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Choi, Youn, Hong, Park, Yuk, Kwon, Noh, Kang, Cho, Ryu, Lee, Park, Choi, Lee, and Song: Intranasal Administration Model for Evaluating Protection Against Influenza Virus in Mice
Original Article

J Bacteriol Virol 2015;45(1):44-50.

Published online: 9 February 2015

DOI: https://doi.org/10.4167/jbv.2015.45.1.44

Intranasal Administration Model for Evaluating Protection Against Influenza Virus in Mice

Soo-Won Choi1, †, Ha-Na Youn1, †, Wootack Hong1, Jae-Keun Park1, Seong-Su Yuk1, Jung-Hoon Kwon1, Jin-Yong Noh1, Jung-Sun Kang2, Kyung-Jin Cho2, Jeoung-Jin Ryu2, Joong-Bok Lee1, Seung-Yong Park1, In-Soo Choi1, Sang-Won Lee1, Chang-Seon Song1, *

1Avian Disease Laboratory, College of Veterinary Medicine, Konkuk University, Seoul, Korea

2GENEBIOTECH CO., LTD, 166 Sinwonsa-ro, Gyeryong-myeon, Gongju-si, Chungcheongnam-do, Korea

*Corresponding author: Chang-Seon Song, Ph.D. Avian Disease Laboratory, College of Veterinary Medicine, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701, Korea. Phone: +82-2-450-3712, Fax: +82-2-455-3712, e-mail: songcs@konkuk.ac.kr

** This research was supported by bio-industry technology development program No. 312059-03, Ministry of Agriculture, Food and Rural Affairs.

† Both authors contributed equally to this work.

Received 25 November 201421 January 2015       Accepted 4 February 2015

Copyright © 2015 Journal of Bacteriology and Virology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Antiviral activity against Influenza virus of 14 Lactobacillus species isolated from food was monitored. Lactobacillus species were isolated from traditional Korean fermented food. Each live Lactobacillus was administered into the nasal cavity of SPF 6-week-old BALB/c mice. After the Lactobacillus treatment, Influenza virus (A/NWS/33/H1N1) was inoculated to each mouse. Clinical signs and mortality was monitored for 21 days. Each Lactobacillus strain showed various level of antiviral activity against Influenza virus. As a result of this study, this mouse experiment model, including intranasal treatment of live Lactobacillus species, could be effective model in evaluating immunomodulatory response of probiotics against respiratory viruses.

Keywords: Animal model, Probiotics, Influenza, Antiviral

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Figure 1.
Schedule for animal experiment. BALB/c mice were treated with intranasally with Lactobacillus for 2 weeks before inoculation with A/NWS/33 virus. Mice were monitored daily for clinical signs for 14 days.
jbv-45-44f1.tif
Figure 2.
Rooted neighbor-joining tree based on 16s rRNA sequences showing relationships between Lactobacillus strains. The scale bar indicates 0.1 nucleotide substitution per nucleotide position.
jbv-45-44f2.tif
Table 1.
Origin of isolated Lactobacillus strains
Lactobacillus strain No. Origin Genetic ClusterA
Lactobacillus strain No. 1–3 Pickled Radish Group 1
Lactobacillus strain No. 2–6 Pickled Pepper Group 1
Lactobacillus strain No. 3–5 Pickled Pepper Group 1
Lactobacillus strain No. 3–6 Dongchimi Group 1
Lactobacillus strain No. 3–8 Pickled Pepper Group 1
Lactobacillus strain No. 3–8-1 Dongchimi Group 1
Lactobacillus strain No. 4–20 Pickled Radish Group 1
Lactobacillus strain No. 5–9 Dongchimi Group 4
Lactobacillus strain No. 5–11 Salted shrimp Group 1
Lactobacillus strain No. 6–5 Pickled Pepper Group 1
Lactobacillus strain No. 6–13 White Kimchi Group 1
Lactobacillus strain No. 7–1 Dongchimi Group 3
Lactobacillus strain No. 7–13 Pickled Pepper
Lactobacillus strain No. 7–25 Salted shrimp Group 2

A Genetic Cluster was determined by phylogenetic tree based on 16s rRNA

Table 2.
The effect of intranasal administration of various Lactobacillus species on challenge with influenza virus. BALB/c mice were treated intranasally with Lactobacillus species for 2 weeks before inoculation with A/NWS/33 virus. Mice were monitored daily for clinical signs for 21 days. Statistical significance was determined by Fisher's exact test. Asterisks indicates significant differences (*p < 0.05, **p < 0.01) compared with results in positive control
Group Administered Lactobacillus species Number of mice Preventive effect after challenge with influenza virusA
 PI14C
Survival MDT (day)B
1 Lactobacillus 1–3 10 6/10 10.0 1.4
2 Lactobacillus 2–6 10 8/10** 9.0 1.2
3 Lactobacillus 3–5 10 8/10** 10.5 1.2
4 Lactobacillus 3–6 10 4/10 10.5 1.6
5 Lactobacillus 3–8 10 8/10** 12.0 1.2
6 Lactobacillus 3-8-1 10 4/10 8.5 1.6
7 Lactobacillus 4–20 10 3/10 10.1 1.7
8 Lactobacillus 5–9 10 4/10 10.8 1.6
9 Lactobacillus 5–11 10 5/10 8.4 1.5
10 Lactobacillus 6–5 10 8/10** 11.0 1.2
11 Lactobacillus 6–13 10 5/10 10.6 1.5
12 Lactobacillus 7-1 10 2/10 10.5 1.8
13 Lactobacillus 7-13 10 5/10 9.0 1.5
14 Lactobacillus 7–25 10 6/10 9.8 1.4
15 Positive control 10 1/10 9.0 1.9
16 Negative control 10 10/10 − 0

A Mice were inoculated intranasally with 104.0EID50 of influenza virus A/NWS/33 (H1N1) strain

B Mean death time

C Pathogenicity index: average score of clinical sign were recorded following challenge and were scored as follow: 0 = normal, 1 = sick, 2 = dead

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