Objective To establish the HHV-6-specific CD4⁺ T cell clones and further study the character of HHV-6-specific CD4⁺ T cells. Methods We acquired CD4⁺ T cell clones by the liquid microwell limiting dilution technique, and selected HHV-6-specific CD4⁺ T cell clones by means of 3H-thymidine uptake. The phenotype of HHV-6-specific T cell clones were analyzed with FACS. We tested the IL-10, IL-4 or IFN-γ secretion from HHV-6-specific CD4⁺ T cell clones by ELISA. Results Of the five HHV-6-specific CD4⁺ T cell clones, four showed proliferative responses to the stimulation with HHV-6 infected JJHAN lysates. Moreover, proliferation responses of HHV-6-specific CD4⁺T cell clones depended on the concentration of the HHV-6-infected JJHAN lysates. The phenotypic analysis with FACS demonstrated that HHV-6-specific T cell clones consisted of CD3⁺ CD4⁺ T cells. CD4⁺ T cell clones-W-2 and W-4 possessed the feature of high levels of IL-10 production, while CD4⁺ T cell clones-W-1 possessed the feature of high levels of IL-10 and IFN-γ production and W-3 possessed the feature of high levels of IFN-γ production. Conclusions The HHV-6-specific CD4⁺ T cell clones, were established in our study so as to further study their pneotype, the character of cytokine secrection, and to provide experimental basis for selecting HHV-6-specific CD4⁺ Treg cell clones.