Abstract

Outer membrane protein C of Edwardsiella tarda is a major cell surface antigen and it was identified to be an immunogenic protein by Western blot using flounder (Paralichthys olivaceus) anti-recombinant OmpC (rOmpC), and anti-E. tarda antibodies. rOmpC tested the immune protective effect against E. tarda challenge in a flounder model and produced a relative percentage of survival rate of 85%. The immune response of flounder induced by rOmpC was investigated, and the results showed that: (1) the levels of specific serum antibodies induced by rOmpC were significantly higher than the control group after the second week after immunization, and the peak level occurred at week five after immunization; (2) rOmpC could induce the proliferation of sIg+ lymphocytes, and the peak levels of sIg+ lymphocytes in blood, spleen, and pronephros occurred at 4–5 weeks after immunization; and (3) the MHCIIα, CD4-1, IL-1β, IL-6 and TNF-α genes were significantly induced after being injected with rOmpC. Taken together, these results demonstrated that rOmpC could evoke highly protective effects against E. tarda challenge and induce strong innate immune response and humoral immune response of flounder, which indicated that OmpC was a promising vaccine candidate against E. tarda infection.

Highlights

  • Edwardsiella tarda is a Gram-negative pathogen and the etiological agent of edwardsiellosis, which affects a wide range of marine and freshwater fish [1,2,3], causing enormous economic losses around the world [4,5]

  • Previous studies showed that a homologue of the 37 kDa protein, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) when in form of recombinant protein could produce immune-protective effects in flounder against edwardsiellosis [20,21]

  • The results showed that recombinant OmpC (rOmpC) could induce the proliferation of sIg+ lymphocytes and the peak levels in blood, spleen, and pronephros occurred at week 5, week 4 and week 5 after immunization

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Summary

Introduction

Edwardsiella tarda is a Gram-negative pathogen and the etiological agent of edwardsiellosis, which affects a wide range of marine and freshwater fish [1,2,3], causing enormous economic losses around the world [4,5]. IItt wwaass aallssoo rreeppoorrtteedd tthhaatt OOmmppCC ooff SSaallmmoonneellllaa rreevveeaalleedd bbeetttteerr pprrootteeccttiioonn aaggaaiinnsstt vviirruulleenntt SSaallmmoonneellllaa cchhaalllleennggee wwhheenn ccoommppaarreedd wwiitthh ffoorrmmaalliinn--kkiilllleedd,,wwhhoollee--cceellllbbaaccteterriaiallvvaaccccinineeininaabbiridrdmmooddelel[1[31,31,41]4.].MMooreroeovvere,ra, aShSigheiglleallflaefxlenxenrier3ia3OamOpmCpeCpietpopiteopweaws raescoregcnoizgendizbeyd hbuymhaunmuamnbuilmicbailliccoarldcsoerrda saenrda aasnsdocaiassteodciwatietdh wpriothtecptrivoeteacctitviveitay,ctmiviictey,immmicueniimzemd uwnitizheOdmwpiCthreOtaminpeCd lroentagi-nlaesdtinlognpgr-olatsetcitniognpargoateincstitoanleatghaailndsot sae loefthbaoltdhohsoemofobloogthouhsomanodlohgeotuersoalnogdohuestesrtoraloingsouosf sthtreaipnastohfotgheenps a[t1h5o].geAndsd[1it5io].nAadllyd,itOiomnaplCly,inOomthpeCr iGnroatmhe-nr eGgraatmiv-enbegaactteivreiab, ascutcehriaa,ssAucerhomasoAnaesrohmyodnroapshhiyladraonpdhilSa.aenndterSi.ceansteerriocvaasrerToyvpahriT, yhpashia, lhsaosbaeleson breepenorrteepdotrotehdatvoehtahvee pthoetepnottieanl tiinalviancvcaincecindeevdeelvoeplmopemnten[9t,[196,]1.6]E. IInntteerreessttiinnggllyy,, bbootthh ooff ttwwoo ddiiffffeerreenntt aannttiibbooddiieess ccoouulldd ssppeecciiffiiccaallllyy bbiinndd tthhee 3377 kkDDaa pprrootteeiinn ((FFiigguurree 22)). MMaassss ssppeeccttrroommeettrriicc rreessuullttss ooff tthhee 3377 kkDDaa pprrootteeiinn.

Flow Cytometric Immunofluorescence Analysis
Discussion
Expression and Purification of Recombinant Outer Membrane Protein C
Analysis of the Immunogenicity of OmpC
Vaccination and Sampling
Analysis of the Expression of Immune-Related Genes by qRT-PCR
Challenge
Statistical Analysis
Conclusions
Full Text
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