Liver environment and HCV replication affect human T-cell phenotype and expression of inhibitory receptors.

BACKGROUND & AIMS: There is an unclear relationship between inhibitory receptor expression on T cells and their ability to control viral infections. Studies of human immune cells have been mostly limited to T cells from blood, which is often not the site of infection. We investigated the relationship between T-cell location, expression of inhibitory receptors, maturation, and viral control using blood and liver T cells from patients with hepatitis C virus (HCV) and other viral infections. METHODS: We analyzed 36 liver samples from HCV antibody-positive patients (30 from patients with chronic HCV infection, 5 from patients with sustained virological responses to treatment, and 1 from a patient with spontaneous clearance) with 19 paired blood samples and 51 liver samples from HCV-negative patients with 17 paired blood samples. Intrahepatic and circulating lymphocytes were extracted; T-cell markers and inhibitory receptors were quantified for total and virus-specific T cells by flow cytometry. RESULTS: Levels of the markers PD-1 and 2B4 (but not CD160, TIM-3, or LAG-3) were increased on intrahepatic T cells from healthy and diseased liver tissues compared with T cells from blood. HCV-specific intrahepatic CD8(+) T cells from patients with chronic HCV infection were distinct in that they expressed TIM-3 along with PD-1 and 2B4. In comparison, HCV-specific CD8(+) T cells from patients with sustained virological responses and T cells that recognized cytomegalovirus lacked TIM-3 but expressed higher levels of LAG-3; these cells also had different memory phenotypes and proliferative capacity. CONCLUSIONS: T cells from liver express different inhibitory receptors than T cells from blood, independent of liver disease. HCV-specific and cytomegalovirus-specific CD8(+) T cells can be differentiated based on their expression of inhibitory receptors; these correlate with their memory phenotype and levels of proliferation and viral control.

Non-tuberculous Mycobacterium Rhinosinusitis in an Immunocompetent Patient.

Non-tuberculous mycobacteria (NTM) are slow-growing opportunistic pathogens that cause a variety of cutaneous, soft tissue, and pulmonary infections. On rare occasions, NTM causes chronic rhinosinusitis, with the majority of cases presenting in immunocompromised individuals. Other potential risk factors include the presence of foreign bodies, previous sinus surgery or chemoradiation, and use of contaminated water in sinus rinses. We report here a rare case of NTM rhinosinusitis in an otherwise immunocompetent 66-year-old female. The patient underwent functional endoscopic sinus surgery where intraoperative acid-fast bacteria cultures grew Mycobacterium abscessus. She received five weeks of broad-spectrum IV antibiotic therapy followed by three months of oral azithromycin, tigecycline, and linezolid. A one-year post-operative visit showed appropriate healing without crusting or visible infection. This case contributes to the small handful of documented presentations of NTM rhinosinusitis in immunocompetent patients. NTM should be considered when patients present with refractory rhinosinusitis as they may require extended courses of antibiotic treatment. Familiarity with risk factors can further expedite making a diagnosis, ensuring prompt initiation of treatment and relief of symptoms for patients.

Quantifying Karenia brevis bloom severity and respiratory irritation impact along the shoreline of Southwest Florida.

Nearly all annual blooms of the toxic dinoflagellate Karenia brevis (K. brevis) pose a serious threat to coastal Southwest Florida. These blooms discolor water, kill fish and marine mammals, contaminate shellfish, cause mild to severe respiratory irritation, and discourage tourism and recreational activities, leading to significant health and economic impacts in affected communities. Despite these issues, we still lack standard measures suitable for assessing bloom severity or for evaluating the efficacy of modeling efforts simulating bloom initiation and intensity. In this study, historical cell count observations along the southwest Florida shoreline from 1953 to 2019 were used to develop monthly and annual bloom severity indices (BSI). Similarly, respiratory irritation observations routinely reported in Sarasota and Manatee Counties from 2006 to 2019 were used to construct a respiratory irritation index (RI). Both BSI and RI consider spatial extent and temporal evolution of the bloom, and can be updated routinely and used as objective criteria to aid future socioeconomic and scientific studies of K. brevis. These indices can also be used to help managers and decision makers both evaluate the risks along the coast during events and design systems to better respond to and mitigate bloom impacts. Before 1995, sampling was done largely in response to reports of discolored water, fish kills, or respiratory irritation. During this timeframe, lack of sampling during the fall, when blooms typically occur, generally coincided with periods of more frequent-than-usual offshore winds. Consequently, some blooms may have been undetected or under-sampled. As a result, the BSIs before 1995 were likely underestimated and cannot be viewed as accurately as those after 1995. Anomalies in the frequency of onshore wind can also largely account for the discrepancies between BSI and RI during the period from 2006 to 2019. These findings highlighted the importance of onshore wind anomalies when predicting respiratory irritation impacts along beaches.

Estimating cyanobacterial bloom transport by coupling remotely sensed imagery and a hydrodynamic model.

The ability to forecast the transport of harmful cyanobacterial blooms in the Laurentian Great Lakes is beneficial to natural resource managers concerned with public health. This manuscript describes a method that improves the prediction of cyanobacterial bloom transport with the use of a preoperational hydrodynamic model and high temporal resolution satellite imagery. Two scenarios were examined from separate cyanobacterial blooms in western Lake Erie, USA. The first scenario modeled bloom position and extent over the span of 13 days. A geographic center, or centroid, was calculated and assigned to the bloom from observed satellite imagery. The bloom centroid was projected forward in time, and the projected position was compared to the final observed bloom centroid. Image pixels flagged as cyanobacterial bloom were compared between the initial image and the final image, and this was assumed as persistence. The second bloom scenario was modeled for a period of 12 days, and the results were framed in an ecological context in an effort to gain further understanding of cyanobacterial bloom dynamics. These modeling techniques can be incorporated into an operational forecasting system.

A large-scale sustained fish kill in the St. Johns River, Florida: A complex consequence of cyanobacteria blooms.

In the summer of 2010, a sustained multispecies fish kill, affecting primarily adult red drum (Sciaenops ocellatus) and Atlantic stingray (Dasyatis sabina), along with various baitfish such as menhaden (Brevoortia spp.) and shad (Dorosoma spp.), was documented for six weeks along 50 km of the Lower St. Johns River (LSJR), Florida. An Aphanizomenon flos-aquae bloom was present in the freshwater reaches before the fish kill. The kill was triggered by a significant reverse-flow event and sudden influx of high-salinity water in late May that contributed to the collapse of the bloom upstream and brought euryhaline fish downstream into the vicinity of the senescing bloom or its by-products. The decomposing bloom led to a sequence of events, including the release of small amounts of cyanotoxins, bacterial lysis of cyanobacterial cells, high organic loading, and changes in the diversity and dominance of the plankton community to include Microcystis spp., Leptolyngbya sp., Pseudanabaena spp., Planktolyngbya spp., and low concentrations of Heterosigma akashiwo. Dissolved oxygen levels were within normal ranges in the reach of the fish kill, although elevated ammonia concentrations and high pH were detected farther upstream. These conditions resulted in complex pathological changes in fish that were not consistent with acute cyanotoxin exposure or with poor water quality but were attributable to chronic lethal hemolysis. Potential sources of hemolytic activity included H. akashiwo, Microcystis spp., and Bacillus cereus, a hemolytic bacterium. The continued presence of A. flos-aquae in the LSJR could have significant environmental repercussions and ideally the causal factors contributing to bloom growth and maintenance should be fully understood and managed.

An extraordinary Karenia mikimotoi "beer tide" in Kachemak Bay Alaska.

In autumn of 2013 an immense dinoflagellate bloom developed in Kachemak Bay, AK, USA. Much of the Bay was discolored a dark amber color and raised public concerns as small scale fish kills were reported in a few locations. Light microscopy revealed a monospecific bloom of gymnodinoid dinoflagellates that were previously unknown from the Bay. Gene sequencing of SSU rDNA from cells collected from the bloom confirmed the causative species to be Karenia mikimotoi. This represents the first report of a K. mikimotoi bloom in Alaska. After the bloom organism was confirmed, a K. mikimotoi species-specific qPCR assay was developed and used to assess K. mikimotoi abundances in DNA extracted from phytoplankton samples from Kachemak Bay and Lower Cook Inlet (LCI) obtained over a six-year period. The K. mikimotoi abundances were compared with corresponding time series of environmental variables (water temperature, salinity, water column stability, nutrients, precipitation and wind speed) to assess the factors contributing to the development of the bloom. The results showed early bloom development occurred in August when snow melt reduced salinities and increased water column stability during a period of calm winds. Peak bloom concentrations occurred in late September (107 cell eq. L-1) even as water temperatures were decreasing. The bloom gradually declined over the winter but persisted until April of 2014. Karenia mikimotoi cells were not detected two years prior or three years following the bloom, suggesting cells were introduced to Kachemak Bay at a time when conditions allowed K. mikimotoi to thrive.

Skill assessment for an operational algal bloom forecast system.

An operational forecast system for harmful algal blooms (HABs) in southwest Florida is analyzed for forecasting skill. The HABs, caused by the toxic dinoflagellate, Karenia brevis, lead to shellfish toxicity and to respiratory irritation. In addition to predicting new blooms and their extent, HAB forecasts are made twice weekly during a bloom event, using a combination of satellite derived image products, wind predictions, and a rule-based model derived from previous observations and research. These forecasts include: identification, intensification, transport, extent, and impact; the latter being the most significant to the public. Identification involves identifying new blooms as HABs and is validated against an operational monitoring program involving water sampling. Intensification forecasts, which are much less frequently made, can only be evaluated with satellite data on mono-specific blooms. Extent and transport forecasts of HABs are also evaluated against the water samples. Due to the resolution of the forecasts and available validation data, skill cannot be resolved at scales finer than 30 km. Initially, respiratory irritation forecasts were analyzed using anecdotal information, the only available data, which had a bias toward major respiratory events leading to a forecast accuracy exceeding 90%. When a systematic program of twice-daily observations from lifeguards was implemented, the forecast could be meaningfully assessed. The results show that the forecasts identify the occurrence of respiratory events at all lifeguard beaches 70% of the time. However, a high rate (80%) of false positive forecasts occurred at any given beach. As the forecasts were made at half to whole county level, the resolution of the validation data was reduced to county level, reducing false positives to 22% (accuracy of 78%). The study indicates the importance of systematic sampling, even when using qualitative descriptors, the use of validation resolution to evaluate forecast capabilities, and the need to match forecast and validation resolutions.

Chronic HCV infection affects the NK cell phenotype in the blood more than in the liver.

Although epidemiological and functional studies have implicated NK cells in protection and early clearance of HCV, the mechanism by which they may contribute to viral control is poorly understood, particularly at the site of infection, the liver. We hypothesized that a unique immunophenotypic/functional NK cell signature exists in the liver that may provide insights into the contribution of NK cells to viral control. Intrahepatic and blood NK cells were profiled from chronically infected HCV-positive and HCV-negative individuals. Baseline expression of activating and inhibitory receptors was assessed, as well as functional responses following stimulation through classic NK cell pathways. Independent of HCV infection, the liver was enriched for the immunoregulatory CD56(bright) NK cell population, which produced less IFNγ and CD107a but comparable levels of MIP1ß, and was immunophenotypically distinct from their blood counterparts. This profile was mostly unaltered in chronic HCV infection, though different expression levels of NKp46 and NKG2D were associated with different grades of fibrosis. In contrast to the liver, chronic HCV infection associated with an enrichment of CD161(low)perforin(high) NK cells in the blood correlated with increased AST and 2B4 expression. However, the association of relatively discrete changes in the NK cell phenotype in the liver with the fibrosis stage nevertheless suggests an important role for the NK response. Overall these data suggest that tissue localization has a more pervasive effect on NK cells than the presence of chronic viral infection, during which these cells might be mostly attuned to limiting immunopathology. It will be important to characterize NK cells during early HCV infection, when they should have a critical role in limiting infection.