CD Cell Lines
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Background of Classification Determinant
Classification determinant, also known as cluster of differentiation (CD), is a series of cell surface molecules used for identifying and analyzing cellular immunophenotypes and classified according to similar nomenclature. The CD nomenclature was first established by the International Symposium on Human Leukocyte Differentiation Antigens in 1982. This protocol was originally used to standardize the classification and nomenclature procedure of monoclonal antibodies to leukocyte surface molecules, and it is now applicable to many other cell types.
Fig. 1 Definition of distinct subsets of hematopoietic cells by specific CD markers.1,2
Functions and Mechanisms of CD Markers
At the physiological level, CD marker molecules can participate in cellular functions in many ways, usually acting as important receptors or ligands and initiating relevant downstream pathways. Some molecules do not play roles in the process of signal transduction but regulate cell adhesion, migration, and other behaviors. However, the most common use of CD markers is for cell identification to distinguish cell sub-populations, which is widely used in flow cytometry.
Applications and Pathology of CD Cell Lines
CD marker molecules participate in many essential processes such as cell signaling and antigen recognition, and are widely utilized in various methods for cell sorting. The disturbance, disfunction, over-expression, and oppression of CD markers always lead to severe pathological effects. Some common cell types and signature CD markers are listed below.
Published Data
| Paper Title | Standardization of workflow and flow cytometry panels for quantitative expression profiling of surface antigens on blood leukocyte subsets: an HCDM CDMaps initiative |
| Journal | Frontiers in Immunology |
| Published | 2022 |
| Abstract | The Human Leukocyte Differentiation Antigen (HLDA) Workshop analyzes, characterizes, and names antibody clusters reactive to specific antigens. These clusters of differentiation (CDs) provide the scientific community with consistent and standardized target nomenclature, reproducible markers of leukocyte subsets, and validated clonal antibodies, providing the feasibility of standardized and reproducible collection of expression patterns for immunology research. However, many procedures in this process still need to be further optimized to construct quantitative reagent benchmarks and CD marker expression profiles at the single-cell level to enable large-scale deployment and iteration of CD Maps resources. Flow cytometry and immunophenotyping are already key methods for assessing the expression of single-cell proteins and specific markers. While constructing a complete CD marker expression profile, each mAb reagent should ideally be titrated appropriately and specifically, with optimal concentrations for accurate molecular quantification and to limit unwanted background staining. In this experiment, the researchers developed and standardized a flow cytometry procedure for quantitative expression profiling of surface antigens on subsets of blood leukocytes. Standardized and semi-automated procedures are available for high-throughput profiling of surface protein expression. The approach was validated globally at multiple centers with HLDA-approved antibody clones against CD3, CD11b, CD31, CD38, and CD40 and demonstrated the feasibility of benchmarking antibody reactivity within this framework. |
| Result |
To standardize the process of defining many functional immune cell subsets in blood, the researchers created a high-content framework to assess phycoerythrin-conjugated mAb reactivity and titers. They design flow cytometry panels for granulocytes/ dendritic cells/ monocytes/ NK cells/ innate lymphocytes and B/T cells respectively, and built backbone reagents capable of processing measurements + automated data annotation programs. Through the standardization and optimization of high-throughput reagent titration procedures, multicolor flow cytometry panel designed for innate and adaptive immune cells, and more efficient data analysis pipelines, researchers pioneered a method for high-throughput reproducible assessment of CD marker expression in human peripheral blood subpopulations. Experimental results show that this procedure has good robustness and standardized properties, and can be used to define new blood cell subsets in blood, detect HLDA-approved CD markers, or compare new antibody clones of interest to existing CD markers. This validated CD marker assay greatly standardizes and optimizes existing antigen benchmark detection capabilities and measurement efficiencies.
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Fig. 2 Universal titration procedure for PE-labeled mAbs.3,4
References
- Piszczatowski, Richard T., et al. "A glycan-based approach to cell characterization and isolation: Hematopoiesis as a paradigm." Journal of Experimental Medicine 219.11 (2022): e20212552.
- Image retrieved from Figure 1 " A panel of HS-specific scFv antibodies defines distinct glycotypes of hematopoietic cells and reveals divergent HS glycotypes between megakaryocyte and erythroid lineages. " Piszczatowski, et al. 2022, used under CC BY 4.0. The original image was modified by extracting and using part a-f and the title was changed to " Definition of distinct subsets of hematopoietic cells by specific CD markers.".
- Kužílková, Daniela, et al. "Standardization of workflow and flow cytometry panels for quantitative expression profiling of surface antigens on blood leukocyte subsets: an HCDM CDMaps initiative." Frontiers in Immunology 13 (2022): 827898.
- Image retrieved from Figure 2 " Universal titration procedure for PE-labeled mAbs. " Kužílková, et al. 2022, used under CC BY 4.0. The original image was modified by extracting and the title was changed to " Universal titration procedure for PE-labeled mAbs.".
