As the Marine fish with the highest aquaculture yield in China, the aquaculture industry of large yellow crocea is facing various diseases in recent years. IL-2 can promote the proliferation and differentiation of T and B lymphocytes and play an important immune function in the immune process of large yellow crocea against foreign invasion diseases. MicroRNAs, as non-coding RNAs with only 18-30 nucleotides, exert their functions by negatively regulating target genes through mRNA shearing and inhibiting protein translation. In order to study the role of microRNA in the regulation of T and B lymphocytes proliferation and differentiation in large yellow croaker, we used purified recombinant large yellow croaker IL-2 (rLcIL-2) to stimulate primary head kidney lymphocytes isolated and cultured in vitro, with illumina high-throughput sequencing technology, microRNA expression profiles of head kidney lymphocytes stimulated by rLcIL-2 were obtained. In the rLcIL-2 stimulation group and the rGFP control group, the three micrornas with the highest expression levels were abu-miR-100, ola-miR-146a-5p, and abu-miR-155. There are 40 microRNAs were differences in expression between the two groups of cells, among which 32 microRNAs were significantly up-regulated, including abu-miR-146a, abu-miR-155, abu-miR-147, etc. 8 types of microRNAs were significantly down-regulated, including abu-miR-100 etc. Combined with miRanda and RNAhybrid software, 16005 target genes with 40 differentially expressed microRNAs were predicted, including STAT3, Bcl-6,Caspase3, Bmf, IL-2 signaling pathway key molecules and downstream effector molecules. These results are of great significance for further understanding the mechanism of fish proliferation and differentiation, and provide theoretical basis for the design of relevant vaccines with IL-2 as immune adjuvant.