Killer-like receptors and GPR56 progressive expression defines cytokine production of human CD4+ memory T cells.

All memory T cells mount an accelerated response on antigen reencounter, but significant functional heterogeneity is present within the respective memory T-cell subsets as defined by CCR7 and CD45RA expression, thereby warranting further stratification. Here we show that several surface markers, including KLRB1, KLRG1, GPR56, and KLRF1, help define low, high, or exhausted cytokine producers within human peripheral and intrahepatic CD4+ memory T-cell populations. Highest simultaneous production of TNF and IFN-γ is observed in KLRB1+KLRG1+GPR56+ CD4 T cells. By contrast, KLRF1 expression is associated with T-cell exhaustion and reduced TNF/IFN-γ production. Lastly, TCRß repertoire analysis and in vitro differentiation support a regulated, progressive expression for these markers during CD4+ memory T-cell differentiation. Our results thus help refine the classification of human memory T cells to provide insights on inflammatory disease progression and immunotherapy development.

Comparative Evaluation of Two Obstetrical/Gynecology Resident "Boot Camps" of Different Lengths: Equivalent Practice Skills Confidence and Knowledge Levels.

CONTEXT: Since the earlier time of master-apprentice type GME relationships, more residency program educators have developed various forms of boot camps to ease incoming learners into their new specialty roles as first-year residents. Such boot camps have ranged from informal informational sessions with faculty using simulation activities, to more formal workshops entailing pre- and post-event skills assessments with simulation exercises, formative feedback and debriefing sessions. The purpose of this pilot project was to examine for relative pre- and post-boot camp changes in Obstetrics/Gynecology (OB/GYN) practice skills confidence and knowledge levels in two consecutive cohorts (2014 and 2015) of first-year residents. METHODS: Boot camps were of two different lengths: a five-day 2014 camp (n = 32 residents) and shortened three-day 2015 boot camp (n = 29 residents). Respondents from both boot camp cohorts were invited to complete the same 25-item OB/GYN practice skills confidence and knowledge survey. The first three authors developed this survey prior to the initial boot camp (2014). Revisions/adjustments were then made to content after the 2014 to pare down from the initial five days' worth of content for the 2014 boot camp to three days for the 2015 boot camp. RESULTS: Each of 45 sample resident respondents who provided complete pre-and post-boot camp data demonstrated improvements in self-rated practice confidence and knowledge levels. Mean per resident pre-post-boot camp survey rating levels for individual items in the shorter 2015 boot camp cohort increased by 1.096 (SD = 0.5487), over a two-fold increase for most individual items in the 2014 residents. Mean cohort differences represented a non¬-significant equivalent increase in pre-post practice confidence and knowledge levels for individual ratings items between the 2014 and 2015 cohorts (p = 0.241). CONCLUSIONS: Based on these preliminary results, the authors conclude that it may be possible to adjust their OB/GYN boot camp from five days to three and still achieve comparable learner outcomes while delivering the same basic content.

Age and gender leucocytes variances and references values generated using the standardized ONE-Study protocol.

Flow cytometry is now accepted as an ideal technology to reveal changes in immune cell composition and function. However, it is also an error-prone and variable technology, which makes it difficult to reproduce findings across laboratories. We have recently developed a strategy to standardize whole blood flow cytometry. The performance of our protocols was challenged here by profiling samples from healthy volunteers to reveal age- and gender-dependent differences and to establish a standardized reference cohort for use in clinical trials. Whole blood samples from two different cohorts were analyzed (first cohort: n = 52, second cohort: n = 46, both 20-84 years with equal gender distribution). The second cohort was run as a validation cohort by a different operator. The "ONE Study" panels were applied to analyze expression of >30 different surface markers to enumerate proportional and absolute numbers of >50 leucocyte subsets. Indeed, analysis of the first cohort revealed significant age-dependent changes in subsets e.g. increased activated and differentiated CD4(+) and CD8(+) T cell subsets, acquisition of a memory phenotype for Tregs as well as decreased MDC2 and Marginal Zone B cells. Males and females showed different dynamics in age-dependent T cell activation and differentiation, indicating faster immunosenescence in males. Importantly, although both cohorts consisted of a small sample size, our standardized approach enabled validation of age-dependent changes with the second cohort. Thus, we have proven the utility of our strategy and generated reproducible reference ranges accounting for age- and gender-dependent differences, which are crucial for a better patient monitoring and individualized therapy. © 2016 International Society for Advancement of Cytometry.

The use of novel diagnostics to individualize immunosuppression following transplantation.

Despite major improvements in short-term survival of organ allografts, long-term graft survival has not changed significantly. It is also known that toxic side effects of current immunosuppressive drugs (IS) especially calcineurin inhibitors (CNI) contribute to the unsatisfactory graft and patient survival following transplantation. Thus, clinicians strive to reduce or wean IS in potentially eligible patients. Research in the last 10 years has focussed on identification of biomarkers suitable for patient stratification in minimization or weaning trials. Most of the described biomarkers have been run retrospectively on samples collected within single-centre trials. Thus, often their performance has not been validated in other potentially multicentre clinical trials. Ultimately, the utility of biomarkers to identify potential weaning candidates should be investigated in large randomized prospective trials. In particular, for testing in such trials, we need more information about the accuracy, reproducibility, stability and limitations of the described biomarkers. Also, data repositories summarizing crucial information on biomarker performance in age- and gender-matched healthy individuals of different ethnicity are missing. This together with improved bioinformatics tools might help in developing better scores for patient stratification. Here, we will summarize the current results, knowledge and limitations on biomarkers for drug minimization or weaning trials.

Optical coherence tomography of bilateral posterior microphthalmos with papillomacular fold and novel features of retinoschisis and dialysis.

PURPOSE: To report a case of retinoschisis and dialysis associated with bilateral posterior microphthalmos and papillomacular fold. DESIGN: Observational case series. METHODS: Complete ophthalmologic examination of three of five siblings presenting with bilateral posterior microphthalmos and papillomacular fold. Optical coherence tomography (OCT) data are presented to confirm the abnormal anatomy. RESULTS: All subjects have bilateral elevated horizontal papillomacular retinal fold with cystoid macular edema and shallow subretinal fluid. Optical coherence tomography was consistent with our examinations. One subject, a 13-year-old Hispanic, initially presented with retinoschisis in the superotemporal quadrant of the left retina that developed 9 years later into a retinal dialysis without subretinal fluid. The right eye of this same patient developed retinoschisis in the far superotemporal retinal periphery during 9 years of observation. CONCLUSION: Retinoschisis and dialysis may occur in patients with posterior microphthalmos with papillomacular fold. Optical coherence tomography may be helpful in assessing these patients.