Antigenic crossreactivity and lipopolysaccharide neutralization properties of bovine immunoglobulin G

We investigated a possible mechanism by which immunization against core and lipid A determinants of lipopolysaccharide reduced clinical cases of mastitis and symptoms commonly associated with heterologous Gram-negative IMI. The IgG fraction of sera from cows immunized with either Escherichia coli J5 bacterin, E. coli J5 lipopolysaccharide conjugate vaccine, or unimmunized controls was purified by precipitation with caprylic acid and ammonium sulfate. The degree of IgG crossreactivity with Gram-negative bacteria that were isolated from clinical quarters was greater than that with Gram-positive isolates of Staphylococcus aureus. The highest magnitude of crossreactivity was against smooth strain E. coli isolates, followed by heterologous species of Enterobacter, Serratia, and Klebsiella isolates. Serum IgG from cows immunized with conjugate was highly crossreactive to E. coli J5, E. coli O111:B4, Serratia marcescens, Klebsiella pneumoniae, and Salmonella typhimurium lipopolysaccharides. The magnitude of antibody crossreactivity with lipopolysaccharides coincided with the ability of IgG to suppress the mitogenic effect of lipopolysaccharides on bovine lymphocytes.     

Effect of vaccination with an Escherichia coli bacterin-toxoid on milk production in dairy cattle

OBJECTIVE: To determine whether vaccination of lactating cattle with an Escherichia coli J5 bacterin-toxoid would produce a significant short-term change in milk production. DESIGN: Randomized, controlled clinical trial. ANIMALS: 84 healthy, lactating cows (42 Holsteins and 42 Jerseys). PROCEDURE: Control and vaccinated cows were paired on the basis of breed, days in milk, daily milk production 1 week prior to vaccination, and parity. One cow in each pair was inoculated IM with a commercially available bacterin-toxoid according to label directions; the other cow was given saline solution. Cows were milked twice daily for 5 days before and 5 days after inoculation. Milk production was compared by ANCOVA. RESULTS: Vaccinated cows produced significantly less milk than did control cows at the second and third milkings after inoculation. At these milkings, milk production in vaccinated cows was approximately 7% less than that of controls. CLINICAL IMPLICATIONS: Vaccination of lactating cattle with an E coli J5 bacterin-toxoid may cause a significant short-term decrease in milk production.     

Multiplication of Staphylococci in vitro in normal and mastitic milk from vaccinated and nonvaccinated cows

Mastitis was induced by injection of cell walls of Staphylococcus aureus into the mammary glands of normal cows and of cows which had been vaccinated parenterally with a staphylococcal bacterin in adjuvant. Multiplication of S aureus in normal milk and in mastitic milk from vaccinated and nonvaccinated cows was determined in constant volume cultures. Growth was significantly inhibited during the 1st 6 hours of incubation, regardless of the nature of the milk or the vaccination status of the cows. Growth was inhibited for 24 hours in normal milk, and the organisms grew exponentially in mastitic milk regardless of the vaccination status of the cows.     

The Pseudomonas aeruginosa outer membrane protein I vaccine: immunogenicity and safe administration in man

After expression in Escherichia coli and purification by Ni++ chelate-affinity chromatography, the outer membrane protein I (OprI) of Pseudomonas aeruginosa was tested in experimental animals for its safety and pyrogenicity. Four groups of 7 adult human volunteers were then vaccinated 3 times at four-weekly intervals with either 500 micrograms, 200 micrograms, 50 micrograms or 20 micrograms of OprI adsorbed onto aluminum hydroxide. The vaccinations were well tolerated and without systemic side effects, but a significant rise of antibody titers against OprI was measured in the serum of those who had received the 500 micrograms, 200 micrograms or 50 micrograms doses. Raised antibody titers against OprI were still present 30 weeks after the final vaccination. It was possible to demonstrate binding of the complement component C1q to the elicited antibodies, and this confirms their ability to promote antibody-mediated complement-dependent opsonization.     

Effects on nutrient and hormonal profile of long-term infusions of glucose or insulin plus glucose in cows treated with recombinant bovine somatotropin before peak milk yield

Ten Holstein cows were treated with 30.9 mg.d-1 of recombinant bST from 15 to 41 d of lactation. The Latin square design included three infusion periods of 6 d each with 3 d of rest between infusion periods. Infusions were physiological saline, glucose (50 g.h-1), and insulin plus glucose (12.5 IU.h-1 + 50 g.h-1). Blood was collected continuously during the last 24 h of each infusion period. Statistical analyses of data for energy balance, milk yield, and DMI were performed on the last 3 d of each infusion period. Production data before and after infusions (i.e., no recombinant bST) estimated that recombinant bST increased milk yield of cows infused with glucose and saline by 3.1 and 3.6 kg.d-1, respectively. Net energy intake was not affected by infusion, but glucose infusion resulted in higher BW loss than did saline infusion (2.33 vs. 0.08 kg.d-1, respectively), and insulin plus glucose infusion resulted in BW gain (0.65 kg.d-1). Milk yield was 39.9, 39.6, and 37.6 kg.d-1 for cows infused with saline, glucose, and insulin plus glucose, respectively. The insulin plus glucose infusion increased milk protein 11 and 14% compared with response to saline and glucose infusions, respectively; no change occurred in the proportion of casein and whey proteins. Serum bST was increased 109% with exogenous recombinant bST. Serum IGF-I was lower for cows infused with glucose than for those infused with saline (21.03 vs. 27.44 ng.ml-1) and increased to 46.55 ng.ml-1 for cows infused with insulin plus glucose. Serum concentrations of insulin and glucose were 13.7 and 56.7, 18.5 and 61.9, and 30.5 muIU.ml-1 and 39.4 mg.dl-1 for cows infused with saline, glucose, and insulin plus glucose, respectively. The results of this study suggest that low concentrations of plasma insulin in early lactation may limit the IGF-I response to recombinant bST (uncoupling). Despite higher IGF-I, milk yield was lower, probably as a result of low blood glucose. These results suggest that, in early lactation, insulin is still anabolic because the BW gain of cows increased. However, milk yield was still higher than that for cows in late lactation with similar insulin concentrations.     

Effects of growth hormone, insulin-like growth factor-I, and cortisol on periparturient antibody response profiles of dairy cattle

The objectives of this study were to determine hormone and antibody response profiles from the prepartum period to peak lactation, and evaluate potential immunomodulatory effects of the classic endocrine hormones, growth hormone (GH), insulin-like growth factor-I (IGF-I) and cortisol. Specifically, 33 Holstein cows were immunized with ovalbumin (OVA) and Escherichia coli J5 at weeks -8 and -3 prior to parturition. At parturition (week 0), cows received an additional immunization of OVA. Blood was collected at weeks -8, -3, 0, 3 and 6 relative to parturition and various samples were used to determine plasma hormone concentration, serum immunoglobulin (Ig), and specific antibody response to OVA and E. coli. Colostrum and milk samples were also collected post-parturition to monitor local immunoglobulin and antibody responses. Results indicated that not all periparturient cows exhibited depressed immune response, and that antibody response to OVA could be used to partition cows into 3 groups recognizing animals with sustained measurable antibody response before and after parturition (Group 1), animals which responded poorly to immunization at parturition (Group 2), and animals which did not respond to immunizations at week -3 or parturition (Group 3). Cows with the highest antibody response to OVA (Group 1) also tended (P < or = 0.10) to have the highest response to E. coli J5 at parturition and had the lowest incidence of disease, particularly mastitis. Antibody response to OVA measured in milk tended to be higher in Group 1 cows, particularly at week 0 (P < or = 0.06) compared to cows of Group 3. IGF-I was higher (P < or = 0.05) in cows of Group 1 than Group 3 at peak lactation (week 6).     

Antiserum against Escherichia coli J5 contains antibodies reactive with outer membrane proteins of heterologous gram-negative bacteria

The binding of IgG in antiserum to Escherichia coli J5 to the surface of Enterobacteriaceae and to cell wall fragments released from serum-exposed bacteria was studied in a search for potentially protective epitopes other than lipopolysaccharide (LPS). IgG titers to multiple heterologous gram-negative smooth bacteria increased following incubation of the bacteria in serum and decreased following absorption with serum-exposed heterologous bacteria. IgG eluted from absorbing bacteria bound to at least three conserved bacterial outer membrane proteins (OMPs), but not LPS, as assessed by immunoblotting. The same OMPs were present in LPS-containing macromolecular cell wall fragments released by incubation of heterologous gram-negative bacteria in human serum. Part of the protection offered by J5 antiserum could be from binding of IgG to conserved OMPs at the bacterial surface or to OMPs in cell-wall fragments released from dying bacteria.     

Selectivity of ferric enterobactin binding and cooperativity of transport in gram-negative bacteria

The ligand-gated outer membrane porin FepA serves Escherichia coli as the receptor for the siderophore ferric enterobactin. We characterized the ability of seven analogs of enterobactin to supply iron via FepA by quantitatively measuring the binding and transport of their 59Fe complexes. The experiments refuted the idea that chirality of the iron complex affects its recognition by FepA and demonstrated the necessity of an unsubstituted catecholate coordination center for binding to the outer membrane protein. Among the compounds we tested, only ferric enantioenterobactin, the synthetic, left-handed isomer of natural enterobactin, and ferric TRENCAM, which substitutes a tertiary amine for the macrocyclic lactone ring of ferric enterobactin but maintains an unsubstituted catecholate iron complex, were recognized by FepA (Kd approximately 20 nM). Ferric complexes of other analogs (TRENCAM-3,2-HOPO; TREN-Me-3,2-HOPO; MeMEEtTAM; MeME-Me-3,2-HOPO; K3MECAMS; agrobactin A) with alterations to the chelating groups and different net charge on the iron center neither adsorbed to nor transported through FepA. We also compared the binding and uptake of ferric enterobactin by homologs of FepA from Bordetella bronchisepticus, Pseudomonas aeruginosa, and Salmonella typhimurium in the native organisms and as plasmid-mediated clones expressed in E. coli. All the transport proteins bound ferric enterobactin with high affinity (Kd /=50 pmol/min/10(9) cells) in their own particular membrane environments. However, the FepA and IroN proteins of S. typhimurium failed to efficiently function in E. coli. For E. coli, S. typhimurium, and P. aeruginosa, the rate of ferric enterobactin uptake was a sigmoidal function of its concentration, indicating a cooperative transport reaction involving multiple interacting binding sites on FepA.     

Receptor-dependent immune responses in pigs after oral immunization with F4 fimbriae

F4 receptor-positive (F4R+) and F4 receptor-negative (F4R-) pigs were orally vaccinated with purified F4 fimbriae of enterotoxigenic Escherichia coli (ETEC). Serum immunoglobulin G (IgG) and IgA responses were readily detected in F4R+ animals, whereas immune responses were not detected in F4R- animals. Even after a subsequent oral infection with virulent F4(+) ETEC and a booster immunization with F4, the F4R- animals remained F4 seronegative whereas the unvaccinated F4R+ pigs exhibited clear IgA and IgG responses. These results clearly demonstrate that F4Rs are a prerequisite for an immune response following oral immunization. Furthermore, indications that oral F4 vaccination can induce mucosal protection were obtained, since the experimental ETEC infection did not induce a systemic booster response or fecal ETEC excretion in orally vaccinated F4R+ pigs, in contrast to the clear immune response and ETEC excretion of unvaccinated F4R+ animals. F4-specific IgA antibodies could be found in the feces of the vaccinated F4R+ pigs. They are secreted at the intestinal mucosal surface and appear to prevent ETEC infection. The F4R-dependent induction of a mucosal immune response can be used as a model to better understand mucosal immunization and mucosal immune responses and can contribute to the development of oral vaccines in veterinary as well as in human medicine.     

Double mutagenesis of a positive charge cluster in the ligand-binding site of the ferric enterobactin receptor, FepA

Siderophores and colicins enter bacterial cells through TonB-dependent outer membrane proteins. Using site-directed substitution mutagenesis, we studied ligand recognition by a prototypic Escherichia coli siderophore receptor, FepA, that binds the iron chelate ferric enterobactin and colicins B and D. These genetic experiments identified a common binding site for two of the three ligands, containing multiple positive charges, within cell surface residues of FepA. Elimination of single residues in this region did not impair the adsorption or transport of ferric enterobactin, but double mutagenesis in the charge cluster identified amino acids (Arg-286 and Arg-316) that participate in siderophore binding and function in FepA-mediated killing by colicins B and D. Ferric enterobactin binding, furthermore, prevented covalent modification of FepA within this domain by either a fluorescent probe or an arginine-specific reagent, corroborating the involvement of this site in ligand recognition. These results identify, for the first time, residues in a TonB-dependent outer membrane protein that participate in ligand binding. They also explain the competition between ferric enterobactin and the colicins on the bacterial cell surface: all three ligands interact with the same arginine residues within FepA during their penetration through the outer membrane.     

Immunogenicity of casein phosphopeptides derived from tryptic hydrolysis of beta-casein

The immunogenicity of phosphopeptides derived from tryptic hydrolysis of beta-casein (CN) was investigated in a rat model system. The titers of specific immunoglobulin (Ig)G and IgE antibodies made in response to intraperitoneal sensitization to beta-CN, casein phosphopeptides, and skim milk proteins were examined using indirect and amplified indirect ELISA, respectively. Serum IgG antibodies from rats injected with beta-CN were significantly more reactive to beta-CN, casein phosphopeptides, and skim milk proteins coated on microtiter plate wells than were the IgG antibodies generated in rats that had been subjected to other treatments. A significant difference in titers because of the time of sampling (14 or 21 d postinjection) was noted for IgE but not for IgG. Rats that were injected with casein phosphopeptides did not produce IgG antibodies that crossreacted with either skim milk proteins or beta-CN. Specific antibody levels for the IgE class rarely exceeded those of unimmunized controls. The findings suggest that immunogenicity of the phosphopeptides was reduced compared with that of native beta-CN and skim milk proteins.     

Further studies on the efficacy of a live vaccine against mastitis caused by Streptococcus uberis

Three groups of dairy cows were immunized by subcutaneous (s.c.) administration of a preparation of live Streptococcus uberis (strain 0140J) and an intramammary infusion of a soluble surface extract derived from same the bacteria. Animals in Groups 1 and 2 received two s.c. vaccinations plus an intramammary inoculation. Animals in Group 3 received two s.c. vaccinations but did not receive the intramammary infusion. In addition to the vaccinated animals, each group also contained two non-vaccinated (control) animals. All animals were challenged experimentally by intramammary infusion (in two quarters per animal) of ca 100 c.f.u. of S. uberis (strain 0140J or C221) and monitored for clinical signs of disease, bacterial numbers in milk, somatic cell count in milk, and daily milk yield for the following 10 days. Animals in Group I were challenged with strain 0140J. Only one out of six challenged quarters of three vaccinated cows developed clinical disease compared to all (four out of four) quarters of non-vaccinated cows. Animals in Group 2 were challenged with strain C221. All challenged quarters of three vaccinated (six out of six) and two non-vaccinated (four out of four) cows developed clinical mastitis. Animals in Group 3 were challenged with strain 0140J. Five out of eight quarters on four vaccinated cows developed clinical mastitis but the onset was delayed in comparison with that in both non-vaccinated cows in which four out of four challenged quarters developed clinical mastitis. These results indicated that vaccination with live S. uberis protects against challenge with the homologous strain but was less effective against a heterologous strain. Reduced protection was also seen when the intramammary booster was omitted.     

Bovine mastitis caused by Listeria monocytogenes: kinetics of antibody responses in serum and milk after experimental infection

The kinetics of antibodies in serum and milk directed against proteins from Listeria monocytogenes were studied using 4 lactating cows after infection was experimentally induced in the udder with four strains of serotypes 4b or 1/2a. Antibodies (IgG and IgA) in samples of composite quarter milk and serum of the cow were measured by indirect ELISA. Microtiter plates were coated with proteins obtained from the culture supernatant of L. monocytogenes 4b. After challenge, an IgG response in serum and milk to listerial infections in the udder occurred for all cows, although the response varied among cows. In sera, the IgG titers reached a peak at 9 to 13 wk after challenge and remained elevated until 21 to 33 wk after challenge. In milk, the IgG titer increased significantly 3 wk after the challenge for all cows. A weak and nonpersistent increase in IgA antibodies also occurred. These results indicate that IMI by L. monocytogenes induced an increase of antibodies in milk, which could be detected with an ELISA test using our antigenic preparation. Therefore, this antigenic preparation could be used for the evaluation of a new method of diagnosis for bovine mastitis caused by L. monocytogenes.     

Milk production and reproductive performance in dairy cows given bovine respiratory syncytial virus vaccine prior to parturition

OBJECTIVE: To assess the effect of vaccination against bovine respiratory syncytial virus on milk production, reproductive performance, and health in lactating dairy cows. DESIGN: Prospective randomized block design. ANIMALS: 385 Holstein dairy cows and heifers. PROCEDURE: Cows were grouped by lactation number, season of calving, and previous mature equivalent 305-day milk production (where appropriate). Prior to parturition, cows and heifers were randomly assigned to be vaccinated i.m. against infectious bovine rhinotracheitis, bovine viral diarrhea, and parainfluenza 3 viruses by use of a three-way vaccine, or to be vaccinated against those viruses as well as bovine respiratory syncytial virus, using a four-way vaccine. Milk production was measured daily through 305 days of lactation. Reproductive and medical records were reviewed to obtain insemination dates and record medical problems of cows in each vaccine treatment group. RESULTS: Compared with the three-way vaccine, administration of the four-way vaccine was associated with higher milk production (1.39 kg [3.06 lb] more milk/d) in first-parity cows during the first 21 weeks of lactation. Vaccination did not have any effect on milk production after the first 21 weeks of lactation in cows of any parity. Conception rates at first insemination were higher for four-way vaccinated first-parity cows than for three-way vaccinated first-parity cows (54.6 vs 32.7%). Compared with second-parity cows that received the three-way vaccine, first insemination conception rate was improved for second-parity cows vaccinated with the four-way vaccine (28.9 vs 47.8%, respectively). In cows of third or greater parity, first insemination conception rate was not different between the 2 vaccine treatment groups. CLINICAL IMPLICATIONS: Vaccination of heifers against bovine respiratory syncytial virus prior to partrition may increase milk production and first insemination conception rates.     

Partial protection against Plasmodium vivax blood-stage infection in Saimiri monkeys by immunization with a recombinant C-terminal fragment of merozoite surface protein 1 in block copolymer adjuvant

Merozoite surface protein 1 is a candidate for blood-stage vaccines against malaria parasites. We report here an immunization study of Saimiri monkeys with a yeast-expressed recombinant protein containing the C terminus of Plasmodium vivax merozoite surface protein 1 and two T-helper epitopes of tetanus toxin (yP2P30Pv20019), formulated in aluminum hydroxide (alum) and block copolymer P1005. Monkeys immunized three times with yP2P30Pv20019 in block copolymer P1005 had significantly higher prechallenge titers of immunoglobulin G (IgG) antibodies against the immunogen and asexual blood-stage parasites than those immunized with yP2P30Pv20019 in alum, antigen alone, or phosphate-buffered saline (PBS) (P < 0.05). Their peripheral blood mononuclear cell proliferative responses to immunogen stimulation 4 weeks after the second immunization were also significantly higher than those from the PBS control group (P < 0.05). Upon challenge with 100,000 asexual blood-stage parasites 5 weeks after the last immunization, monkeys immunized with yP2P30Pv20019 in block copolymer P1005 had prepatent periods longer than those for the control alone group (P > 0.05). Three of the five animals in this group also had low parasitemia (peak parasitemia, 相似文献   

Immunization of LDL receptor-deficient mice with homologous malondialdehyde-modified and native LDL reduces progression of atherosclerosis by mechanisms other than induction of high titers of antibodies to oxidative neoepitopes

We and others previously showed that immunization of rabbits with different forms of oxidized low density lipoprotein (LDL) significantly reduced atherogenesis. We now investigated the effect of continued immunization on atherosclerosis in LDL receptor-deficient (LDLR-/-) mice to determine whether a similar reduction of atherosclerosis occurred in murine models and whether this was due to humoral immune responses, ie, formation of high titers of antibodies to oxidation-specific epitopes. Three groups of LDLR-/- mice were repeatedly immunized with homologous malondialdehyde-modified LDL (MDA-LDL), native LDL, or phosphate-buffered saline (PBS) for 7 weeks. Extensive hypercholesterolemia and accelerated atherogenesis were then induced by feeding a cholesterol-rich diet for 17 weeks, during which immunizations were continued. Binding of immunoglobulin (Ig) M and IgG antibodies, as well as IgG1 and IgG2a isotypes, to several epitopes of oxidized LDL were followed throughout the study. After 24 weeks of intervention, atherosclerosis in the aortic origin was significantly reduced by 46.3% and 36.9% in mice immunized with MDA-LDL and native LDL, respectively, compared with PBS (133 558 and 157 141 versus 248 867 microm2 per section, respectively). However, the humoral immune response to oxidative neoepitopes in the MDA-LDL group was very different from that of the LDL or PBS group. IgG antibody binding to MDA-LDL and other epitopes of oxidized LDL, such as oxidized phospholipid (cardiolipin), oxidized cholesterol, or oxidized cholesteryl linoleate, but not native LDL, increased markedly in mice immunized with MDA-LDL, but not in mice immunized with native LDL or PBS. In the MDA-LDL group, both T helper cell (Th)2-dependent IgG1 antibody and Th1-dependent IgG2a antibody binding to oxidative neoepitopes increased significantly over time. The fact that mice immunized with both MDA-LDL and native LDL had a significant reduction in atherosclerosis, whereas only the MDA-LDL group developed very high titers of antibodies to oxidation-specific epitopes, suggests that the antiatherogenic effect of immunization is not primarily dependent on very high titers of antibodies to oxidation-specific epitopes but is more likely to result from the activation of cellular immune responses.     

Oral immunization with a Salmonella typhimurium vaccine vector expressing recombinant enterotoxigenic Escherichia coli K99 fimbriae elicits elevated antibody titers for protective immunity

Bovine enterotoxigenic Escherichia coli (ETEC) continues to cause mortality in piglets and newborn calves. In an effort to develop a safe and effective vaccine for the prevention of F5(+) ETEC infections, a balanced lethal asd+ plasmid carrying the complete K99 operon was constructed and designated pMAK99-asd+. Introduction of this plasmid into an attenuated Salmonella typhimurium Deltaaro Deltaasd strain, H683, resulted in strain AP112, which stably expresses E. coli K99 fimbriae. A single oral immunization of BALB/c and CD-1 mice with strain AP112 elicited significant mucosal immunoglobulin A (IgA) titers that remained elevated for >11 weeks. IgA and IgG responses in serum specific for K99 fimbriae were also induced, with a prominent IgG1, as well as IgG2a and IgG2b, titer. To assess the derivation of these antibodies, a K99 isotype-specific B-cell ELISPOT analysis was conducted by using mononuclear cells from the lamina propria of the small intestines (LP), Peyer's patches (PP), and spleens of vaccinated and control BALB/c mice. This analysis revealed elevated numbers of K99 fimbria-specific IgA-producing cells in the LP, PP, and spleen, whereas elevated K99 fimbria-specific IgG-producing cells were detected only in the PP and spleen. These antibodies were important for protective immunity. One-day-old neonates from dams orally immunized with AP112 were provided passive protection against oral challenge with wild-type ETEC, in contrast to challenged neonates from unvaccinated dams or from dams vaccinated with a control Salmonella vector. These results confirm that oral Salmonella vaccine vectors effectively deliver K99 fimbriae to mucosal inductive sites for sustained elevation of IgA and IgG antibodies and for eliciting protective immunity.     

Main cardiotoxic drugs

The potential to increase passive serum antibody titres to a polysaccharide antigen in neonates, by preparturient vaccination of the dams was investigated. Dairy cows in five private herds were vaccinated with a commercial Pasteurella haemolytica culture supernatant vaccine (Presponse, Langford Inc.), at 6 and 3 weeks before their calculated due dates. Dams' sera, colostral whey, and post-colostral calf sera were assayed for antibodies of the IgG1 isotype binding purified capsular polysaccharide of P. haemolytica A1, using an enzyme immunoassay. Antibody titres were analyzed using the General Linear Model procedure (Statistical Analysis Systems Institute Inc.). Vaccinated dams had a significant increase in serum antibody titre after vaccination compared with non-vaccinates (P <0.01), and their antibody titres in colostral whey were significantly higher (P <0.05). Calves of vaccinated dams had significantly higher passive antibody titres than those of non-vaccinates (P <0.01) in all herds.     

Further observations on the use of a bivalent bacterin against Haemophilus gallinarum

Chickens vaccinated with two doses of a bivalent Haemophilus gallinarum bacterin were protected against seven strains of the organism. Vaccinated and unvaccinated birds with antibody to Mycoplasma gallisepticum had increased HI titers when challenged with H. gallinarum. Birds positive for antibody to Mycoplasma gallisepticum which were not challenged showed no increase in HI titer.     

Humoral and cellular immune responses in the murine respiratory tract following oral immunization with cholera toxin or Escherichia coli heat-labile enterotoxin

Cholera toxin (CT) and Escherichia coli heat-labile enterotoxin (LT) are the strongest mucosal immunogens identified to date and are also good adjuvants when given orally together in combination with unrelated antigens. We used these potent immunogens to monitor local and systemic immune responses following oral immunization of BALB/c mice, and compared their action on the following: (a) immunoglobulin production rates (IgG, IgM and IgA) in mucosal inductive (Peyer's patches-PPs), effector (intestinal lamina propria-LP, respiratory tract) and systemic (spleen) sites; (b) analysis of systemic antigen-specific antibodies (IgG subclasses, IgA and IgE); (c) time monitoring of fecal anti-CT and anti-LT antibodies, and (d) in vivo relevance of interleukin-6 (IL-6) to mucosal responses. Both mucosal immunogens elicited specific antibody responses (IgA, IgG) not only in the gastrointestinal tract (PP's and intestinal LP), but also in the respiratory tract and spleens of orally immunized mice. These mucosal responses were accompained by elevated secretion of IL-6 in all investigated tissues, indicating involvement of this cytokine in B-cell maturation processes. Furthermore, oral immunization with CT and LT induced elevated serum titers of IgG1 followed by IgG2a, IgG2b, IgG3 and IgA, while high antigen-specific IgA and IgG1 responses were found in fecal extracts. These findings illustrate the action of orally administered CT and LT, respectively, on several humoral and cellular immune responses not only at the gastrointestinal tract, the application site, but also in distant mucosal effector sites such as the respiratory tract. These data suggest the potential use of these mucosal adjuvants in oral immunization strategies to improve the local immune response in remote mucosal tissues, in accordance with the concept of a common mucosa-associated immune system.