Technique / Immunology / Assays for lymphocyte function

Some important assays for measuring T and B lymphocyte functions in vitro are listed here. Those include methods for B cell antibody production assay, lymphokine production assay, T and B cell proliferation assay, fluorescence activated cell sorting based on lymphocyte cell-surface antigens and multiparameter flow cytometric analysis for immunophenotyping, as well as in vivo lymphocyte function assays.

LYMPHOCYTE PROLIFERATION ASSAY (LPA) recommended protocol
Protocol written by Fred Valentine and Howard Lederman. Lymphocyte proliferation assay (LPA) measures the ability of lymphocytes placed in shortterm tissue culture to undergo a clonal proliferation when stimulated in vitro by a foreign molecule, antigen ...

ISOLATION OF T CELLS FROM SYNOVIAL MEMBRANES protocol
It includes adherent and nonadherent cell isolation. (Dr. Bart Frank, Arthritis and Immunology Program, OMRF) ...

Protein Kinase C Assay protocol
The following is a modification of Kikkawa et al., (1982) J. Biol. Chem. 257, 13341. This assay measures phosphorylation (32P) of a protein kinase C-peptide substrate, which is subsequently separated by binding to phosphocellulose paper. (Newton lab, Departmen ...

C7 Isolation of mouse peripheral blood mononuclear cells by flotation protocol
Isolation of PBMC using Axis Shield Density Gradient Media. (Axis Shield inc.) Standard human peripheral blood mononuclear cell (PBMC) isolation media such as lymphoprep or Nycoprep 1.077 are less effective for the isolation of these cells from the blood of ...

Measuring human T-lymphocyte function review
Written by Julian K. Hickling, Expert Reviews in Molecular Medicine http://www.expertreviews.org PDF file is available in this page. T lymphocytes (T cells) play critical roles in the regulation of immune responses, and are responsible for mediating many ...

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Preparation and culture of human lymphocytes.
(Curr Protoc Cell Biol. 2001 May;Chapter 2:Unit 2.2.)
Biddison WE.

National Institute of Neurological Disorders and Stroke/NIH, Bethesda, Maryland,
USA.

This unit presents protocols for preparation of lymphocytes from peripheral
(whole) blood or leucopherisis samples, first by differential centrifugation in
Ficoll-Hypaque to separate them from erythrocytes and granulocytes. Repeated
centrifugation removes the plasma and platelets. Monocyte/macrophage cells and
dendritic cells can be purified from the T and B cell population by exploiting
their adhesion to serum-coated plates in the presence of recombinant IL-3.
Different subpopulations of lymphocytes can be isolated by specific antibodies
bound to magnetic beads.

Figure. Lymphocyte separation by using antibodies coupled to magnetic beads
(Immunobiology)
Appendix I. Immunologists' Toolbox Isolation of lymphocytes

Characterization of lymphocyte specificity, frequency, and function
(Immunobiology)
Appendix I. Immunologists' Toolbox

Isolation of lymphocytes
(Immunobiology)
Appendix I. Immunologists' Toolbox