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Updated: Mar 9, 2026
Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs
Published on: December 27, 2016
Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs
Jennifer N Gibson1, Pavani Beesetty1, Courtney Sulentic2
1Department of Neuroscience, Cell Biology and Physiology, Boonshoft School of Medicine, Wright State University.
Insights
This study presents a rapid automated cell counter assay to quantify T-lymphocyte blastogenesis. This method offers precise measurements of cell diameter for assessing immunomodulatory drugs and mitogens.
Area of Science:
- Immunology
- Cell Biology
- Drug Discovery
Background:
- Lymphocyte proliferation is crucial for immune responses and assessing immunomodulatory compounds.
- Early mitogenesis involves cell enlargement (blastogenesis) before division, detectable in T-lymphocytes.
- Existing proliferation assays are often laborious and assess population-level effects.
Purpose of the Study:
- To develop and describe a rapid method for quantifying T-lymphocyte blastogenesis.
- To utilize an automated cell counter for precise measurement of cell diameter.
- To provide an alternative to laborious proliferation assays for evaluating immunomodulatory agents.
Main Methods:
- Isolation of T lymphocytes from mouse spleens and human peripheral blood mononuclear cells (PBMCs).
- Stimulation of lymphocytes with mitogens.
- Quantification of cell diameter changes using an automated cell counter.
Main Results:
- The automated cell counter assay provides rapid and direct measurements of cell diameters.
- This method precisely quantifies blastogenesis in T lymphocytes.
- The assay is effective for assessing various mitogens and immunomodulatory drugs in vitro.
Conclusions:
- An automated cell counter assay offers a rapid, precise, and direct method for quantifying T-lymphocyte blastogenesis.
- This technique overcomes limitations of traditional laborious proliferation assays.
- The assay is valuable for in vitro assessment of immunomodulatory compounds and mitogens.
Abstract:
Lymphocyte proliferation in response to antigenic or mitogenic stimulation is a readily quantifiable phenomenon useful for testing immunomodulatory (i.e., immunosuppressive or immunostimulatory) chemical compounds and biologics. One of the earliest steps during mitogenesis is cell enlargement or blastogenic transformation, whereupon the cell volume increases before division. It is usually detectable in the first several hours of T-lymphocyte stimulation. Here, we describe a rapid method to quantify blastogenesis in T lymphocytes isolated from mouse spleens and human peripheral blood mononuclear cells (PBMCs) using an automated cell counter. Various commonly used proliferation assays for the most part are laborious and only reflect the overall population effect rather than individual cellular effects within a population. In contrast, the presented automated cell counter assay provides rapid, direct, and precise measurements of cell diameters that can be used for assessing the effectiveness of various mitogens and immunomodulatory drugs in vitro.
