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Published on: March 10, 2017
Cellular assays to evaluate B-cell function
1Children's Hospital Los Angeles and Keck School of Medicine, USC, 4650 Sunset Blvd, Los Angeles, CA 90027, United States of America.
Insights
Cellular assays like lymphocyte proliferation, opsonophagocytic, and ELISPOT assays are crucial for evaluating B-cell function in diagnosing antibody deficiencies and assessing immune responses. These methods aid in understanding B-cell function for better patient care.
Area of Science:
- Immunology
- Clinical Diagnostics
Background:
- Inborn errors of immunity (IEI) often manifest as recurrent infections due to antibody deficiencies.
- Evaluating the B-cell and antibody compartment is essential for diagnosing IEI.
Purpose of the Study:
- To provide an overview of cellular assays for studying B-cell function.
- To discuss the clinical applications and limitations of lymphocyte proliferation assays (LPAs), opsonophagocytic assays (OPAs), and ELISPOT assays.
Main Methods:
- Lymphocyte Proliferation Assay (LPA): Assesses ex-vivo cell proliferation in response to stimuli.
- Opsonophagocytic Assay (OPA): Evaluates the ability of IgG antibodies to induce phagocytosis.
- Enzyme-linked Immunosorbent Spot Assay (ELISPOT): Assesses antibody production at a cellular level.
Main Results:
- LPAs can assess T-cell proliferation clinically, with research settings offering more B-cell specific stimuli.
- OPAs are used to assess vaccine potency and IgG quality.
- B-cell ELISPOT assays can characterize specific antibody responses and are useful in patients on IgG therapy or immunomodulatory drugs.
Conclusions:
- Cellular assays like LPA, OPA, and ELISPOT are vital tools for assessing B-cell function in various clinical and research settings.
- These assays provide critical insights into antibody deficiencies and immune responses, aiding in diagnosis and treatment evaluation.
Abstract:
Inborn errors of immunity (IEI) that present with recurrent infections are largely due to antibody (Ab) deficiencies. Therefore, assessment of the B-cell and Ab compartment is a major part of immunologic evaluation. Here we provide an overview about cellular assays used to study B-cell function and focus on lymphocyte proliferation assay (LPA), opsonophagocytic assay (OPA), and the Enzyme-linked Immunosorbent Spot Assay (ELISPOT) including clinical applications and limitations of these techniques. LPAs assess ex-vivo cell proliferation in response to various stimuli. Clinically available LPAs utilize peripheral blood mononuclear cells and mostly assess T-cell proliferation with pokeweed mitogen considered the most B-cell specific stimulus. In the research setting, isolating B cells or using more B-cell specific stimuli such as CD40L with IL-4/IL-21 or the TLR9 ligand CpG can more specifically capture the proliferative ability of B cells. OPAs are functional in-vitro killing assays used to evaluate the ability of IgG Ab to induce phagocytosis applied when assessing the potency of vaccine candidates or along with avidity assays to evaluate the quality of secreted IgG. The B-cell ELISPOT assesses Ab production at a cellular level and can characterize the Ab response of particular B-cell subtypes. It can be used in patients on IgG therapy by capturing specific Abs produced by individual B cells, which is not affected by exogenous IgG from plasma donors, and when assessing the vaccine response in patients on immunomodulatory drugs that can affect memory B-cell function. Emerging approaches that are only available in research settings are also briefly introduced.
