Sains Malaysiana 45(5)(2016): 787–793

Replication of a Malaysian Strain Avian Influenza A Virus H5N1 in Madin-Darby Canine Kidney and African Green Monkey Kidney Cells

(Replikasi Virus Influenza Avian A Jenis H5N1 dalam Sel Ginjal Kanin Madin-Darby dan Monyet Hijau Afrika)

TOONG SENG TAN¹, SHARIFAH SYED HASSAN² & WEI BOON YAP¹*

¹Biomedical Science Programme, School of Diagnostic and Applied Health Sciences, Faculty of Health Science, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia

²School of Medicine and Health Sciences, Monash University Malaysia, Jalan Lagoon Selatan

47500 Bandar Sunway, Selangor Darul Ehsan, Malaysia

Received: 1 September 2015/Accepted: 17 November 2015

ABSTRACT

The use of cell lines such as Madin-Darby Canine Kidney (MDCK) and African Green Monkey Kidney (Vero) cells in influenza vaccine production is much advocated presently as a safer alternative to chicken embryonated eggs. It is thus essential to understand the influenza virus replication patterns in these cell lines prior to utilizing them in vaccine production. The infectivity of avian influenza A virus (A/Chicken/Malaysia/5858/2004) H5N1 in MDCK and Vero cell lines was first assessed by comparing the cytopathic effect (CPE) caused by the virus infection. The viral loads in both of the infected media and cells were also compared. The results showed that both of the MDCK and Vero cells began to exhibit significant CPE (p<0.05) after 48 h post-infection (h p.i). The MDCK cell line was more susceptible to the virus infection compared to Vero cell line throughout the incubation period. A higher viral load was also detected in the host cells compared to their respective culturing media. Interestingly, after reaching its maximum titer at 48 h p.i, the viral load in MDCK cells declined meanwhile the viral load in Vero cells increased gradually and peaked at 120 h p.i. Overall, both cell lines support efficient H5N1 virus replication. While the peak viral loads measured in the two cell lines did not differ much, a more rapid replication was observed in the infected MDCK samples. The finding showed that MDCK cell line might serve as a more time-saving and cost-effective cell culture-based system compared to Vero cell line for influenza vaccine production.

Keywords: Cytopathic effect; H5N1; MDCK; Vero; viral load

ABSTRAK

Penggunaan kultur sel seperti sel ginjal kanin Madin-Darby (MDCK) dan monyet hijau Afrika (Vero) dalam penghasilan vaksin influenza adalah lebih selamat berbanding telur ayam berembrio yang disarankan pada masa ini. Maka, adalah penting untuk kita memahami corak replikasi virus influenza dalam sel-sel tersebut sebelum digunakan dalam penghasilan vaksin. Keboleh-jangkitan virus influenza jenis H5N1 (A/Chicken/Malaysia/5858/2004) dalam sel MDCK dan Vero dinilai dengan membandingkan kesan sitopatik (CPE) yang diakibatkan oleh jangkitan virus influenza. Titer virus dalam media dan sel terjangkit turut dibandingkan. Hasil kajian menunjukkan kedua-dua sel MDCK dan Vero mula mempamerkan CPE yang signifikan (p < 0.05) selepas 48 jam pasca-infeksi (h p.i). Sel MDCK adalah lebih rentan kepada jangkitan virus sepanjang tempoh eraman virus berbanding dengan sel Vero. Titer virus yang lebih tinggi diperoleh dalam sel perumah terjangkit berbanding dengan media kultur. Selepas mencapai titer maksimum pada 48 h p.i, titer virus dalam sel MDCK menurun manakala titer virus dalam sel Vero kekal meningkat secara perlahan dan memuncak pada 120 h p.i. Secara keseluruhan, kedua-dua sel perumah didapati mampu menyokong replikasi virus H5N1 dengan cekap. Meskipun titer virus maksimum yang dicapai antara kedua-dua sel perumah tidak berbeza, replikasi yang lebih pantas diperhatikan dalam sel MDCK. Hal ini mencadangkan penggunaan sel MDCK dalam penghasilan vaksin influenza adalah lebih jimat daripada segi masa dan kos berbanding dengan sel Vero.

Kata kunci: H5N1; kesan sitopatik; MDCK; titer virus; Vero

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*Corresponding author; email: yapweiboon@ukm.edu.my