Allomyrina dichotoma larvae are a promising alternative protein resources for foods. Although nutrients are abundant in A. dichotoma larvae and exhibit various physiological functions, nothing is known of the immune activities of A. dichotoma larvae extracts at low temperatures (5°C). This study investigated the immune-enhancing activity of A. dichotoma larvae extract (ADL) at low-temperatures in RAW264.7 cells, a reprensentative macrophage cell line. ADL (5°C) was not cytotoxic at 100 or 200 μg/mL and increased interleukin-6, interleukin-1β production at both concentrations. Furthermore, these immunomodulatory effects were mediated by the phosphorylations of MAPKs (p38, JNK, and ERK) and the translocation of NK-ĸB. Consequently, the study shows that ADL at low-temperature can be used as an immunity-enhancing ingredient.