cells could represent an alternative treatment applicable to all patients. We investigated whether WAS gene transfer with MMLV-based oncoretroviral and HIV-based lentiviral vectors could restore normal functions of patients' T cells. T cells transduced either with lentiviral vectors expressing the WAS protein (WASP) from the ubiquitous PGK promoter or the tissue-specific WASP promoter or with an oncoretroviral vector expressing WASP from the LTR, reached normal levels of WASP with correction of functional defects, including proliferation, IL-2 production, and lipid raft upregulation. Lentiviral vectors transduced T cells from WAS patients at higher rates, compared to oncoretroviral vectors, and efficiently transduced both activated and naive WAS T cells. Furthermore, a selective growth advantage of T cells corrected with the lentiviral vectors was demonstrated. The observation that lentiviral vector-mediated gene transfer results in correction of T cell defects in vitro supports their application for gene therapy in WAS patients

Machine learning outperformed logistic regression classification even with limit sample size: A model to predict pediatric HIV mortality and clinical progression to AIDS

Logistic regression (LR) is the most common prediction model in medicine. In recent years, supervised machine learning (ML) methods have gained popularity. However, there are many concerns about ML utility for small sample sizes. In this study, we aim to compare the performance of 7 algorithms in the prediction of 1-year mortality and clinical progression to AIDS in a small cohort of infants living with HIV from South Africa and Mozambique. The data set (n = 100) was randomly split into 70% training and 30% validation set. Seven algorithms (LR, Random Forest (RF), Support Vector Machine (SVM), K-Nearest Neighbor (KNN), Naive Bayes (NB), Artificial Neural Network (ANN), and Elastic Net) were compared. The variables included as predictors were the same across the models including sociodemographic, virologic, immunologic, and maternal status features. For each of the models, a parameter tuning was performed to select the best-performing hyperparameters using 5 times repeated 10-fold cross-validation. A confusion-matrix was built to assess their accuracy, sensitivity, and specificity. RF ranked as the best algorithm in terms of accuracy (82,8%), sensitivity (78%), and AUC (0,73). Regarding specificity and sensitivity, RF showed better performance than the other algorithms in the external validation and the highest AUC. LR showed lower performance compared with RF, SVM, or KNN. The outcome of children living with perinatally acquired HIV can be predicted with considerable accuracy using ML algorithms. Better models would benefit less specialized staff in limited resources countries to improve prompt referral in case of high-risk clinical progression

The continuum of care from HIV exposure to early diagnosis, early treatment and sustained viral suppression in infants, in southern Mozambique

A seguito del Piano Globale per l'eliminazione dell'HIV pediatrico e del Start Free Stay Free AIDS Free framework, sono stati fatti notevoli progressi nella prevenzione della trasmissione verticale dell’HIV (PMTCT), nell’accesso al trattamento antiretrovirale (ART) e nella riduzione dei decessi dovuti all'AIDS pediatrico. Tuttavia nel 2018, 160.000 nuove infezioni pediatriche si sono verificate a livello globale, il 52% dei bambini ha avuto accesso ad un test per la diagnosi precoce e solo la metà dei 1,7 milioni di bambini infetti ha ricevuto il trattamento. Dati recenti della valutazione dell'impatto dell'HIV nella popolazione hanno riportato una soppressione virale (SV) del 37% tra i bambini. Nell'attuale sforzo globale per ottenere l'eliminazione di nuove infezioni pediatriche, garantire l'accesso universale precoce all'ART e ottenere una SV sostenuta tra i bambini, è necessaria una migliore comprensione del continuum di cure del PMTCT fino alla diagnosi finale, all’accesso ai servizi di cura e il pattern della risposta virologica nei bambini, per disegnare una risposta su misura ed efficace. In Mozambico, la prevalenza dell'HIV tra le donne in età fertile è del 15,4% e nonostante più del 95% delle donne in gravidanza che accedono alle cure prenatali abbiano fatto il test dell’HIV e ricevuto il trattamento antiretrovirale, il tasso di trasmissione verticale è del 10%, si sono verificate 16.000 nuove infezioni pediatriche nel 2018; la soppressione virale tra i bambini è del 46%. Ci sono pochi dati sulle lacune del programma PMTCT e sulla risposta virale all'ART nei bambini che iniziano il trattamento ad 1 mese di vita. L'obiettivo principale di questo progetto È migliorare le attuali conoscenze sulle lacune nel continuum di assistenza alle mamme e ai bambini esposti all'HIV fino alla diagnosi, al trattamento precoce e alla soppressione virale nel Mozambico meridionale, dove la prevalenza dell'HIV È del 22,8%. In primo luogo, è stato condotto uno studio analitico trasversale utilizzando i dati dell'indagine nazionale sull'HIV 2015 per studiare i fattori associati allo stato di esposizione all'HIV in bambini di età inferiore ai 2 anni. Lo studio ha rilevato il 32% di bambini con stato di esposizione all’HIV sconosciuto; i fattori associati erano: residenza nella regione settentrionale e centrale del Mozambico, nelle zone rurali, madre senza istruzione e avere come capofamiglia l’uomo. In secondo luogo, uno studio di coorte retrospettivo ha analizzato la proporzione dei bambini esposti all'HIV che non hanno completato il follow-up (LTFU) fino alla diagnosi definitiva e i fattori correlati. Ã stato trovato un tasso del 16% di LTFU. I fattori associati erano l'età inferiore ai 2 mesi all'entrata nel servizio postnatale, l'allattamento al seno non exclusivo, la scarsa aderenza alla profilaxia con cotrimoxazolo. In terzo luogo, uno studio di coorte ha descritto la risposta virale durante 18 mesi di follow-up dei bambini che hanno iniziato il trattamento a 1 mese di età. La soppressione virale (SV) (<1000 copie/ml) È stata raggiunta dal 60% dei bambini, il 47% ha mantenuto la SV; la probabilità cumulativa di SV era del 43% a 6 mesi, 56% a 12 mesi, 73% a 18 mesi; il principale ostacolo per ottenere la SV era l'adesione all’ ART. Infine, un'analisi qualitativa ha valutato se il pacchetto standard di supporto psicosociale (APSS) usato con i genitori dei bambini in trattamento, identifica i fattori correlati con il tipo di risposta virale. I risultati hanno dimostrato che l'approccio di routine APSS non ha identificato in modo chiaro e tempestivo i bambini con scarsa risposta virale. Nel complesso, questo progetto di ricerca ha migliorato la comprensione delle principali lacune del programma PMTCT in Mozambico come l'accesso non ottimale e l'abbandono elevato della coppia mamma-bambino dal programma e i fattori associati; ha prodotto dati sulla risposta virale dei bambini che iniziano il trattamento ad 1 mese di vita e la necessità di un pacchetto di APSS su misura per i genitori di questi bambini. I risultati sono importanti per informare il programma dell’HIV e PMTCT nazionale al fine della progettazione di un modello di assistenza differenziata per le madri e le esigenze specifiche dei loro bambini. Sono necessarie ulteriori ricerche per studiare l'entità della perdita dei bambini esposti all’HIV dal follow-up e i determinanti socio-economici sottostanti, in particolare le dinamiche di genere all'interno della famiglia e il ruolo maschile nell'eliminazione dell'HIV pediatrico. Inoltre, ulteriori studi sono cruciali per valutare il continuum delle cure APSS durante l'infanzia per ottenere evidenze più solide sui fattori che influenzano l'aderenza dei genitori e come questi possono essere affrontati in modo efficace

Accelerated CD8+ T cell maturation in infants with perinatal HIV infection

In perinatal HIV infection, early antiretroviral therapy (ART) initiation is recommended but questions remain regarding infant immune responses to HIV and its impact on immune development. Using single cell transcriptional and phenotypic analysis we evaluated the T cell compartment at pre-ART initiation of infants with perinatally acquired HIV from Maputo, Mozambique (Towards AIDS Remission Approaches cohort). CD8+ T cell maturation subsets exhibited altered distribution in HIV exposed infected (HEI) infants relative to HIV exposed uninfected infants with reduced naive, increased effectors, higher frequencies of activated T cells, and lower frequencies of cells with markers of self-renewal. Additionally, a cluster of CD8+ T cells identified in HEI displayed gene profiles consistent with cytotoxic T lymphocytes and showed evidence for hyper expansion. Longitudinal phenotypic analysis revealed accelerated maturation of CD8+ T cells was maintained in HEI despite viral control. The results point to an HIV-directed immune response that is likely to influence reservoir establishment. [Display omitted] •Effector: naive CD8 T cell ratios are elevated in infants with perinatal HIV•scRNA-seq revealed clonally expanded CTL-like cells in HEI prior to ART initiation•Correlates of pre-ART virus load include frequencies of circulating GC-like Tfh•CD38+HLADR+ CD8 T cells are strong correlate of virus load in perinatal HIV Immunology; Virolog

Caregivers' psychosocial assessment for identifying HIV-infected infants at risk of poor treatment adherence: an exploratory study in southern Mozambique

Psychosocial support (PSS) to caregivers of HIV-infected infants on antiretroviral treatment (ART) is crucial to ensure ART adherence and sustained long-term viral suppression in children. A specific approach including tools to monitor and understand adherence behavior and risk factors that prevent optimal treatment compliance are urgently needed. This qualitative exploratory study, conducted in southern Mozambique, monitored the infants' viral response trajectories during 18 months follow-up, as a measure of adherence, reviewed the caregiver's PSS session notes and the answers to a study questionnaire, to analyze whether the standard PSS checklist applied to infants' caregivers can identify barriers influencing their adherence. Only 9 of 31 infants had sustained virologic response. Reported factors affecting adherence were: difficulties in drugs administration, shared responsibility to administer treatment; disclosure of child's HIV status to family members but lack of engagement; mother's ART interruption and poor viral response. In conclusion, we found that the standard PSS approach alone, applied to caregivers, was lacking focus on many relevant matters that were identified by the study questionnaire. A comprehensive patient-centered PSS package of care, including an adherence risk factor monitoring tool, tailored to caregivers and their children must be developed

Erratum to |[ldquo]|Lentiviral Vector-Mediated Gene Transfer in T Cells from Wiskott|[ndash]|Aldrich Syndrome Patients Leads to Functional Correction|[rdquo]|

The label on the bottom right panel of Fig. 3D on page 907 should be "WGFP/cyto" instead of "WGFP/TCR." The publisher regrets the error

Longitudinal analysis of innate immune system in infants with perinatal HIV infection until 18 months of age

With the advent of antiretroviral therapy (ART), perinatal HIV infection is declining globally but prevalence in Sub-Saharan Africa is still greater than other nations. The relationship of HIV replication in early infancy and the developing immune system is not well understood. In this study, we investigated cellular components of the innate immune system including Natural Killer (NK) cells, monocytes, and Dendritic Cells (DC) in a cohort of HIV exposed infected (HEI) and age-matched HIV exposed uninfected (HEU) infants from Mozambique. Study entry was at the first visit after delivery at age 1-2 months for HIV diagnosis and initiation of ART. Phenotypic analysis by multi-parameter flow cytometry revealed an expansion of total NK cells and the dysfunctional, CD56 CD16 , NK cell subset; increased activation in monocytes and DC; and higher levels of inflammatory homing receptor CCR5 on circulating DC subsets in the HEI infants. NKG2A, an inhibitory receptor for NK cytolytic function, was reduced in HEI compared to HEU and positively correlated with pre-ART viral load (VL) while expression of CCR2, the inflammatory homing receptor, on NK was negatively correlated with VL. Other subsets exhibited positive correlations with VL including the frequency of intermediate monocytes amongst total monocytes. Longitudinal analysis of VL indicated suboptimal ART adherence in HEI. Regardless of level of viral suppression achieved, the frequencies of specific innate immune subsets in HEI were normalized to HEU by 18m. These data support the notion that in early life, NK cells play a role in virus control and should be explored for functional attributes that are effective against HIV at this time during development. Overall, our study provides high resolution overview of the innate immune system during perinatal HIV infection

B-cell immunity and vaccine induced antibody protection reveal the inefficacy of current vaccination schedule in infants with perinatal HIV-infection in Mozambique, AfricaResearch in context

Summary: Background: Despite antiretroviral treatment (ART), immune dysfunction persists in children with perinatal HIV infection (HEI). Here we investigated the impact of HIV status on maternal antibody (Ab) passage, long-term vaccine induced immunity and B-cell maturation. Methods: 46 HIV Exposed Uninfected (HEU), 43 HEI, and 15 HIV unexposed uninfected (HUU) infants were vaccinated with 3 doses of DTaP-HepB-Hib-PCV10-OP at 2, 3, and 4 months at Matola Provincial Hospital, Maputo, Mozambique. Tetanus toxoid specific (TT) IgG, HIV Ab and B-cell phenotype characteristics were evaluated at entry, pre-ART, 5, 10, and 18 months in this longitudinal cohort study. Findings: Baseline (maternal) plasma TT Ab levels were significantly lower in HEI compared to both HEU and HUU and a faster decay of TT Ab was observed in HEI compared to HEU with significantly lower TT Ab levels at 10 and 18 months of age. TT unprotected (UP) (≤0.1 IU/mL) HEI showed higher HIV-RNA at entry and higher longitudinal HIV viremia (Area Under the Curve) compared to TT protected (P) HEI. A distinct HIV-Ab profile was found at entry in HEI compared to HEU. B-cell phenotype showed a B-cell perturbation in HEI vs HEU infants at entry (mean age 40.8 days) with lower transitional CD10+CD19+ B-cells and IgD+CD27− naive B-cells and an overall higher frequency of IgD−CD27− double negative B-cell subsets in HEI. Interpretation: B-cell perturbation, presenting with higher double negative IgD−CD27− B-cells was observed in neonatal age and may play a major role in the B-cell exhaustion in HEI. The ability to maintain TT protective Ab titers over time is impaired in HEI with uncontrolled viral replication and the current vaccination schedule is insufficient to provide long-term protection against tetanus. Funding: This work was supported by: NIH grant to SP (5R01AI127347-05); Children’s Hospital Bambino Gesú (Ricerca corrente 2019) to NC, and Associazione Volontari Bambino Gesù to PP

Viral Response among Early Treated HIV Perinatally Infected Infants: Description of a Cohort in Southern Mozambique

Early initiation of antiretroviral therapy and adherence to achieve viral load suppression (VLS) are crucial for reducing morbidity and mortality of perinatally HIV-infected infants. In this descriptive cohort study of 39 HIV perinatally infected infants, who started treatment at one month of life in Mozambique, we aimed to describe the viral response over 2 years of follow up. VLS &le; 400 copies/mL, sustained VLS and viral rebound were described using a Kaplan&ndash;Meier estimator. Antiretroviral drug transmitted resistance was assessed for a sub-group of non-VLS infants. In total, 61% of infants reached VLS, and 50% had a rebound. Cumulative probability of VLS was 36%, 51%, and 69% at 6, 12 and 24 months of treatment, respectively. The median duration of VLS was 7.4 months (IQR 12.6) and the cumulative probability of rebound at 6 months was 30%. Two infants had resistance biomarkers to drugs included in their treatment regimen. Our findings point to a low rate of VLS and high rate of viral rebound. More frequent viral response monitoring is advisable to identify infants with rebound and offer timely adherence support. It is urgent to tailor the psychosocial support model of care to this specific age group and offer differentiated service delivery to mother&ndash;baby pairs

Novel Approaches to Postnatal Prophylaxis to Eliminate Vertical Transmission of HIV

Despite progress in providing antiretroviral therapy to pregnant women living with HIV, a substantial number of vertical transmissions continue to occur. Novel approaches leveraging modern potent, safe, and well-tolerated antiretroviral drugs are urgently needed