Intrarectal transmission, systemic infection, and CD4+ T cell depletion in humanized mice infected with HIV-1.

Intrarectal infection between men who have sex with men represents a predominant form of human immunodeficiency virus (HIV) transmission in developed countries. Currently there are no adequate small animal models that recapitulate intrarectal HIV transmission. Here we demonstrate that human lymphocytes generated in situ from hematopoietic stem cells reconstitute the gastrointestinal tract of humanized mice with human CD4(+) T cells rendering them susceptible to intrarectal HIV transmission. HIV infection after a single intrarectal inoculation results in systemic infection with depletion of CD4(+) T cells in gut-associated lymphoid tissue and other pathologic sequela that closely mimics those observed in HIV infected humans. This novel model provides the basis for the development and evaluation of novel approaches aimed at immune reconstitution of human gut-associated lymphoid tissue and for the development, testing, and implementation of microbicides to prevent intrarectal HIV-1 transmission.

Humanized mice mount specific adaptive and innate immune responses to EBV and TSST-1.

Here we show that transplantation of autologous human hematopoietic fetal liver CD34+ cells into NOD/SCID mice previously implanted with human fetal thymic and liver tissues results in long-term, systemic human T-cell homeostasis. In addition, these mice show systemic repopulation with human B cells, monocytes and macrophages, and dendritic cells (DCs). T cells in these mice generate human major histocompatibility complex class I- and class II-restricted adaptive immune responses to Epstein-Barr virus (EBV) infection and are activated by human DCs to mount a potent T-cell immune response to superantigens. Administration of the superantigen toxic shock syndrome toxin 1 (TSST-1) results in the specific systemic expansion of human Vbeta2+ T cells, release of human proinflammatory cytokines and localized, specific activation and maturation of human CD11c+ dendritic cells. This represents the first demonstration of long-term systemic human T-cell reconstitution in vivo allowing for the manifestation of the differential response by human DCs to TSST-1.

One percent tenofovir applied topically to humanized BLT mice and used according to the CAPRISA 004 experimental design demonstrates partial protection from vaginal HIV infection, validating the BLT model for evaluation of new microbicide candidates.

Recent iPrEx clinical trial results provided evidence that systemic preexposure prophylaxis (PrEP) with emtricitabine (FTC) and tenofovir disoproxil fumarate (TDF) can partially prevent rectal HIV transmission in humans. Similarly, we have previously demonstrated that systemic administration of the same FTC-TDF combination efficiently prevented rectal transmission in humanized bone marrow/liver/thymus (BLT) mice. The CAPRISA 004 trial recently demonstrated that topical application of the tenofovir could partially prevent vaginal HIV-1 transmission in humans. To further validate the usefulness of the BLT mouse model for testing HIV prevention strategies, we evaluated the topical administration of tenofovir as used in CAPRISA 004 to prevent vaginal HIV transmission in BLT mice. Our results demonstrate that vaginally administered 1% tenofovir significantly reduced HIV transmission in BLT mice (P = 0.002). Together with the results obtained after systemic antiretroviral PrEP, these topical inhibitor data serve to validate the use of humanized BLT mice to evaluate both systemic and topical inhibitors of HIV transmission. Based on these observations, we tested six additional microbicide candidates for their ability to prevent vaginal HIV transmission: a C-peptide fusion inhibitor (C52L), a membrane-disrupting amphipathic peptide inhibitor (C5A), a trimeric d-peptide fusion inhibitor (PIE12-Trimer), a combination of reverse transcriptase inhibitors (FTC-TDF), a thioester zinc finger inhibitor (TC247), and a small-molecule Rac inhibitor (NSC23766). No protection was seen with the Rac inhibitor NSC23766. The thioester compound TC247 offered partial protection. Significant protection was afforded by FTC-TDF, and complete protection was offered by three different peptide inhibitors tested. Our results demonstrate that these effective topical inhibitors have excellent potential to prevent vaginal HIV transmission in humans.

Prevention and management of oronasal fistulas.

PURPOSE OF REVIEW: The aim of this article is to present the current opinion on the prevention and management of oronasal fistulas in cleft palate patients. RECENT FINDINGS: Though cleft palate repair has seen numerous modifications and improvements, oronasal fistulas remain one of the most common complications of palatoplasty. There are various techniques available for preventing and managing this complication. SUMMARY: Oronasal fistulas can be minimized by employing proper principles for palatoplasty. Once a fistula occurs, the repair technique should be appropriate for the fistula type. Oronasal fistula classifications, various repair techniques, tissue adjuncts, and biomaterials used in both the primary palate repair and oronasal fistula repair are discussed in this review.

Impaired eating-related quality of life in chronic rhinosinusitis.

BACKGROUND: Despite the tremendous burden of smell and taste dysfunction in patients with chronic rhinosinusitis (CRS), objective measures of smell and taste fail to fully account for eating-related disruptions in CRS patient quality of life (QOL). In this study we sought to investigate the driving force behind impaired eating-related QOL in CRS patients. METHODS: Adult CRS patients were prospectively enrolled and answered a series of surveys relating to smell, taste, overall sinus-specific QOL, and depression. Patients with both smell-related and taste-related eating complaints were considered to have impaired eating-related QOL. Clinical demographics, objective chemosensory scores, and endoscopy scores were collected. RESULTS: Seventy patients were enrolled and 23% showed impaired eating-related QOL. In multivariable analyses, patients with aspirin-exacerbated respiratory disease (AERD) showed 10.7 times higher odds of impaired eating-related QOL (odds ratio [OR] 10.72; 95% confidence interval [CI], 1.09 to 105.09; p = 0.042); meanwhile, for every 1-point increase in depression scores, the odds of impaired eating-related QOL increased by 1.3 (OR 1.31; 95% CI, 1.10 to 1.57; p = 0.003). For every 1-point decrease in orthonasal olfactory threshold, the odds of impaired eating-related QOL increased by 1.9 times (OR 1.85; 95% CI, 1.14 to 3.00; p = 0.013). Symptom scores, polyp status, endoscopic scores, and other olfactory measures did not remain significant after adjusting for other variables in forward-selection multivariable modeling. CONCLUSION: Disruptions in eating-related QOL cannot be fully explained by objective smell or taste testing alone. We identified AERD and depression as independent risk factors for greater odds of impaired eating-related QOL in CRS. Improved orthonasal threshold scores were independently associated with better eating-related QOL.

Antiretroviral pre-exposure prophylaxis prevents vaginal transmission of HIV-1 in humanized BLT mice.

BACKGROUND: Worldwide, vaginal transmission now accounts for more than half of newly acquired HIV-1 infections. Despite the urgency to develop and implement novel approaches capable of preventing HIV transmission, this process has been hindered by the lack of adequate small animal models for preclinical efficacy and safety testing. Given the importance of this route of transmission, we investigated the susceptibility of humanized mice to intravaginal HIV-1 infection. METHODS AND FINDINGS: We show that the female reproductive tract of humanized bone marrow-liver-thymus (BLT) mice is reconstituted with human CD4+ T and other relevant human cells, rendering these humanized mice susceptible to intravaginal infection by HIV-1. Effects of HIV-1 infection include CD4+ T cell depletion in gut-associated lymphoid tissue (GALT) that closely mimics what is observed in HIV-1-infected humans. We also show that pre-exposure prophylaxis with antiretroviral drugs is a highly effective method for preventing vaginal HIV-1 transmission. Whereas 88% (7/8) of BLT mice inoculated vaginally with HIV-1 became infected, none of the animals (0/5) given pre-exposure prophylaxis of emtricitabine (FTC)/tenofovir disoproxil fumarate (TDF) showed evidence of infection (Chi square = 7.5, df = 1, p = 0.006). CONCLUSIONS: The fact that humanized BLT mice are susceptible to intravaginal infection makes this system an excellent candidate for preclinical evaluation of both microbicides and pre-exposure prophylactic regimens. The utility of humanized mice to study intravaginal HIV-1 transmission is particularly highlighted by the demonstration that pre-exposure prophylaxis can prevent intravaginal HIV-1 transmission in the BLT mouse model.

Retronasal olfaction in chronic rhinosinusitis.

OBJECTIVE: The goals of this study were to assess retronasal olfaction in patients with chronic rhinosinusitis (CRS) and describe clinical factors that influence retronasal olfaction. This study sought to investigate the influence of retronasal olfaction on patient-perceived outcomes and examine the relationship between retronasal and orthonasal olfaction. METHODS: Retronasal olfactory function was tested using odorized powders in the oral cavity, whereas Sniffin' Sticks test (Burghart Instruments, Wedel, Germany) were used to assess orthonasal function prospectively in 69 adult CRS patients. Endoscopic evaluation of the olfactory cleft was scored using the Olfactory Cleft Endoscopy Scale (OCES). Several quality-of-life (QOL) instruments relating to sinonasal, olfactory, and chemosensory functions were used to assess the interactions between patient-reported outcome measures and retronasal olfaction. RESULTS: There was strong correlation between retronasal and total orthonasal olfaction scores (r = 0.77, P < 0.001) as well as retronasal scores with orthonasal subscores. Retronasal scores were worse in patients with nasal polyposis (P = 0.002), asthma (P = 0.04), and aspirin-exacerbated respiratory disease (AERD) (P = 0.02), whereas OCES was the only independent predictor of retronasal olfaction (r = -0.42, P < 0.001). Significant correlation existed between retronasal olfaction and olfactory-specific QOL and chemosensory smell scores. CONCLUSION: Few studies have examined retronasal olfaction in CRS patients. In this cohort, CRS patients demonstrated deficits in retronasal olfaction, with worse scores in patients with nasal polyposis, asthma, and AERD. Retronasal olfaction scores correlate with degree of inflammation of the olfactory cleft. Retronasal olfaction correlated strongly with orthonasal olfaction and patient-reported smell and taste metrics, although orthonasal olfaction may have a stronger correlation with these metrics. LEVEL OF EVIDENCE: NA. Laryngoscope, 2437-2442, 2018.

Impact of bitter taste receptor phenotype upon clinical presentation in chronic rhinosinusitis.

BACKGROUND: Genetic variation of the bitter taste receptor T2R38 has been associated with recalcitrant chronic rhinosinusitis (CRS). Specific T2R38 polymorphisms, correlating with bitter taste sensitivity to phenylthiocarbamide (PTC), have been identified as an independent risk factor for surgical intervention in CRS patients without polyps; however, the exact role of PTC tasting ability in clinical practice remains unknown. In this investigation we characterize PTC taste sensitivity in a tertiary care rhinology practice with pertinent clinical measures of disease and quality of life (QOL). METHODS: Adult CRS patients were prospectively assessed for their ability to taste PTC and categorized as nontasters, tasters, or supertasters. Objective taste was assessed with strips for bitter, sweet, sour, and salty, whereas olfactory testing was measured with Sniffin' Sticks. Correlation was performed between PTC tasting ability and patient demographics, endoscopy scores, validated QOL surveys, and both subjective and objective measures of taste and olfaction. RESULTS: Sixty-seven patients were enrolled. Fifty-two percent were identified as nontasters, 34% as tasters, and 13% as supertasters. Nontasters were more likely to be non-Hispanic (p = 0.018), white (p = 0.027), without nasal polyposis (p = 0.004), and nonasthmatics (p = 0.019). There were no other statistical differences in patients' demographics, QOL measures, and subjective or objective olfactory and taste scores when compared with patients' oral PTC-sensing ability. CONCLUSION: Oral PTC-sensing ability may serve as a convenient marker of increased disease severity in white CRS patients without polyps and vary among regional populations. PTC tasting ability appears to provide unique phenotypic information not obtained using other subjective or objective measures of smell and taste.

Variability in Retronasal Odor Identification Among Patients With Chronic Rhinosinusitis.

Background Retronasal olfaction is important in flavor detection and enjoyment. The ability to identify specific individual retronasal odors may play a role in quality of life for patients with chronic rhinosinusitis (CRS). Objective To identify patterns and improve understanding of retronasal identification of individual odors in CRS patients. Methods Patients diagnosed with CRS underwent retronasal and orthonasal (Sniffin' Sticks) olfactory testing and taste testing (taste strips). Retronasal identification was tested with presentation of flavored powders on the posterior tongue. Retronasal identification for individual odors was compared with results of orthonasal and taste testing. Results Seventy participants were evaluated. Retronasal identification correlated with orthonasal identification and discrimination for most individual odors. Among all patients, cinnamon and apple were identified better retronasally and banana better orthonasally ( P < .05). Anosmics identified retronasal orange, cinnamon, mushroom, coffee, smoked ham, peach, ginger, grape, and cheese more than would be expected by chance for a forced-choice paradigm with 3 distractor items ( P < .05), and this was independent of objective taste function for most odors. Conclusion Retronasal and orthonasal identification of most odors correlate in CRS patients; however, patients with anosmia can still identify certain retronasal odors more often than expected. These odors do not appear to stimulate gustatory pathways and may involve trigeminal stimulation. Understanding preserved retronasal neural stimuli may allow providers to improve eating-related quality of life in these patients.

Taste impairment in chronic rhinosinusitis.

BACKGROUND: Though many patients with chronic rhinosinusitis (CRS) describe disturbances in smell and taste, there have been no studies specifically assessing taste impairment in CRS. This study sought to objectively assess taste dysfunction in CRS patients and determine whether taste impairment correlates with olfactory dysfunction. Additionally, this investigation sought to determine the impact of taste dysfunction on quality of life (QOL) in CRS and identify the clinical factors that influence taste. METHODS: Sixty-eight CRS patients were prospectively enrolled and completed several QOL surveys in relation to taste, smell, overall sinus-specific QOL, and depression. Validated taste strips were used to determine gustatory dysfunction pertaining to sweet, sour, salty, and bitter. Olfactory testing was assessed using the Sniffin' Sticks Test while both Lund-Kennedy and Olfactory Cleft Endoscopy Scoring (OCES) systems were used for endoscopic evaluation. RESULTS: The overall prevalence of dysgeusia was 28%, with scores significantly lower for sour compared to other subgroups. No correlation was observed between taste scores and objective olfactory metrics including olfaction tests and OCES. Taste scores were better in younger patients (r = 0.28, p = 0.02), female patients (p = 0.004), and never smokers compared to former smokers (p = 0.01). Taste scores did not correlate with patient-reported outcome measures or CRS disease severity metrics. CONCLUSION: Taste dysfunction is a common complaint in CRS. This cohort shows prevalence of gustatory loss to be about 28% using ideal normative values. This dysfunction correlated with male gender, smoking history, and older age. Taste dysfunction did not correlate with measured olfactory outcomes.

Granulocyte-colony stimulating factor reactivates human cytomegalovirus in a latently infected humanized mouse model.

Human cytomegalovirus (HCMV) is a significant cause of morbidity and mortality in organ transplant recipients. The use of granulocyte-colony stimulating factor (G-CSF)-mobilized stem cells from HCMV seropositive donors is suggested to double the risk of late-onset HCMV disease and chronic graft-versus-host disease in recipients when compared to conventional bone marrow transplantation with HCMV seropositive donors, although the etiology of the increased risk is unknown. To understand mechanisms of HCMV transmission in patients receiving G-CSF-mobilized blood products, we generated a NOD-scid IL2Rγ(c)(null)-humanized mouse model in which HCMV establishes latent infection in human hematopoietic cells. In this model, G-CSF induces the reactivation of latent HCMV in monocytes/macrophages that have migrated into organ tissues. In addition to establishing a humanized mouse model for systemic and latent HCMV infection, these results suggest that the use of G-CSF mobilized blood products from seropositive donors pose an elevated risk for HCMV transmission to recipients.