Objective: Toll-like receptors (TLRs) are highly or lowly expressed in a wide variety of tumors and exhibit either pro-tumor or anti-tumor activities. In the present study, we investigate whether there are relationships between the expressions of TLRs and the occurrence of radiation pneumonia in advanced NSCLC patients treated with radiotherapy. Materials and methods: 76 patients diagnosed with NSCLC and 50 healthy controls were recruited from Oct 2012 to Jan 2014. The expressions of serum TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, and TLR9 were detected by ELISA techniques. Fisher exact test, χ² test, ROC working curve and Cox regression model were applied to analyze all data. Results: serum TLR1, TLR2, TLR4, and TLR9 exhibited a relative high expression level in NSCLC patients compared with healthy controls. Importantly, pre-neutrophil granulocyte ratio was associated with the expression of TLR1, TLR2, and TLR4. Moreover, the patients with high ratio of neutrophil granulocyte significantly increased the occurrence of fever in comparison to normal neutrophil ratio in NSCLC patients during the course of radiotherapy. We further evaluated the containing of TLRs when patients had temperatures and found serum TLR1, TLR2 and TLR4 were over-expressed. Finally, 26 of 76 patients were diagnosed with different stages of radiation induced pneumonia; as a result, the contents of TLR1 and TLR4 before radiotherapy were identified as independent significances with pneumonia occurrence. Conclusions: The pretreatment levels of TLR1 and TLR4 have the predictive value to be clinically potential biomarkers of pneumonia risk in locally advanced NSCLC.