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Immunization routes in cattle impact the levels and neutralizing capacity of antibodies induced against S. aureus immune evasion proteins.

Boerhout E, Vrieling M, Benedictus L, Daemen I, Ravesloot L, Rutten V, Nuijten P, van Strijp J, Koets A, Eisenberg S - Vet. Res. (2015)

Bottom Line: It is unknown, however, whether this route is the most suitable to specifically increase intramammary immunity to combat S. aureus at the site of infection.The experimental vaccine comprised the recombinant S. aureus immune evasion proteins extracellular fibrinogen-binding protein (Efb) and the leukotoxin subunit LukM in an oil-in-water adjuvant combined with a hydrogel and alginate.Furthermore, the harmful effects of Efb and leukotoxin LukMF' on host-defense were neutralized by serum antibodies in a route-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Ruminant Research and Development, MSD Animal Health, Wim de Körverstraat 35, 5830, AA, Boxmeer, The Netherlands. eveline.boerhout@merck.com.

ABSTRACT
Vaccines against S. aureus bovine mastitis are scarce and show limited protection only. All currently available vaccines are applied via the parenteral (usually intramuscular) route. It is unknown, however, whether this route is the most suitable to specifically increase intramammary immunity to combat S. aureus at the site of infection. Hence, in the present study, immunization via mucosal (intranasal; IN), intramuscular (triangle of the neck; IM), intramammary (IMM) and subcutaneous (suspensory ligament; SC) routes were analyzed for their effects on the quantity of the antibody responses in serum and milk as well as the neutralizing capacity of the antibodies within serum. The experimental vaccine comprised the recombinant S. aureus immune evasion proteins extracellular fibrinogen-binding protein (Efb) and the leukotoxin subunit LukM in an oil-in-water adjuvant combined with a hydrogel and alginate. The highest titer increases for both Efb and LukM specific IgG1 and IgG2 antibody levels in serum and milk were observed following SC/SC immunizations. Furthermore, the harmful effects of Efb and leukotoxin LukMF' on host-defense were neutralized by serum antibodies in a route-dependent manner. SC/SC immunization resulted in a significant increase in the neutralizing capacity of serum antibodies towards Efb and LukMF', shown by increased phagocytosis of S. aureus and increased viability of bovine leukocytes. Therefore, a SC immunization route should be considered when aiming to optimize humoral immunity against S. aureus mastitis in cattle.

No MeSH data available.


Related in: MedlinePlus

LukMF’ neutralization assay. The presence of LukMF’ neutralizing antibodies in serum of cows from different immunization groups were analyzed in a LukMF’ neutralization assay. For each group, the time of lysis onset between pre- (▲) and post- (■) immunization serum was calculated. Results are expressed as time of lysis onset ± SEM of three independent neutralization experiments. Differences in the time of lysis onset pre- and post-immunization within each immunization group were analyzed by repeated measures analysis. LukMF’ neutralization post-immunization was significantly increased in cows immunized via SC/SC (p = 0.009) and IM/IM (p = 0.012) administration when compared to pre- immunization.
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Fig4: LukMF’ neutralization assay. The presence of LukMF’ neutralizing antibodies in serum of cows from different immunization groups were analyzed in a LukMF’ neutralization assay. For each group, the time of lysis onset between pre- (▲) and post- (■) immunization serum was calculated. Results are expressed as time of lysis onset ± SEM of three independent neutralization experiments. Differences in the time of lysis onset pre- and post-immunization within each immunization group were analyzed by repeated measures analysis. LukMF’ neutralization post-immunization was significantly increased in cows immunized via SC/SC (p = 0.009) and IM/IM (p = 0.012) administration when compared to pre- immunization.

Mentions: Serum antibodies were also tested for their ability to neutralize the effects of LukM on PBL lysis. Figure 4 shows that LukM specific antibodies in serum of SC/SC immunized cows significantly blocked the pore forming ability of LukMF’ when compared to pre-immunization serum (p = 0.009). LukM specific antibodies in serum of IM/IM immunized cows also significantly decreased the pore formation by LukMF’ compared to pre-immunization serum (p = 0.012). However, the differences in neutralization pre- and post-immunization were limited compared to the increase observed in SC/SC immunized cows. A trend in LukMF’ neutralization was observed in serum of IMM/SC immunized cows (p = 0.058), whereas no differences in neutralization of LukMF’ were observed in serum of cows immunized via the IN/IN route.Figure 4


Immunization routes in cattle impact the levels and neutralizing capacity of antibodies induced against S. aureus immune evasion proteins.

Boerhout E, Vrieling M, Benedictus L, Daemen I, Ravesloot L, Rutten V, Nuijten P, van Strijp J, Koets A, Eisenberg S - Vet. Res. (2015)

LukMF’ neutralization assay. The presence of LukMF’ neutralizing antibodies in serum of cows from different immunization groups were analyzed in a LukMF’ neutralization assay. For each group, the time of lysis onset between pre- (▲) and post- (■) immunization serum was calculated. Results are expressed as time of lysis onset ± SEM of three independent neutralization experiments. Differences in the time of lysis onset pre- and post-immunization within each immunization group were analyzed by repeated measures analysis. LukMF’ neutralization post-immunization was significantly increased in cows immunized via SC/SC (p = 0.009) and IM/IM (p = 0.012) administration when compared to pre- immunization.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4584483&req=5

Fig4: LukMF’ neutralization assay. The presence of LukMF’ neutralizing antibodies in serum of cows from different immunization groups were analyzed in a LukMF’ neutralization assay. For each group, the time of lysis onset between pre- (▲) and post- (■) immunization serum was calculated. Results are expressed as time of lysis onset ± SEM of three independent neutralization experiments. Differences in the time of lysis onset pre- and post-immunization within each immunization group were analyzed by repeated measures analysis. LukMF’ neutralization post-immunization was significantly increased in cows immunized via SC/SC (p = 0.009) and IM/IM (p = 0.012) administration when compared to pre- immunization.
Mentions: Serum antibodies were also tested for their ability to neutralize the effects of LukM on PBL lysis. Figure 4 shows that LukM specific antibodies in serum of SC/SC immunized cows significantly blocked the pore forming ability of LukMF’ when compared to pre-immunization serum (p = 0.009). LukM specific antibodies in serum of IM/IM immunized cows also significantly decreased the pore formation by LukMF’ compared to pre-immunization serum (p = 0.012). However, the differences in neutralization pre- and post-immunization were limited compared to the increase observed in SC/SC immunized cows. A trend in LukMF’ neutralization was observed in serum of IMM/SC immunized cows (p = 0.058), whereas no differences in neutralization of LukMF’ were observed in serum of cows immunized via the IN/IN route.Figure 4

Bottom Line: It is unknown, however, whether this route is the most suitable to specifically increase intramammary immunity to combat S. aureus at the site of infection.The experimental vaccine comprised the recombinant S. aureus immune evasion proteins extracellular fibrinogen-binding protein (Efb) and the leukotoxin subunit LukM in an oil-in-water adjuvant combined with a hydrogel and alginate.Furthermore, the harmful effects of Efb and leukotoxin LukMF' on host-defense were neutralized by serum antibodies in a route-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Ruminant Research and Development, MSD Animal Health, Wim de Körverstraat 35, 5830, AA, Boxmeer, The Netherlands. eveline.boerhout@merck.com.

ABSTRACT
Vaccines against S. aureus bovine mastitis are scarce and show limited protection only. All currently available vaccines are applied via the parenteral (usually intramuscular) route. It is unknown, however, whether this route is the most suitable to specifically increase intramammary immunity to combat S. aureus at the site of infection. Hence, in the present study, immunization via mucosal (intranasal; IN), intramuscular (triangle of the neck; IM), intramammary (IMM) and subcutaneous (suspensory ligament; SC) routes were analyzed for their effects on the quantity of the antibody responses in serum and milk as well as the neutralizing capacity of the antibodies within serum. The experimental vaccine comprised the recombinant S. aureus immune evasion proteins extracellular fibrinogen-binding protein (Efb) and the leukotoxin subunit LukM in an oil-in-water adjuvant combined with a hydrogel and alginate. The highest titer increases for both Efb and LukM specific IgG1 and IgG2 antibody levels in serum and milk were observed following SC/SC immunizations. Furthermore, the harmful effects of Efb and leukotoxin LukMF' on host-defense were neutralized by serum antibodies in a route-dependent manner. SC/SC immunization resulted in a significant increase in the neutralizing capacity of serum antibodies towards Efb and LukMF', shown by increased phagocytosis of S. aureus and increased viability of bovine leukocytes. Therefore, a SC immunization route should be considered when aiming to optimize humoral immunity against S. aureus mastitis in cattle.

No MeSH data available.


Related in: MedlinePlus