Distinct regions of frequent loss of heterozygosity of chromosome 5p and 5q in human esophageal cancer.

Loss of heterozygosity (LOH) studies reported thus far suggest that tumor suppressor loci on chromosome 5q are important in esophageal cancer (EC) while little is known about the involvement of chromosome 5p. To investigate the potential existence of tumor suppressor gene(s) on chromosome 5 contributing to the development of EC, we performed LOH studies using a total of 24 polymorphic markers spanning the entire chromosome 5. Seventy primary esophageal cancers were microdissected and allelic deletions were detected by polymerase chain reaction (PCR)-single strand conformation polymorphism or by microsatellite analysis. LOH was observed in at least 1 of the loci in 47 of 70 (67%) esophageal tumors. Initially, 40 tumors [24 squamous cell carcinomas (SCC) and 16 adenocarcinomas (ADC)], each with matched histologically normal esophageal mucosa, were analyzed at 15 marker loci on 5p and 5q. A novel locus, D5S667 on 5p15.2, exhibited the highest frequency of LOH (44%) in these tumors along with another previously reported region of frequent deletion, irf-1 (5q31.1). In a series of 30 additional EC tumors (11 SCC and 19 ADC), a detailed LOH analysis of chromosome 5p15.2 region was conducted using 10 additional polymorphic markers, which mapped the frequently deleted region within 1 cM. Overall, LOH at the D5S667 locus was observed more frequently in SCC than in ADC (62% vs. 23%, p = 0.01). This significant rate of LOH of a distinct region of chromosome 5p implicates the existence of a putative tumor suppressor gene locus involved in EC.

Oral passive immunization against experimental salmonellosis in mice using chicken egg yolk antibodies specific for Salmonella enteritidis and S. typhimurium.

The efficacy of chicken egg yolk homotypic antibodies specific for outer membrane proteins (OMP), lipopolysaccharide (LPS) or flagella (Fla) in controlling experimental salmonellosis in mice was investigated. Mice challenged orally with 2 x 10(9) c.f.u. of Salmonella enteritidis or 2 x 10(7) c.f.u. of S. typhimurium were orally treated with 0.2 ml anti-OMP, -LPS or -Fla yolk antibody three times a day for three consecutive days. In mice challenged with S. enteritidis, antibody treatment resulted in a survival rate of 80%, 47% and 60% using OMP, LPS or Fla specific antibodies respectively, in contrast to only 20% in control mice. In the S. typhimurium trial, survival rate was 40%, 30% and 20% using OMP, LPS or Fla specific antibodies respectively in contrast to 0% in control mice. In vitro adhesion of S. enteritidis and S. typhimurium to HeLa cells was significantly reduced by anti-OMP, -LPS, and -Fla homotypic antibodies. Results suggest that egg yolk antibodies specific for Salmonella OMP, LPS, and Fla may protect mice from experimental salmonellosis when passively administered orally. Of these antibodies, anti-OMP exhibited the highest level of protection in vivo and in vitro.

Prevention of fatal salmonellosis in neonatal calves, using orally administered chicken egg yolk Salmonella-specific antibodies.

OBJECTIVE: To protect neonatal calves against fatal salmonellosis within the first 2 weeks after birth, using chicken egg yolk antibodies specific against Salmonella typhimurium or S dublin. ANIMALS: 38 neonatal Holstein calves from Salmonella-free farms. PROCEDURE: After removal of the lipid components with hydroxypropylmethylcellulose phthalate, egg yolk antibodies were spray dried. At 4 days of age, calves were challenge exposed by oral inoculation with 10(11) virulent S typhimurium (experiment 1) or S dublin (experiment 2). Starting from the challenge-exposure day, egg yolk antibody preparations were administered orally 3 times a day for 7 to 10 days. RESULTS: In passive immunization trials, the orally administered antibodies conferred dose-dependent protection against infection with each of the homologous strains of Salmonella. Within 7 to 10 days after challenge exposure, all control calves died, whereas low-titer antibody-treated calves had 60 to 100% mortality. Only fever and diarrhea, but no deaths (P < 0.01), were observed in calves given the highest titer of antibody. CONCLUSIONS AND CLINICAL RELEVANCE: Compared with that in control calves, survival was significantly higher among calves given antibodies with titers of 500 (P < 0.05) and 1,000 (P < 0.01) homotypic for S typhimurium and with titer of 5,000 (P < 0.01) for S dublin. Egg yolk antibodies specific for whole cell S typhimurium or S dublin are protective against fatal salmonellosis when given in sufficiently high concentration, and may be clinically useful during a salmonellosis outbreak.

Passage of chicken egg yolk antibody treated with hydroxypropyl methylcellulose phthalate in the gastrointestinal tract of calves.

Two types of chicken egg yolk antibody samples for oral passage trials in calves were prepared: (1) hydroxypropyl methylcellulose phthalate (HPMCP) antibody powder (HAP)--a powder produced by spray-drying a supernatant obtained after precipitation of lipids from egg yolk with HPMCP and (2) control antibody power (CAP)--a powder produced from an antibody solution with HPMCP. Antibody activity and pattern of distribution of both antibody preparations in the gastrointestinal tract of calves were compared by enzyme-linked immunosorbent assay. At 2 hr post administration, anti-K99 fimbrial antibodies from both the CAP and the HAP were detected in the abomasum of calves with titers of 1:128 and 1:256, respectively. However, at 4 hr, anti-K99 fimbrial titers of the CAP and the HAP were reduced to 1:2 and 1:64, respectively, due to digestion in the abomasum. These results indicated that the egg yolk antibody powder with HPMCP was more resistant against gastric juice in the stomach, thereby, ensuring a transfer of functional antibodies to the small intestine of calves after oral administration.

Passive immunisation against experimental salmonellosis in mice by orally administered hen egg-yolk antibodies specific for 14-kDa fimbriae of Salmonella enteritidis.

Chickens were immunised with a preparation of purified 14-kDa fimbriae of Salmonella serotype Enteritidis (SEF 14) to raise egg-yolk antibodies for protection trials in mice against subsequent challenge-exposure with the homologous strain of Enteritidis. A pronounced specificity of egg-yolk antibodies against the 14-kDa fimbrial antigen was demonstrated by Western blotting analysis. Passive antibody protection was evaluated in a mouse model of experimental salmonellosis: 79 mice (CD 1 strain) were challenged orally with 2 x 10(10) cfu of Enteritidis. Test mice treated with SEF-14 antibodies (titre = 128) had a survival rate of 77.8% compared to 32% survival in control mice fed normal egg-yolk antibodies (titre < 10) (p < 0.01). In-vitro adhesion of Enteritidis to mouse intestinal epithelial cells was reduced by anti-fimbrial antibodies. An indirect immunofluorescence method demonstrated the localisation of Enteritidis along the villous margins of the small intestine of control mice, whereas in test mice adherent bacteria were not detected. Results suggest that 14-kDa fimbriae may influence, enhance or contribute to the overall adhesive properties of Enteritidis and that egg-yolk antibodies directed against these fimbriae may have played a substantial role in protection, possibly by minimising bacterial colonisation and invasion during the early stages of infection.

Passive protection against bovine rotavirus in calves by specific immunoglobulins from chicken egg yolk.

The efficacy of chicken egg yolk immunoglobulins (yIg) from hens immunized with bovine rotavirus (BRV) serotype G6 (strain Shimane) or serotype G10 (strain KK-3) for protection against homologous BRV in calves was investigated. A significant protection by anti-BRV yIg having 6400 neutralizing antibody titer per dose could be achieved in calves (P < 0.01).

Research note: avidity of chicken yolk antibodies to enterotoxigenic Escherichia coli fimbriae.

The yolk antibodies from chickens and the serum and colostrum antibodies from cows were obtained after immunization of these animals with inactivated bacterin or purified K99 fimbriae from enterotoxigenic Escherichia coli (ETEC). The avidity of anti-K99 fimbriae antibodies produced from either chickens or cows was measured by competitive binding assay of ELISA. The yolk antibodies competed strongly with the serum and colostrum antibodies from immunized cows and inhibited 40 to 80% of the binding of these antibodies. Results demonstrate that the avidity of antibodies obtained from immunized chickens compares with that obtained from immunized cows. Thus, the yolk antibody from immunized chickens, aside from its use for prophylaxis against some infectious diseases, may also serve as effective ligand for purification of biologically active substances such as fimbrial antigens by affinity chromatographic procedures.

Passive protection against bovine rotavirus-induced diarrhea in murine model by specific immunoglobulins from chicken egg yolk.

Chicken egg yolk immunoglobulins (yIg) specific against bovine rotavirus (BRV) serotypes 6 (strain Shimane) and 10 (strain KK-3) were used for oral passive immunization of suckling mice against experimental BRV challenge. The protective capacity of the antibody preparation was tested using different concentrations of yIg against a challenge dose of 10(7.5) TCID50 for Shimane and 10(7.0) TCID50 for KK-3 strain. There was a significant homotypic (P < 0.05) and heterotypic (P < 0.01) protection using 160 anti-Shimane or 160 anti-KK-3 neutralizing antibody titer (NAT) compared to control mice given yIg derived from eggs of mock-immunized (control) hens. The titer of infectious BRV recovered from intestinal tissue or luminal chyme decreased with increasing homotypic yIg NAT. A decrease in degree and duration of BRV antigen localization in the villus epithelial lining was observed in mice treated with homotypic yIg at optimum dose for prevention of diarrhea. The NAT in sera of challenged mice increased with decreasing NAT in the yIg given before challenge suggesting that protection was dose-dependent. The present findings indicate that a passive protection could be achieved by the use of yIg against BRV-induced diarrhea in this murine model.

Detection of passage and absorption of chicken egg yolk immunoglobulins in the gastrointestinal tract of pigs by use of enzyme-linked immunosorbent assay and fluorescent antibody testing.

Chicken egg yolk IgG can be absorbed and transferred as efficiently as colostral antibodies in the blood of neonatal pigs. Egg yolk IgG has a half-life of 1.85 days in newborn pig serum. This is shorter than the reported half-life (12 to 14 days) of homologous IgG in serum of pigs. Similar to colostral antibodies, egg yolk IgG absorption from intestine ceased at about 34 hours of age, after a logarithmic decrease in absorption rate from birth. Egg yolk IgG absorption inhibition time in the gastrointestinal tract took 1.73 hours to decrease by half. Egg yolk IgG was protective against experimentally induced diarrhea in pigs when it was administered at high dose, and multiple dosing was instituted. Adverse effects were not observed when chicken egg yolk IgG was administered orally to pigs.

Protection of neonatal calves against fatal enteric colibacillosis by administration of egg yolk powder from hens immunized with K99-piliated enterotoxigenic Escherichia coli.

The protective effects of egg yolk powder prepared from hens vaccinated with heat-extracted antigens from K99-piliated enterotoxigenic Escherichia coli (ETEC) strain 431 were evaluated in a colostrum-fed calf model of ETEC-induced diarrhea caused by a heterologous strain (B44). The antibody powder was obtained by spray-drying the water-soluble protein fraction of egg yolks after removing the lipid and fatty components by precipitation with hydroxypropylmethylcellulose phthalate. A total of 16 colostrum-fed calves were studied to determine whether the orally administered antibody powder would prevent fatal bovine colibacillosis caused by a virulent ETEC strain. Clinical response of individual calves was monitored and evaluated in the context of these variables: fecal consistency score, intestinal colonization, weight loss, and mortality. Control calves that were treated with vehicle (milk with egg yolk powder from nonimmunized hens) had severe diarrhea and dehydration and died within 72 hours after infection was manifested. In contrast, calves fed milk containing egg yolk powder with antipili agglutinin titers of 1:800 and 1:1,600 had transient diarrhea, 100% survival, and good body weight gain during the course of the study. Results indicate that the orally administered egg yolk powder protected against ETEC-induced diarrhea in neonatal calves and that the protective components may have been the antibodies raised by vaccination of chickens against ETEC.

Passive protective effect of chicken egg yolk immunoglobulins against experimental enterotoxigenic Escherichia coli infection in neonatal piglets.

Passive protection of neonatal piglets against fatal enteric colibacillosis was achieved with powder preparations of specific antibodies against K88, K99, and 987P fimbrial adhesins of enterotoxigenic Escherichia coli. The antibody powders were obtained by spray drying the water-soluble protein fraction of egg yolks from immunized hens after the lipid components were precipitated with an aqueous dispersion of acrylic resins (Eudragit L30D-55; Rohm pharma). The anti-K88, -K99, and -987P antibody preparations reacted specifically against the corresponding fimbrial antigens in an enzyme-linked immunosorbent assay. The orally administered antibodies protected in a dose-dependent fashion against infection with each of the three homologous strains of E. coli in passive immunization trials with a colostrum-deprived piglet model of enterotoxigenic E. coli diarrhea. Scanning electron microscopy revealed adherence of enterotoxigenic E. coli in intestinal epithelial surfaces of control piglets, whereas in treated piglets treated with high-titer antibodies, a resistance to bacterial adhesion was observed. An enzyme immunoassay with avidin-biotin complex demonstrated specific local antibody activity in target areas of the small intestines. In vitro, E. coli K88+, K99+, and 987P+ strains adhered equally to porcine duodenal and ileal epithelial cells but failed to do so in the presence of homologous anti-fimbrial antibodies. Absorption of egg yolk antibodies with fimbrial immunosorbent removed the anti-fimbrial antibody fraction and reduced significantly the protective nature of the antibody preparation in a passive immunization experiment, suggesting that anti-fimbrial antibodies were the active components.