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Largemouth bass iridovirus (LMBV) can cause high mortality and lead to heavy economic loss in the cultivation of largemouth bass, but there was no effective treatment. Here, the present study constructed a recombinant  Pichia pastoris  expressing LMBV major capsid protein ( MCPD ). The recombinant GS115-pW317- MCPD  was then used to immunize largemouth bass  via  oral administration, and mucosal immune response mediated by immunoglobulins (Igs) was measured after oral immunization. Serum antibody levels were measured by ELISA, neutralizing antibody titers were determined by serum neutralization test (SNT), antigen presentation-related gene expressions were detected by RT-PCR, and the histopathological characteristics of immunized fish were assessed after challenging with 0.1 ml 10 7.19  TCID 50 /ml LMBV. The relative percentage survival (RPS) was also determined. Our results showed that the serum antibody titers of immunized fish were significantly higher than that of control groups (P &amp;lt; 0.05). IgT and IgM expressions in gut were increased significantly after vaccination with GS115-pW317- MCPD ; however, much stronger response in gut was observed as compared with gill. The expression levels of major histocompatibility complex ( MHC ) II,  CD 8, and T-cell receptor ( TCR ) were significantly elevated in GS115-pW317- MCPD  group (P &amp;lt; 0.05), while  CD 4 and  MHC  I transcription levels remained unchanged after oral immunization ( P  &amp;gt; 0.05). The RPS of fish orally immunized with 1.0 × 10 8  CFU/g GS115-pW317- MCPD  was reached up to 41.6% after challenge with 0.1 ml 10 9.46  TCID 50 /ml LMBV. Moreover, orally immunizing with GS115-pW317- MCPD  can relieve the pathological damage caused by LMBV. Therefore, GS115-pW317- MCPD  showed a promising potential against LMBV.

frontiers in immunology

Oral Vaccination With Recombinant Pichia pastoris Expressing Iridovirus Major Capsid Protein Elicits Protective Immunity in Largemouth Bass (Micropterus salmoides)