Cox, Eric

Goddeeris, Bruno

Goetstouwers, Tiphanie

Melkebeek, Vesna

Nguyen Van, Ut

Peelman, Luc

Van Poucke, Mario

English

Ghent University Academic Bibliography

Title: ELISPOT ON ENRICHED IgA+ B-LYMPHOCYTES CAN BE USED TO DETECT THE IgA RESPONSE FOLLOWING ORAL IMMUNIZATION WITH F4 FIMBRIAE OF PIGS WITH COLOSTRAL F4-SPECIFIC SERUM ANTIBODIES  Authors & affiliations: NGUYEN V. U.(1), MELKEBEEK V.(1), GOETSTOUWERS T.(2), VAN POUCKE M.(2), PEELMAN L.(2), GODDEERIS B.M.(1,3), COX E(1). (1) Laboratory of Immunology, Faculty of Veterinary medicine, Ghent University Salisburylaan 133, Merelbeke 9820, Belgium (2) Laboratory of Animal Genetics, Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, 9820 Merelbeke, Belgium (3) Department of Biosystems, Faculty of Bioscience Engineering, KULeuven, Kasteelpark Arenberg 30, B-3001 Leuven, Belgium Nguyen.vanut@ugent.be   Diarrhoea	  and	  mortality	  in	  piglets	  are	  mainly	  caused	  by	  F4	  enterotoxigenic	  Escherichia	  coli	  (ETEC)	  and	  lead	  to	  serious	  economical	  losses.	  	  Oral	  immunization	  of	  F4	  receptor	  positive	  F4-­‐seronegative	  pigs	  with	  F4	  fimbriae	  can	  induce	  a	  protective	  intestinal	  immune	  response	  evidenced	  by	  an	  F4	  specific	  serum	  and	  intestinal	  mucosal	  IgA	  response.	  However,	  on	  farms	  most	  pigs	  obtain	  maternal	  F4-­‐specific	  colostral	  IgG	  at	  birth	  and	  F4-­‐specific	  IgA	  from	  milk	  during	  the	  suckling	  period	  resulting	  in	  the	  presence	  of	  F4-­‐specific	  serum	  antibodies.	  The	  aim	  of	  the	  present	  study	  was	  to	  determine	  if	  piglets	  with	  F4-­‐specific	  passive	  immunity	  could	  be	  orally	  immunized	  with	  F4.	  Whereas	  the	  F4-­‐specific	  immune	  response	  of	  orally	  immunized	  F4-­‐seronegative	  pigs	  can	  be	  easily	  measured	  in	  serum,	  maternal	  antibodies	  will	  obscure	  the	  response	  in	  orally	  immunized	  pigs	  with	  colostral	  F4-­‐specific	  serum	  antibodies.	  In	  the	  current	  experiment,	  a	  protocol	  was	  optimized	  to	  detect	  the	  F4-­‐specific	  IgA	  response	  following	  oral	  immunization	  of	  3-­‐	  to	  4-­‐week-­‐old	  pigs	  with	  passive	  immunity.	  As	  controls,	  pigs	  that	  were	  intramuscularly	  immunized	  and	  pigs	  that	  are	  not	  immunized	  were	  included.	  To	  determine	  the	  humoral	  F4-­‐specific	  immune	  response,	  blood	  was	  taken	  weekly	  until	  3	  weeks	  post	  immunization	  and	  ELISA’s	  were	  performed	  to	  determine	  the	  F4-­‐specific	  serum	  IgA	  antibody	  titres	  as	  well	  as	  ELIspot	  assays	  to	  enumerate	  the	  induced	  F4-­‐specific	  antibody	  secreting	  cells.	  The	  ELIspot	  assays	  were	  determined	  on	  peripheral	  blood	  mononuclear	  cells	  (PBMCs)	  following	  density	  gradient	  centrifugation	  and	  on	  the	  IgA+	  B	  cell	  population	  enriched	  from	  the	  PBMCs	  by	  magnetic-­‐activated	  cell	  sorting	  (MACs).	  	  Whereas	  the	  intramuscularly	  induced	  IgA	  response	  could	  be	  measured	  with	  both	  ELISA	  and	  ELIspot	  assays,	  active	  immune	  responses	  in	  the	  orally	  immunized	  pigs	  could	  only	  be	  detected	  by	  the	  ELIspot	  assay	  performed	  21	  days	  post	  immunization	  on	  the	  enriched	  IgA+	  B-­‐cell	  populations.	  These	  results	  demonstrate	  that	  piglets	  with	  maternal	  antibodies	  against	  F4	  still	  can	  be	  immunized	  orally	  with	  F4	  fimbriae,	  most	  likely	  protecting	  these	  piglets	  against	  infection	  with	  F4	  ETEC.	  Furthermore,	  our	  ELIspot	  assay	  on	  IgA+	  B-­‐cells	  has	  the	  potential	  to	  be	  used	  as	  a	  rapid	  procedure	  to	  optimize	  immunization	  protocols	  in	  pigs	  with	  passive	  immunity	  in	  comparison	  with	  the	  much	  more	  labour	  intense	  challenge	  assays.	  	     

Elispot on enriched IgA+ B-lymphocytes can be used to detect the IgA response following oral immunization with F4 fimbriae of pigs with colostral F4-specific serum antibodies

https://biblio.ugent.be/publication/4160553/file/4160593.pdf

Elispot on enriched IgA+ B-lymphocytes can be used to detect the IgA response following oral immunization with F4 fimbriae of pigs with colostral F4-specific serum antibodies

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