Khác biệt trong sự mẫn cảm với Novirhabdo virus của Zebrafish dòng hoang dại và đột biến và vai trò của lymphocytes

t quả tốt hơn) hoặc thời gian lấy 
mẫu hơi sớm so với đáp ứng miễn dịch ở cá.
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DIFFERENTIAL SUSCEPTIBILITY OF WILD-TYPE AND T AND 
B LYMPHOCYTE DEFICIENT ZEBRAFISH INFECTED WITH A 
NOVIRHABDO VIRUS REVEALS THE ROLE OF LYMPHOCYTES
Nguyen Ngoc Du1*, Lorelei Ford2, Lora Petri- Hanson2, Larry Hanson2
ABSTRACT
In order to study the contribution of innate defenses and lymphocyte based immunity in protection 
against a primary viral infection in fish, wild-type and Rag1 mutant zebrafish (T and B lymphocyte 
deficient) were infected with a novirhabdovirus, Snakehead Rhabdovirus (SHRV), by intraperitoneal 
(IP) injection at different ages and temperatures. Both strains of fish were highly susceptible to 
SHRV. Diseased fish demonstrated exophthalmia, hemorrhaged fin bases and protruding scales. The 
susceptibilities of the wild-type decreased as the fish aged; 2-month post hatch fish had significantly 
more losses than 4 month old and 7 month old fish. Also, temperature had a significant effect on 
the susceptibility of the wild-type. The wild-type zebrafish held at 24°C had significantly higher 
mortality than those held at 28°C. In cell culture, SHRV had similar growth kinetics at 20°C, 
24°C, and 28°C suggesting that reduced virus replication was not a factor in the lower mortality. 
The age and temperature effects were not seen with the Rag1 mutants. There was a difference in 
mortality pattern between the wild-type and Rag1 mutant. All mortalities in the wild-type occurred 
between day 3 and 10 post-infection, whereas the Rag1 mutant fish continued to die for at least one 
more week. To assess differences in the response of NK and T-cells of Rag1 mutant and wild-type 
zebrafish we evaluated transcript levels of the interferon gamma (IFNγ) gene in the posterior kidney 
by quantitative reverse transcriptase PCR (qRT-PCR). Expression was low in un-exposed fish but 
increased substantially in both strains at day 2 after exposure to SHRV. IFNγ over 400-fold and 
100-fold in Rag1 mutant at 24°C and 28°C, respectively; and over 40-fold and 80-fold in wild-type 
at 24°C and 28°C, respectively. The strong IFNγ response in lymphocyte deficient fish suggests 
Natural Killer cell activation. MxA (Myxovirus Resistance gene A) expression also increased in 
non- infected zebrafish (at day 8) and in SHRV- infected fish (both strains) at day 2 and day 8. We 
attempted to evaluate the initial activation of the acquired responses by evaluating of Blimp-1 and 
CD40 ligand expression at day 8, and at day 1 and day 7 after a second challenge, but no significant 
induction was detected at this early time point. 
The infection data suggest that innate immunity is important in controlling virus infection early 
during the infection, but then the acquired immune system clears the infection. Furthermore, 
age and the environmental temperature have a strong effect on this acquired immune response. 
These results may help explain the temperature and age associated resistance observed in other 
novirhabdovirus diseases in fish. 
Keywords: Zebrafish, Rag1, T and B lymphocyte deficient, Snakehead Rhabdovirus, interferon 
gamma, MxA, Blimp-1, CD40 ligand.
Người phản biện: TS. Lê Hồng Phước
Ngày nhận bài: 11/12/2017
Ngày thông qua phản biện: 25/12/2017
Ngày duyệt đăng: 30/12/2017
1 Southern Monitoring Center for Aquaculture Environment and Epidemic, Research Institute for Aquaculture No.2. 
2 Mississippi State University
*Email: ngocduaqua@yahoo.com