The natural polysaccharides extracted from the pollen of Pinus massoniana (TPPPS) have been shown to be a promising immune adjuvant against several viral chicken diseases. However, the exact mechanism through which TPPPS enhances the host immune response in chicken remains poorly understood. In the current study, chicken peripheral blood lymphocytes were treated with varying concentrations of TPPPS and pro-inflammatory cytokines such as IFN-γ, iIL-2 and IL-6 were measured to determine the optimal dose of the polysaccharide. A comparative analysis was subsequently performed between the proteome of lymphocytes subjected to the best treatment conditions and that of untreated cells. Protein identification and quantitation revealed a panel of three up-regulated and seven down-regulated candidates in TPPPS-treated chicken peripheral blood lymphocytes. Further annotation and functional analysis suggested that a number of those protein candidates were involved in the regulation of host innate immune response, inflammation and other immune-related pathways. We believe that our results could serve as a stepping stone for further research on the immune-enhancing properties of TPPPS and other polysaccharide-based immune adjuvants.