Determinants of virological failure and antiretroviral drug resistance in Mozambique

Objectives The objective of this study was to inform public health actions to limit first-line ART failure and HIV drug resistance in Mozambique. Methods This was a cross-sectional study. HIV-1-infected adults on first-line ART for at least 1 year attending routine visits in the Manhiça District Hospital, in a semi-rural area in southern Mozambique with no HIV-1 RNA monitoring available, were evaluated for clinical, socio-demographic, therapeutic, immunological and virological characteristics. Factors associated with HIV-1 RNA ≥1000 copies/mL and HIV drug resistance were determined using multivariate logistic regression. Results The study included 334 adults on first-line ART for a median of 3 years, of which 65% (214/332) had suppressed viraemia, 11% (37/332) had low-level viraemia (HIV-1 RNA 150-999 copies/mL) and 24% (81/332) had overt virological failure (HIV-1 RNA ≥1000 copies/mL). HIV drug resistance was detected in 89% of subjects with virological failure, but in none with low-level viraemia. Younger age [OR = 0.97 per additional year (95% CI = 0.94-1.00), P = 0.039], ART initiation at WHO stage III/IV [OR = 2.10 (95% CI = 1.23-3.57), P = 0.003] and low ART adherence [OR = 2.69 (95% CI = 1.39-5.19), P = 0.003] were associated with virological failure. Longer time on ART [OR = 1.55 per additional year (95% CI = 1.00-2.43), P = 0.052] and illiteracy [OR = 0.24 (95% CI = 0.07-0.89), P = 0.033] were associated with HIV drug resistance. Compared with HIV-1 RNA, clinician's judgement of ART failure, based on clinical and immunological outcomes, only achieved 29% sensitivity and misdiagnosed 1 out of every 4.5 subjects. Conclusions Public health programmes in Mozambique should focus on early HIV diagnosis, early ART initiation and adherence support. Virological monitoring drastically improves the diagnosis of ART failure, enabling a better use of resource

Systems-Level Immunomonitoring from Acute to Recovery Phase of Severe COVID-19

Peer reviewe

HIV-1 Tropism Testing in Subjects Achieving Undetectable HIV-1 RNA : Diagnostic Accuracy, Viral Evolution and Compartmentalization

Technically, HIV-1 tropism can be evaluated in plasma or peripheral blood mononuclear cells (PBMCs). However, only tropism testing of plasma HIV-1 has been validated as a tool to predict virological response to CCR5 antagonists in clinical trials. The preferable tropism testing strategy in subjects with undetectable HIV-1 viremia, in whom plasma tropism testing is not feasible, remains uncertain. We designed a proof-of-concept study including 30 chronically HIV-1-infected individuals who achieved HIV-1 RNA <50 copies/mL during at least 2 years after first-line ART initiation. First, we determined the diagnostic accuracy of 454 and population sequencing of gp120 V3-loops in plasma and PBMCs, as well as of MT-2 assays before ART initiation. The Enhanced Sensitivity Trofile Assay (ESTA) was used as the technical reference standard. 454 sequencing of plasma viruses provided the highest agreement with ESTA. The accuracy of 454 sequencing decreased in PBMCs due to reduced specificity. Population sequencing in plasma and PBMCs was slightly less accurate than plasma 454 sequencing, being less sensitive but more specific. MT-2 assays had low sensitivity but 100% specificity. Then, we used optimized 454 sequence data to investigate viral evolution in PBMCs during viremia suppression and only found evolution of R5 viruses in one subject. No de novo CXCR4-using HIV-1 production was observed over time. Finally, Slatkin-Maddison tests suggested that plasma and cell-associated V3 forms were sometimes compartmentalized. The absence of tropism shifts during viremia suppression suggests that, when available, testing of stored plasma samples is generally safe and informative, provided that HIV-1 suppression is maintained. Tropism testing in PBMCs may not necessarily produce equivalent biological results to plasma, because the structure of viral populations and the diagnostic performance of tropism assays may sometimes vary between compartments. Thereby, proviral DNA tropism testing should be specifically validated in clinical trials before it can be applied to routine clinical decision-making

Deep Molecular Characterization of HIV-1 Dynamics under Suppressive HAART

In order to design strategies for eradication of HIV-1 from infected individuals, detailed insight into the HIV-1 reservoirs that persist in patients on suppressive antiretroviral therapy (ART) is required. In this regard, most studies have focused on integrated (proviral) HIV-1 DNA forms in cells circulating in blood. However, the majority of proviral DNA is replication-defective and archival, and as such, has limited ability to reveal the dynamics of the viral population that persists in patients on suppressive ART. In contrast, extrachromosomal (episomal) viral DNA is labile and as a consequence is a better surrogate for recent infection events and is able to inform on the extent to which residual replication contributes to viral reservoir maintenance. To gain insight into the diversity and compartmentalization of HIV-1 under suppressive ART, we extensively analyzed longitudinal peripheral blood mononuclear cells (PBMC) samples by deep sequencing of episomal and integrated HIV-1 DNA from patients undergoing raltegravir intensification. Reverse-transcriptase genes selectively amplified from episomal and proviral HIV-1 DNA were analyzed by deep sequencing 0, 2, 4, 12, 24 and 48 weeks after raltegravir intensification. We used maximum likelihood phylogenies and statistical tests (AMOVA and Slatkin-Maddison (SM)) in order to determine molecular compartmentalization. We observed low molecular variance (mean variability ≤0.042). Although phylogenies showed that both DNA forms were intermingled within the phylogenetic tree, we found a statistically significant compartmentalization between episomal and proviral DNA samples (P<10−6 AMOVA test; P = 0.001 SM test), suggesting that they belong to different viral populations. In addition, longitudinal analysis of episomal and proviral DNA by phylogeny and AMOVA showed signs of non-chronological temporal compartmentalization (all comparisons P<10−6) suggesting that episomal and proviral DNA forms originated from different anatomical compartments. Collectively, this suggests the presence of a chronic viral reservoir in which there is stochastic release of infectious virus and in which there are limited rounds of de novo infection. This could be explained by the existence of different reservoirs with unique pharmacological accessibility properties, which will require strategies that improve drug penetration/retention within these reservoirs in order to minimise maintenance of the viral reservoir by de novo infection

Clinical Value of Massive Parallel HIV-1 Sequencing in Antiretroviral Treatment-Experienced Subjects

Els test de resistència al antiretrovirals són utilitzats en la pràctica clínica per la personalització del tractament antiretroviral (TARV) de les persones infectades pel VIH-1. La seqüenciació poblacional (SP) és la tècnica més emprada pel genotipat del VIH-1 obtenint una bona correlació amb la resposta clínica dels pacients. No obstant, la baixa sensibilitat de la SP pel que fa a la detecció de mutacions de resistència als antiretrovirals (DRM) o soques X4-tròpiques presents en baixes prevalences pot comprometre l’eficàcia del TARV. L’aplicació de noves tècniques de seqüenciació massiva pel genotipat del VIH-1 (genotipat ultrasensible) permetria una caracterització de la població viral amb major resolució optimitzant el TARV. L’objectiu d’aquesta tesi era avaluar el valor clínic del genotipat ultrasensible pel maneig clínic de pacients infectats pel VIH-1 que han estat exposats a TARV. Durant la utilització de 454 sequencing per la determinació del tropisme viral, 454 sequencing va obtenir una major concordança amb ESTA (Enhanced Sensitivity TrofileTM Assay) en l’anàlisi de l’ARN viral. Tot i que la SP tenia una menor sensibilitat respecte 454 sequencing, la seva elevada especificitat li va permetre obtenir una millor precisió. Anàlisis filogenètics a partir de dades obtingudes per 454 sequencing van revelar que, encara que les determinacions eren equivalents entre amdós compartiments, en alguns pacients es va observar compartimentalització de seqüències pel que fa als haplotips i a la seva prevalença. Aquesta compartimentalització podria comprometre la determinació del tropisme viral en funció del compartiment que s’analitzi. Durant aquest treball també es va fer la validació de la SP per determinacions del tropisme a partir d’ADN proviral en un assaig clínic prospectiu i aleatoritzat. Aquest assaig va demostrar ser útil per guiar canvis de tractament antiretroviral que incloguin antagonistes de CCR5. De tota manera, 454 sequencing va ser capaç de detectar variants X4 tròpiques en dos pacients prèviament classificats com a R5 tròpics per SP. Pel que fa a la detecció de DRM en el gen pol, 454 sequencing va demostrar ser no-inferior a la SP durant l’avaluació de la susceptibilitat al tractament antiretroviral. Cal destacar que 454 sequencing va aportar informació genotípica addicional en la majoria dels pacients, encara que aquesta modifiqués poques prediccions de susceptibilitat. A més, aquesta tecnologia va ser utilitzada per explorar l’origen de les DRM en el gen de la integrasa del VIH-1, demostrant que virus mutants detectats durant el fracàs virològic poden ésser originats a partir de virus mutants minoritaris preexistents. En resum, aquesta tesi ha mostrat la utilitat clínica del genotipat ultrasensible del VIH-1 en pacients experimentats al TARV. 454 sequencing ha permès una profunda caracterització de la diversitat viral dins l’hoste, facilitant l’estudi l’evolució viral i la seva patogènesi del VIH-1. A més, l’estandarització i automatització dels protocols de 454 sequencing permetria la implementació d’aquesta tecnologia pel diagnòstic del VIH-1.Los ensayos de resistencia a los antiretrovirales son utilizados en la práctica clínica para la personalización del tratamiento antirretroviral (TARV) de las personas infectadas por el VIH-1. La secuenciación poblacional (SP) es la técnica más utilizada para el genotipado del VIH-1 obteniendo una buena correlación con la respuesta clínica de los pacientes. Asimismo, la baja sensibilidad de la SP para la detección de mutaciones de resistencia a los antiretrovirales o variantes X4-trópicas presentes en baja prevalencia puede comprometer la eficacia de la TARV. El objetivo de esta tesis era evaluar el valor clínico del genotipado ultrasensible para el manejo clínico de pacientes infectados por el VIH-1 que han estado expuestos a TARV. Durante la utilización de 454 sequencing para la determinación del tropismo viral, 454 sequencing obtuvo una mayor concordancia con ESTA (Enhanced Sensitivity TrofileTM Assay) en el análisis del ARN viral. Aunque la SP mostró una menor sensibilidad que 454 sequencing, su elevada especificidad le permitió obtener una mayor precisión. La información genotípica generada por 454 sequencing fue utilizada para evaluar la compartimentalización. Aunque las determinaciones del tropismo viral eran equivalentes entre los compartimentos, en algunos pacientes se observó compartimentalización de secuencias, lo que puede comprometer la determinación del tropismo viral en función del compartimento que analizado. Durante este trabajo también se hizo la validación de la SP para determinaciones del tropismo a partir de ADN proviral en un ensayo clínico prospectivo y aleatorizado. Este ensayo demostró ser útil para guiar cambios de tratamiento antirretroviral que incluyan antagonistas de CCR5. De todos modos, 454 sequencing fue capaz de detectar variantes X4 trópicas en dos pacientes previamente clasificados como R5 trópicos por SP. En cuanto a la detección de DRM en el gen pol, 454 sequencing demostró ser no inferior a la SP durante la evaluación de la susceptibilidad al tratamiento antirretroviral. Cabe mencionar que 454 sequencing aportó información genotípica adicional en la mayoría de los pacientes, aunque esta modificara pocas predicciones de susceptibilidad. Además, esta tecnología fue útil para explorar el origen de las DRM en el gen de la integrasa del VIH-1, mostrando que los mutantes detectados durante el fracaso virológico pueden ser originados a partir de mutantes pre-existentes. En resumen, esta tesis ha mostrado la utilidad clínica del genotipado ultrasensible del VIH-1 en pacientes experimentados al TARV. 454 sequencing ha permitido una profunda caracterización de la diversidad viral dentro del huésped, facilitando el estudio de la evolución viral i la patogénesis del VIH-1. Además, la estandarización i automatización de los protocolos de 454 sequencing permitiría la implementación de esta tecnología para el diagnóstico clínico del VIH-1.Drug resistance testing is utilized in the clinical routine to personalize ART of HIV-1 infected people. Population sequencing is employed for HIV-1 genotyping obtaining good correlations with clinical outcomes. However, its lack of sensitivity to detect minority DRM mutations or minor CXCR4-using viruses can compromise the efficacy of antiretroviral therapy (ART). The use of next generation sequencing technologies for ultrasensitive HIV-1 genotyping might allow deep characterization of the viral population optimizing the ART. The objective of this work was to evaluate the clinical value of ultrasensitive HIV-1 genotyping obtained by 454 sequencing for the management of ART-experienced HIV- 1 subjects in different treatment situations. During the utilization of 454 sequencing for viral tropism determinations, this technology achieved the closest diagnostic accuracy to ESTATM in plasma ARN. Although population sequencing had lower sensitivity than 454 sequencing, its highest specificity led to obtain closest accuracy to ESTA. Even though tropism determinations were equivalent between plasma and cellular compartments, phylogenetic analyses from data generated by 454 sequencing revealed sequence compartmentalization in some subjects. We also validated triplicate population HIV-1 genotyping from PBMC-associated DNA for viral tropism determinations in a prospective and randomized clinical trial. Here, genotypic tropism testing in proviral DNA demonstrated to become a suitable tool to guide treatment switches to CCR5 antagonists in aviremic individuals. In this study, 454 sequencing was capable to detect non-R5 viral strains in two subjects previously classified as harboring R5 HIV-1 strains by population sequencing. This compartmentalization might compromise viral tropism determinations depending on the compartment analyzed. Furthermore, the absence of evolution observed during prolonged periods of aviremia suggested that testing of stored plasma sample would be generally safe and informative. Regarding the detection of minority DRM in pol gene, 454 sequencing for ultrasensitive HIV-1 genotyping was technically non-inferior than population HIV-1 genotyping during the assessment of antiretroviral drug susceptibility. However, although this technology provided additional genotypic information beyond population sequencing in most of heavily pre-treated individuals tested, only few antiretroviral susceptibility predictions were modified. This technology was also useful to explore the origin of DRM in integrase gene, revealing that mutants detected at the time of virological failure can be originated from pre-existing minority drug resistance variants. In summary, this thesis has shown the clinical utility of ultrasensitive HIV-1 genotyping in ART-experienced HIV-1 infected individuals. However, further studies should extend our findings. 454 sequencing allowed a deep characterization of the HIV-1 diversity within a host helping to understand viral evolution and HIV-1 pathogenesis. Moreover, the standardization and automation of 454 sequencing protocols for HIV-1 sequencing would allow the implementation of this technology for HIV-1 diagnosis

Clinical value of massive Parallel HIV-1 sequencing in antiretroviral treatment-experienced subjects

Els test de resistència al antiretrovirals són utilitzats en la pràctica clínica per la personalització del tractament antiretroviral (TARV) de les persones infectades pel VIH-1. La seqüenciació poblacional (SP) és la tècnica més emprada pel genotipat del VIH-1 obtenint una bona correlació amb la resposta clínica dels pacients. No obstant, la baixa sensibilitat de la SP pel que fa a la detecció de mutacions de resistència als antiretrovirals (DRM) o soques X4-tròpiques presents en baixes prevalences pot comprometre l'eficàcia del TARV. L'aplicació de noves tècniques de seqüenciació massiva pel genotipat del VIH-1 (genotipat ultrasensible) permetria una caracterització de la població viral amb major resolució optimitzant el TARV. L'objectiu d'aquesta tesi era avaluar el valor clínic del genotipat ultrasensible pel maneig clínic de pacients infectats pel VIH-1 que han estat exposats a TARV. Durant la utilització de 454 sequencing per la determinació del tropisme viral, 454 sequencing va obtenir una major concordança amb ESTA (Enhanced Sensitivity TrofileTM Assay) en l'anàlisi de l'ARN viral. Tot i que la SP tenia una menor sensibilitat respecte 454 sequencing, la seva elevada especificitat li va permetre obtenir una millor precisió. Anàlisis filogenètics a partir de dades obtingudes per 454 sequencing van revelar que, encara que les determinacions eren equivalents entre amdós compartiments, en alguns pacients es va observar compartimentalització de seqüències pel que fa als haplotips i a la seva prevalença. Aquesta compartimentalització podria comprometre la determinació del tropisme viral en funció del compartiment que s'analitzi. Durant aquest treball també es va fer la validació de la SP per determinacions del tropisme a partir d'ADN proviral en un assaig clínic prospectiu i aleatoritzat. Aquest assaig va demostrar ser útil per guiar canvis de tractament antiretroviral que incloguin antagonistes de CCR5. De tota manera, 454 sequencing va ser capaç de detectar variants X4 tròpiques en dos pacients prèviament classificats com a R5 tròpics per SP. Pel que fa a la detecció de DRM en el gen pol, 454 sequencing va demostrar ser no-inferior a la SP durant l'avaluació de la susceptibilitat al tractament antiretroviral. Cal destacar que 454 sequencing va aportar informació genotípica addicional en la majoria dels pacients, encara que aquesta modifiqués poques prediccions de susceptibilitat. A més, aquesta tecnologia va ser utilitzada per explorar l'origen de les DRM en el gen de la integrasa del VIH-1, demostrant que virus mutants detectats durant el fracàs virològic poden ésser originats a partir de virus mutants minoritaris preexistents. En resum, aquesta tesi ha mostrat la utilitat clínica del genotipat ultrasensible del VIH-1 en pacients experimentats al TARV. 454 sequencing ha permès una profunda caracterització de la diversitat viral dins l'hoste, facilitant l'estudi l'evolució viral i la seva patogènesi del VIH-1. A més, l'estandarització i automatització dels protocols de 454 sequencing permetria la implementació d'aquesta tecnologia pel diagnòstic del VIH-1.Los ensayos de resistencia a los antiretrovirales son utilizados en la práctica clínica para la personalización del tratamiento antirretroviral (TARV) de las personas infectadas por el VIH-1. La secuenciación poblacional (SP) es la técnica más utilizada para el genotipado del VIH-1 obteniendo una buena correlación con la respuesta clínica de los pacientes. Asimismo, la baja sensibilidad de la SP para la detección de mutaciones de resistencia a los antiretrovirales o variantes X4-trópicas presentes en baja prevalencia puede comprometer la eficacia de la TARV. El objetivo de esta tesis era evaluar el valor clínico del genotipado ultrasensible para el manejo clínico de pacientes infectados por el VIH-1 que han estado expuestos a TARV. Durante la utilización de 454 sequencing para la determinación del tropismo viral, 454 sequencing obtuvo una mayor concordancia con ESTA (Enhanced Sensitivity TrofileTM Assay) en el análisis del ARN viral. Aunque la SP mostró una menor sensibilidad que 454 sequencing, su elevada especificidad le permitió obtener una mayor precisión. La información genotípica generada por 454 sequencing fue utilizada para evaluar la compartimentalización. Aunque las determinaciones del tropismo viral eran equivalentes entre los compartimentos, en algunos pacientes se observó compartimentalización de secuencias, lo que puede comprometer la determinación del tropismo viral en función del compartimento que analizado. Durante este trabajo también se hizo la validación de la SP para determinaciones del tropismo a partir de ADN proviral en un ensayo clínico prospectivo y aleatorizado. Este ensayo demostró ser útil para guiar cambios de tratamiento antirretroviral que incluyan antagonistas de CCR5. De todos modos, 454 sequencing fue capaz de detectar variantes X4 trópicas en dos pacientes previamente clasificados como R5 trópicos por SP. En cuanto a la detección de DRM en el gen pol, 454 sequencing demostró ser no inferior a la SP durante la evaluación de la susceptibilidad al tratamiento antirretroviral. Cabe mencionar que 454 sequencing aportó información genotípica adicional en la mayoría de los pacientes, aunque esta modificara pocas predicciones de susceptibilidad. Además, esta tecnología fue útil para explorar el origen de las DRM en el gen de la integrasa del VIH-1, mostrando que los mutantes detectados durante el fracaso virológico pueden ser originados a partir de mutantes pre-existentes. En resumen, esta tesis ha mostrado la utilidad clínica del genotipado ultrasensible del VIH-1 en pacientes experimentados al TARV. 454 sequencing ha permitido una profunda caracterización de la diversidad viral dentro del huésped, facilitando el estudio de la evolución viral i la patogénesis del VIH-1. Además, la estandarización i automatización de los protocolos de 454 sequencing permitiría la implementación de esta tecnología para el diagnóstico clínico del VIH-1.Drug resistance testing is utilized in the clinical routine to personalize ART of HIV-1 infected people. Population sequencing is employed for HIV-1 genotyping obtaining good correlations with clinical outcomes. However, its lack of sensitivity to detect minority DRM mutations or minor CXCR4-using viruses can compromise the efficacy of antiretroviral therapy (ART). The use of next generation sequencing technologies for ultrasensitive HIV-1 genotyping might allow deep characterization of the viral population optimizing the ART. The objective of this work was to evaluate the clinical value of ultrasensitive HIV-1 genotyping obtained by 454 sequencing for the management of ART-experienced HIV- 1 subjects in different treatment situations. During the utilization of 454 sequencing for viral tropism determinations, this technology achieved the closest diagnostic accuracy to ESTATM in plasma ARN. Although population sequencing had lower sensitivity than 454 sequencing, its highest specificity led to obtain closest accuracy to ESTA. Even though tropism determinations were equivalent between plasma and cellular compartments, phylogenetic analyses from data generated by 454 sequencing revealed sequence compartmentalization in some subjects. We also validated triplicate population HIV-1 genotyping from PBMC-associated DNA for viral tropism determinations in a prospective and randomized clinical trial. Here, genotypic tropism testing in proviral DNA demonstrated to become a suitable tool to guide treatment switches to CCR5 antagonists in aviremic individuals. In this study, 454 sequencing was capable to detect non-R5 viral strains in two subjects previously classified as harboring R5 HIV-1 strains by population sequencing. This compartmentalization might compromise viral tropism determinations depending on the compartment analyzed. Furthermore, the absence of evolution observed during prolonged periods of aviremia suggested that testing of stored plasma sample would be generally safe and informative. Regarding the detection of minority DRM in pol gene, 454 sequencing for ultrasensitive HIV-1 genotyping was technically non-inferior than population HIV-1 genotyping during the assessment of antiretroviral drug susceptibility. However, although this technology provided additional genotypic information beyond population sequencing in most of heavily pre-treated individuals tested, only few antiretroviral susceptibility predictions were modified. This technology was also useful to explore the origin of DRM in integrase gene, revealing that mutants detected at the time of virological failure can be originated from pre-existing minority drug resistance variants. In summary, this thesis has shown the clinical utility of ultrasensitive HIV-1 genotyping in ART-experienced HIV-1 infected individuals. However, further studies should extend our findings. 454 sequencing allowed a deep characterization of the HIV-1 diversity within a host helping to understand viral evolution and HIV-1 pathogenesis. Moreover, the standardization and automation of 454 sequencing protocols for HIV-1 sequencing would allow the implementation of this technology for HIV-1 diagnosi

The submental island flap is a viable reconstructive option for a variety of head and neck ablative defects

The submental island flap (SIF) is a pedicled flap based upon the submental artery and vein. Its utility in reconstruction following ablative head and neck procedures has been applied to various subsites including skin, lip, buccal mucosa, retromolar trigone, parotidectomy defects, and tongue. We review our experience using the SIF for reconstruction following tumor ablation.This prospective case series with medical record review includes consecutive patients undergoing SIF reconstruction following ablative surgery for malignancy at a single tertiary care facility between November 2014 and November 2016. We examined preoperative variables, surgical procedures, and postoperative outcomes.Thirty-seven patients met inclusion criteria. Twenty-nine were male; the average age was 64.3 (±12.4) years. Seventeen cancers involved the oral cavity, 11 involved the skin, 8 were in the oropharynx, and 1 was in the paranasal sinus. The average size of the SIF was 38.8 cm(±17.6 cm). Four partial flap losses occurred; none required revision surgery. The average length of stay for these patients was 7.2 (±6.1) days.The SIF is a robust flap that can be reliably used for a variety of head and neck defects following tumor ablation with an acceptable rate of donor- and flap-related complications

Inpatient Decongestive Therapy for Lymphedema in Acute Postsurgical Head and Neck Cancer Patients*

Introduction Head and neck lymphedema is an omnipresent morbidity related to head and neck cancer therapies. Studies on therapy for these patients in the acute postsurgical population have not been published to date. Objective To assess changes in the measurements of lymphedema in surgical head and neck cancer patients during the hospital stay with implementation of modified decongestive therapy (MDT). Methods Patients aged > 18 years undergoing neck dissection with or without primary-site resection or laryngectomy between 2016 and 2019 were included. Facial measurements were obtained prior to beginning MDT and again prior to discharge. A total facial composite measurement was calculated and used to assess change over time. Rates ≥ 2% of change were considered significant. Results A total of 38 patients were included (subsites: larynx = 27; thyroid = 4; oral cavity = 3; and neck = 4). The mean number of days between surgery and the start of lymphedema therapy was 3.0 days. The mean number of days between measurements was 5.2 days. Reduction in the total composite score was observed in 37 (97%) patients, and 35 (92%) patients had a total composite reduction score > 2%. Tumor subsite and surgery type did not portend toward greater percent change, except for those patients treated with total laryngectomy, regional flap reconstruction, and neck dissection (p = 0.02). Conclusion Acute postsurgical inpatient MDT was associated with reduced total composite measurements in patients after head and neck surgery. As the first published study on lymphedema therapy in this acute postsurgical period, further prospective case-control studies are warranted to explore further benefits of acute therapy

Enhanced TLR3 responsiveness in hepatitis C virus resistant women from the Irish anti-D cohort

Cytokine data presented in this manuscript is available at https://dx.doi.org/10.17632/m65wh4vc9z.1. NanoString data is available at https://data.mendeley.com/datasets/z4nr7p7ry7. Additional data reported in this paper will be shared by the lead contact upon request.International audienceNatural resistance to infection is an overlooked outcome after hepatitis C virus (HCV) exposure. Between 1977 and 1979, 1,200 Rhesus D-negative Irish women were exposed to HCV-contaminated anti-D immunoglobulin. Here, we investigate why some individuals appear to resist infection despite exposure (exposed seronegative [ESN]). We screen HCV-resistant and -susceptible donors for anti-HCV adaptive immune responses using ELISpots and VirScan to profile antibodies against all know human viruses. We perform standardized ex vivo whole blood stimulation (TruCulture) assays with antiviral ligands and assess antiviral responses using NanoString transcriptomics and Luminex proteomics. We describe an enhanced TLR3-type I interferon response in ESNs compared with seropositive women. We also identify increased inflammatory cytokine production in response to polyIC in ESNs compared with seropositive women. These enhanced responses may have contributed to innate immune protection against HCV infection in our cohort