Cytokines, a class of small molecular proteins with a wide range of biological activities, are secreted mainly by immune cells and function by binding to the corresponding receptors to regulate cell growth, differentiation and effects. CD4 T cells can be defined into different lineages based on the unique set of signature cytokines and transcription factors, including helper T cells (Th1, Th2, Th17) and regulatory T cells (Treg). In teleost, CD4 T cells have been identified in a variety of fish species, thought to play roles as Th cells, and shown to be involved in the immune response following specific antigen stimulation. With the update of sequencing technologies, a variety of cytokines and transcription factors capable of characterizing CD4 T cell subsets also have been described in fish, including hallmark cytokines such as IFN-γ, TNF-α, IL-4, IL-17, IL-10, TGF-β and unique transcription factors such as T-bet, GATA3, RORγt, and Foxp3. Hence, there is increasing evidence that the subpopulation of Th and Treg cells present in mammals may also exist in teleost fish. However, the differentiation, plasticity and precise roles of Th cell subsets in mammals remain controversial. Research on the identification and differentiation of fish Th cells is still in its infancy and requires more significant effort. Here we will review recent research advances in characterizing the differentiation of fish CD4 T cells by cytokines and transcription factors, mainly including the identification of Th and Treg cell hallmark cytokines and transcription factors, the regulatory role of cytokines on Th cell differentiation, and the function of Th and Treg cells in the immune response. The primary purpose of this review is to deepen our understanding of cytokine networks in characterizing the differentiation of CD4 T cells in teleost.