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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.


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IgG1 and IgG2 antibody titer increases in serum and milk. Isotype specific antibody titers in serum (A-D) and milk (E-H) following IN/IN (●), IM/IM (■IMM/SC (▲) or SC/SC (▼) immunization with Efb (A-B, E-F) and LukM (C-D, G-H) were measured by ELISA. Results are expressed as the mean per group ± SEM. Differences in total titer increases following prime plus booster immunizations were analyzed using the One-way ANOVA. Letters indicate significant differences between IN/IN and IM/IM (a), IN/IN and IMM/SC (b), IN/IN and SC/SC (c), IM/IM and SC/SC (d), and IMM/SC and SC/SC (e) immunization routes.
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Fig2: IgG1 and IgG2 antibody titer increases in serum and milk. Isotype specific antibody titers in serum (A-D) and milk (E-H) following IN/IN (●), IM/IM (■IMM/SC (▲) or SC/SC (▼) immunization with Efb (A-B, E-F) and LukM (C-D, G-H) were measured by ELISA. Results are expressed as the mean per group ± SEM. Differences in total titer increases following prime plus booster immunizations were analyzed using the One-way ANOVA. Letters indicate significant differences between IN/IN and IM/IM (a), IN/IN and IMM/SC (b), IN/IN and SC/SC (c), IM/IM and SC/SC (d), and IMM/SC and SC/SC (e) immunization routes.

Mentions: Figure 2 shows the IgG1 and IgG2 antibody titer increases in serum and milk. Efb specific antibody titers in serum increased from 100 up to 2000 (IgG1) and from 1000 up to 15 000 (IgG2). LukM specific antibody titers in serum increased from 300 up to 7000 (IgG1) and from 6000 up to over 70 000 (IgG2). Milk titer increases were minimal for Efb specific IgG1 and IgG2 antibodies, whereas milk titer increases for LukM specific antibodies ranges from 50 to 600 (IgG1) and from 20 up to 130 (IgG2). No significant route specific increases in total antibody titers were observed over time. However, between routes statistically significant different titer increases were observed. Following priming and booster immunizations, the increase in Efb-specific IgG1 levels in serum was significantly higher in IM/IM, IMM/SC and SC/SC immunized animals than in IN/IN immunized cows (p = 0.037, p = 0.001 and p = 0.000, respectively; Figure 2A). The increase in Efb-specific IgG2 serum levels were higher in SC/SC immunized animals compared to IN/IN and IM/IM immunized cows (both p < 0.001; Figure 2B). Elevations in serum levels of LukM-specific IgG1 were also significantly higher following SC/SC immunizations compared to IN/IN immunizations (p = 0.041; Figure 2C). In contrast, route specific increases in LukM-specific IgG2 serum levels were not observed (Figure 2D). In milk, the increase in Efb-specific IgG1 was higher following SC/SC immunization than following IMM/SC immunization (p = 0.029; Figure 2E). These differences were not observed for Efb-specific IgG2 levels (Figure 2F). LukM-specific antibody titers in milk were only slightly affected by immunization via the different administration routes with the highest increases following SC/SC immunizations (Figure 2G–H). However, for all groups, increases in milk antibody levels compared to serum were only moderate and pre- and post-immunization titers were not statistically different.Figure 2


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)

IgG1 and IgG2 antibody titer increases in serum and milk. Isotype specific antibody titers in serum (A-D) and milk (E-H) following IN/IN (●), IM/IM (■IMM/SC (▲) or SC/SC (▼) immunization with Efb (A-B, E-F) and LukM (C-D, G-H) were measured by ELISA. Results are expressed as the mean per group ± SEM. Differences in total titer increases following prime plus booster immunizations were analyzed using the One-way ANOVA. Letters indicate significant differences between IN/IN and IM/IM (a), IN/IN and IMM/SC (b), IN/IN and SC/SC (c), IM/IM and SC/SC (d), and IMM/SC and SC/SC (e) immunization routes.
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Related In: Results  -  Collection

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Fig2: IgG1 and IgG2 antibody titer increases in serum and milk. Isotype specific antibody titers in serum (A-D) and milk (E-H) following IN/IN (●), IM/IM (■IMM/SC (▲) or SC/SC (▼) immunization with Efb (A-B, E-F) and LukM (C-D, G-H) were measured by ELISA. Results are expressed as the mean per group ± SEM. Differences in total titer increases following prime plus booster immunizations were analyzed using the One-way ANOVA. Letters indicate significant differences between IN/IN and IM/IM (a), IN/IN and IMM/SC (b), IN/IN and SC/SC (c), IM/IM and SC/SC (d), and IMM/SC and SC/SC (e) immunization routes.
Mentions: Figure 2 shows the IgG1 and IgG2 antibody titer increases in serum and milk. Efb specific antibody titers in serum increased from 100 up to 2000 (IgG1) and from 1000 up to 15 000 (IgG2). LukM specific antibody titers in serum increased from 300 up to 7000 (IgG1) and from 6000 up to over 70 000 (IgG2). Milk titer increases were minimal for Efb specific IgG1 and IgG2 antibodies, whereas milk titer increases for LukM specific antibodies ranges from 50 to 600 (IgG1) and from 20 up to 130 (IgG2). No significant route specific increases in total antibody titers were observed over time. However, between routes statistically significant different titer increases were observed. Following priming and booster immunizations, the increase in Efb-specific IgG1 levels in serum was significantly higher in IM/IM, IMM/SC and SC/SC immunized animals than in IN/IN immunized cows (p = 0.037, p = 0.001 and p = 0.000, respectively; Figure 2A). The increase in Efb-specific IgG2 serum levels were higher in SC/SC immunized animals compared to IN/IN and IM/IM immunized cows (both p < 0.001; Figure 2B). Elevations in serum levels of LukM-specific IgG1 were also significantly higher following SC/SC immunizations compared to IN/IN immunizations (p = 0.041; Figure 2C). In contrast, route specific increases in LukM-specific IgG2 serum levels were not observed (Figure 2D). In milk, the increase in Efb-specific IgG1 was higher following SC/SC immunization than following IMM/SC immunization (p = 0.029; Figure 2E). These differences were not observed for Efb-specific IgG2 levels (Figure 2F). LukM-specific antibody titers in milk were only slightly affected by immunization via the different administration routes with the highest increases following SC/SC immunizations (Figure 2G–H). However, for all groups, increases in milk antibody levels compared to serum were only moderate and pre- and post-immunization titers were not statistically different.Figure 2

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