Recent Progress in Understanding Host Mucosal Response to Avian Coccidiosis and Development of Alternative Strategies to Mitigate the Use of Antibiotics in Poultry Production

Hyun Soon Lillehoj,Sung Hyen Lee,Seung Ik Jang,Duk Kyung Kim,Kyung Woo Lee 한국가금학회 2011 韓國家禽學會誌 Vol.38 No.4

In ovo vaccination using Eimeria profilin and Clostridium perfringens NetB proteins in Montanide IMS adjuvant increases protective immunity against experimentally-induced necrotic enteritis

Lillehoj, Hyun Soon,Jang, Seung Ik,Panebra, Alfredo,Lillehoj, Erik Peter,Dupuis, Laurent,Arous, Juliette Ben,Lee, Seung Kyoo,Oh, Sung Taek Asian Australasian Association of Animal Productio 2017 Animal Bioscience Vol.30 No.10

Objective: The effects of vaccinating 18-day-old chicken embryos with the combination of recombinant Eimeria profilin plus Clostridium perfringens (C. perfringens) NetB proteins mixed in the Montanide IMS adjuvant on the chicken immune response to necrotic enteritis (NE) were investigated using an Eimeria maxima (E. maxima)/C. perfringens co-infection NE disease model that we previously developed. Methods: Eighteen-day-old broiler embryos were injected with $100{\mu}L$ of phosphate-buffered saline, profilin, profilin plus necrotic enteritis B-like (NetB), profilin plus NetB/Montanide adjuvant (IMS 106), and profilin plus Net-B/Montanide adjuvant (IMS 101). After post-hatch birds were challenged with our NE experimental disease model, body weights, intestinal lesions, serum antibody levels to NetB, and proinflammatory cytokine and chemokine mRNA levels in intestinal intraepithelial lymphocytes were measured. Results: Chickens in ovo vaccinated with recombinant profilin plus NetB proteins/IMS106 and recombinant profilin plus NetB proteins/IMS101 showed significantly increased body weight gains and reduced gut damages compared with the profilin-only group, respectively. Greater antibody response to NetB toxin were observed in the profilin plus NetB/IMS 106, and profilin plus NetB/IMS 101 groups compared with the other three vaccine/adjuvant groups. Finally, diminished levels of transcripts encoding for proinflammatory cytokines such as lipopolysaccharide-induced tumor necrosis $factor-{\alpha}$ factor, tumor necrosis factor superfamily 15, and interleukin-8 were observed in the intestinal lymphocytes of chickens in ovo injected with profilin plus NetB toxin in combination with IMS 106, and profilin plus NetB toxin in combination with IMS 101 compared with profilin protein alone bird. Conclusion: These results suggest that the Montanide IMS adjuvants potentiate host immunity to experimentally-induced avian NE when administered in ovo in conjunction with the profilin and NetB proteins, and may reduce disease pathology by attenuating the expression of proinflammatory cytokines and chemokines implicated in disease pathogenesis.

Recent Progress in Understanding Host Mucosal Response to Avian Coccidiosis and Development of Alternative Strategies to Mitigate the Use of Antibiotics in Poultry Production

Lillehoj, Hyun-Soon,Lee, Sung-Hyen,Jang, Seung-Ik,Kim, Duk-Kyung,Lee, Kyung-Woo The Korean Society of Poultry Science 2011 韓國家禽學會誌 Vol.38 No.4

As the world population grows and developing countries become more affluent, the global consumption of meat will increase by more than 50% within the next 10 years. Confronting the increased demand for poultry food products are emerging field diseases, increasing regulatory bans of antimicrobial growth promoters, high-density growth conditions, and waste management. Although biotechnology offers solutions to some of these challenges, basic studies are needed to better understand the complex interaction between the intestinal microbiome, host immunity and the environment. This presentation will focus on emerging strategies to enhance gut immunity and to decrease economic losses due to poultry diseases. This presentation will highlight recent developments in coccidiosis research and provide information on host immunity, immunomodulation, and the latest advances in dietary and nutritional approaches against coccidiosis. Such information will magnify our understanding of host-parasite biology, mucosal immunology, and design of future nutritional interventions and vaccination strategies for coccidiosis.

Avian leucocyte common antigens : molecular weight determination and flow cytometric analysis using new monoclonal antibodies

Chung, Kyeong-Soo,Hyun Soon Lillehoj,Mark Christopher Jenkins 충남대학교부설 생명공학연구소 1992 생물공학연구지 Vol.2 No.-

Chung, K.S., Lillehoj, H.S. and Jenkins, M.C., 1991. Avian leucocyte common antigens: molecular weight determination and flow cytometric analysis using new monoclonal antibodies. Vet. Immunol. Immunopathol., 28: 259-273. New leucocyte common antigens expressed on all the normal chicken leucocytes have been characterized using two monoclonal antibodies designated as K-11 and K-55. These monoclonal antibodies stained virtually 100% of the leucocytes derived from various lymphoid organs including the spleen, thymus, bursa of Fabricius, caecal tonsil and peripheral blood, as well as a monocytic cell line (MC29), aB cell line (LSCC-RP9), and a T cell line (CU12). However, they did not stain mature erythrocytes, intestinal epithelial cells, or chicken embryonic fibroblasts. The two monoclonal antibodies showed different staining patterns and detected non-overlapping epitopes on MC29 cells in two color immunofluorescence analysis. Western blot analysis under non-reducing conditions showed that the monoclonal antibody K-11 recognized three splenic leucocyte proteins with molecular weights of 92, 42 and 41 kDa, whereas the monoclonal antibody K-55 recognized two proteins with molecular weights of 97 and 42 kDa. The data indicate that the monoclonal antibodies K-11 and K-55 recognize novel leucocyte-common antigens which have lower molecular weights than the previously reported leucocyte-common antigen family.

Development and functional characterization of monoclonal antibodies recognizing chicken lymphocytes with natural killer cell activity

Chung, Kyeong-Soo,Hyun Soon Lillehoj 충남대학교부설 생명공학연구소 1992 생물공학연구지 Vol.2 No.-

Chung, K.S. and Lillehoj, H.S., 1991. Development and functional characterization of monoclonal antibodies recognizing chicken lymphocytes with natural killer cell activity. Vet. Immunol. Immunopathol, 28: 351-363. We have developed two monoclonal antibodies which detect cell surface antigens present on chicken lymphocytes mediating natural killer (NK) cell activity against the avian tumor cell target. The monoclonal antibodies, K-14 and K-108, stained 17 and 6% of splenic lymphocytes, and 11 and 14% of peripheral blood lymphocytes (PBL), respectively, and fewer than 5% of thymic and bursal lymphocytes. Neither of these monoclonal antibodies stained adherent macrophages or the MC29-virus transformed monocytic cell line. Both monoclonal antibodies significantly inhibited Nk cell activity in a standard 4 h ^51Cr-release cytotoxicity assay using the LSCC-RP9 tumor cell line as target cells at an effector to target ratio of 50:1. Pretreatment of splenocytes with either monoclonal antibody in the presence of rabbit complement (C) resulted in a significant reduction in NK cell activity. However, the monoclonal antibody K-1 which detects normal chicken macrophages did not interfere with NK cell activity. The monoclonal antibody K-108 significantly blocked Fc receptor-mediated rosette formation of sheep red blood cells coated with IgG antibodies (EA) by 56% while the monoclonal antibody K-14 did not show a significant blocking. These results indicate that the monoclonal antibodies K-108 and K-14 identify different epitopes present on the surface of chicken splenic lymphocytes which mediate spontaneous NK cytotoxicity.

Development and functional characterization of monoclonal antibodies recognizing chicken lymphocytes with natural killer cell activity

Chung, Kyeong-Soo,Lillehoj, Hyun Soon 충남대학교 약학대학 의약품개발연구소 1991 藥學論文集 Vol.7 No.-

Chung, K.S. and Lillehoj, H.S., 1994. Development and functional characterization of monoclonal antibodies recognizing chicken lymphocytes with natural killer cell activity. Vet. Immunol. Immunopathol., 28:351-363. We have developed two monoclonal antibodies which detect cell surface antigens present on chicken lymphocytes mediating natural killer (NK) cell activity against the avian tumor cell target. The monoclonal antibodies, K-14 and K-108, stained 17 and 6% of splenic lymphocytes, and 11 and 14% of peripheral blood lymphocytes (PBL), respectively, and fewer than 5% of thymic and bursal lymphocytes. Neither of these monoclonal antibodies stained adherent macrophages or the MC29-virus transformed monocytic cell line. Both monoclonal antibodies significantly inhibited NK cell activity in a standard 4h ^51Cr-release cytotoxicity assay using the LSCC-RP9 tumor cell line as target cells at an effector to target ratio of 50:1. Pretreatment of splenocytes with either monoclonal antibody in the presence of rabbit complement (C) resulted in a significant reduction in NK cell activity. However, the monoclonal antibody K-1 which detects normal chicken macrophages did not interfere with NK cell activity. The monoclonal antibody K-108 significantly blocked Fc receptor-mediated rosette formation of sheep red blood cells coated with IgG antibodies (EA) by 56% while the monoclonal antibody K-14 did not show a significant blocking. These results indicate that the monoclonal antibodies K-108 and K-14 identify different epitopes present on the surface of chicken splenic lymphocytes which mediate spontaneous NK cytotoxicity.

Protective immunity against Eimeria infection using dendritic cells or exosomes

Emilio del Cacho,Margarita Gallego,Hyun Soon Lillehoj 한국가금학회 2012 한국가금학회 심포지움 Vol.2012 No.5

Coccidiosis is a complex intestinal disease of major economic importance in chickens that is caused by multiple species of the protozoan, Eimeria. Conventional disease control methods have relied on prophylactic administration of drugs with anticoccidial activity, or on vaccination with live or attenuated parasites. However, alternative methods of disease mitigation are needed due to increasing government restrictions on the use of coccidiostats. An immunization strategy against Eimeria tenella infection was for the first time undertaken using parasite antigen(Ag)-loaded dendritic cells(DCs), or their derived exosomes, in the absence of free Ag. In a first clinical trial, chickens were immunized with E.tenella Ag-loaded DCs or exosomes and subsequently given a live parasite challenge infection to determine which immunization was most efficient in preventing disease. Immune parameters demonstrated that increased protective immunity against E.tenella infection was induced in chickens by immunization with Ag-loaded exosomes, compared with chickens immunized with Ag-loaded DCs or Ag alone. These findings led us to undertake a second clinical trial to evaluate Ag-loaded exososomes as potential vaccines against coccidiosis. Chickens were immunized with exosomes loaded with Ags from E.tenella, E.acervulina, and E.maxima. Subsequently, the animals were coinfected with oocysts from these three eimerian species. Cecal tonsils, Peyer's patches and spleens of immunized and infected chickens had increased numbers of cells secreting IL-16, IL-2, and IFN-γ, greater Ag-stimulated proliferative responses, and higher numbers of Ag-reactive IgG- and IgA-producing cells following in vitro stimulation with the Ags, compared with the non-immunized/non-infected and non-immunized/infected controls. In addition, immunized and infected chickens had increased body weight gains, reduced feed conversion ratios, diminished fecal oocyst shedding, lessened intestinal lesion scores, and reduced mortality compared with the non-immunized/infected controls. These results suggest that successful vaccination may be possible against avian coccidiosis using exosomes derived from DCs incubated with Ags isolated from Eimeria species.