Abstract: To establish the in vitro immunotoxicity of cigarette smoke, mouse lymphoma cell EL-4 and mouse macrophage cell Ana-1 were treated with the total particular matter (TPM) in mainstream smoke of Kentucky 3R4F reference cigarette, then the viability of cells was measured by CCK-8 assay. Luminex liquid chip was used to determine the levels of 20 cytokines (FGF-basic, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, KC, IP-10, MCP-1, MIP-1 α, VEGF, MIG, TNF-α) in the supernatant. The results showed that after exposure to TPM, GM-CSF, IL-10, VEGF were inhibited in EL-4, while MCP-1, MIP-1 α, TNF-α were induced in Ana-1. The levels of the other 17 cytokines in two cell types did not change significantly with the increase of TPM concentration. The cell reactions of different types or sources to the toxicological effect of TPM of cigarette smoke differed, therefore adopting two kinds of cells is helpful to the better evaluation of in vitro immunotoxicity of cigarette smoke. The method is rapid, sensitive, high-throughput and suitable for assessing in vitro immunotoxicity of cigarette smoke.