Sains Malaysiana 45(6)(2016): 933–940
Development
of Salmonella typhimurium Ghost Vaccine
using Asd-Based Vector System with E
Gene and the Immune Responses Evoked in BALB/c Mice
(Pembangunan
Vaksin Kosong Salmonella typhimurium menggunakan
Sistem Vektor Berasaskan Asd dengan Gen E dan Meningkatkan
Tindak Balas Imun pada Tikus BALB/c)
SE WON KIM1 , JEONG MAN ROH2, JOHN HWA LEE3, SUNG SILL LEE4, JEONG DONG BAHK1, TAE WAN KIM5, IL-SUK KIM5 & SAM WOONG KIM5*
1Division
of Applied Life Sciences (BK21 & EBNCRC), Graduate School of Gyeongsang, National
University, Jinju 660-701, South Korea
2Cheonryeong-Pork
Genetics, 588-3 Yieun-ri, Hamyang-eup, Hamyang-gun, Gyeongnam
South
Korea
3College
of Veterinary Medicine, Chonbuk National University, Chon-ju 561-756, South
Korea
4Department
of Animal Science and Biotechnology, Gyeongsang National University, Jinju
660-701
South Korea
5Swine Science and
Technology Center, Gyeongnam National University of Science & Technology, Jinju
660-758, South Korea
Diserahkan: 13 November 2015/Diterima: 21 Januari 2016
ABSTRACT
Salmonella typhimurium MMP13 harboring pMMP99,
a recombinant plasmid derived by cloning a ghost cassette into T-vector, were
employed for production of ghost cells. Growth of MMP13
(pMMP99) showed an initial increase after a shift in
temperature from 28 to 42°C, and then decreased gradually with ghost formation
being complete within 3 h. Ghost yield of MMP13
(pMMP99) were 99.99% less than 104 CFU/mL. MMP13
(pMMP101-1) showed a ghost yield similar to MMP13
(pMMP99). Immune responses of BALB/c
mice against ghost cells originated from MMP13
(pMMP101-1) were assessed by measuring total IgG, IgG1,
IgG2a, and secretory IgA levels in each sample. Levels of total IgG, IgG1 and
IgG2a and vaginal IgA, increased abruptly after 4 weeks post vaccination,
whereas the fecal IgA level did not induce significant change. Splenocyte
proliferation was observed at the cellular level owing to stimulation of ghost
cells. Ghost vaccination protected 25-59% of mice against wild-type S.
typhimurium more than those of controls.
Keywords: Ghost cassette; ghost vaccine; immune response; non-antibiotic
marker; protection; Salmonella typhimurium
ABSTRAK
Salmonella typhimurium MMP13
melindungi pMMP99, plasmid rekombinan yang diperoleh melalui
pengklonan kaset kosong ke dalam T-vektor, telah digunakan untuk pengeluaran
sel kosong. Pertumbuhan MMP13
(pMMP99) menunjukkan peningkatan awal selepas berlaku
perubahan dalam suhu daripada 28 kepada 42°C dan kemudian menurun secara
beransur-ansur dengan pembentukan kosong yang lengkap dalam masa 3 jam. Hasil kosong MMP13 (pMMP99)
adalah 99.99% kurang daripada 104 CFU/mL. MMP13
(pMMP101-1) menunjukkan hasil kosong adalah sama dengan MMP13 (pMMP99). Tindak balas imun tikus BALB/c
terhadap sel kosong daripada MMP13 (pMMP101-1)
dinilai dengan mengukur jumlah IgG, IgG1, IgG2a dan tahap IgA rembes dalam
setiap sampel. Tahap jumlah IgG, IgG1 dan IgG2a serta IgA faraj
meningkat secara mendadak selepas 4 minggu vaksinasi, manakala tahap IgA tahi
tidak menunjukkan perubahan ketara. Percambahan splenocyte
diperhatikan pada tahap sel kerana rangsangan sel kosong. Vaksinasi
kosong melindungi 25-59% tikus daripada S. typhimurium jenis liar lebih daripada kawalan.
Kata kunci: Kaset kosong; penanda bukan antibiotik; perlindungan; Salmonella typhimurium; tindak balas imun; vaksin kosong
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*Pengarang
untuk surat-menyurat; email: swkim@gntech.ac.kr
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