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

Abstract

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, IgG(1)and IgG(2) antibodies. Two immunizations increased mean serum IgM and the IgG(1)antibody profiles above preimmunization levels, but 5 immunizations were required to detect significant IgG(2) 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 IgG(2) antibodies.

Association ofEscherichia coliJ5-Specific Serum Antibody Responses with Clinical Mastitis Outcome for J5 Vaccinate and Control Dairy Cattle

“…A threshold of J5-specific IgG1 above which cows are protected against coliform mastitis has been reported (17), but an optimal range of J5-specific IgG1 or IgG2 antibody production above or below which protection was poor has not been described previously for bovine J5. The present results support previous evidence that OD values of Ͻ0.3 and Ͼ1.0 for IgG1 represent extreme values (2,16), but those studies did not measure milk production change following disease. These findings are similar to those of a recent intramammary E. coli challenge trial (25), in which cows with the highest production of milk IgM or IgG2 antibodies against J5 following challenge had significantly greater milk production loss.…”

“…Significantly increased J5-specific IgM, IgG1, and IgG2 antibodies were observed following the third of 12 doses of J5 bacterin administered over 170 days in five Holstein steers. Compared to the response after three doses, antibody increased significantly again following the 12th immunization for IgM, the sixth immunization for IgG1, and the seventh immunization for IgG2 (2). In the current study, protection with J5 vaccine appeared to decline as lactation (and the time since vaccination) progressed, disappearing by approximately 75 DIM.…”

“…Antibody was measured by ELISA as described previously (2,25). Briefly, flat-bottom 96-well ELISA plates (BD Falcon, Bedford, MA) were coated with 100 l of J5 E. coli (1 ϫ 10 9 CFU/ml in sterile saline, killed with 1% phenol), covered with an adhesive plate sealer, and left on a flat surface at room temperature for 12 h to enable adherence of the bacteria to the plate wells.…”

“…The commercially available vaccines are often called J5 core antigen vaccines, but the antigen(s) and mechanism(s) of J5 immunization have not been clearly identified (2,23). Associations between J5 vaccination of cattle, various measures of clinical severity of CM, and J5-specific antibodies have rarely been reported for naturally occurring cases of CM (2,17,19,23). Use of survival (time-to-event) analysis to evaluate the relationships between J5 vaccination, antibody production, and time until culling or death has recently been reported for the first time (23).…”

Bovine IgG1 antibodies againstMycobacterium aviumsubsp.paratuberculosisprotein p34-cx improve association of bacteria and macrophages

“…In fact, this Th2 response had been previously described as characteristic to protein antigens in cattle [7]. Recently, Chaiyotwittayakun et al [8] evaluated a J5 E. coli bacterin and described that at least six doses of vaccine were needed to detect an increment of specific IgG2 in cattle. We were not able to establish a predominant isotype response in Map whole bacteria vaccinated cattle with two doses of vaccine.…”