Identification of functional domains of chicken interleukin 2.

Interleukin 2 (IL-2) is an essential cytokine that plays a pivotal role in the replication, maturation and differentiation of lymphocytes. In this study, the functional domains of chicken IL-2 (chIL-2) were mapped with monoclonal antibodies (mAb), a synthetic peptide, and a phage display peptide library. Nine neutralizing mAbs to chIL-2 were produced using the recombinant chIL-2 monomer expressed in prokaryotic cells as an immunogen and used to finely map the functional domains of the chIL-2 protein. The mimotopes of nine anti-chIL-2 mAbs, including KIELPSL, EHLDXNDSLYL, NHLXGXY, WHLPPSL, EFKASXL, TENPFPE, SGLYL, AHGYWEL and HHGYWEL, were respectively identified by phage display and peptide-competitive ELISA. These mimotopes constitute three conformational functional domains in the chIL-2 molecule, that is, N(26)K(27)I(28)H(29)L(30)E(31)L(32)P(35)Q(43)Q(44)T(45)L(46)Q(47)C(48)Y(49)L(50) (domain I), E(68)E(69)F(70)K(79)K(82)S(83)L(84)T(85)G(86)L(87) (domain II) and N(88)H(89)G(91)K(104)F(105)P(106)D(107)E(111)L(112)Y(118)L(119) (domain III). The neutralizing mAbs to chIL-2 inhibited the in vitro lymphocyte proliferation stimulated by three peptide domains of chIL-2. The predicted tertiary structure of chIL-2 reveals that domain I was positioned in the long A-B loop and the N terminal of Helix B, domain II was mostly situated in Helix C, and domain III was distributed in the C-D loop and Helix D. These data demonstrate the functional domains of chIL-2 and provide a clue for elucidating the interaction between chIL-2 and its receptor.