Immunization Of Dams Research Articles

Evolution of protection after maternal immunization for respiratory syncytial virus in cotton rats.

Maternal anti-respiratory syncytial virus (RSV) antibodies acquired by the fetus through the placenta protect neonates from RSV disease through the first weeks of life. In the cotton rat model of RSV infections, we previously reported that immunization of dams during pregnancy with virus-like particles assembled with mutation stabilized pre-fusion F protein as well as the wild type G protein resulted in robust protection of their offspring from RSV challenge. Here we describe the durability of those protective responses in dams, the durability of protection in offspring, and the transfer of that protection to offspring of two consecutive pregnancies without a second boost immunization. We report that four weeks after birth, offspring of the first pregnancy were significantly protected from RSV replication in both lungs and nasal tissues after RSV challenge, but protection was reduced in pups at 6 weeks after birth. However, the overall protection of offspring of the second pregnancy was considerably reduced, even at four weeks of age. This drop in protection occurred even though the levels of total anti-pre-F IgG and neutralizing antibody titers in dams remained at similar, high levels before and after the second pregnancy. The results are consistent with an evolution of antibody properties in dams to populations less efficiently transferred to offspring or the less efficient transfer of antibodies in elderly dams.

Immunizing adult female mice with a TcpA-A2-CTB chimera provides a high level of protection for their pups in the infant mouse model of cholera.

Vibrio cholerae expresses two primary virulence factors, cholera toxin (CT) and the toxin-coregulated pilus (TCP). CT causes profuse watery diarrhea, and TCP (composed of repeating copies of the major pilin TcpA) is required for intestinal colonization by V. cholerae. Antibodies to CT or TcpA can protect against cholera in animal models. We developed a TcpA holotoxin-like chimera (TcpA-A2-CTB) to elicit both anti-TcpA and anti-CTB antibodies and evaluated its immunogenicity and protective efficacy in the infant mouse model of cholera. Adult female CD-1 mice were immunized intraperitoneally three times with the TcpA-A2-CTB chimera and compared with similar groups immunized with a TcpA+CTB mixture, TcpA alone, TcpA with Salmonella typhimurium flagellin subunit FliC as adjuvant, or CTB alone. Blood and fecal samples were analyzed for antigen-specific IgG or IgA, respectively, using quantitative ELISA. Immunized females were mated; their reared offspring were challenged orogastrically with 10 or 20 LD50 of V. cholerae El Tor N16961; and vaccine efficacy was assessed by survival of the challenged pups at 48 hrs. All pups from dams immunized with the TcpA-A2-CTB chimera or the TcpA+CTB mixture survived at both challenge doses. In contrast, no pups from dams immunized with TcpA+FliC or CTB alone survived at the 20 LD50 challenge dose, although the anti-TcpA or anti-CTB antibody level elicited by these immunizations was comparable to the corresponding antibody level achieved by immunization with TcpA-A2-CTB or TcpA+CTB. Taken together, these findings comprise strong preliminary evidence for synergistic action between anti-TcpA and anti-CTB antibodies in protecting mice against cholera. Weight loss analysis showed that only immunization of dams with TcpA-A2-CTB chimera or TcpA+CTB mixture protected their pups against excess weight loss from severe diarrhea. These data support the concept of including both TcpA and CTB as immunogens in development of an effective multivalent subunit vaccine against V. cholerae.

Passive protection to bovine rotavirus (BRV) infection induced by a BRV VP8* produced in plants using a TMV-based vector.

We have previously reported on the use of a tobacco mosaic virus (TMV) vector TMV-30B to express foreign viral antigens for use as experimental immunogens. Here we describe the development of an improved TMV-30B vector that adds a sequence of 7 histidine residues to the C-terminus of recombinant proteins expressed in the vector. We used this TMV-30B-HISc vector to express the VP8* fragment of the VP4 protein from bovine rotavirus (BRV) strain C-486 in plants. Recombinant VP8* protein was purified from N. benthamiana leaves at 7 days post-inoculation by immobilized metal affinity chromatography. The plant-produced VP8* was initially detected using anti-His tag mAb and its antigenic nature was confirmed using both monoclonal and polyclonal specific antisera directed against BRV. Adult female mice, inoculated by the intraperinoteal route with an immunogen containing 4 microg of recombinant VP8*, developed a specific and sustained response to the native VP8* from the homologous BRV. Eighty five percent of suckling mice from immunized dams that were challenged with the homologous virus at the fifth day of age were protected from virus as compared to 35% of the pups from mothers immunized with a control protein. These results demonstrate that the plant-produced VP8* was able to induce passive protection in the new born through the immunization of dams. This suggests that the technology presented here provides a simple method for using plants as an inexpensive alternative source for production of recombinant anti-rotavirus antigens.

Prevention of Vertical Transfer of Neospora caninum in BALB/c Mice by Vaccination

Neosporosis is an important cause of abortion and neonatal morbidity in dairy cattle. The disease is caused by Neospora caninum, an intracellular protozoan parasite. In this report, we describe the use of a mouse model in the preliminary evaluation of vaccination as a means to prevent vertical transfer of N. caninum. Parasites present in the tissues of the offspring were detected using an N. caninum-specific polymerase chain reaction assay. Immunization of dams with a single inoculation of a crude lysate of N. caninum tachyzoites appeared to induce complete protection against infection of the offspring.

Protection of kids against Cryptosporidium parvum infection after immunization of dams with CP15-DNA

In this study the effectiveness of a DNA vaccine to confer protection against cryptosporidiosis, an enteric infection of lifestock and humans, was evaluated. A vaccination protocol using a recombinant plasmid encoding the 15 kDa surface sporozoite protein of Cryptosporidium parvum was developed in adult pregnant goats. The present study reports that nasal immunization of pregnant goats with CP15-DNA led to a transfer of immunity to offspring conferring protection against C. parvum infection. Kids from CP15-DNA-vaccinated dams shed significantly fewer oocysts and over a shorter period than did kids from unvaccinated goats. The low level of parasite development in protected kids did not affect their growth whereas unprotected kids grew much slowly. There was still a significant difference in the weights of protected and unprotected kids after complete recovery. Anti-CP15 antibodies were present in serum and colostrum from vaccinated goats. Nevertheless, the precise immune mechanism of protection has still to be determined. This vaccine should reduce the economic losses due to cryptosporidiosis in ruminants, specially in small ruminants (calves, lambs, kids). It has also the potential to reduce environmental contamination by reducing oocyst shedding.

Protection against colibacillosis in neonatal piglets by immunization of dams with procholeragenoid.

Protection against colibacillosis in neonatal piglets was obtained by immunization of pregnant dams with procholeragenoid. Procholeragenoid is a stable high-molecular-weight aggregate of cholera toxin formed during the heating of cholera toxin. Procholeragenoid retained approximately 1% of the toxicity of native toxin as determined in the rabbit ileal loop and Y-1 adrenal cell assays and 5% of the activity in the rabbit skin assay. Immunization of pregnant dams with 50 micrograms of procholeragenoid 5 and 2 weeks before the expected delivery date elicited high titers of antitoxic immunoglobulin G and toxin-neutralizing antibody in both the colostrum and serum. In three independent field trials, immunization with procholeragenoid resulted in a substantial decrease in diarrhea (73% in controls versus 11% in immunized) and death (4.7% in controls versus 0.77% in immunized) in neonatal piglets. The protection rate in the immunized population was approximately 85% for both diarrhea and death. In the following gestation period, reimmunization of dams with a single dose of procholeragenoid (50 micrograms) 2 weeks before delivery elicited titers of antitoxic immunoglobulin G and toxin-neutralizing antibody comparable to those obtained during the primary immunization. The death rate in neonatal piglets (0.86%) was comparable to that seen after immunization during the first gestation period (0.77%). These results indicate that substantial protection of neonatal piglets against colibacillosis can be obtained by immunization of dams with procholeragenoid. Protection was found to be based solely on antitoxic immunity.