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Trends and Applications of Immunohistochemistry in Research and Clinical Practice

Scientist on August 14, 2024

This installment of Tech Snapshot® was written by Applied Pathology Systems, a CLIA certified laboratory located in Greater Boston that supports projects at different stages. They offer a full spectrum of histopathology services, including histology, immunohistochemistry, multiplex IF, imaging, image analysis, antibody screening, TMA construction and pathology evaluations covering tissues from different animal species to human. Their services are available on the Scientist.com marketplace.

Immunohistochemistry (IHC) has emerged as a powerful technique in biomedical research, revolutionizing our understanding of cellular and tissue biology. By combining the principles of immunology and histology, IHC allows researchers to visualize and analyze the distribution, localization and abundance of specific proteins within tissues. Its versatile application spans various domains including cancer biology, neuroscience, immunology and developmental biology. In this overview, we outline the dynamic trends and broad-ranging applications of IHC across clinical settings and its multiple purposes within biomedical research.

Trends in Immunohistochemistry

Multiplexing: One of the prominent trends in IHC is multiplexing, which involves the simultaneous detection of multiple antigens within a single tissue section. This allows researchers to analyze complex biological processes and interrelationships between different biomolecules within the same sample.

Automation: Automation has streamlined the IHC workflow, reducing manual errors and increasing throughput. Automated platforms for slide staining, image acquisition and analysis have improved the reproducibility and efficiency of IHC experiments, making them more accessible to researchers.

Digital Pathology: The integration of IHC with digital pathology platforms enables high-resolution imaging and quantitative analysis of stained tissue sections. Digital pathology facilitates data sharing, collaboration and the development of algorithms to analyze and compare tissue specimens.

Figure 1: Current trends in immunohistochemistry study. Left: multiplexed staining of CD4, CD8, FoxP3 and GrzyB in mouse spleen. Middle: PhenoCycler Fusion automation and Imaging systems in APS laboratory. Digital pathology system at APS allow us to analyze the spatial relationship between PD-L1 positive cells and Ki-67 positive cells

Application of IHC as a Versatile Tool in Clinical Practice and Research

Medical Diagnostics: IHC staining of tissue sections allows pathologists to confirm the presence or absence of specific diagnostic markers, aiding in the accurate diagnosis of diseases and guiding patient management decisions. Additionally, IHC can be used to detect infectious agents, identify tissue origins and differentiate between benign and malignant lesions in pathological specimens.

Molecular Profiling and Subtyping: By assessing the expression of specific proteins associated with distinct molecular subtypes or pathological features, researchers can classify tumors into different subgroups with distinct clinical characteristics and treatment responses. This information guides personalized treatment strategies and improves patient outcomes in oncology and other fields.

Protein Spatial Localization and Cellular Distribution: IHC is commonly used to determine the cellular localization and distribution of specific proteins within tissues. By employing antibodies tagged with fluorescent dyes or enzyme labels, researchers can visualize and analyze the spatial distribution of proteins within individual cells or across tissue sections. This information provides insights into protein function, subcellular localization and cellular interactions within complex biological systems.

Quantification of Protein Expression Levels: IHC can be utilized to quantify the expression levels of target proteins in tissue samples. Image analysis software allows researchers to measure the intensity of staining or the proportion of positively stained cells, providing quantitative data on protein expression levels. This quantitative analysis is valuable for comparing protein expression between different experimental conditions, disease states or patient samples.

Figure 2: . Applications of IHC in clinical practice and biomedical research. 2A: Detection of ERα in a breast tissue guides the diagnosis and treatment of breast cancer. 2B: PD-L1 as a companion diagnostic marker was validated in APS laboratory. 2C: Expression and deposition of Amyloid-β in hippocampus play a central role in the pathogenesis of Alzheimer disease. 2D: A drug candidate (green) was shown to be associated with a tumor marker expression in mouse brain (yellow)

Biomarker Discovery and Validation: IHC plays a critical role in the discovery and validation of biomarkers for various diseases and biological processes. By screening tissue samples with panels of antibodies targeting different proteins, researchers can identify candidate biomarkers associated with specific diseases, such as cancer, neurodegenerative disorders or autoimmune diseases. Once identified, these biomarkers can be further validated using larger patient cohorts to assess their diagnostic, prognostic or predictive value.

Pharmacodynamic and Drug Target Analysis: In drug development and preclinical research, IHC techniques are employed to evaluate pharmacodynamic effects and assess drug target engagement in tissue samples. Particularly the binding of a therapeutic antibody to different types of cells in specific tissues can be used to predict the efficacy or side effect of the lead antibody. By measuring changes in protein expression or activation following drug treatment, researchers can assess the efficacy and mechanism of action of therapeutic agents. This information is crucial for optimizing drug dosing, predicting treatment responses and identifying potential biomarkers of drug response or resistance.

Investigation of Disease Pathogenesis and Mechanisms: IHC is widely utilized to investigate disease pathogenesis and underlying molecular mechanisms in various pathological conditions. By studying the expression patterns of specific proteins associated with disease processes, researchers can elucidate the molecular pathways involved in disease initiation, progression and tissue damage. This knowledge informs the development of targeted therapies and interventions for treating or preventing disease progression.

Antibody clone screening for IHC: IHC detects the native epitope of the target protein. However, not all antibodies are suitable for IHC purposes. IHC staining can be utilized under specific conditions to screen antibodies for active clones during antibody development.

Conclusion

IHC continues to be an essential technique in biomedical research, facilitating discoveries across diverse fields of study. With ongoing technological innovations and expanding applications, IHC becomes increasingly indispensable for elucidating intricate spatial relationships among protein markers and furthering our comprehension of disease development.

APS Comprehensive Support to Various Types of Study
Due to the inherent complexity of IHC experiments and the growing need to understand the expression of multiple protein markers and their spatial relationships, obtaining consistent and high-quality data can pose significant challenges. As a leading histopathology laboratory, APS has dedicated extensive efforts to validate essential markers across various research areas. With cutting-edge imaging and staining equipment, coupled with our expertise in antibody validation, multiplex staining and imaging analysis, APS scientists are well-prepared to assist your team in achieving its research goals. For more information about our validated antibody list, platforms and technologies, please visit our website at www.appliedpathology.com or contact us via email at info@appliedpathology.com.