Newcastle disease vaccine virus I-2 fails to acquire virulence during repeated passage in vivo

Shahn P R Bisschop; Andrew Peters; Gil Domingue; Michael C Pearce; Jeanette Verwey; Petrus Poolman

doi:10.12688/gatesopenres.13212.3

Newcastle disease vaccine virus I-2 fails to acquire virulence during repeated passage in vivo

Gates Open Res. 2021 Dec 23:5:76. doi: 10.12688/gatesopenres.13212.3. eCollection 2021.

Authors

Shahn P R Bisschop^{1

2}, Andrew Peters^{3

4}, Gil Domingue^{3

5}, Michael C Pearce^{3

6}, Jeanette Verwey⁷, Petrus Poolman^{2

8}

Affiliations

¹ Avimune (Pty.) Ltd, Queenswood, Pretoria, South Africa.
² Poultry Reference Centre, Department of Production Animal Studies, Faculty of Veterinary Science, University of Pretoria, Pretoria, South Africa.
³ GALVmed, Pentlands Science Park, Midlothian, Scotland, UK.
⁴ Supporting Evidence based Interventions-Livestock, University of Edinburgh, Easter Bush Campus, Midlothian, Scotland, UK.
⁵ GD Associates, Fairmilehead, Edinburgh, Scotland, UK.
⁶ Epidemiological Research Unit, Scotland's Rural College, An Lòchran, Inverness, UK.
⁷ Deltamune, Roodeplaat, Pretoria, South Africa.
⁸ Kuipers Group (Pty.) Ltd, Zeekoeigat, Pretoria, South Africa.

Abstract

Background This study determined whether the naturally attenuated, thermotolerant Newcastle disease vaccine virus I-2 could acquire virulence after five in vivo passages through SPF chickens. Methods Study design was to international requirements including European Pharmacopoeia, Ph. Eur., v9.0 04/2013:0450, 2013. I-2 Working Seed (WS) was compared with five-times-passaged I-2 WS (5XP WS) in intracerebral pathogenicity index (ICPI), F _o cleavage site sequencing and Safety tests. Results The first passage series used a 50% brain: 50% tracheal tissue challenge homogenate and was unsuccessful as I-2 was not detected after the fourth passage. A second passage series used 10% brain: 90% tracheal tissue homogenates. I-2 was isolated from tracheal tissue in each passage. However harvested titres were below the minimum challenge level (10 ⁷ EID ₅₀) specified for the ICPI and Safety tests, possibly reflecting I-2's inherently low pathogenicity (interestingly caecal tonsils yielded significant titres). Given this the WS and 5XP WS comparisons proceeded. ICPI values were 0.104 and 0.073 for the WS group and the 5XP WS group respectively confirming that I-2, whether passaged or not, expressed low pathogenicity. F ₀ amino-acid sequences for both WS and 5XP WS were identified as ¹¹²R-K-Q-G-R-↓-L-I-G ¹¹⁹ and so compatible with those of avirulent ND viruses. In safety, no abnormal clinical signs were observed in both groups except for two chicks in the 5XP WS group, where one bird was withdrawn due to a vent prolapse, and another bird died with inconclusive necropsy results. Conclusions: These data, the issue of low passage titres with little or no virus isolation from brain tissues and the genomic copy approach suggest a need to amend Ph. Eur. v9.0 04/2013:0450, 2013 for naturally attenuated, low pathogenicity vaccine viruses such as I-2. From an international regulatory perspective, the study provides further definitive data demonstrating that Newcastle disease vaccine virus I-2 is safe for use.

Keywords: European Pharmacopoeia; I-2; passage; Newcastle Disease; acquired virulence; village chickens.

Grants and funding

We thank the Bill and Melinda Gates Foundation (BMGF) and the Department for International Development (DFID) UK whose joint funding (Grant No. 49064) facilitated the sponsorship of these studies by GALVmed (Global Alliance for Veterinary Medicines). The views expressed in the submitted article are those of the authors and not an official position of the institution or funder.