In lymphoid and myeloid cells, membrane hyperpolarization by the opening of K⁺ channels increases the activity of Ca²⁺ release-activated Ca²⁺ (CRAC) channels and transient receptor potential (TRP) Ca²⁺ channels. The intermediate-conductance Ca²⁺-activated K⁺ channel K_Ca3.1 plays an important role in cell proliferation, differentiation, migration, and cytokine production in innate and adaptive immune systems. K_Ca3.1 is therefore an attractive therapeutic target for allergic, inflammatory, and autoimmune disorders. In the past several years, studies have provided new insights into 1) K_Ca3.1 pharmacology and its auxiliary regulators; 2) post-transcriptional and proteasomal regulation of K_Ca3.1; 3) K_Ca3.1 as a regulator of immune cell migration, cytokine production, and phenotypic polarization; 4) the role of K_Ca3.1 in the phosphorylation and nuclear translocation of Smad2/3; and 5) K_Ca3.1 as a therapeutic target for cancer immunotherapy. In this review, we have assembled a comprehensive overview of current research on the physiological and pathophysiological significance of K_Ca3.1 in the immune system.