title: Cytosine-phosphate-guanine oligodeoxynucleotides containing GACGTT motifs enhance the immune responses elicited by keyhole limpet hemocyanin antigen in dairy cattle.
abstract: Adjuvants are important components of vaccine formulations. Effective adjuvants line innate and adaptive immunity by signaling through pathogen recognition receptors. Synthetic cytosine-phosphate-guanine (CpG) oligodeoxynucleotides (ODNs) have been shown to have potentials as adjuvants for vaccines. However, the immunostimulatory effect of CpG is species-specific and depends on the sequence of CpG motifs. A CpG ODN (2135), containing 3 identical copies of GTCGTT motif, was previously reported to have the strongest effects on bovine peripheral blood mononuclear cells (PBMC). Based on the sequence of 2135, we replaced the GTCGTT motif with 11 other sequences containing CG and investigated their effects on bovine lymphocyte proliferation. Results showed that the CpG ODNs containing 3 copies of GACGTT motif had the highest lymphocyte stimulation index (7.91±1.18), which was significantly (P<0.05) higher than that of 2135 (4.25±0.56). The CpG ODNs containing 3 copies of GACGTT motif also significantly increased the mRNA expression of interferon (IFN)-α, interleukin (IL)-12, and IL-21 in bovine PBMC. When dairy cows were immunized with the keyhole limpet hemocyanin (KLH) antigen formulated with CpG ODNs containing 3 copies of GACGTT, production of KLH-specific antibodies in serum and in milk whey was significantly (P<0.05) enhanced. IFN-γ in whole blood stimulated by KLH was also significantly (P<0.05) increased in cows immunized with KLH plus CpG ODNs. Our results indicate that CpG ODNs containing 3 copies of the GACGTT motifs is a potential adjuvant for bovine vaccines.
authors.affiliation: Graduate Institute of Animal Vaccine Technology, National Pingtung University of Science and Technology, Neipu, Taiwan.
authors.forename: Chun-Yen
authors.initials: CY
authors.lastname: Chu
authors.forename: Shang-Chun
authors.initials: SC
authors.lastname: Lee
authors.forename: Shyh-Shyan
authors.initials: SS
authors.lastname: Liu
authors.forename: Yu-Ming
authors.initials: YM
authors.lastname: Lin
authors.forename: Perng-Chi
authors.initials: PC
authors.lastname: Shen
authors.forename: Chi
authors.initials: C
authors.lastname: Yu
authors.forename: Kuo-Hua
authors.initials: KH
authors.lastname: Lee
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.forename: Jai-Wei
authors.initials: JW
authors.lastname: Lee
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D000941:Antigens / Q000276:immunology*; D001483:Base Sequence; D002417:Cattle / Q000276:immunology*; D049109:Cell Proliferation; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D015226:Dinucleoside Phosphates / Q000494:pharmacology; D005260:Female; D005620:Freund's Adjuvant / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D006433:Hemocyanins / Q000276:immunology*; D007106:Immune Sera / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D007075:Immunoglobulin M / Q000378:metabolism; D007371:Interferon-gamma / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects / Q000502:physiology; D008892:Milk / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D011247:Pregnancy; D051217:Toll-Like Receptor 9 / Q000235:genetics / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D000941:Antigens; D016207:Cytokines; D015226:Dinucleoside Phosphates; D005620:Freund's Adjuvant; D006433:Hemocyanins; D007106:Immune Sera; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007371:Interferon-gamma; D009838:Oligodeoxyribonucleotides; D051217:Toll-Like Receptor 9; C015772:CpG dinucleotide; C032808:keyhole-limpet hemocyanin