Creative Biolabs provides services for the identification and characterization of professional antigen-presenting cells (APCs) based on a flow cytometry-based immune analysis platform. This platform enables characterize specific subsets of cells along the dendritic, monocyte/macrophage, and B cell lineages. This service offers valuable and accurate information on the frequency of APC subsets to understand the efficacy of immune response and cancer treatment.
The draining lymph node (dLN) serves an important innate barrier function and is also the anatomical site in which adaptive immune responses are initiated following vaccination and infections. Upon encountering a pathogen or a vaccine in peripheral tissues, antigen-presenting cells (APC) take up antigen, mature, and start to migrate via afferent lymphatic vessels to dLNs, where they present the antigen to T cells for the induction of adaptive immune responses. The size of the DLNs increases dramatically during an immune response owing to the massive trapping of naive lymphocytes and the proliferation of antigen-specific lymphocytes.
Fig.1 Activation of antigen-specific T cells by APCs. (Gu, et al., 2020)
Professional antigen-presenting cells include dendritic cells (DCs), macrophages, and B cells, mediating the cellular immune response by processing and presenting antigens for recognition by certain lymphocytes such as T cells. DCs are the most capable APC and they possess a strong T-cell stimulatory capacity. They are a heterogeneous cell population in terms of locations, phenotypes, and immunological functions. Different types of APC interact with different lymphocyte populations and stimulate distinct types of immune responses. Therefore, identifying the APC populations in human lymph nodes will provide a better understanding of how immune responses are initiated and may also help improve vaccine design and delivery strategies.
Fig.2 Major subsets of DC (Dalod, et al., 2014); the complex network of subsets. (Caminschi, et al., 2012)
Fluorescence flow cytometry is the most commonly used platform for identifying APCs. Differential expression of cell surface markers is key to phenotyping subsets of APCs with unique surface profiles. For example, conventional DCs (cDCs) express the surface markers CD11c and MHCII; the two human cDC subsets are distinguished by the expression of CD141 (cDC1) or CD1 (cDC2). For the phenotyping of APC subsets, fluorochrome-labeled antibodies against numerous cell-specific proteins can be used. Therefore, the selection and rigorous testing of monoclonal antibodies and fluorochromes are crucial to successful polychromatic flow cytometry. Creative Biolabs provides state-of-the-art flow cytometry services for cell counting and sorting from homogeneous or mixed cell populations. We offer guidance at each experimental stage, from initial study design and staining protocols, to cell sorting and data interpretation.
Sample processing is at the beginning of all good flow cytometry data. We can process lymph node samples into a single-cell suspension before flow analysis by using enzymatic digestion or mechanical dissociation of the tissue. We select appropriate surface markers, monoclonal antibodies and fluorochromes to ensure optimal signal-to-noise ratio for our cell types of interest. Our rigorous and reproducible workflow will help accelerate your project goals.
Creative Biolabs provides comprehensive services for analysis of APCs subsets in the dLNs. Our scientists with deep experience and flow cytometry expertise are available to help determine the most appropriate assay design and validation strategy. We customize our service for each client to provide the solutions and expertise they require to address their unique research requirements. If you are interested in our services, please feel free to contact us to talk about your project.
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For Research Use Only | Not For Clinical Use
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