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Apr 2020 • The Journal of Clinical Investigation 130 (4), 1595-1607, 2020

Personal tumor antigens in blood malignancies: genomics-directed identification and targeting

Livius Penter, Catherine J Wu

Hematological malignancies have long been at the forefront of the development of novel immune-based treatment strategies. The earliest successful efforts originated from the extensive body of work in the field of allogeneic hematopoietic stem cell transplantation. These efforts laid the foundation for the recent exciting era of cancer immunotherapy, which includes immune checkpoint blockade, personal neoantigen vaccines, and adoptive T cell transfer. At the heart of the specificity of these novel strategies is the recognition of target antigens presented by malignant cells to T cells. Here, we review the advances in systematic identification of minor histocompatibility antigens and neoantigens arising from personal somatic alterations or recurrent driver mutations. These exciting efforts pave the path for the implementation of personalized combinatorial cancer therapy.

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Feb 2020 • Adaptive Optics and Wavefront Control for Biological Systems VI 11248, 57-63, 2020

Creating correct aberrations: why blur isn’t always bad in the eye

Gordon D Love, Martin S Banks, Steven A Cholewiak, Abigail P Finch

In optics in general, a sharp aberration-free image is normally the desired goal, and the whole field of adaptive optics has developed with the aim of producing blur-free images. Likewise, in ophthalmic optics we normally aim for a sharp image on the retina. But even with an emmetropic, or well-corrected eye, chromatic and high order aberrations affect the image. We describe two different areas where it is important to take these effects into account and why creating blur correctly via rendering can be advantageous. Firstly we show how rendering chromatic aberration correctly can drive accommodation in the eye and secondly report on matching defocus-l generated using rendering with conventional optical defocus.

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Jan 2020 • Clinical Cancer Research

Cross-site concordance evaluation of tumor DNA and RNA sequencing platforms for the CIMAC-CIDC network

Zexian Zeng, Jingxin Fu, Carrie Cibulskis, Aashna Jhaveri, Curtis Gumbs, Biswajit Das, Beatriz Sanchez-Espiridion, Sylvie Janssens, Len Taing, Jin Wang, James Lindsay, Tomas Vilimas, Jianhua Zhang, Collin Tokheim, Avinash Das Sahu, Peng Jiang, Chunhua Yan, Dzifa Y Duose, Ethan Cerami, Li Chen, David Cohen, Qing-Rong Chen, Rebecca A Enos, Xin Huang, J Jack Lee, Yang Liu, Donna S Neuberg, Cu Nguyen, Candace Patterson, Sachet A Shukla, Ming Tang, Junko Tsuji, Mohamed Uduman, Xiaoman Wang, Jason L Weirather, Jijun Yu, Joyce Yu, Jianjun Zhang, Jiexin Zhang, Daoud Meerzaman, Magdalena Thurin, P Andrew Futreal, Chris Karlovich, Stacey B Gabriel, Ignacio I Wistuba, Xiaole Shirley Liu, Catherine Wu

Purpose: Whole-exome (WES) and RNA sequencing (RNA-seq) are key components of cancer immunogenomic analyses. To evaluate the consistency of tumor WES and RNA-seq profiling platforms across different centers, the Cancer Immune Monitoring and Analysis Centers (CIMAC) and the Cancer Immunologic Data Commons (CIDC) conducted a systematic harmonization study.Experimental Design: DNA and RNA were centrally extracted from fresh frozen and formalin-fixed paraffin-embedded non–small cell lung carcinoma tumors and distributed to three centers for WES and RNA-seq profiling. In addition, two 10-plex HapMap cell line pools with known mutations were used to evaluate the accuracy of the WES platforms.Results: The WES platforms achieved high precision (> 0.98) and recall (> 0.87) on the HapMap pools when evaluated on loci using > 50× common coverage. Nonsynonymous mutations …

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Jan 2020 • Cancer Research

Enhanced Efficacy of Simultaneous PD-1 and PD-L1 Immune Checkpoint Blockade in High-Grade Serous Ovarian Cancer

Changxin Wan, Matthew P Keany, Han Dong, Linah F Al-Alem, Unnati M Pandya, Suzan Lazo, Karsten Boehnke, Katherine N Lynch, Rui Xu, Dominique T Zarrella, Shengqing Gu, Paloma Cejas, Klothilda Lim, Henry W Long, Kevin M Elias, Neil S Horowitz, Colleen M Feltmate, Michael G Muto, Michael J Worley, Ross S Berkowitz, Ursula A Matulonis, Marisa R Nucci, Christopher P Crum, Bo R Rueda, Myles Brown, Xiaole Shirley Liu, Sarah J Hill

Immune therapies have had limited efficacy in high-grade serous ovarian cancer (HGSC), as the cellular targets and mechanism(s) of action of these agents in HGSC are unknown. Here we performed immune functional and single-cell RNA sequencing transcriptional profiling on novel HGSC organoid/immune cell co-cultures treated with a unique bispecific anti-programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) antibody compared with monospecific anti-PD-1 or anti-PD-L1 controls. Comparing the functions of these agents across all immune cell types in real time identified key immune checkpoint blockade (ICB) targets that have eluded currently available monospecific therapies. The bispecific antibody induced superior cellular state changes in both T and natural killer (NK) cells. It uniquely induced NK cells to transition from inert to more active and cytotoxic phenotypes, implicating NK …

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Jan 2020

In vivo CRISPR Screens Identify E3 Ligase Cop1 as a Modulator of Macrophage Infiltration and Cancer Immunotherapy Target

Xiaoqing Wang, Collin Tokheim, Binbin Wang, Shengqing Stan Gu, Qin Tang, Yihao Li, Nicole Traugh, Yi Zhang, Ziyi Li, Boning Zhang, Jingxin Fu, Tengfei Xiao, Wei Li, Clifford A Meyer, Peng Jiang, Jun Chu, Paloma Cejas, Klothilda Lim, Henry Long, Myles Brown, Xiaole Shirley Liu

Despite remarkable clinical efficacies of immune checkpoint blockade (ICB) in cancer treatment, ICB benefits in triple-negative breast cancer (TNBC) remain limited. Through pooled in vivo CRISPR knockout (KO) screens in syngeneic TNBC mouse models, we found that inhibition of the E3 ubiquitin ligase Cop1 in cancer cells decreases the secretion of macrophage-associated chemokines, reduces tumor macrophage infiltration, and shows synergy in anti-tumor immunity with ICB. Transcriptomics, epigenomics, and proteomics analyses revealed Cop1 functions through proteasomal degradation of the C/ebpδ protein. Cop1 substrate Trib2 functions as a scaffold linking Cop1 and C/ebpδ, which leads to polyubiquitination of C/ebpδ. Cop1 inhibition stabilizes C/ebpδ to suppress the expression of macrophage chemoattractant genes. Our integrated approach implicates Cop1 as a target for improving cancer immunotherapy efficacy by regulating chemokine secretion and macrophage levels in the TNBC tumor microenvironment.

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Jan 2020 • Cancer Research

Enhanced Efficacy of Simultaneous PD-1 and PD-L1 Immune Checkpoint Blockade in High-Grade Serous Ovarian Cancer

Changxin Wan, Matthew P Keany, Han Dong, Linah F Al-Alem, Unnati M Pandya, Suzan Lazo, Karsten Boehnke, Katherine N Lynch, Rui Xu, Dominique T Zarrella, Shengqing Gu, Paloma Cejas, Klothilda Lim, Henry W Long, Kevin M Elias, Neil S Horowitz, Colleen M Feltmate, Michael G Muto, Michael J Worley, Ross S Berkowitz, Ursula A Matulonis, Marisa R Nucci, Christopher P Crum, Bo R Rueda, Myles Brown, Xiaole Shirley Liu, Sarah J Hill

Immune therapies have had limited efficacy in high-grade serous ovarian cancer (HGSC), as the cellular targets and mechanism(s) of action of these agents in HGSC are unknown. Here we performed immune functional and single-cell RNA sequencing transcriptional profiling on novel HGSC organoid/immune cell co-cultures treated with a unique bispecific anti-programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) antibody compared with monospecific anti-PD-1 or anti-PD-L1 controls. Comparing the functions of these agents across all immune cell types in real time identified key immune checkpoint blockade (ICB) targets that have eluded currently available monospecific therapies. The bispecific antibody induced superior cellular state changes in both T and natural killer (NK) cells. It uniquely induced NK cells to transition from inert to more active and cytotoxic phenotypes …

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Jan 2020 • Clinical Cancer Research

Inhibition of MAN2A1 enhances tumor response to anti-PD-L1

Sailing Shi, Shengqing Gu, Tong Han, Wubing Zhang, Lei Huang, Ziyi Li, Deng Pan, Jingxin Fu, Jun Ge, Myles Brown, Peng Zhang, Peng Jiang, Kai W Wucherpfennig, Xiaole Shirley Liu


Jan 2020 • Clinical Cancer Research

Cross-site concordance evaluation of tumor DNA and RNA sequencing platforms for the CIMAC-CIDC network

Zexian Zeng, Jingxin Fu, Carrie Cibulskis, Aashna Jhaveri, Curtis Gumbs, Biswajit Das, Beatriz Sanchez-Espiridion, Sylvie Janssens, Len Taing, Jin Wang, James Lindsay, Tomas Vilimas, Jianhua Zhang, Collin Tokheim, Avinash Das Sahu, Peng Jiang, Chunhua Yan, Dzifa Y Duose, Ethan Cerami, Li Chen, David Cohen, Qing-Rong Chen, Rebecca A Enos, Xin Huang, J Jack Lee, Yang Liu, Donna S Neuberg, Cu Nguyen, Candace Patterson, Sachet A Shukla, Ming Tang, Junko Tsuji, Mohamed Uduman, Xiaoman Wang, Jason L Weirather, Jijun Yu, Joyce Yu, Jianjun Zhang, Jiexin Zhang, Daoud Meerzaman, Magdalena Thurin, P Andrew Futreal, Chris Karlovich, Stacey B Gabriel, Ignacio I Wistuba, Xiaole Shirley Liu, Catherine Wu

Purpose: Whole-exome (WES) and RNA-sequencing (RNA-seq) are key components of cancer immunogenomic analyses. To evaluate the consistency of tumor WES and RNA-seq profiling platforms across different centers, the Cancer Immune Monitoring and Analysis Centers (CIMACs) and the Cancer Immunologic Data Commons (CIDC) conducted a systematic harmonization study. Experimental Design: DNA and RNA were centrally extracted from fresh frozen (FF) and formalin-fixed paraffin-embedded (FFPE) non-small cell lung carcinoma (NSCLC) tumors and distributed to three centers for WES and RNA-seq profiling. In addition, two 10-plex HapMap cell-line pools with known mutations were used to evaluate the accuracy of the WES platforms. Results: The WES platforms achieved high precision (> 0.98) and recall (> 0.87) on the HapMap pools when evaluated on loci using > 50X common coverage. Non …

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Jan 2020 • Clinical Cancer Research

Cross-site concordance evaluation of tumor DNA and RNA sequencing platforms for the CIMAC-CIDC network

Zexian Zeng, Jingxin Fu, Carrie Cibulskis, Aashna Jhaveri, Curtis Gumbs, Biswajit Das, Beatriz Sanchez-Espiridion, Sylvie Janssens, Len Taing, Jin Wang, James Lindsay, Tomas Vilimas, Jianhua Zhang, Collin Tokheim, Avinash Das Sahu, Peng Jiang, Chunhua Yan, Dzifa Y Duose, Ethan Cerami, Li Chen, David Cohen, Qing-Rong Chen, Rebecca A Enos, Xin Huang, J Jack Lee, Yang Liu, Donna S Neuberg, Cu Nguyen, Candace Patterson, Sachet A Shukla, Ming Tang, Junko Tsuji, Mohamed Uduman, Xiaoman Wang, Jason L Weirather, Jijun Yu, Joyce Yu, Jianjun Zhang, Jiexin Zhang, Daoud Meerzaman, Magdalena Thurin, P Andrew Futreal, Chris Karlovich, Stacey B Gabriel, Ignacio I Wistuba, Xiaole Shirley Liu, Catherine Wu

Purpose: Whole-exome (WES) and RNA sequencing (RNA-seq) are key components of cancer immunogenomic analyses. To evaluate the consistency of tumor WES and RNA-seq profiling platforms across different centers, the Cancer Immune Monitoring and Analysis Centers (CIMAC) and the Cancer Immunologic Data Commons (CIDC) conducted a systematic harmonization study. Experimental Design: DNA and RNA were centrally extracted from fresh frozen and formalin-fixed paraffin-embedded non–small cell lung carcinoma tumors and distributed to three centers for WES and RNA-seq profiling. In addition, two 10-plex HapMap cell line pools with known mutations were used to evaluate the accuracy of the WES platforms. Results: The WES platforms achieved high precision (> 0.98) and recall (> 0.87) on the HapMap pools when evaluated on loci …

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Jan 2020 • Clinical Cancer Research

Inhibition of MAN2A1 enhances tumor response to anti-PD-L1

Sailing Shi, Shengqing Gu, Tong Han, Wubing Zhang, Lei Huang, Ziyi Li, Deng Pan, Jingxin Fu, Jun Ge, Myles Brown, Peng Zhang, Peng Jiang, Kai W Wucherpfennig, Xiaole Shirley Liu

Purpose: Immune checkpoint blockade (ICB) has shown remarkable efficacy, but in only a minority of cancer patients, suggesting the need to develop additional treatment strategies. Aberrant glycosylation in tumors, resulting from the dysregulated expression of key enzymes in glycan biosynthesis, modulates the immune response. However, the role of glycan biosynthesis enzymes in anti-tumor immunity is poorly understood. We aimed to study the immunomodulatory effects of these enzymes. Experimental Design: We integrated transcriptional profiles of treatment-naïve human tumors and functional CRISPR screens to identify glycometabolism genes with immunomodulatory effects. We further validated our findings using in vitro co-culture and in vivo syngeneic tumor growth assays. Results: We identified MAN2A1, encoding an enzyme in N-glycan maturation, as a key immunomodulatory gene. Analyses of public …

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2020 • Frontiers in genetics

ABC-GWAS: Functional Annotation of Estrogen Receptor-Positive Breast Cancer Genetic Variants

Mohith Manjunath, Yi Zhang, Shilu Zhang, Sushmita Roy, Pablo Perez-Pinera, Jun S Song

Over the past decade, hundreds of genome-wide association studies (GWAS) have implicated genetic variants in various diseases, including cancer. However, only few of these variants have been functionally characterized to date, mainly owing to the fact that the majority of the variants reside in noncoding regions of the human genome with unknown function. A comprehensive functional annotation of the candidate variants is thus necessary to fill the gap between the correlative findings of GWAS and the development of therapeutic strategies. By integrating large-scale multi-omics datasets such as the Cancer Genome Atlas (TCGA) and the Encyclopedia of DNA Elements (ENCODE), we performed multivariate linear regression analysis of expression quantitative trait loci, sequence permutation test of transcription factor binding perturbation, and modeling of three-dimensional chromatin interactions to analyze the potential molecular functions of 2,813 single nucleotide variants in 93 genomic loci associated with estrogen receptor-positive breast cancer. To facilitate rapid progress in functional genomics of breast cancer, we have created “Analysis of Breast Cancer GWAS” (ABC-GWAS), an interactive database of functional annotation of estrogen receptor-positive breast cancer GWAS variants. Our resource includes expression quantitative trait loci, long-range chromatin interaction predictions, and transcription factor binding motif analyses to prioritize putative target genes, causal variants and transcription factors. An embedded genome browser also facilitates convenient visualization of the GWAS loci in genomic and epigenomic context. ABC …

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2020 • Frontiers in Genetics

ABC-GWAS: Functional Annotation of Estrogen Receptor-Positive Breast Cancer Genetic Variants

Mohith Manjunath, Yi Zhang, Shilu Zhang, Sushmita Roy, Pablo Perez-Pinera, Jun S Song

Over the past decade, hundreds of genome-wide association studies (GWAS) have implicated genetic variants in various diseases, including cancer. However, only few of these variants have been functionally characterized to date, mainly owing to the fact that the majority of the variants reside in noncoding regions of the human genome with unknown function. A comprehensive functional annotation of the candidate variants is thus necessary to fill the gap between the correlative findings of GWAS and the development of therapeutic strategies. By integrating large-scale multi-omics datasets such as the Cancer Genome Atlas (TCGA) and the Encyclopedia of DNA Elements (ENCODE), we performed multivariate linear regression analysis of expression quantitative trait loci, sequence permutation test of transcription factor binding perturbation, and modeling of three-dimensional chromatin interactions to analyze the potential molecular functions of 2,813 single nucleotide variants in 93 genomic loci associated with estrogen receptor-positive breast cancer. To facilitate rapid progress in functional genomics of breast cancer, we have created “Analysis of Breast Cancer GWAS” (ABC-GWAS), an interactive database of functional annotation of estrogen receptor-positive breast cancer GWAS variants. Our resource includes expression quantitative trait loci, long-range chromatin interaction predictions, and transcription factor binding motif analyses to prioritize putative target genes, causal variants and transcription factors. An embedded genome browser also facilitates convenient visualization of the GWAS loci in genomic and epigenomic context. ABC …

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2020 • Genomics Proteomics Bioinformatics

CRISPR screens identify essential cell growth mediators in BRAF inhibitor-resistant melanoma

Liu XS Li Z, Wang B, Gu S, Peng J, Sahu A, Chen C, Han T, Shi S, Wang X, Traugh N, Liu H, Liu Y, Wu Q, Brown M, Xiao T, Boland G

BRAF is a serine/threonine kinase that harbors activating mutations in ∼7% of human malignancies and ∼60% of melanomas. Despite initial clinical responses to BRAF inhibitors, patients frequently develop drug resistance. To identify candidate therapeutic targets for BRAF inhibitor resistant melanoma, we conduct CRISPR screens in melanoma cells harboring an activating BRAF mutation that had also acquired resistance to BRAF inhibitors. To investigate the mechanisms and pathways enabling resistance to BRAF inhibitors in melanomas, we integrate expression, ATAC-seq, and CRISPR screen data. We identify the JUN family transcription factors and the ETS family transcription factor ETV5 as key regulators of CDK6, which together enable resistance to BRAF inhibitors in melanoma cells. Our findings reveal genes contributing to resistance to a selective BRAF inhibitor PLX4720, providing new insights into …

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2020 • Nat Commun

Determinants of transcription factor regulatory range

Liu XS*# Chen C#, Zheng R, Tokheim C#, Dong X, Fan J, Wan C, Tang Q, Brown M, Liu JS, Meyer C


2020 • Genome Medicine

Large-scale public data reuse to model immunotherapy response and resistance

Liu XS* 215. Fu J, Li K, Zhang W, Wan C, Zhang J*, Jiang P*


2020 • Cell Res

ERCC6L2 promotes DNA orientation-specific recombination in mammalian cells

Meng F Liu X, Liu T, Shang Y, Dai P, Zhang W, Lee BJ, Huang M, Yang D, Wu Q, Liu DL, Zheng X, Zhou BO, Dong J, Yeap L, Hu J, Xiao T, Zha S, Casellas R, Liu XS

Programmed DNA recombination in mammalian cells occurs predominantly in a directional manner. While random DNA breaks are typically repaired both by deletion and by inversion at approximately equal proportions, V (D) J and class switch recombination (CSR) of immunoglobulin heavy chain gene overwhelmingly delete intervening sequences to yield productive rearrangement. What factors channel chromatin breaks to deletional CSR in lymphocytes is unknown. Integrating CRISPR knockout and chemical perturbation screening we here identify the Snf2-family helicase-like ERCC6L2 as one such factor. We show that ERCC6L2 promotes double-strand break end-joining and facilitates optimal CSR in mice. At the cellular levels, ERCC6L2 rapidly engages in DNA repair through its C-terminal domains. Mechanistically, ERCC6L2 interacts with other end-joining factors and plays a functionally redundant role …

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2020 • iScience

Loss of H3K36 Methyltransferase SETD2 Impairs V(D)J Recombination during Lymphoid Development

Armstrong SA 214. Chu SH, Chabon JR, Matovina CN, Chen BR, Zhang J, Kumar V, Xiong Y, Callen E, Hung PJ, Feng Z, Koche RP, Liu XS, Chaudhuri J, Nussenzweig A, Sleckman BP

Repair of DNA double-stranded breaks (DSBs) during lymphocyte development is essential for V(D)J recombination and forms the basis of immunoglobulin variable region diversity. Understanding of this process in lymphogenesis has historically been centered on the study of RAG1/2 recombinases and a set of classical non-homologous end-joining factors. Much less has been reported regarding the role of chromatin modifications on this process. Here, we show a role for the non-redundant histone H3 lysine methyltransferase, Setd2, and its modification of lysine-36 trimethylation (H3K36me3), in the processing and joining of DNA ends during V(D)J recombination. Loss leads to mis-repair of Rag-induced DNA DSBs, especially when combined with loss of Atm kinase activity. Furthermore, loss reduces immune repertoire and a severe block in lymphogenesis as well as causes post-mitotic neuronal apoptosis …

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2020 • Genome Biology

Integrative analyses of single-cell transcriptome and regulome using MAESTRO

Liu XS Wang C, Sun D, Huang X, Wan C, Li Z, Han Y, Qin Q, Fan J, Qiu X, Xie Y, Meyer C, Brown M, Tang M, Long H, Liu T


2020 • Bioinformatics for Cancer Immunotherapy, 249-262, 2020

Computational deconvolution of tumor-infiltrating immune components with bulk tumor gene expression data

Bo Li, Taiwen Li, Jun S Liu, X Shirley Liu

Tumor-infiltrating immune cells play critical roles in immune-mediated tumor rejection and/or progression, and are key targets of immunotherapies. Estimation of different immune subsets becomes increasingly important with the decreased cost of high-throughput molecular profiling and the rapidly growing amount of cancer genomics data. Here, we present Tumor IMmune Estimation Resource (TIMER), an in silico deconvolution method for inference of tumor-infiltrating immune components. TIMER takes bulk tissue gene expression profiles measured with RNA-seq or microarray to evaluate the abundance of six immune cell types in the tumor microenvironment: B cell, CD4+ T cell, CD8+ T cell, neutrophil, macrophage, and dendritic cell. We further introduce its associated webserver for convenient, user-friendly analysis of tumor immune infiltrates across multiple cancer types.

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2020 • Mol Cell

Synthetic lethal and resistance interaction with BET Bromodomain inhibitors in triple-negative breast cancer

BET bromodomain inhibitors (BBDIs) are candidate therapeutic agents for triple-negative breast cancer (TNBC) and other cancer types, but inherent and acquired resistance to BBDIs limits their potential clinical use. Using CRISPR and small-molecule inhibitor screens combined with comprehensive molecular profiling of BBDI response and resistance, we identified synthetic lethal interactions with BBDIs and genes that, when deleted, confer resistance. We observed synergy with regulators of cell cycle progression, YAP, AXL, and SRC signaling, and chemotherapeutic agents. We also uncovered functional similarities and differences among BRD2, BRD4, and BRD7. Although deletion of BRD2 enhances sensitivity to BBDIs, BRD7 loss leads to gain of TEAD-YAP chromatin binding and luminal features associated with BBDI resistance. Single-cell RNA-seq, ATAC-seq, and cellular barcoding analysis of BBDI …

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Oct 2019 • European journal of immunology

Guidelines for the use of flow cytometry and cell sorting in immunological studies

Andrea Cossarizza, Hyun‐Dong Chang, Andreas Radbruch, Andreas Acs, Dieter Adam, Sabine Adam‐Klages, William W Agace, Nima Aghaeepour, Mübeccel Akdis, Matthieu Allez, Larissa Nogueira Almeida, Giorgia Alvisi, Graham Anderson, Immanuel Andrä, Francesco Annunziato, Achille Anselmo, Petra Bacher, Cosima T Baldari, Sudipto Bari, Vincenzo Barnaba, Joana Barros‐Martins, Luca Battistini, Wolfgang Bauer, Sabine Baumgart, Nicole Baumgarth, Dirk Baumjohann, Bianka Baying, Mary Bebawy, Burkhard Becher, Wolfgang Beisker, Vladimir Benes, Rudi Beyaert, Alfonso Blanco, Dominic A Boardman, Christian Bogdan, Jessica G Borger, Giovanna Borsellino, Philip E Boulais, Jolene A Bradford, Dirk Brenner, Ryan R Brinkman, Anna ES Brooks, Dirk H Busch, Martin Büscher, Timothy P Bushnell, Federica Calzetti, Garth Cameron, Ilenia Cammarata, Xuetao Cao, Susanna L Cardell, Stefano Casola, Marco A Cassatella, Andrea Cavani, Antonio Celada, Lucienne Chatenoud, Pratip K Chattopadhyay, Sue Chow, Eleni Christakou, Luka Čičin‐Šain, Mario Clerici, Federico S Colombo, Laura Cook, Anne Cooke, Andrea M Cooper, Alexandra J Corbett, Antonio Cosma, Lorenzo Cosmi, Pierre G Coulie, Ana Cumano, Ljiljana Cvetkovic, Van Duc Dang, Chantip Dang‐Heine, Martin S Davey, Derek Davies, Sara De Biasi, Genny Del Zotto, Gelo Victoriano Dela Cruz, Michael Delacher, Silvia Della Bella, Paolo Dellabona, Günnur Deniz, Mark Dessing, James P Di Santo, Andreas Diefenbach, Francesco Dieli, Andreas Dolf, Thomas Dörner, Regine J Dress, Diana Dudziak, Michael Dustin, Charles‐Antoine Dutertre, Friederike Ebner, Sidonia BG Eckle, Matthias Edinger, Pascale Eede, Götz RA Ehrhardt, Marcus Eich, Pablo Engel, Britta Engelhardt, Anna Erdei, Charlotte Esser, Bart Everts, Maximilien Evrard, Christine S Falk, Todd A Fehniger, Mar Felipo‐Benavent, Helen Ferry, Markus Feuerer, Andrew Filby, Kata Filkor, Simon Fillatreau, Marie Follo, Irmgard Förster, John Foster, Gemma A Foulds, Britta Frehse, Paul S Frenette, Stefan Frischbutter, Wolfgang Fritzsche, David W Galbraith, Anastasia Gangaev, Natalio Garbi, Brice Gaudilliere, Ricardo T Gazzinelli, Jens Geginat, Wilhelm Gerner, Nicholas A Gherardin, Kamran Ghoreschi, Lara Gibellini, Florent Ginhoux, Keisuke Goda, Dale I Godfrey, Christoph Goettlinger, Jose M González‐Navajas, Carl S Goodyear, Andrea Gori, Jane L Grogan, Daryl Grummitt, Andreas Grützkau, Claudia Haftmann, Jonas Hahn, Hamida Hammad, Günter Hämmerling, Leo Hansmann, Goran Hansson, Christopher M Harpur, Susanne Hartmann, Andrea Hauser, Anja E Hauser, David L Haviland

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer‐reviewed by leading experts in the field, making this an essential research companion.

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