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Hyperimmunization of Steers with J5 Escherichia coli Bacterin: Effects on Isotype-Specific Serum Antibody Responses and Cross Reactivity with Heterogeneous Gram-Negative Bacteria

¹ Department of Large Animal Clinical Sciences and
² Department of Animal Science, Michigan State University, East Lansing, 48824

Corresponding author: R. J. Erskine; e-mail: erskine{at}cvm.msu.edu.

Isotype-specific antibody responses and cross reactivity were profiled following hyperimmunization of steers with J5 Escherichia coli bacterin. The vaccine was administered at time 0, 30 d later, and every 2 wk for 10 subsequent immunizations. Blood was collected preimmunization and multiple times following each immunization. Isotype-specific anti-J5 Escherichia coli antibody response profiles in diluted sera harvested from each sample were assayed by ELISA and recorded as optical density. Selected sera were assayed for anti-J5 Escherichia coli antibody titers and used to determine cross reactivity against a variety of gram-negative bacteria. Immunization number and day postimmunization influenced response profiles for anti-J5 E. coli IgM, IgG₁and IgG₂ antibodies. Two immunizations increased mean serum IgM and the IgG₁antibody profiles above preimmunization levels, but 5 immunizations were required to detect significant IgG₂ antibody responses that were above preimmunization levels. Isotype-specific cross reactivity of the serum antibodies with a variety of heterologous gram-negative bacteria was also increased by hyperimmunization. However, no cross reactivity was observed for Staphylococcus aureus, purified lipopolysaccharide, or lipid A. Our results indicate that multiple booster doses of J5 E. coli bacterin may be required to elicit high levels of cross-reactive serum IgG₂ antibodies.

Key Words: J5 Escherichia coli • hyperimmunization • antibody response • cross reactivity

Abbreviation key: FBS = fetal bovine serum, INF- = interferon-gamma, OD = optical density, OMP = outer membrane protein, TSB = trypticase soy broth.

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