Following traumatic brain injury (TBI), the neuroinflammation may cause acute secondary injury which leads to neuropsychiatric problems and neurodegenerative pathologies. Through single-cell transcriptomics, target cell types and pathways of immune response to TBI have been explored in recent years. However, global transcriptional changes reflecting cellular responses through cell-cell interactions remain elusive in physical space and time. In this study, we performed the spatio-temporal analysis using Spatial Transcriptomics to understand the immune response across sectioned TBI tissues in different conditions and time points, including transgenic mice with microglia depletion. We confirmed the microglia cells, the resident immune cells of the brain, rapidly change states and communicate with other cell types during the response to their spatial environment. We found in samples collected post-TBI at 6 hours and 3 days that microglia were distributed at a remarkably higher density at hippocampal regions and injured sites compared to in the naive brain and sham control. In the post-TBI 3 days sample, we found a significant cellular signaling event mediated by astrocytic Serpin Family G Member 1 (Serping1) and microglial Type 1 Transmembrane Receptor (Lrp1) as the target ligand-receptor pair between astrocyte and microglia. This interaction change was specific in the hippocampal region. Furthermore, our data suggest significant changes in the distribution of other immune, glial, and neuron cell types, contributing to the overall spatial heterogeneity of responses to TBI. Our spatial transcriptomics analysis of immune responses in TBI brain samples across space and time confirmed and added new insights into the pathogenic pathways in TBI and related neuroinflammation disorders.