Intestinal intussusception (ISS) commonly causes intestinal obstruction in children. One proposed mechanism of ISS is that inflammation-induced alterations of intestinal motility cause ISS. We investigated whether innate inflammatory factors or altered motility are required for induction of ISS by LPS. We compared rates of ISS among BALB/c and C57Bl/6 mice, mice lacking lymphocytes or depleted of phagocytes, or mice with defects in the TLR4 signaling pathway following administration of LPS or the calcium analog MnCl₂. Six or two hours after administration of LPS or MnCl₂ respectively, mice underwent image analysis to assess intestinal contraction rate or laparotomy to identify ISS. LPS-induced ISS (LPS-ISS) was observed in BALB/c mice but not in C57BL/6 mice or in any BALB/c mice with disruptions in TLR4 signaling. LPS-induced serum TNF-α, IL-6, and NO and intestinal NO levels were similar between BALB/c and C57BL/6 mice. The rate of LPS-ISS was significantly reduced in phagocyte-depleted but not in lymphocyte-deficient mice. Intestinal contraction rates were reduced in LPS-ISS susceptible BALB/c but not in LPS-ISS resistant C57BL/6 or TLR4 mutant mice, suggesting a role for reduced intestinal contraction rate in LPS-ISS susceptibility. This was tested with MnCl₂, a calcium antagonist that reduced intestinal contraction rates and induced ISS irrespective of mouse strain. Therefore, LPS-ISS is initiated by innate immune signaling that requires TLR4 and phagocytes but may be independent from TNF-α, IL-6, and NO levels. Further, alteration in intestinal motility, specifically reduced intestinal contraction rate, is a key factor in the development of ISS.