Objective: Lung cancer remains the leading cause of cancer death; metastatic non-small cell lung cancer is clinically difficult to treat and immunotherapy is a novel treatment of lung cancer like PD1 or PD-l1 immune checkpoint inhibitors (ICIs). However, it still needs an approach for killing tumor cell and active of immune cell like tumor lysates vaccine thorough hapten (hydrazine) enhance intratumoral chemotherapy (HEIC. Method: scRNA-Seq and Olink proteomics were employed to study the initiation of immune response at the cell level of untreated and the treated major tumor after HEIC, also analyzed cytokines of expression of genes following the initiation of immune response. Result: The immunity reaction was activated like MPs, mature DC cells and T and NK cells including CD8+ effector T cells, CD8Trm, NK cells, also awaking memory T cells, CD8+ effector T cells, CD8Trm and Naive T increased in untreated tumor after the major tumor treated. It confirmed that HEIC can kill tumor and modify associate tumor antigens (ATAs) to be neo ATAs for initiation of immune response. It resulted in 30 % high expression over 92 genes detecting for longer survival group with 2.5 years than short survival group with less 1.5 years. Conclusion: We have demonstrated that immune response can be induced by hapten associated therapy of HEIC with up to 30% expression of genes in long survival group. This significant difference may be related to the heterogeneity of immune cell individuals and tumor environment between the long and short survival groups.