The effects of several types of crowding on immune functions were studied in mice. This study consisted of two experiments. Experiment one: Male BALB/c mice were initially housed in groups of four mice per cage. After fourteen days of acclimation, the mice were randomly divided into three groups, Control (four mice per cage, control group), Crowd-I (four mice per small space) and Crowd-II (sixteen mice per cage). These conditions were maintained for seven days. The results of experiment one were as follows: (1) The percentage of lymphocytes in the blood of Crowd-II was significantly lower than that of Control (p < 0.05). (2) The percentage of neutrophils and the absolute number of neutrophils in blood of Crowd-II were significantly higher than in Control (p < 0.05). (3) Superoxide production activity (NBT reduction activity) of blood neutrophils in Crowd-II tended to be depressed, and phagocytic activity of neutrophils was significantly depressed in Crowd-II as compared with Control (p < 0.01). These results suggest that the complexity of interrelationships among mice caused by an increase in the number of animals per cage is a very important stress factor. Experiment two: Male BALB/c mice were initially housed in groups of five mice per cage. After fourteen days of acclimation, the mice were divided into three groups, Control (five mice per cage, control group), Crowd-1 (five mice per small space) and Crowd-2 (twenty mice per cage). In Control and Crowd-1, the same mice were used as in the acclimation period. These conditions were maintained for seven days. In this period, on the second day, all the mice were injected intraperitoneally with sheep red blood cells (SRBC). The results of experiment two were as follows: (1) The specific humoral immune response to SRBC was investigated in terms of the number of PFC in the spleens and hemagglutination in sera, but significant differences were not found among the groups. (2) Plasma IgG levels in the Crowd 1 were significantly higher than those in Control (p < 0.05). (3) Both superoxide production activity and phagocytic activity of neutrophils were significantly depressed in Crowd-2 as compared with Control (p < 0.01, respectively), whereas each neutrophil function of Crowd-1 tended to be enhanced as compared with control. These results suggest that when group members do not change after initial acclimation, a decrease of personal space may cause enhancement of immune functions in mice, and it is considered that changing the mice in a cage itself affects their immune functions.
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