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

Received: 13 November 2015/Accepted: 21 January 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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*Corresponding author; email: swkim@gntech.ac.kr