Summary
An immunohistochemical method for assessing the level of tumour necrosis factor-α in alveolar macrophages obtained by brochoalveolar lavage is described. Cytospins of mixed populations of lung cells were incubated first with a monoclonal antibody to CD68 and then with a specific peroxidase-labelled second antibody in a two-step reaction for the detection of the macrophage marker CD68. A second similarly based two-step reaction for the detection of tumour necrosis factor-α followed. Both reactions were visualized, on completion, using different coloured peroxidase substrates which produced a third colour in the event of dual deposition of the substrates. Dual substrate deposition was indicative of alveolar macrophages positive for tumour necrosis factor-α. This method has provided a specific and reproducible semi-quantitative test for the presence of tumour necrosis factor-α in human activated alveolar macrophages, which can be performed retrospectively on clinical material. A range of concentrations of the cytokine has been demonstrated in individual samples. This dual detection method has the potential for detection of any cell-associated protein product by minor modification of the described method.
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Hamilton, S., Healy, M., Corris, P. et al. An immunohistochemical method for the detection of tumour necrosis factor alpha in cytospins of human bronchoalveolar lavage cells. Histochem J 27, 487–493 (1995). https://doi.org/10.1007/BF00173715
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DOI: https://doi.org/10.1007/BF00173715