Peripheral Populations Research Articles

Herstory of the Periphery: Approaching Placemaking through Feminist Autoethnography

This article elaborated on the possibilities of feminist interference to the reading of informal settlements. In so doing, we focused on a squatter settlement (Limontepe) in a metropolitan municipality (İzmir Metropolitan Municipality) in Turkey, and tried to interrogate the institutional, official history, based on city registers, parliamentary decisions and state and municipality archives. We argue that the way peripheral populations and/or marginalized groups are treated in official histories of nation-states leads to partial knowledge of the place. We propose that the knowledge collected, accumulated and exchanged through everyday lives of the inhabitants, past and present might be a viable option to check the official history writing and fill in the blanks therein. The article is an attempt to walk through feminist (auto)ethnography to tie the knowledge of the past to today’s placemaking practices. As feminist researchers we consider engaging in the everyday life practices of inhabitants as a way to participate in the dynamic knowledge production processes of the place.

Differential Effects of Regulatory T Cells in the Meninges and Spinal Cord of Male and Female Mice with Neuropathic Pain.

Immune cells play a critical role in promoting neuroinflammation and the development of neuropathic pain. However, some subsets of immune cells are essential for pain resolution. Among them are regulatory T cells (Tregs), a specialised subpopulation of T cells that limit excessive immune responses and preserve immune homeostasis. In this study, we utilised intrathecal adoptive transfer of activated Tregs in male and female mice after peripheral nerve injury to investigate Treg migration and whether Treg-mediated suppression of pain behaviours is associated with changes in peripheral immune cell populations in lymphoid and meningeal tissues and spinal microglial and astrocyte reactivity and phenotypes. Treatment with Tregs suppressed mechanical pain hypersensitivity and improved changes in exploratory behaviours after chronic constriction injury (CCI) of the sciatic nerve in both male and female mice. The injected Treg cells were detected in the choroid plexus and the pia mater and in peripheral lymphoid organs in both male and female recipient mice. Nonetheless, Treg treatment resulted in differential changes in meningeal and lymph node immune cell profiles in male and female mice. Moreover, in male mice, adoptive transfer of Tregs ameliorated the CCI-induced increase in microglia reactivity and inflammatory phenotypic shift, increasing M2-like phenotypic markers and attenuating astrocyte reactivity and neurotoxic astrocytes. Contrastingly, in CCI female mice, Treg injection increased astrocyte reactivity and neuroprotective astrocytes. These findings show that the adoptive transfer of Tregs modulates meningeal and peripheral immunity, as well as spinal glial populations, and alleviates neuropathic pain, potentially through different mechanisms in males and females.

Insights into the genetic makeup of French Polynesian peripheral populations of the small giant clam Tridacna maxima

Abstract The small giant clam, Tridacna maxima, is distributed from the Red Sea and East African coast to French Polynesia. Across this widespread Indo‐Pacific range, T. maxima shows strong population structure, in agreement with its limited dispersal abilities. Peripheral populations may have smaller effective population sizes, increasing their vulnerability under any environmental changes. Understanding evolutionary processes at play in such regions located at the edges of T. maxima distribution is a prerequisite in the context of transfers and restocking programmes. In this study, giant clams were sampled from 14 atolls and islands within four archipelagos in the peripheral region of French Polynesia, in 2001–2002 and/or in 2012–2013, then genotyped at the COI gene and at nine microsatellite loci. Mitochondrial lineages of T. maxima from French Polynesia diverged from those sampled in Micronesia, Melanesia, the Coral Triangle and the Red Sea by 6.6–7.3%. Within French Polynesia, significant genetic structure was found, indicating restricted gene flow, and it was stable through time. Most of the genetic variation at microsatellite loci was between archipelagos. The most differentiated archipelago was the most geographically isolated (the Austral Islands). The current patterns of genetic structuring of T. maxima in French Polynesia probably result from long‐term genetic isolation with limited dispersal ability. In addition, these results underlined that sufficiently large populations of T. maxima have persisted in the Central Pacific during the last sea‐level regression. Strategies to optimize transfers and restocking programmes should be designed to preserve the genetic diversity and structure observed here, to avoid the risks of altering the genetic structure, allele loss and/or introduction of maladapted alleles in the receiving populations.

Antigen-specific decidual CD8+ T cells include distinct effector memory and tissue-resident memory cells.

Maternal decidual CD8+ T cells must integrate the antithetical demands of providing immunity to infection while maintaining immune tolerance for fetal and placental antigens. Human decidual CD8+ T cells were shown to be highly differentiated memory T cells with mixed signatures of dysfunction, activation, and effector function. However, no information is present on how specificity for microbial or fetal antigens relates to their function or dysfunction. In addition, a key question, whether decidual CD8+ T cells include unique tissue-resident memory T cells (Trm) or also effector memory T cell (Tem) types shared with peripheral blood populations, is unknown. Here, high-dimensional flow cytometry of decidual and blood CD8+ T cells identified 2 Tem populations shared in blood and decidua and 9 functionally distinct Trm clusters uniquely found in decidua. Interestingly, fetus- and virus-specific decidual CD8+ Trm cells had similar features of inhibition and cytotoxicity, with no significant differences in their expression of activation, inhibitory, and cytotoxic molecules, suggesting that not all fetus-specific CD8+ T cell responses are suppressed at the maternal-fetal interface. Understanding how decidual CD8+ T cell specificity relates to their function and tissue residency is crucial in advancing understanding of their contribution to placental inflammation and control of congenital infections.

Evaluating the use of non-invasive hair sampling and ddRAD to characterize populations of endangered species: Application to a peripheral population of the European mink.

The application of next-generation sequencing (NGS) to non-invasive samples is one of the most promising methods in conservation genomics, but these types of samples present significant challenges for NGS. The European mink (Mustela lutreola) is critically endangered throughout its range. However, important aspects such as census size and inbreeding remain still unknown in many populations, so it is crucial to develop new methods to monitor this species. In this work, we placed hair tubes along riverbanks in a border area of the Iberian population, which allowed the genetic identification of 76 European mink hair samples. We then applied a reduced representation genomic sequencing (ddRAD) technique to a subset of these samples to test whether we could extract sufficient genomic information from them. We show that several problems with the DNA, including contamination, fragmentation, oxidation, and possibly sample mixing, affected the samples. Using various bioinformatic techniques to reduce these problems, we were able to unambiguously genotype 19 hair samples belonging to six individuals. This small number of individuals showed that the demographic status of the species in this peripheral population is worse than expected. The data obtained also allowed us to perform preliminary analyses of relatedness and inbreeding. Although further improvements in sampling and analysis are needed, the application of the ddRAD technique to non-invasively obtained hairs represents a significant advance in the genomic study of endangered species.

Disease activity in chronic inflammatory demyelinating polyneuropathy: association between circulating B-cell subsets, cytokine levels, and clinical outcomes.

Chronic inflammatory demyelinating polyneuropathy (CIDP), a common and treatable autoimmune neuropathy, is frequently misdiagnosed. The aim of this study is to evaluate the relationship between immunological markers and clinical outcome measures in a mixed cohort of patients with typical CIDP and CIDP variants at different disease stages. Twenty-three typical, 16 multifocal and five distal CIDP patients were included. Twenty-five sex and age-matched healthy controls and 12 patients with Charcot-Marie-Tooth type 1A (CMT1A) disease served as controls. Peripheral B-cell populations were analyzed by flow cytometry. IL6, IL10, TNFA mRNA and mir-21, mir-146a, and mir-155-5p expression levels were evaluated by real-time polymerase chain reaction in peripheral blood mononuclear cells (PBMC) and/or skin biopsy specimens. Results were then assessed for a possible association with clinical disability scores and intraepidermal nerve fiber densities (IENFD) in the distal leg. We detected a significant reduction in naive B cells (P ≤ 0.001), plasma cells (P ≤ 0.001) and regulatory B cells (P < 0.05), and an elevation in switched memory B cells (P ≤ 0.001) in CIDP compared to healthy controls. CMT1A and CIDP patients had comparable B-cell subset distribution. CIDP cases had significantly higher TNFA and IL10 gene expression levels in PBMC compared to healthy controls (P < 0.05 and P ≤ 0.01, respectively). IENFDs in the distal leg showed a moderate negative correlation with switched memory B-cell ratios (r = -0.51, P < 0.05) and a moderate positive correlation with plasma cell ratios (r = 0.46, P < 0.05). INCAT sum scores showed a moderate positive correlation with IL6 gene expression levels in PBMC (r = 0.54, P < 0.05). Altered B-cell homeostasis and IL10 and TNFA gene expression levels imply chronic antigen exposure and overactivity in the humoral immune system, and seem to be a common pathological pathway in both typical CIDP and CIDP variants.

Circulating and intratumoral immune determinants of response to atezolizumab plus bevacizumab in patients with variant histology or sarcomatoid renal cell carcinoma

Abstract Background Renal cell carcinoma of variant histology (vRCC) encompasses approximately 20% of RCC diagnoses. Due to a poor understanding of the different biologies of vRCCs, there is currently no standard of care for this type of tumor and guidelines are extrapolated from clear-cell RCC trials. A phase II trial of atezolizumab plus bevacizumab in patients with vRCC or sarcomatoid differentiation had an acceptable safety profile and evidence of clinical activity, particularly in tumors with positive PD-L1 expression or sarcomatoid differentiation. To better understand the determinants of immunotherapy response in this population, we characterized blood- and tissue-based immune markers for patients with vRCC, or any RCC histology with sarcomatoid differentiation, enrolled in this phase II trial of atezolizumab and bevacizumab. Methods We used patient data and biospecimens (blood and tumor) from a prospective phase-II clinical trial of the anti-PD-L1 antibody atezolizumab and the anti-VEGF-A antibody bevacizumab in patients with metastatic RCC with variant histology and/or sarcomatoid differentiation (NCT02724878). Blood samples were collected at baseline, prior to therapy (C1D1: cycle one day one), and on-treatment, following two cycles of therapy (C3D1: cycle three day one). Plasma was assayed for the levels of soluble factors (e.g., cytokines and growth factors), and peripheral blood mononuclear cells (PBMCs) were immunophenotyped using flow cytometry. In addition, formalin-fixed, paraffin-embedded (FFPE) archival or pre-treatment study tumor biopsies were analyzed by immunohistochemistry and immunofluorescence for enrolled subjects with available biospecimen. Baseline demographics, clinical characteristics, and treatment outcomes, including progression-free survival (PFS), overall survival (OS), and clinical benefit, were collected, and analyzed from the trial database. This study was approved by each participating institution's institutional review board, and all patients provided written informed consent. We used R (version 4.1.1) to carry out our analyses. We assessed the relationships between biomarker variables were analyzed using Pearson correlations and hierarchical clustering using Euclidean distance. All biomarker variables were Z-score normalized. We calculated an inflammatory module, the systemic inflammation score (SIS), which corresponds to the average score of the five highly correlated inflammatory cytokines: MIP-1b, IL-1, MCP-1, IL-6, and IL-13. We applied pair-wise Wilcoxon tests to evaluate the association of biomarkers with clinical benefit groups and log-rank-tests and univariate Cox regression models to evaluate the relationship of biomarkers with PFS and OS. Results With the exception of two angiogenic cytokines (VEGF-A and Angiopoietin 2) that were more abundant in patients with non-sarcomatoid tumors, baseline circulating cytokines (plasma) had similar distributions between sarcomatoid and non-sarcomatoid subgroups. Baseline circulating inflammatory cytokines were highly correlated with one another, forming an “inflammatory module” that was increased in IMDC-poor risk patients and was associated with worse PFS (p=0.028) and with resistance to therapy (p=0.033). At baseline, an elevated circulating VEGF-A level was associated with a lack of response (p=0.03) and worse PFS (p=0.021). However, a larger increase in on-treatment levels of circulating VEGF-A was associated with clinical benefit (p=0.01) and improved overall survival (p=0.0058). Among peripheral immune cell populations, an on-treatment decrease in circulating PD-L1+ T cells was associated with improved outcomes, with a reduction in CD4+ PD-L1+ (HR:0.62[0.49-0.91], p=0.016) and CD8+ PD-L1+ T cells (HR:0.59[0.39-0.87], p=0.009) correlated with improved PFS. Within the tumor itself, a higher percentage of terminally exhausted (PD-1+ and either TIM-3+ or LAG-3+) CD8+ T cells was associated with worse PFS (p=0.028). We observed similar trends for all of these results in different subgroups according to sarcomatoid status and histology. Conclusions Overall, these findings support the value of tumor and blood-based immune assessments in determining therapeutic benefit for patients with RCC receiving atezolizumab plus bevacizumab. Furthermore, they provide a foundation for future biomarker studies for patients with variant histology RCC receiving immunotherapy-based combinations.

Range Contraction and Population Decline of the European Dupont’s Lark Population

The Dupont’s lark (Chersophilus duponti) is an endangered passerine typical of Mediterranean shrub-steppes, whose European distribution is restricted to Spain. Here, we update the population size and distribution range of the species at a European scale and evaluate (i) the current status; (ii) the change in population size and distribution range of the species from 2004 to 2009 to the current period (2017–2022); and (iii) the effectiveness of the current network of special protection areas (SPAs) for protecting the Dupont’s lark. The European Dupont’s lark population showed a decrease of 29.9%, declining from ca. 3267 to 2289 territorial males from 2004 to 2009 to the current period. Moreover, the species has suffered a contraction in its distribution range of 35.9%, with only 39.3% of the species’ territories located within the current network of SPAs. Our findings agree with the previously described decline of the Dupont’s lark in Europe. The population decline was even larger in peripheral regions, which suggests that the species is suffering a centripetal process of contraction and extinction. These results indicate that if there is no change in present-day declining forces, several peripheral populations will reach extinction in a few decades and the overall population size of the species will continue decreasing. Our study should be considered as a last call for action and used for implementing urgent conservation measures to protect the species and its habitat. Future studies should focus on analyzing and managing the factors driving the species’ extinction and future actions for the conservation of the species should focus on increasing the percentage of the Dupont’s lark territories within protected areas, since the data are alarmingly low for a species that is facing clear risk of extinction.

Changes in peripheral blood immune cell population in thyroid cancer patients treated with lenvatinib

This study evaluated changes in the peripheral blood immune cell population in patients with advanced thyroid cancer receiving lenvatinib treatment to confirm the immune-modulatory effect of lenvatinib. After obtaining informed consent from patients, we prospectively collected 20 ml of whole blood at 2–3 months intervals 2–4 times from each patient; peripheral blood mononuclear cells (PBMCs) were separated, and the Maxpar Direct Immune Profiling Assay was performed. A total of 10 patients were enrolled, and 31 blood samples were obtained. The median age of patients was 65 years, and all patients showed durable responses to the lenvatinib treatment. When we compared the PBMC profiles between the pre-treatment, on-treatment, and off-treatment samples, the peripheral natural killer (NK) cell proportion differed significantly. The proportion of NK cells among total live cells significantly increased from 9.3 ± 4.5 (%) in the pre-treatment samples to 20.8 ± 7.9 (%) in the on-treatment samples (P = 0.009) and decreased to 13.3 ± 3.1 (%) in the off-treatment samples (P = 0.07). There was a significant increase in the peripheral NK cell population with lenvatinib treatment in advanced thyroid cancer patients. This finding confirms the immune-modulatory effect of lenvatinib.

Next generation 3D-printed intravaginal ring for prevention of HIV and unintended pregnancy

Globally, there are 20 million adolescent girls and young women living with HIV who have limited access to long-acting, effective, women-controlled preventative methods. Additionally, although there are many contraceptive methods available, globally, half of all pregnancies remain unintended. Here we report the first 3D-printed multipurpose prevention technology (MPT) intravaginal ring (IVR) for HIV prevention and contraception. We utilized continuous liquid interface production (CLIP™) to fabricate MPT IVRs in a biocompatible silicone-based resin. Etonogestrel (ENG), ethinyl estradiol (EE), and islatravir (ISL) were loaded into the silicone poly(urethane) IVR in a controlled single step drug loading process driven by absorption. ENG/EE/ISL IVR promoted sustained release of drugs for 150 days in vitro and 14 days in sheep. There were no adverse MPT IVR-related findings of cervicovaginal toxicity or changes in vaginal biopsies or microbiome community profiles evaluated in sheep. Furthermore, ISL IVR in macaques promoted sustained release for 28 days with ISL-triphosphate levels above the established pharmacokinetic benchmark of 50–100 fmol/106 PBMCs. The ISL IVR was found to be safe and well tolerated in the macaques with no observed mucosal cytokine changes or alterations in peripheral CD4 T-cell populations. Collectively, the proposed MPT IVR has potential to expand preventative choices for young women and girls.

Older forests function as energetic and demographic refugia for a climate-sensitive species.

More frequent and extreme heat waves threaten climate-sensitive species. Structurally complex, older forests can buffer these effects by creating cool microclimates, although the mechanisms by which forest refugia mitigate physiological responses to heat exposure and subsequent population-level consequences remain relatively unexplored. We leveraged fine-scale movement data, doubly labeled water, and two decades of demographic data for the California spotted owl (Strix occidentalis occidentalis) to (1) assess the role of older forest characteristics as potential energetic buffers for individuals and (2) examine the subsequent value of older forests as refugia for a core population in the Sierra Nevada and a periphery population in the San Bernardino Mountains. Individuals spent less energy moving during warmer sampling periods and the presence of tall canopies facilitated energetic conservation during daytime roosting activities. In the core population, where tall-canopied forest was prevalent, temperature anomalies did not affect territory occupancy dynamics as warmer sites were both less likely to go extinct and less likely to become colonized, suggesting a trade-off between foraging opportunities and temperature exposure. In the peripheral population, sites were more likely to become unoccupied following warm summers, presumably because of less prevalent older forest conditions. While individuals avoided elevated energetic expenditure associated with temperature exposure, behavioral strategies to conserve energy may have diverted time and energy from reproduction or territory defense. Conserving older forests, which are threatened due to fire and drought, may benefit individuals from energetic consequences of exposure to stressful thermal conditions.

Extent of intraspecific trait variability in ecologically central and marginal populations of a dominant alpine plant across European mountains.

Studying trait variability and restricted gene flow between populations of species can reveal species dynamics. Peripheral populations commonly exhibit lower genetic diversity and trait variability due to isolation and ecological marginality, unlike central populations experiencing gene flow and optimal conditions. This study focused on Carex curvula, the dominant species in alpine acidic meadows of European mountain regions. The species is sparser in dry areas such as the Pyrenees and Balkans, compared to the Central-Eastern Alps and Carpathians. We hypothesized that distinct population groups could be identified based on their mean functional trait values and their correlation with the environment; we predicted that ecologically marginal populations would have stronger trait correlations, lower within-population trait variability (intraspecific trait variability, ITV) and lower genetic diversity than populations of optimal habitats. Sampling was conducted in 34 populations that spanned the entire distribution range of C. curvula. We used hierarchical clustering to identify emergent functional groups of populations, defined by combinations of multiple traits associated with nutrient economy and drought tolerance (e.g. specific leaf area, anatomy). We contrasted the geographical distribution of these groups in relation to environment and genetic structure. We compared pairwise trait relationships, within-population trait variation (ITV) and neutral genetic diversity between groups. Our study identified emergent functional groups of populations. Those in the southernmost ranges, specifically the Pyrenees and Balkan region, showed drought-tolerant trait syndromes and correlated with indicators of limited water availability. While we noted a decline in population genetic diversity, we did not observe any significant changes in ITV in ecologically marginal (peripheral) populations. Our research exemplifies the relationship between ecological marginality and geographical peripherality, which in this case study is linked to genetic depauperation but not to reduced ITV. Understanding these relationships is crucial for understanding the biogeographical factors shaping trait variation.

Habitat suitability and centrality—not peripherality—predict demographic performance in a Neotropical lizard

AbstractAimThe centre–periphery hypothesis (CPH) sustains that peripheral populations are subjected to harsher environmental conditions, which reduces demographic performance manifested, among others, by lower abundance, higher inbreeding and higher developmental instability. However, range centrality/peripherality is not necessarily a good proxy for environmental suitability since geographical distributions are often irregular in shape, barriers may limit dispersal towards suitable areas, and ancestral range shifts may have occurred unevenly in each direction. We evaluated predictions of the CPH for populations of the Neotropical lizard Notomabuya frenata by modelling the effects of environmental suitability and geographical centrality/peripherality upon demographic performance.LocationBrazilian Cerrado.TaxonNotomabuya frenata (Squamata, Scincidae).MethodsWe used populational values of body size, body mass, body condition, tail autotomy and fluctuating asymmetry (FA; a proxy of developmental instability) to estimate demographic performance. Then, we built correlative models to estimate environmental suitability for each sampled population. Finally, we used hierarchical Bayesian models based on a combination of stochastic partial differential equations and integrated nested Laplace approximations to model the relationships between demographic performance and predictor variables (geographical centrality/peripherality and environmental suitability) and make predictions across the geographical distribution of N. frenata.ResultsGeneralized boosting produced the best environmental suitability models. The total suitability area spanned more than 4 million km2 in cis‐Andean South America. All scalation counts presented some degree of asymmetry. Only three response variables—all related to FA—responded significantly to predictor changes. Contrary to the predictions of the CPH, one scalation count presented higher asymmetry closer to the range center and another in more suitable environments. Peripherality did not significantly influence any of the response variables.Main ConclusionsPopulations may respond differently to geographical centrality/peripherality and environmental suitability, as these characteristics do not necessarily coincide. Studies testing the CPH should ideally incorporate these different predictors of demographic performance.

Morphological variability decreases in populations living in less suitable environments and close to the range edges

AbstractAimGeographical range expansion depends on ecological and evolutionary processes that may hamper local adaptations in populations living at range edges by constraining phenotypic variability. This study investigates the spatial patterns of the intraspecific variability of skull traits throughout the geographical range of a marsupial from the Brazilian Atlantic Forest. We aimed to answer whether the distance from the range edge and the environmental suitability explain the geographical variation of morphological variability of the species.LocationAtlantic Forest, Brazil.TaxonMarmosops incanus (Didelphimorphia, Didelphidae).MethodsWe analysed adult specimens deposited in the main biological collections in southeastern Brazil. The morphological variability extent and integration within populations were characterised by 13 linear measures of the skull, using a multivariate approach. Environmental suitability for the species' occurrence was estimated by ecological niche models, using climate variables and vegetation productivity as predictors and three different modelling methods. Distance from the range edge was calculated based on the minimum linear distance between populations and the closest range limit. We fitted linear regressions and selected the best models that explained the spatial variation of morphological variability based on the Akaike information criterion.ResultsThe extent of morphological variability of M. incanus is positively correlated with morphological integration and increases with local environmental suitability and distance from the range edges. However, the relationship between morphological variability and environmental suitability depends on the niche modelling method.Main ConclusionsUnfavourable environmental conditions constrain morphological variability within populations of the Gray slender opossum and may hamper local adaptation in peripheral populations living in less suitable environments. In addition to environmental conditions, geographical location of populations also plays an important role in phenotypic variability among populations. We stress the boundary effect of the species range on local adaptation of peripheral populations and its possible consequences for the range expansion dynamics.

LIVING AT THE EDGE: A COMPARATIVE STUDY OF THE CENTRAL AND MARGINAL POPULATIONS OF DACTYLORHIZA TRAUNSTEINERI (ORCHIDACEAE) IN EUROPEAN NORTHEAST OF RUSSIA

Peripheral populations are considered more vulnerable than central ones, but recent studies have questioned the existence of clear patterns within species ranges. We studied reproductive and demographic characteristics of the rare orchid Dactylorhiza traunsteineri at the northeastern limit of its distribution range (the Subpolar Urals) and compared them with the data of four populations located closer to the center of the range (south of the Komi Republic). In peripheral populations, number of individuals per population, plant sizes, number of owers per individual and seeds per fruit were signi cantly lower than those in populations located closer to the center of the area. Contrary to expectations, the peripheral populations were renewed better than the central ones, which may be due to the high level of moisture availability in the Subpolar Urals. The proportion of juvenile individuals in the peripheral populations was 18.7%, in populations in the south of the Komi Republic - 6.3-6.9%. In addition, we found that in the direction from the center to the periphery of the species range, the proportion of empty space in seeds increased (from 63.9 to 75.6%). This provides the seeds with better volatility and increases the chances of reaching places favorable for germination, which are becoming fewer at the edge of the range.

Natural killer cell subsets in endometrial fluid: a pilot study of their association with the endometrial cycle and reproductive parameters.

To investigate if there are natural killer (NK) cells in endometrial fluid (EF) and their relationship with the endometrial cycle and reproductive parameters. The population under study consisted of 43 women aged 18-40 undergoing infertility workup at our University Hospital in 2021-2022. The EF samples were obtained at the first visit to our unit, on occasion of the mock embryo transfer. The day of the cycle was considered only in cycles of 27-29 days. An immunophenotype study of NK in EF was performed by flow cytometry analysis. In a subgroup of women, on the same day, NK was studied in EF and peripheral blood. Our study is the first to evidence NK cells in EF. None of the NK cells observed corresponded to a mature peripheral blood NK cell population (stages 4-5), and neither endometrial nor decidual uNK cells were detected. Nevertheless, we found 2 patient groups with an NK cell subset with a higher expression of CD16+, which could belong to an intermediate or transient stage between the uNK and pbNK NK cell population in the EF. We found that CD16 was significantly increased in the mid-late luteal phase and its correlation with the day of the cycle. The NK immunophenotype was different in EF and peripheral blood. We described a new component of the EF, the NK cells, whose CD16 activity is closely correlated with the day of the cycle. These cells could play a role in implantation/implantation failure.

Youth and Abuse of Substances

The following comprehensive and thorough literature review focuses on Youth and Abuse of Substances. More specifically, this study describes in twenty first century youth are more into abuse of substances, the types of substances described in the currently available literature, the various existing prevention and treatment programs, factors which have been attributed to youth and abuse of substances, and focuses on areas in this field that need further research. More complaint of this study is to focuses on youth mental health and motivation towards abuse of substances. Secondary sources were utilized, providing a complete picture of the ever-growing challenge of youth and abuse of substances. This study shall then conclude with the author’s personal integration of this information, and how this information can potentially benefit professionals working with peripheral population.

201-OR: Characterization of Type 1 Diabetes (T1D) Endotypes

To better understand how T1D heterogeneity relates to disease progression and therapeutic responsiveness, we studied 566 recent onset subjects (mean age at onset: 16.5 +/- 8.2) that had participated in the following TrialNet trials: MMF-DZB, anti-CD20; GAD-alum; CTLA4-Ig; Canakinumab; and ATG-GCSF. Our analysis began with a bioassay where plasma collected pre-intervention and within 100 days of diagnosis was used to induce transcription in a well-controlled peripheral blood mononuclear cell population. Microarray analysis identified 2,854 transcripts exhibiting high variation in at least 5 out of 6 trials. Hierarchical clustering and principal component analysis of these transcripts concordantly divided the participants into 2 major groups: Group 1 and Group 2. Analysis of post-baseline samples found group assignments conserved over time. Proportional analyses of the 406 subjects aged &amp;lt;18 years found Groups 1 and 2 distinct: subjects aged &amp;gt;14years segregated within Group 2 (39.9% vs 29.3%, p=0.03); subjects possessing neutral/low-risk HLA haplotypes segregated within Group 1 (20.5% vs 11.8%, p=0.04); placebo arm participants with the most rapid C-peptide rate of decline (&amp;lt;-0.15) segregated within Group 1 (20.3% vs 7.1%, p=0.04); and the subjects most responsive to anti-CD20 (p=0.01) and ATG-GCSF (p=0.07) segregated within Group 1. A local validation cohort (n=66, mean age at onset: 11.1 +/- 4.0) was analyzed by plasma induced transcription. A Random Forest model, which utilized the TrialNet data as a training set, was used to assign the local subjects to Group 1 or 2 (ROC: AUC=0.95; p=7.7E-9). Consistent with pathway analysis of the TrialNet transcriptomic data, ELISA analysis identified elevated plasma IFNγ, TNFα, IL-1β, IL-6, IL-12p70 levels (p&amp;lt;0.05) in Group 1. Flow cytometry measured higher percentages CD4+ CD45RO+ memory T cells (among total CD4 T cells, p=0.01) in Group 1. These analyses support the existence of two major T1D endotypes that exhibit differing immune phenotypes and therapeutic responsiveness. Disclosure A.Bedrat: None. T.Pant: None. S.Jia: None. M.Roethle: None. N.A.Truchan: None. S.M.Cabrera: Research Support; Abbott Diabetes. Y.Chen: None. C.Lin: None. M.Hessner: None. Funding National Institutes of Health (R01DK121528)

BIOL-12. ENHANCING IMMUNE-MEDIATED TUMOR CONTROL IN MURINE MODELS OF GROUP 3 MEDULLOBLASTOMA

Abstract Stagnation in clinical outcomes as well as severe side-effects of current treatment approaches for Group 3 medulloblastoma have prompted the search for novel therapies. To date, immunotherapies have not demonstrated success in medulloblastoma. Due to the highly aggressive nature of relapsed medulloblastoma, we suggest that it would be advantageous to introduce new immunotherapies first-line, alongside conventional therapies. To design immunotherapeutic strategies that are compatible in combination with standard therapies, we first determined the effect of standard-of-care craniospinal irradiation (CSI) and chemotherapy on the immunology of these tumours in immune competent and deficient murine models. We show that standard treatment approaches deplete adaptive immune cells, while macrophage and brain-resident microglia are not depleted. Furthermore, medulloblastomas are myeloid dominant tumors, and blocking the anti-phagocytic CD47-SIRPa interaction has shown preclinical efficacy. Therefore, we tested the combination of anti-CD47 immunotherapy with radiotherapy in a patient-derived xenograft model of Group 3 medulloblastoma. We found that anti-CD47 monotherapy had limited efficacy, while CSI results in only a temporary reduction in tumor size. In the cerebellum of relapsed tumors post-CSI, flow cytometry revealed increased peripheral myeloid populations. Remarkably, the combination of anti-CD47 and CSI was effective in clearing tumors and maintaining long-term tumor-free survival. Using RNA sequencing and immunohistochemistry, we found that the combination therapy results in enhanced cytokine signalling and phagocytic myeloid cell infiltration into the tumors compared to the monotherapies. We have also developed and characterized novel age-appropriate preclinical models for medulloblastoma and have observed altered myeloid cell phenotypes in these age-appropriate models. We are now testing this novel combination therapy in our new neonatal murine models to determine efficacy in age-appropriate hosts.

Circulating and Intratumoral Immune Determinants of Response to Atezolizumab plus Bevacizumab in Patients with Variant Histology or Sarcomatoid Renal Cell Carcinoma.

Renal cell carcinoma (RCC) of variant histology comprises approximately 20% of kidney cancer diagnoses, yet the optimal therapy for these patients and the factors that impact immunotherapy response remain largely unknown. To better understand the determinants of immunotherapy response in this population, we characterized blood- and tissue-based immune markers for patients with variant histology RCC, or any RCC histology with sarcomatoid differentiation, enrolled in a phase II clinical trial of atezolizumab and bevacizumab. Baseline circulating (plasma) inflammatory cytokines were highly correlated with one another, forming an "inflammatory module" that was increased in International Metastatic RCC Database Consortium poor-risk patients and was associated with worse progression-free survival (PFS; P = 0.028). At baseline, an elevated circulating vascular endothelial growth factor A (VEGF-A) level was associated with a lack of response (P = 0.03) and worse PFS (P = 0.021). However, a larger increase in on-treatment levels of circulating VEGF-A was associated with clinical benefit (P = 0.01) and improved overall survival (P = 0.0058). Among peripheral immune cell populations, an on-treatment decrease in circulating PD-L1+ T cells was associated with improved outcomes, with a reduction in CD4+PD-L1+ [HR, 0.62; 95% confidence interval (CI), 0.49-0.91; P = 0.016] and CD8+PD-L1+ T cells (HR, 0.59; 95% CI, 0.39-0.87; P = 0.009) correlated with improved PFS. Within the tumor itself, a higher percentage of terminally exhausted (PD-1+ and either TIM-3+ or LAG-3+) CD8+ T cells was associated with worse PFS (P = 0.028). Overall, these findings support the value of tumor and blood-based immune assessments in determining therapeutic benefit for patients with RCC receiving atezolizumab plus bevacizumab and provide a foundation for future biomarker studies for patients with variant histology RCC receiving immunotherapy-based combinations.