Although many recent studies have suggested that CD4(+) helper T cell (Th-cell) functions are well conserved among teleost fishes and mammals, there is little evidence that CD4(+) Th-cells in fish are actually involved in both humoral and cell-mediated immunity during a secondary immune response. In the present study, adoptive transfer using clonal ginbuna crucian carp and crucian carp hematopoietic necrosis virus (CHNV) was used to investigate the functions of CD4(+) cells during humoral and cell-mediated immunity. With regard to humoral immunity, transplanting CHNV-sensitized donor cells, containing CD4(+) cells, into naive fish induced more rapid and stronger antibody production than by transplanting non-sensitized donor cells or sensitized donor cells lacking CD4(+) cells. During cell-mediated immunity, no significant differences were found in recipients that received sensitized cells regardless of whether the donor cells contained CD4(+) cells, although recipients that received both sensitized donor cells (with and without CD4(+) cells) exhibited more efficient cell-mediated cytotoxicity than those that received non-sensitized donor cells. These findings suggest that inducing a secondary antibody response requires CD4(+) cell help, and secondary cell-mediated immunity can be induced both by CD4(+) cells and leukocytes other than CD4(+) cells.