Whole blood assay for assessment of the mixed lymphocyte reaction

• Published in: Journal of immunological methods 2001, Vol.247 (1), p.1-8
• Main Authors: Bromelow, K.V, Hirst, W, Mendes, R.L, Winkley, A.R, Smith, I.E, O’Brien, M.E.R, Souberbielle, B.E
• Format: Article
• Language: English
• Subjects: Aged; Biological and medical sciences; Female; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Human mycoses; Humans; Immunobiology; Infectious diseases; Leukocytes, Mononuclear - cytology; Lymphocyte Count; Lymphocyte Culture Test, Mixed - methods; Lymphocyte Culture Test, Mixed - standards; Lymphocytes - immunology; Lymphoid cells: ontogeny, maturation, markers, receptors, circulation and recirculation; Male; Medical sciences; Mesothelioma - blood; Mesothelioma - immunology; Microbiology; Middle Aged; Mycoses; Mycotic sepsis
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/S0022-1759(00)00311-2

When lymphocytes from genetically different individuals are mixed together in tissue culture blast transformation occurs, a reaction known as the mixed lymphocyte reaction (MLR). The MLR is a clinically relevant in vitro assay where lymphocytes from one individual (effector, E) are incubated with the lymphocytes of another individual (stimulator, S) which have been previously rendered incapable of blast transformation by gamma-irradiation. We have standardised a whole blood (WB) MLR assay where the E lymphocytes were provided by 20 μl of WB and the S lymphocytes were provided by irradiated peripheral blood mononuclear cells (PBMCs) either as a mixed pool of 20 donor PBMCs or as single donor PBMC. The optimum number of S lymphocytes needed was comparatively higher than in the standard PBMC MLR: the optimum calculated E:S ratio was 1:20 compared to a E:S ratio of 1:1 or 3:2 in the standard PBMC MLR. In ten normal individuals the WB/PBMC MLR was similar to the standard PBMC/PBMC MLR. As a clinical example, the WB/PBMC MLR proliferative capacity of 13 patients with malignant mesothelioma was no different from the proliferative capacity of their age–sex matched controls. This standardised WB/PBMC MLR assay is a simple and more practical assay than the standard MLR assay and can be incorporated easily in clinical studies with biological end-points.