Recombinant interleukin 6 with M cell-targeting moiety produced in Lactococcus lactis IL1403 as a potent mucosal adjuvant for peroral immunization.
Development and application of safe and effective mucosal adjuvants are important to improve immunization efficiency in oral vaccine. Here, we report a novel mucosal adjuvant, IL-6-CKS9, a recombinant cytokine generated by conjugating an M cell-targeting peptide (CKS9) with c-terminus of the murine interleukin 6 (IL-6), which facilitated enhancement of mucosal immune response. Lactococcus lactis IL1403, a food-grade strain of lactic acid bacteria (LAB) which is widely used in dairy industry, was used as a host cell to express and secrete the IL-6-CKS9 for a mucosal vaccine adjuvant. The recombinant L. lactis IL1403 secreting IL-6-CKS9 was orally administered with a model antigen protein, M-BmpB (Brachyspira membrane protein B conjugated with CKS9), to BALB/c mice for mucosal immunization. ELISA analyses showed consistent enhancement tendencies in induction of anti-M-BmpB antibody levels both with mucosal (IgA) and systemic (IgG) immune responses in IL-6-CKS9-LAB treated group compared with other groups tested by conducting two separated mice immunization assays. In addition, we characterized that the oral administration of model protein antigen with live LAB producing IL-6-CKS9 could induce both Th1 and Th2 type immune responses by analysis of the specific anti-BmpB IgG1 and IgG2a isotypes in the sera and also investigated possible oral tolerance in our vaccine strategy. Collectively, our results showed successful production and secretion of recombinant murine IL-6 with M cell-targeting moiety (IL-6-CKS9) from L. lactis IL1403 and demonstrated the live recombinant LAB producing IL-6-CKS9 could have a potential to be used as an efficient adjuvant for peroral vaccination.
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea.
Hui-Shan
HS
Li
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea.
Da-Chuan
DC
Piao
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea.
Tao
T
Jiang
Institute of Green-Bio Science & Technology, Seoul National University, Pyeongchang-gun 232-916, Republic of Korea.
Jin-Duck
JD
Bok
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea.
Chong-Su
CS
Cho
Institute of Green-Bio Science & Technology, Seoul National University, Pyeongchang-gun 232-916, Republic of Korea.
Yoon-Seok
YS
Lee
Institute of Green-Bio Science & Technology, Seoul National University, Pyeongchang-gun 232-916, Republic of Korea. Electronic address: kangsk01@snu.ac.kr.
Sang-Kee
SK
Kang
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea. Electronic address: cyjcow@snu.ac.kr.
Yun-Jaie
YJ
Choi
D000276:Adjuvants, Immunologic; D000284:Administration, Oral; D000818:Animals; D005260:Female; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors; D007108:Immune Tolerance; D018928:Immunity, Mucosal; D007114:Immunization; D015850:Interleukin-6 / Q000235:genetics / Q000276:immunology*; D007413:Intestinal Mucosa / Q000276:immunology; D013294:Lactococcus lactis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D010455:Peptides / Q000235:genetics / Q000276:immunology*; D010581:Peyer's Patches / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000276:immunology*
D000276:Adjuvants, Immunologic; D015850:Interleukin-6; D010455:Peptides; D011993:Recombinant Fusion Proteins
Adjuvant; Interleukin-6; Lactococcus lactis IL1403; M-cell targeting peptide; Mucosal immune response; Peroral vaccination