Int J Clin Exp Pathol. 2020 Jul 1;13(7):1867-1872. eCollection 2020.
OBJECTIVE: To observe alveolar macrophages (AMs) in the microenvironment of patients with non-small cell lung cancer (NSCLC).
MATERIALS AND METHODS: 20 NSCLC patients received bronchoalveolar lavage, and the bronchial alveolar lavage fluid (BALF) was collected. The phenotypes of AMs were detected by the opal multiplex immunofluorescence assay (mIF), flow cytometry, and western blot.
RESULTS: AMs could easily be made into paraffin sections after agar pre-embedding. The mIF results showed that AMs highly expressed M1-type marker CD86, and M2-type marker CD163 under PerkinElmer Vectra microscope, while there was a significant difference between the expression of CD86 and CD163 (**P<0.01), consistent with the flow cytometry results. Western blot revealed that the other markers of M1-type (CD16 and iNOS) expression in the AMs were compared with M2-type markers CD206 and ARG (*P<0.05).
CONCLUSIONS: Our results showed that AMs simultaneously expressed M1-type markers and M2-type markers, while the M2 markers still dominated. This suggests agar pre-embedding is a very convenient method to embed cells to paraffin tissue, so that cell membrane or nuclear antigens are very easily detected by mIF.