Cerebral malaria (CM) is a serious and often fatal complication of Plasmodium falciparum infections; however, the precise mechanisms leading to CM is poorly understood. Mouse malaria models have provided insight into the key events in pathogenesis of CM. T cell immune response is known to play an important role in malaria infection, and members of the T-cell immunoglobulin– and mucin–domain–containing molecule (TIM) family have roles in T-cell–mediated immune responses. Tim-1 and Tim-3 are expressed on terminally differentiated Th2 and Th1 cells, respectively, and participate in the regulation of Th immune response. Until now, the role of Tim family proteins in Plasmodium infection remains unclear. In the present study, the mRNA levels of Tim-1, Tim-3, and some key Th1 and Th2 cytokines in the spleen of female Kunming outbred mice infected with P. berghei ANKA (PbANKA) were determined using real-time polymerase chain reaction (qRT-PCR). Tim-1 expression was significantly decreased at day 10 postinfection (p.i.) in infected mice with CM, and significantly increased at day 22 p.i. in infected mice with non-CM, compared with uninfected controls (P < 0.01); in contrast, Tim-3 expression was significantly increased in both CM and non-CM infected mice at days 10 and 22 p.i., respectively. Furthermore, the expression of Tim-1 and Tim-3 mRNA in spleen was significantly positively correlated with the level of Th2 and Th1 cytokine mRNA in the spleens, respectively. PbANKA infection could inhibit the differentiation of T lymphocytes toward Th2 cells, promote the Th1 cell differentiation, and induce Th1-biased immune response in the early infective stage in infected mice with CM; whereas the infection could promote Th2 cell differentiation, and induce Th2-biased immune response in the late infective stage, in infected mice with non-CM. Our data suggest that both Tim-1 and Tim-3 may play an important role in the pathogenesis of P. berghei infection, which may represent a potential therapeutic target.