Temperature pretreatment alters the polarization response of human neutrophils to the chemoattractant N-formyl-Met-Leu-Phe.

Neutrophils present a polarized morphology upon stimulation of chemoattractants, which play a vital role in host-defense mechanisms. Many studies have been published on neutrophil polarization, in which three different temperatures pretreatment (4 degrees C, 25 degrees C and 37 degrees C) have been used. However, no study has investigated whether different temperature pretreatments affect neutrophil polarization. In the current study, we examined the effects of 4 degrees C, 25 degrees C and 37 degrees C pretreatment temperatures on short-term (1 or 3 min) chemoattractant-induced polarization. Human neutrophils were polarized upon the stimulation of N-formyl-Met-Leu-Phe (fMLP) after pretreated by different temperature. The morphological changes of the neutrophils were investigated under the microscopy. The F-actin polymerization was determined by immunological histological chemistry. There were more head-tail polarized cells (>50% of the cells) in the 25 degrees C and 37 degrees C pretreatment groups than in the 4 degrees C group (32.4%). The average lengths of the pseudopod were 3.2 +/- 1.1 microm (n = 17), 5.3 +/- 2.1 microm (n = 12) and 7.4 +/- 2.7 microm (n = 21) in the 4 degrees C, 25 degrees C and 37 degrees C pretreatment groups, respectively; the 4 degrees C and 37 degrees C pretreatment groups were statistically different (P < 0.05). Additionally, there was a statistically significant difference in the pseudopod extension rate among the three groups, as well as the Lamellipod percentage between the 4 degrees C group and the other two groups within 1 min of stimulation with fMLP. This study demonstrates that different temperature pretreatments affect neutrophil polarization upon short-term stimulation with fMLP.