Expansions of adaptive-like NK cells with a tissue-resident phenotype in human lung and blood

Human adaptive-like “memory” CD56^dimCD16⁺ natural killer (NK) cells in peripheral blood from cytomegalovirus-seropositive individuals have been extensively investigated in recent years and are currently explored as a treatment strategy for hematological cancers. However, treatment of solid tumors remains limited due to insufficient NK cell tumor infiltration, and it is unknown whether large expansions of adaptive-like NK cells that are equipped for tissue residency and tumor homing exist in peripheral tissues. Here, we show that human lung and blood contains adaptive-like CD56^brightCD16⁻ NK cells with hallmarks of tissue residency, including expression of CD49a. Expansions of adaptive-like lung tissue-resident NK (trNK) cells were found to be present independently of adaptive-like CD56^dimCD16⁺ NK cells and to be hyperresponsive toward target cells. Together, our data demonstrate that phenotypically, functionally, and developmentally distinct subsets of adaptive-like NK cells exist in human lung and blood. Given their tissue-related character and hyperresponsiveness, human lung adaptive-like trNK cells might represent a suitable alternative for therapies targeting solid tumors.

Natural killer (NK) cells are a crucial part of the innate immune system by eliminating virus-infected and malignant cells and boosting immunity through the production of proinflammatory cytokines including IFN-γ and TNF. The concept of adaptive-like or “memory” NK cells has emerged from studies in mice (1–4) and humans (5–10). Adaptive-like NK cells have a distinct phenotype and increased target cell responsiveness, as well as features of longevity and superior recall potential reminiscent of memory T cells (11).Most studies of human adaptive-like NK cells have focused on subsets of NKG2C⁺(KIR⁺)CD56^dimCD16⁺ NK cells found to be expanded and stably maintained in blood of ∼30 to 40% of human cytomegalovirus (HCMV)-seropositive individuals (5, 10). Adaptive-like CD56^dimCD16⁺ NK cells in human peripheral blood (herein defined as adaptive-like CD56^dimCD16⁺ peripheral blood NK [pbNK] cells) have a distinctive phenotypic (5, 10), epigenetic (8, 9), and functional (8–10) profile compared to conventional NK cells and have been suggested to contribute to immunity against HCMV (1, 12). Importantly, adaptive-like CD56^dimCD16⁺ pbNK cells are currently explored for improving NK cell-mediated cancer therapies. While adoptive NK cell transfer showed optimistic results in the treatment of hematological malignancies, targeting solid tumors was less successful due to poor migration to and infiltration into the tumor (reviewed in ref. 13). In these cases, adaptive-like NK cells with an increased capacity to infiltrate tissues, e.g., through coexpression of tissue-specific ligands, might be desirable.We and others recently identified a subset of tissue-resident CD49a⁺CD56^brightCD16⁻ NK cells in the human lung (14, 15). The human lung is a frequent site of infection with viruses such as influenza virus and HCMV, as well as a reservoir for latent HCMV (16). Although human CD56^dimCD16⁺ lung NK cells are hyporesponsive to ex vivo target cell stimulation (17), exposure of human lung NK cells to virus-infected cells is likely to result in functional NK cell priming and expansion of distinct NK cell subsets. However, whether there are expansions of adaptive-like tissue-resident NK (trNK) cells in the human lung is to date unknown.Here, we identify and examine a CD49a⁺KIR⁺NKG2C⁺CD56^brightCD16⁻ NK cell population (herein defined as adaptive-like CD49a⁺ NK cells) with features of tissue residency in human lung and blood, which is distinct from adaptive-like CD56^dimCD16⁺ pbNK cells. In donors with expansions of adaptive-like CD49a⁺ NK cells in the lung, a small but detectable population of adaptive-like CD49a⁺ NK cells was observed in paired peripheral blood. While adaptive-like CD56^dimCD16⁺ pbNK cells (as commonly identified in peripheral blood of HCMV-seropositive donors) and adaptive-like CD49a⁺ NK cells in lung and blood shared a common core gene signature, we identified several unique features of each population, indicating that they may represent developmentally distinct populations. Notably, NK cells from donors with an adaptive-like CD49a⁺ NK cell expansion in the lung were hyperresponsive toward target cells. Thus, we provide evidence indicating that adaptive-like CD49a⁺ NK cells in the human lung represent a population distinct from adaptive-like CD56^dimCD16⁺ pbNK cells with potential implications in lung surveillance and future therapies against solid tumors.     

Targeted nalmefene with simple medical management in the treatment of heavy drinkers: a randomized double-blind placebo-controlled multicenter study

BACKGROUND: Clinical studies with opioid antagonists for treatment of problem drinking have mainly been conducted in specialized alcohol treatment centers, included structured psychosocial treatment, and have focused on maintaining abstinence after a period of abstinence from alcohol. METHODS: This multisite, randomized double-blind study investigated targeted nalmefene in reducing heavy drinking. Specialized alcohol treatment centers and private general practices enrolled 403 subjects (328 men, 75 women). Subjects were instructed to take nalmefene 10 to 40 mg (n=242) or placebo (n=161) when they believed drinking to be imminent. After 28 weeks, 57 subjects from the nalmefene group continued into a 24-week randomized withdrawal extension. Concomitant psychosocial intervention was minimal and no treatment goals were imposed. Alcohol consumption was recorded using the time-line follow-back method. Biochemical indicators of alcohol use were also measured. RESULTS: The mean monthly number of heavy drinking days (HDDs) during the 12-week period before inclusion was 15.5 (SD 6.9) in the nalmefene group and 16.2 (SD 6.9) in the placebo group. During treatment, the mean numbers of HDDs were 8.6 to 9.3 in the nalmefene group and 10.6 to 12.0 in the placebo group (p=0.0065). The levels of serum alanine aminotransferase and gamma-glutamyl transferase decreased in the nalmefene group compared with the placebo group (p=0.0088 and 0.0023). During the randomized withdrawal period, subjects randomized to placebo apparently returned to heavier drinking. Subjects receiving nalmefene reported more nausea, insomnia, fatigue, dizziness, and malaise than subjects on placebo. CONCLUSIONS: Nalmefene appears to be effective and safe in reducing heavy drinking, even when accompanied by minimal psychosocial support.     

Vitamin D status as a determinant of peak bone mass in young Finnish men

Severe vitamin D deficiency causes rickets, but scarce data are available about the extent to which vitamin D status determines the development of the peak bone mass in young adults. Our aim was to evaluate the prevalence of vitamin D deficiency [serum 25-hydroxyvitamin D (25-OHD) less than the lower limit of the reference range of 20-105 nmol/liter] and the relationship between vitamin D status and peak bone mass among young Finnish men. A cross-sectional study of determinants of peak bone mass with data on lifestyle factors collected retrospectively was performed in 220 young men, aged 18.3-20.6 yr. One hundred and seventy men were recruits of the Finnish Army, and 50 were men of similar age who had postponed their military service for reasons not related to health. Bone mineral content, bone mineral density, and scan area were measured in lumbar spine and upper femur by dual energy x-ray absorptiometry. Serum 25-OHD concentrations were followed prospectively for 1 yr. In July 2000, only 0.9% of the men had vitamin D deficiency, but 6 months later, in the winter, the respective percentage was 38.9%. After adjusting for age, height, weight, exercise, smoking, calcium, and alcohol intake, there existed a positive correlation between serum 25-OHD and bone mineral content at lumbar spine (P = 0.057), femoral neck (P = 0.041), trochanter (P = 0.010), and total hip (P = 0.025). The correlation coefficients for the bone mineral densities at the four measurement sites were 0.035, 0.061, 0.056, and 0.068, respectively. No correlation was found to scan area. We conclude that vitamin D deficiency is very common in Finnish young men in the winter, and it may have detrimental effects on the acquisition of maximal peak bone mass. As in Finland vitamin D supplementation to infants is now stopped at the age of 3 yr, it can be asked whether at our latitude it should be continued from that age onward, not for the prevention of rickets, but as prophylaxis for osteoporosis.     

Awareness of hypertension and depressive symptoms: a cross-sectional study in a primary care population

Objective: To investigate the association of hypertension awareness and depressive symptoms, and to analyse factors predisposing aware hypertensives to depressive symptoms.

Design: Cross-sectional study in a primary care population.

Setting: Cardiovascular risk factor survey in two semi-rural towns in Finland.

Subjects: Two thousand six hundred seventy-six middle-aged risk persons without an established cardiovascular or renal disease or type 2 diabetes.

Main outcome measures: Depressive symptoms, previous and new diagnosis of hypertension.

Results: Hypertension was diagnosed in 47.9% of the subjects, of whom 34.5% (442/1 282) had previously undetected hypertension. Depressive symptoms were reported by 14% of the subjects previously aware of their hypertension, and by 9% of both unaware hypertensives and normotensive subjects. In the logistic regression analysis, both the normotensive (OR 0.62, 95% CI 0.45–0.86) (p?=?0.0038) and the unaware hypertensive subjects (OR 0.54, 95% CI 0.35–0.84) (p?=?0.0067) had lower risk for depressive symptoms than the previously diagnosed hypertensives. Among these aware hypertensives, female gender (OR 3.61, 95% CI 2.06–6.32), harmful alcohol use (OR 2.55, 95% CI 1.40–4.64) and obesity (OR 2.50, 95% CI 1.01–6.21) predicted depressive symptoms. Non-smoking (OR 0.57, 95% Cl 0.33–0.99) and moderate leisure-time physical activity compared to low (OR 0.53, 95% CI 0.33–0.84) seemed to buffer against depressive symptoms.

Conclusion: Depressive symptoms are common in hypertensive persons even without comorbidities, if the person is already aware of his/her hypertension. Many modifiable, lifestyle associated factors may contribute to the association of hypertension and depressive symptoms.

• Key Points

• Hypertension and depressive symptoms are known to form a toxic combination contributing even to all-cause mortality.

• Comorbidities or the labelling effect of the diagnosis of hypertension can confound their association.

• Our study shows that depressive symptoms are common in hypertensive persons even without comorbidities, if the person is already aware of his/her hypertension.

• Many modifiable, lifestyle-associated factors may contribute to the association of hypertension and depressive symptoms.

• When treating hypertensive patients, consideration of depressive symptoms is important in order to promote favorable lifestyle and control of hypertension.

     

Electric field navigated transcranial magnetic stimulation for chronic tinnitus: A pilot study

Objective: Repetitive transcranial magnetic stimulation (rTMS) has shown potential in reducing tinnitus symptoms. We evaluated effects of electric field (E-field) navigated rTMS targeted neuroanatomically according to tinnitus pitch. Design: In this open methodological pilot study, the patients received E-field navigated 1-Hz rTMS in daily treatment sessions to the left superior temporal gyrus, targeted according to tonotopic representation of their individual tinnitus pitch. Patients rated their tinnitus intensity and annoyance with a numeric rating scale (NRS) from 0 to 10 at the baseline and after each rTMS session. They also rated their global impression of change (scale ? 3 to + 3) after the treatment. Study sample: Thirteen patients (mean age 53 years; 10 men, 3 women) with chronic, intractable tinnitus. Results: The mean intensity was 7.1 (SD 1.8) at the baseline, decreasing to 4.5 (SD 2.2) after the rTMS (p < 0.0001). The mean annoyance 7.0 (SD 1.8) at the baseline decreased to 4.0 (SD 2.4) after the treatment (p < 0.0001). Intensity diminished at least 30% in 8/13 patients and annoyance in 9/13 patients. A total of 10/13 patients felt subjective benefit from the treatment. Conclusions: These preliminary observations suggest that E-field-rTMS may improve the current treatment options for intractable tinnitus.     

Long-term effectiveness of enzyme replacement therapy in Fabry disease with the p.Arg227Ter variant: Fabry disease in Ostrobothnia (FAST) study

Fabry disease (FD) is an X chromosome-linked, life-threatening lysosomal disease caused by one of more than 1000 currently known variants in the α-galactosidase A (GLA) gene. The follow-up part of the Fabry Disease in Ostrobothnia (FAST) study reports the long-term effect of enzyme replacement therapy (ERT) on a prospectively collected cohort of 12 patients, 4 males and 8 females, mean age 46 years (SD 16), with the classical variant c.679C > T p.Arg227Ter, which is one of the most common FD variants worldwide. In the natural history period of the FAST study, half of the patients in both sexes had at least one major event, of which 80% were of cardiac origin. During 5 years of ERT, four patients had a total of six major clinical events consisting of one silent ischemic stroke, three ventricular tachycardias and two increased left ventricular mass indexes. In addition, four patients developed minor cardiac events, four patients minor renal events, and one patient a minor neurological event. ERTs may delay but not prevent the progression of the disease in most patients with the variant Arg227Ter. This variant might be suitable for investigating the efficacy of second-generation ERTs compared to the currently used ERTs regardless of sex.     

Eosinophilia is a favorable prognostic marker for oral cavity and lip squamous cell carcinoma

Eosinophils are frequently encountered with squamous cell carcinomas (SCC) and it has been proposed that tumor‐associated tissue eosinophilia (TATE) could be of prognostic significance in oral SCC. The aim was to evaluate TATE in 83 oral cavity and 16 lip SCCs as well as the best possible use of TATE as a prognostic marker. The number of eosinophils was counted per high power fields (HPF, ×400) in three different representative areas of the tumor and its stroma. The degree of TATE was analyzed in relation to clinicopathological features of tumors and patients’ survival (follow‐up mean 40.7 months) using Fisher's exact test. TATE was detected in 58 (70%) oral and 8 (50%) lip SCC samples. The median number of eosinophils between oral and lip SCC was different (p = 0.028) but TATE was similar per HPF (p = 0.085). Totally, 6% of lip and 21% of oral SCC patients died during the follow‐up. The patients with the higher TATE had significantly better survival than the patients with the lower TATE (p = 0.0136). The best cut‐off value predicting the survival was 4 eosinophils/HPF. TATE is a prognostic marker for oral and lip SCC: more than 4 eosinophils/HPF may predict more favorable prognosis.     

Drug transport in corneal epithelium and blood-retina barrier: emerging role of transporters in ocular pharmacokinetics

Corneal epithelium and blood-retina barrier (i.e. retinal capillaries and retinal pigment epithelium (RPE)) are the key membranes that regulate the access of xenobiotics into the ocular tissues. Corneal epithelium limits drug absorption from the lacrimal fluid into the anterior chamber after eyedrop administration, whereas blood-retina barrier restricts the entry of drugs from systemic circulation to the posterior eye segment. Like in general pharmacokinetics, the role of transporters has been considered to be quite limited as compared to the passive diffusion of drugs across the membranes. As the functional role of transporters is being revealed it has become evident that the transporters are widely important in pharmacokinetics. This review updates the current knowledge about the transporters in the corneal epithelium and blood-retina barrier and demonstrates that the information is far from complete. We also show that quite many ocular drugs are known to interact with transporters, but the studies about the expression and function of those transporters in the eye are still sparse. Therefore, the transporters probably have greater role in ocular pharmacokinetics than we currently realise.