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Germinal centers (GCs) are structures formed within B cell follicles critical for the generation of high affinity antibodies. The evaluation of GCs in secondary lymphoid tissues has emerged as a valuable means for understanding the immunological activity in vaccine responses, autoimmunity and cancer. The analysis has been facilitated by advances in sampling techniques, including non-invasive lymph node collection and fine needle aspiration. In this study, we performed a systematic comparison between immunohistology and flow cytometry for analysis of GCs with the major aim to identify strategies for data analysis that would allow to relate data acquired by the two methods. Lymph nodes from rhesus macaques were divided in half and analysed as either cryosections or cell suspensions. Using human markers such as PD-1 and Ki67 to identify T follicular helper (T) cells and GC B cells, we developed a method for GC analysis by immunohistology using CellProfiler™ software and a flow cytometry panel with relatively limited numbers of antibodies to be scalable and feasible for most laboratories to perform. While some discrepancies between the two methods were identified, T cells and GC B cells normalized by total CD3 T cell or CD20 B cell numbers, respectively, in immunohistology correlated well with matched data from flow cytometry. GC area normalized by section area in immunohistology also correlated well with T cells per total live cells from flow cytometry. Performing this type of data analysis would therefore facilitate comparison of results generated between the two methods.
This article was published in the following journal.
Name: Journal of immunological methods
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The activated center of a lymphoid follicle in secondary lymphoid tissue where B-LYMPHOCYTES are stimulated by antigens and helper T cells (T-LYMPHOCYTES, HELPER-INDUCER) are stimulated to generate memory cells.
Malignant lymphoma in which the lymphomatous cells are clustered into identifiable nodules within the LYMPH NODES. The nodules resemble to some extent the GERMINAL CENTER of lymph node follicles and most likely represent neoplastic proliferation of lymph node-derived follicular center B-LYMPHOCYTES.
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A center in the PUBLIC HEALTH SERVICE which is primarily concerned with the collection, analysis, and dissemination of health statistics on vital events and health activities to reflect the health status of people, health needs, and health resources.
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