Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function

Upon initial immune challenge, dendritic cells (DCs) migrate to lymph nodes and interact with fibroblastic reticular cells (FRCs) via C-type lectin-like receptor 2 (CLEC-2). CLEC-2 binds to the membrane glycoprotein podoplanin (PDPN) on FRCs, inhibiting actomyosin contractility through the FRC network and permitting lymph node expansion. The hyaluronic acid receptor CD44 is known to be required for FRCs to respond to DCs but the mechanism of action is not fully elucidated. Here, we use DNA-PAINT, a quantitative single molecule super-resolution technique, to visualize and quantify how PDPN clustering is regulated in the plasma membrane of FRCs. Our results indicate that CLEC-2 interaction leads to the formation of large PDPN clusters (i.e. more than 12 proteins per cluster) in a CD44-dependent manner. These results suggest that CD44 expression is required to stabilize large pools of PDPN at the membrane of FRCs upon CLEC-2 interaction, revealing the molecular mechanism through which CD44 facilitates cellular crosstalk between FRCs and DCs

Immune function and dysfunction are determined by lymphoid tissue efficacy

Lymphoid tissue returns to a steady state once each immune response is resolved, and although this occurs multiple times throughout life, its structural integrity and functionality remain unaffected. Stromal cells orchestrate cellular interactions within lymphoid tissue, and any changes to the microenvironment can have detrimental outcomes and drive disease. A breakdown in lymphoid tissue homeostasis can lead to a loss of tissue structure and function that can cause aberrant immune responses. This Review highlights recent advances in our understanding of lymphoid tissue function and remodelling in adaptive immunity and in disease states. We discuss the functional role of lymphoid tissue in disease progression and explore the changes to lymphoid tissue structure and function driven by infection, chronic inflammatory conditions and cancer. Understanding the role of lymphoid tissues in immune responses to a wide range of pathologies allows us to take a fuller systemic view of disease progression

Ways to increase equity, diversity and inclusion

The eLife Early-Career Advisory Group (ECAG), an international group of early-career researchers committed to improving research culture, calls for radical changes at eLife and other journals to address racism in the scientific community and to make science more diverse and inclusive.Fil: Mehta, Devang. University of Alberta; CanadáFil: Bediako, Yaw. University Of Ghana; GhanaFil: De Winde, Charlotte M.. Colegio Universitario de Londres; Reino UnidoFil: Ebrahimi, Hedyeh. No especifíca;Fil: Fernández, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Ilangovan, Vinodh. University Aarhus; DinamarcaFil: Paz Quezada, Carolina. Universidad Bernardo O'higgins; ChileFil: Riley, Julia L.. Dalhousie University Halifax; CanadáFil: Saladi, Shyam M.. California Institute of Technology; Estados UnidosFil: Tay, Andy. No especifíca;Fil: Weissgerber, Tracey. No especifíca

Mitigating the impact of conference and travel cancellations on researchers’ futures

The need to protect public health during the current COVID-19 pandemic has necessitated conference cancellations on an unprecedented scale. As the scientific community adapts to new working conditions, it is important to recognize that some of our actions may disproportionately affect early-career researchers and scientists from countries with limited research funding. We encourage all conference organizers, funders and institutions who are able to do so to consider how they can mitigate the unintended consequences of conference and travel cancellations and we provide seven recommendations for how this could be achieved. The proposed solutions may also offer long-term benefits for those who normally cannot attend conferences, and thus lead to a more equitable future for generations of researchers

Towards inclusive funding practices for early career researchers

Securing research funding is a challenge faced by most scientists in academic institutions worldwide. Funding success rates for all career stages are low, but the burden falls most heavily on early career researchers (ECRs). These are young investigators in training and new principal investigators who have a shorter track record. ECRs are dependent on funding to establish their academic careers. The low number of career development awards and the lack of sustained research funding result in the loss of ECR talent in academia. Several steps in the current funding process, from grant conditions to review, play significant roles in the distribution of funds. Furthermore, there is an imbalance where certain research disciplines and labs of influential researchers receive more funding. As a group of ECRs with global representation, we examined funding practices, barriers, and facilitators to the current funding systems. We also identified alternatives to the most common funding distribution practices, such as diversifying risk or awarding grants on a partly random basis. Here, we detail recommendations for funding agencies and grant reviewers to improve ECR funding prospects worldwide and promote a fairer and more inclusive funding landscape for ECRs.Instituto de VirologíaFil: de Winde, Charlotte M. University College London. MRC Laboratory for Molecular Cell Biology; Reino UnidoFil: de Winde, Charlotte M. Amsterdam University Medical Center. Department of Molecular Cell Biology & Immunology; Países BajosFil: Sarabipour, Sarvenaz. Johns Hopkins University. Department of Biomedical Engineering. Institute for Computational Medicine; Estados UnidosFil: Carignano, Hugo Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; ArgentinaFil: Davla, Sejal. City University of New York. Advanced Science Research Center; Estados UnidosFil: Eccles, David. Malaghan Institute of Medical Research; Nueva ZelandaFil: Hainer, Sarah J. University of Pittsburgh. Department of Biological Sciences; Estados UnidosFil: Haidar, Mansour. Hasselt University; BélgicaFil: Ilangovan, Vinodh. Aarhus University; DinamarcaFil: Jadavji, Nafisa M. Midwestern University. Department of Biomedical Sciences; Estados UnidosFil: Jadavji, Nafisa M. Carleton University. Department of Neuroscience; CanadáFil: Kritsiligkou, Paraskevi. German Cancer Research Center; AlemaniaFil: Lee, Tai-Ying. University of Oxford; Reino UnidoFil: Ólafsdóttir, H. Freyja. Radboud University. Donders Institute for Brain, Cognition and Behaviour; Países Bajo

Guidelines Toward Inclusive Practices in Academics by eLife Community Ambassadors

The academic world has become increasingly diverse over the past few decades. However, many of us have unconscious biases against certain groups of people, and common practices in evaluating merit have flaws that can perpetuate these biases. We, a group of eLife Community Ambassadors within the Intersectionality Initiative of the 2019-2020 program, have identified five current practices that are fundamental to academic science that may be hindering inclusivity in science. These practices include hiring, mentoring, writing reference letters, and reviewing grants and manuscripts. To try and level the playing field and increase inclusivity in science, we have compiled a guideline for each of these practices that are intended to help inform scientists, mentors, funders, and journals on ways to mitigate biases. The overall goal of these guidelines is to make science more inclusive and to limit bias in these core features of academic science

Quantitative single molecule analysis of podoplanin clustering in fibroblastic reticular cells uncovers CD44 function

Upon initial immune challenge, dendritic cells (DCs) migrate to lymph nodes and interact with fibroblastic reticular cells (FRCs) via C-type lectin-like receptor 2 (CLEC-2). CLEC-2 binds to the membrane glycoprotein podoplanin (PDPN) on FRCs, inhibiting actomyosin contractility through the FRC network and permitting lymph node expansion. The hyaluronic acid receptor CD44 is known to be required for FRCs to respond to DCs but the mechanism of action is not fully elucidated. Here, we use DNA-PAINT, a quantitative single molecule super-resolution technique, to visualize and quantify how PDPN clustering is regulated in the plasma membrane of FRCs. Our results indicate that CLEC-2 interaction leads to the formation of large PDPN clusters (i.e. more than 12 proteins per cluster) in a CD44-dependent manner. These results suggest that CD44 expression is required to stabilize large pools of PDPN at the membrane of FRCs upon CLEC-2 interaction, revealing the molecular mechanism through which CD44 facilitates cellular crosstalk between FRCs and DCs

Lab Manual Template by eLife Community Ambassadors

We, a group of eLife Community Ambassadors within the Intersectionality Initiative (2019-2020), have compiled this template for a Lab Expectations Document, or Lab Manual, to share with researchers worldwide. The purpose of this document is for lab heads (and/or lab members) to generate a clear guide for their current and new group members. Please feel free to download and modify this template for your own research group/lab