We applied quantitative high-resolution proteomics to systematically evaluate how the human PMN proteome is shaped in response to two LPS preparations that differ primarily in the number of acyl chains associated with their lipid A (E. coli and F. tularensis LPS). To capture both stimulus-specific effects and response kinetics, PMNs were analyzed at multiple time points, allowing assessment of treatment-dependent differences and proteomic changes over time within each condition. We characterized the PMN secretome following exposure to these agents, enabling the comparison of process between intracellular proteomic and protein release. Together, this integrated cellular proteome and secretome approach provides a comprehensive framework to define how structurally distinct LPS molecules modulate PMN functional programs over time. While PMN responses to LPS have been extensively investigated, their temporal proteomic and secretory landscapes in primary human PMNs, particularly in response to LPS with divergent lipid A architectures, remain incompletely characterized.