Review

Authors: Weijun Wei, Zachary T Rosenkrans, Jianjun Liu...

Immuno-positron emission tomography (immunoPET) is a paradigm-shifting molecular imaging modality combining the superior targeting specificity of monoclonal antibody (mAb) and the inherent sensitivity of PET technique. A variety of radionuclides and mAbs have been exploited to develop immunoPET probes, which has been driven by the development and optimization of radiochemistry and conjugation strategies. In addition, tumor-targeting vectors with a short circulation time (e.g., Nanobody) or with an enhanced binding affinity (e.g., bispecific antibody) are being used to design novel immunoPET probes. Accordingly, several immunoPET probes, such as Zr-Df-pertuzumab and Zr-atezolizumab, have been successfully translated for clinical use. By noninvasively and dynamically revealing the expression of heterogeneous tumor antigens, immunoPET imaging is gradually changing the theranostic landscape of several types of malignancies. ImmunoPET is the method of choice for imaging specific tumor markers, immune cells, immune checkpoints, and inflammatory processes. Furthermore, the integration of immunoPET imaging in antibody drug development is of substantial significance because it provides pivotal information regarding antibody targeting abilities and distribution profiles. Herein, we present the latest immunoPET imaging strategies and their preclinical and clinical applications. We also emphasize current conjugation strategies that can be leveraged to develop next-generation immunoPET probes. Lastly, we discuss practical considerations to tune the development and translation of immunoPET imaging strategies.

Topics: Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Cell Line, Tumor; Humans; Immunologic Techniques; Neoplasms; Positron-Emission Tomography; Radiopharmaceuticals; Single-Domain Antibodies

PubMed: 32202104
DOI: 10.1021/acs.chemrev.9b00738

Authors: Cansu Cimen Bozkus, Ana Belen Blazquez, Tomohiro Enokida...

Determining the antigen specificities of the endogenous T-cell repertoire is important for screening naturally occurring or therapy-induced T-cell immunity and may help identify novel targets for T-cell-based therapies. Here, we describe a rapid, sensitive, and high-throughput protocol for expanding antigen-specific T cells from human peripheral blood mononuclear cells following peptide stimulation and detecting antigen-specific effector cytokine formation by flow cytometry. Our approach can be applied to examining specific T-cell subsets from various tissues. For complete details on the use and execution of this protocol, please refer to Roudko et al. (2020) and Cimen Bozkus et al. (2019).

Topics: Cell Culture Techniques; Cryopreservation; Cytokines; Flow Cytometry; Humans; Immunologic Techniques; Leukocytes, Mononuclear; T-Lymphocyte Subsets

PubMed: 34458873
DOI: 10.1016/j.xpro.2021.100758

Authors:

Topics: Animals; Gene Expression Regulation; Gene Regulatory Networks; Genomics; History, 21st Century; Immune System; Immunologic Techniques; Mice

PubMed: 32577013
DOI: 10.1038/s41590-020-0687-4

Review

Authors: Yifei Hu, Jun Huang

Chimeric antigen receptor-T (CAR-T) cell therapy is a promising frontier of immunoengineering and cancer immunotherapy. Methods that detect, quantify, track, and visualize the CAR, have catalyzed the rapid advancement of CAR-T cell therapy from preclinical models to clinical adoption. For instance, CAR-staining/labeling agents have enabled flow cytometry analysis, imaging applications, cell sorting, and high-dimensional clinical profiling. Molecular assays, such as quantitative polymerase chain reaction, integration site analysis, and RNA-sequencing, have characterized CAR transduction, expression, and CAR-T cell expansion kinetics. visualization methods, including confocal and total internal reflection fluorescence microscopy, have captured the molecular details underlying CAR immunological synapse formation, signaling, and cytotoxicity. tracking methods, including two-photon microscopy, bioluminescence imaging, and positron emission tomography scanning, have monitored CAR-T cell biodistribution across blood, tissue, and tumor. Here, we review the plethora of CAR detection methods, which can operate at the genomic, transcriptomic, proteomic, and organismal levels. For each method, we discuss: (1) what it measures; (2) how it works; (3) its scientific and clinical importance; (4) relevant examples of its use; (5) specific protocols for CAR detection; and (6) its strengths and weaknesses. Finally, we consider current scientific and clinical needs in order to provide future perspectives for improved CAR detection.

Topics: Humans; Immunologic Techniques; Immunotherapy, Adoptive; Receptors, Chimeric Antigen

PubMed: 32849635
DOI: 10.3389/fimmu.2020.01770

Authors: J Unsworth

Topics: Allergy and Immunology; Animals; History, 20th Century; Humans; Immunologic Techniques; Immunotherapy; Transplantation Immunology

PubMed: 16049277
DOI: 10.1136/jcp.2005.030619

Review

Authors: Haide Chen, Fang Ye, Guoji Guo...

The immune system is composed of a complex hierarchy of cell types that protect the organism against disease and maintain homeostasis. Identifying heterogeneity of immune cells is the key to understanding the immune system. Advanced single-cell RNA sequencing (scRNA-seq) technologies are revolutionizing our ability to study immunology. By measuring transcriptomes at the single-cell level, scRNA-seq enables identification of cellular heterogeneity in far greater detail than conventional methods. In this review, we introduce the existing scRNA-seq technologies and present their strengths and weaknesses. We also discuss potential applications and future innovations of scRNA-seq in immunology.

Topics: Allergy and Immunology; Animals; Gene Expression Profiling; Humans; Immunologic Techniques; Sequence Analysis, RNA; Single-Cell Analysis; Transcriptome

PubMed: 30796351
DOI: 10.1038/s41423-019-0214-4

Review

Authors: Haleigh B Eppler, Christopher M Jewell

The immune system has remarkable capabilities to combat disease with exquisite selectivity. This feature has enabled vaccines that provide protection for decades and, more recently, advances in immunotherapies that can cure some cancers. Greater control over how immune signals are presented, delivered, and processed will help drive even more powerful options that are also safe. Such advances will be underpinned by new tools that probe how immune signals are integrated by immune cells and tissues. Biomaterials are valuable resources to support this goal, offering robust, tunable properties. The growing role of biomaterials as tools to dissect immune function in fundamental and translational contexts is highlighted. These technologies can serve as tools to understand the immune system across molecular, cellular, and tissue length scales. A common theme is exploiting biomaterial features to rationally direct how specific immune cells or organs encounter a signal. This precision strategy, enabled by distinct material properties, allows isolation of immunological parameters or processes in a way that is challenging with conventional approaches. The utility of these capabilities is demonstrated through examples in vaccines for infectious disease and cancer immunotherapy, as well as settings of immune regulation that include autoimmunity and transplantation.

Topics: Animals; Antigen Presentation; Antigens; Biocompatible Materials; Humans; Immune System; Immunity; Immunologic Techniques; Immunotherapy

PubMed: 31782844
DOI: 10.1002/adma.201903367

Review

Authors: Pratip K Chattopadhyay, Todd M Gierahn, Mario Roederer...

The complex heterogeneity of cells, and their interconnectedness with each other, are major challenges to identifying clinically relevant measurements that reflect the state and capability of the immune system. Highly multiplexed, single-cell technologies may be critical for identifying correlates of disease or immunological interventions as well as for elucidating the underlying mechanisms of immunity. Here we review limitations of bulk measurements and explore advances in single-cell technologies that overcome these problems by expanding the depth and breadth of functional and phenotypic analysis in space and time. The geometric increases in complexity of data make formidable hurdles for exploring, analyzing and presenting results. We summarize recent approaches to making such computations tractable and discuss challenges for integrating heterogeneous data obtained using these single-cell technologies.

Topics: Animals; Computational Biology; Humans; Immune System; Immunologic Techniques; Monitoring, Immunologic; Single-Cell Analysis; Statistics as Topic

PubMed: 24448570
DOI: 10.1038/ni.2796

Review

Authors: Amir Giladi, Ido Amit

The immunology field has invested great efforts and ingenuity to characterize the various immune cell types and elucidate their functions. However, accumulating evidence indicates that current technologies and classification schemes are limited in their ability to account for the functional heterogeneity of immune processes. Single-cell genomics hold the potential to revolutionize the way we characterize complex immune cell assemblies and study their spatial organization, dynamics, clonal distribution, pathways, function, and crosstalks. In this Perspective, we consider recent and forthcoming technological and analytical advances in single-cell genomics and the potential impact of those advances on the future of immunology research and immunotherapy.

Topics: Animals; Genomics; Humans; Immunologic Techniques; Immunotherapy; Single-Cell Analysis

PubMed: 29328909
DOI: 10.1016/j.cell.2017.11.011

Review

Authors: Mark M Davis, Cristina M Tato, David Furman...

Systems-biology approaches in immunology take various forms, but here we review strategies for measuring a broad swath of immunological functions as a means of discovering previously unknown relationships and phenomena and as a powerful way of understanding the immune system as a whole. This approach has rejuvenated the field of vaccine development and has fostered hope that new ways will be found to combat infectious diseases that have proven refractory to classical approaches. Systems immunology also presents an important new strategy for understanding human immunity directly, taking advantage of the many ways the immune system of humans can be manipulated.

Topics: Allergy and Immunology; Humans; Immune System; Immunologic Techniques; Signal Transduction; Systems Biology; Vaccines

PubMed: 28632713
DOI: 10.1038/ni.3768