Effect of immunoglobulin Y formulations on oral microorganisms in human subjects - A systematic review and meta-analysis of clinical trials

Background: Passive immunization using egg yolk-based antibodies has been tested against oral microorganisms. Our study assessed the effect of immunoglobulin Y (IgY) formulations on Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans in human subjects. Highlights: VS and UT independently searched articles using keyword combinations in four search engines; studies in English were selected. Either parallel-arm or split-mouth randomized controlled trials on healthy human subjects were considered. Ten studies remained in the selection; six studies compared the effect of IgY formulations on S. mutans, three on P. gingivalis, and one on C. albicans. Five studies (422 subjects) compared the effect of IgY formulations on S. mutans. When fixed-effect model (FEM) was applied, the risk ratio (RR) (confidence interval [CI]) was found to be 7.81 (6.00, 10.18). Three studies (167 subjects) compared the effect of IgY formulations on P. gingivalis. When FEM was applied, the RR (CI) was found to be 0.06 (?0.03, 0.15) in relation to reduction in probing depth. When FEM was applied, for percentage reduction in bleeding on probing (BOP), the RR (CI) was 1.99 (1.64, 2.41). Only one study (26 subjects) was available of IgY formulation and C. albicans; hence meta-analysis was not performed. The search was extended using Google Scholar, Semantic Scholar, cross-references and by contacting authors and researchers in the field which further yielded five articles. . Conclusions: IgY formulations were effective in the reduction of S. mutans. They were not effective on P. gingivalis in relation to probing depth but were effective in relation to reduction in BOP. No harms were reported. Evidence is of low quality due to high heterogeneity. The ROB was moderate and publication bias was low.

An immunoglobulin y that specifically binds to an in silico-predicted unique epitope of Zika virus non-structural 1 antigen

Objective: To identify unique immunogenic epitopes of Zika virus non-structural 1 (NS1) antigen and produce immunoglobulin Y (IgY) for potential use in he diagnosis of of Zika virus infection. Methods: Immunogenic epitopes were identified using in silico B-cell epitope prediction. A synthetic peptide analog of the predicted epitope was used to induce antipeptide IgY production in hens which was purified using affinity chromatography. Presence of purified IgY and its binding specificity were performed by gel electrophoresis and ELISA, respectively. Results: Out of the nine continuous epitopes identified, the sequence at position 193-208 (LKVREDYSLECDPAVI) was selected and used to produce anti-peptide IgY. The produced IgY was found to bind to the synthetic analog of the Zika virus NS1 immunogenic epitope but not to other flaviviruses and random peptides from other pathogens. Conclusions: In this study, we identified an immunogenic epitope unique to Zika virus that can be used to develop a serodiagnostic tool that specifically detect Zika virus infection.

Microencapsulation of immunoglobulin Y: optimization with response surface morphology and controlled release during simulated gastrointestinal digestion / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Immunoglobulin Y (IgY) is an effective orally administered antibody used to protect against various intestinal pathogens, but which cannot tolerate the acidic gastric environment. In this study, IgY was microencapsulated by alginate (ALG) and coated with chitooligosaccharide (COS). A response surface methodology was used to optimize the formulation, and a simulated gastrointestinal (GI) digestion (SGID) system to evaluate the controlled release of microencapsulated IgY. The microcapsule formulation was optimized as an ALG concentration of 1.56% (15.6 g/L), COS level of 0.61% (6.1 g/L), and IgY/ALG ratio of 62.44% (mass ratio). The microcapsules prepared following this formulation had an encapsulation efficiency of 65.19%, a loading capacity of 33.75%, and an average particle size of 588.75 µm. Under this optimum formulation, the coating of COS provided a less porous and more continuous microstructure by filling the cracks on the surface, and thus the GI release rate of encapsulated IgY was significantly reduced. The release of encapsulated IgY during simulated gastric and intestinal digestion well fitted the zero-order and first-order kinetics functions, respectively. The microcapsule also allowed the IgY to retain 84.37% immune-activity after 4 h simulated GI digestion, significantly higher than that for unprotected IgY (5.33%). This approach could provide an efficient way to preserve IgY and improve its performance in the GI tract.

Microencapsulation of immunoglobulin Y: optimization with response surface morphology and controlled release during simulated gastrointestinal digestion / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Immunoglobulin Y (IgY) is an effective orally administered antibody used to protect against various intestinal pathogens, but which cannot tolerate the acidic gastric environment. In this study, IgY was microencapsulated by alginate (ALG) and coated with chitooligosaccharide (COS). A response surface methodology was used to optimize the formulation, and a simulated gastrointestinal (GI) digestion (SGID) system to evaluate the controlled release of microencapsulated IgY. The microcapsule formulation was optimized as an ALG concentration of 1.56% (15.6 g/L), COS level of 0.61% (6.1 g/L), and IgY/ALG ratio of 62.44% (mass ratio). The microcapsules prepared following this formulation had an encapsulation efficiency of 65.19%, a loading capacity of 33.75%, and an average particle size of 588.75 μm. Under this optimum formulation, the coating of COS provided a less porous and more continuous microstructure by filling the cracks on the surface, and thus the GI release rate of encapsulated IgY was significantly reduced. The release of encapsulated IgY during simulated gastric and intestinal digestion well fitted the zero-order and first-order kinetics functions, respectively. The microcapsule also allowed the IgY to retain 84.37% immune-activity after 4 h simulated GI digestion, significantly higher than that for unprotected IgY (5.33%). This approach could provide an efficient way to preserve IgY and improve its performance in the GI tract.

Isolation of cariogenic Streptococcus mutans and the inhibitory effect of egg yolk antibody on caries / 中华口腔医学杂志

Objective@#To isolate the cariogenic Streptococcus mutans (Sm) strains and study the therapeutical effect of egg yolk antibody (IgY) of the Sm on dental caries development.@*Methods@#Sm strains were isolated from the children's dental plaque samples. Morphological, biochemical and molecular biological methods were applied to identify the serotype, acid producing and adhesion abilities of isolated Sm strains. After inactivation one of the Sm strains was used as antigen to immune laying hens to collect and extract the specific anti-Sm IgY. The rats were infected with Sm (serotype e). After 16 weeks of infection, all the rats were found developing dental caries. The rats were then randomly divided into two groups. The rats in experimental group were supplied with diet containing anti-Sm IgY while the rats in control group with normal IgY. All rats were sacrificed after another 8 weeks' observation. The degree of caries for each rat was assessed using Keyes' method.@*Results@#We isolated 7 Sm strains from the children's dental plaque samples in the present study. The numbers of serotype c, e, f, k were 3, 2, 0 and 2, respectively. All strains showed similar morphological and biochemical characters as standard UA159 Sm strain, and possessed strong capabilities of acid production and adherence. Interestingly, even the same serotypec strains, such as No.3 and No.7 strains, demonstrated significant difference on acid producing and adherence capabilities. After 16 weeks infection with serotype e strain, the rats' mandibular teeth were apparently decayed, and treatment with specific anti-Sm IgY obviously attenuated the development of caries in the experiment group rats (16.4±2.0) compared with that in the control group rats (30.2±9.3) (P<0.05) determined by Keyes' method.@*Conclusions@#Seven cariogenic Sm strains of different serotypes were isolated, which possesses similar morphology and biochemical characters. Although belonging to the same serotype strains they always show significant difference in acid-producing and adherencec apabilities. Further experiment provides evidences that the serotype e strain could obviously induce caries independently, and employment of specific anti-Sm IgY as passive immunotherapy additive might effectively inhibit the further development of dental caries.

Development and characterization of IgY antibodies against synthetic peptides from Brucella abortus OMP25 and BP26 proteins / Desarrollo y caracterización de anticuerpos IgY contra péptidos sintéticos de las proteínas OMP25 y BP26 de Brucella abortus

Brucellosis is a zoonose produced by bacterial species from the Brucella genus. Its isolation and identification in food using classical microbiological techniques is not practical due to its slow growth rate. Therefore, it is necessary to establish fast and specific methods for the detection of the bacteria in food. The goal of this work was the production and characterization of monospecific polyclonal antibodies in chicken (IgY) against synthetic peptides from Brucella abortus OMP25 and BP26 proteins, suitable for an antigen-capture assay. Conformational as well as antigenic predictions were performed using the ANTHEPROT package. Chemical synthesis was carried out by the multiple manual synthesis using the t-boc strategy. The peptides were used as antigens for the preparation of polyclonal antibodies in chicken. Experimental animals produced specific antibodies against the OMP25 and BP26 peptides constructs determined by ELISA and MABA assays showing correspondence between the predictive study and the immunogenicity obtained in chicken. The IgY proved to be able to recognize B. abortus by MABA assays. The binding activity and specificity of antibodies was determined by Western blot with cell extract from B. abortus. In this study, we demonstrated that OMP25 and BP26 peptides constructs are good candidates for production of specific IgY antipeptide antibodies capable of recognizing proteins from sonicated B. abortus strain S19, indicating the potential usefulness of the IgY antibody for development of immunoassays for detection of Brucella abortus.

La brucelosis es una zoonosis producida por especies del género Brucella. El aislamiento e identificación de la bacteria en alimentos usando las técnicas clásicas de microbiología no es práctico debido a su lenta tasa de crecimiento. Por lo tanto, es necesario establecer métodos rápidos para la detección de la bacteria en alimentos. En el presente trabajo se desarrollaron y caracterizaron anticuerpos policlonales monoespecíficos en gallinas (IgY) contra péptidos sintéticos de las proteínas OMP25 y BP26 de Brucella abortus, que puedan ser utilizados en un ensayo de captura. Para ello, se realizaron estudios conformacionales y de predicción de epítopes en la selección de los péptidos, los cuales se utilizaron como antígenos para la producción de las IgY. Los animales desarrollaron anticuerpos específicos contra los péptidos, mostrando correspondencia entre los estudios predictivos y la inmunogenicidad obtenida. Las IgY reconocieron a B. abortus en un ensayo de MABA y la actividad de unión y especificidad fue determinada por western blot con extracto celular de B. abortus. En este estudio, demostramos que los péptidos de las proteínas OMP25 y BP26 de B. abortus son buenos candidatos para la producción de anticuerpos IgY especificos capaces de reconocer proteínas de extracto de B. abortus cepa S19, indicando el potencial uso de anticuerpos IgY para el desarrollo de inmunoensayos para la detección de Brucella abortus.

Using immunoglobulin Y as an alternative antibody for the detection of hepatitis A virus in frozen liver sections

An increasing amount of research has been conducted on immunoglobulin Y (IgY) because the use of IgY offers several advantages with respect to diagnostic testing, including its easy accessibility, low cost and translatability to large-scale production, in addition to the fact that it can be ethically produced. In a previous work, immunoglobulin was produced and purified from egg yolks (IgY) reactive to hepatitis A virus (HAV) antigens. In the present work, this anti-HAV-specific IgY was used in an indirect immunofluorescence assay to detect viral antigens in liver biopsies that were obtained from experimentally infected cynomolgus monkeys. Fields that were positive for HAV antigen were detected in liver sections using confocal microscopy. In conclusion, egg yolks from immunised hens may be a reliable source for antibody production, which can be employed for immunological studies.

Production and characterization protein of anti HIV specific immunoglobulin Y for Immunotherapy.

In this study, an approach for large-scale production of immunoglobulin Y ( Ig Y) specific anti Human Immunodeficiency Virus (HIV) for use as a potential immunotherapy for HIV disease. Lohman laying hens were immunized intramuscularly with HIV virus that had been inactivated using formaline. The immunizations were repeated two times with dose of each 1 ml of HIV antigen (20 copy/ml) with an interval of two week. The first immunizations were HIV antigen mixed with Fruend Adjuvant Complete and subsequently mixed with Freund Adjuvant Incomplete. Egg yolk was separated from egg white and immunoglobulin Y (IgY) antibody was then purified by multiple polyethylene glycol (PEG) 6000 extraction and ammonium sulfate purification steps. Antibody response in yolk was detected by agar gel precipitation test (AGPT) and the protein pattern was detected using polyacrilamid gel electrophoresis (SDS-PAGE). Specific activity of the antibody was tested using commercial ELISA. Antibody of HIV was detected and produce a specific line of precipitation in AGPT beginning the second week after the first immunization. Analysis of results obtained with ELISA showed significant increase in the HIV-specific antibodies after two weeks from the primary immunization. Through the effect of boostering; the anti-HIV antibody levels reached a plateau at six weeks from the primary immunization and remained significantly higher till the end of observation period. SDS-PAGE revealed the IgY preparation to be pure and dissociated into protein bands with molecular weights of 164; 87; 68, and 37 kDa. These results suggested that chicken IgY could be a practical strategy in large-scale production of Ig Y specific anti HIV for immunotherapy and diagnostic KIT of HIV disease.

Avian IgY antibodies: characteristics and applications in immunodiagnostic / Anticorpos IgY aviário: características e aplicações em imunodiagnóstico

Immunoglobulin Y (IgY) is the major antibody isotype in birds, reptiles, amphibia, and lungfish, playing a similar biological role as mammal IgG. Due to its phylogenetic distance, immune diversification and presence in the egg yolk, IgY provide a number of advantages in immunodiagnostic compared to IgG from mammals. Moreover, IgY production is in agreement with international efforts to reduce, refine and if possible, to replace animals in experimentation, contributing substantially in favor of animal welfare. This article presents an overview about structural and functional features, production and applications of IgY in immunodiagnostic, as well as the advantages of chicken antibodies use.

A imunoglobulina Y (IgY) é a classe de anticorpos de maior importância em aves, répteis, anfíbios e peixes pulmonados, desempenhando um papel semelhante a IgG de mamíferos. Devido a sua distância filogenética, mecanismos de diversificação imune e presença na gema do ovo, IgY proporciona uma série de vantagens em imunodiagnóstico, quando comparada a IgG de mamíferos. Além disso, esse método alternativo de produção de anticorpo está de acordo com os esforços internacionais para reduzir, refinar e, se possível, substituir animais em experimentação, contribuindo substancialmente a favor do bem-estar animal. Este artigo apresenta uma revisão sobre as características estruturais e funcionais da IgY, bem como os métodos de produção, vantagens e aplicações em imunodiagnóstico, além das vantagens da sua utilização.

Production of Egg Yolk Antibodies Specific to House Dust Mite Proteins

PURPOSE: House dust mites (HDMs) are an important source of indoor allergens associated with asthma, rhinitis and atopic dermatitis. Chicken immunoglobulin (Ig) Y is known to be a good alternative to mice and rabbit antibody production. In this study, we produced IgYs specific to HDMs and investigated their IgE immunoreactivities. MATERIALS AND METHODS: Total IgYs were isolated from the yolks of White Leghorn hens immunized with either Dermatophagoides pteronyssinus or D. farinae protein extract. Control antibodies were separated from the yolks of immunized hens with phosphate buffered saline. IgYs specific to HDMs were analyzed using enzyme-linked immunosorbent assay and Western blotting analysis. RESULTS: The concentration of egg IgY specific to D. farinae in an immunized hen increased and the highest achieved was 661.3 ug/mg (per an egg) on day 47, compared with 760 ug/mg IgY specific to D. pteronyssinus on day 16. The D. pteronyssinus or D. farinae-specific IgY was detected by binding of each mite proteins, and their immunoreactivities were elevated dependent of the specific IgY concentration. CONCLUSION: IgY specific to HDMs may be a promising antibody for immunological diagnosis as well as identification of possible resistance relating to HDM allergy.

Produção e purificação de imunoglobulinas Y policlonais anti-Leptospira spp / Production and purification of polyclonal anti-Leptospira immunoglobulin Y

Objetivou-se verificar se galinhas imunizadas com uma solução de Leptospira interrogans inativadas e proteínas de membrana externa do sorovar Hardjo, poderiam produzir anticorpos policlonais específicos anti-leptospiras, detectáveis em testes ELISA. Foram imunizados oito galinhas com 25 semanas de idade, da raça White Leghorn, sendo três imunizadas com uma suspensão de leptospiras inativadas, três com uma solução de proteínas de membrana externa extraída do sorovar Hardjo e duas controle. Coletas de sangue foram realizadas quinzenalmente e de ovos diariamente. A IgY foi purificada a partir da gema dos ovos utilizando para a delipidação o método de diluição em água ácida e a precipitação com sulfato de amônio. Nos testes ELISA realizados para verificar a especificidade da IgY, foi demonstrada a produção de anticorpos anti-Leptospira, tanto no soro quanto nas gemas purificadas. O pico de produção de anticorpos específicos ocorreu na 5º semana após a primeira imunização. Ficou demonstrada a possibilidade da indução da produção de anticorpos específicos em galinhas imunizadas com leptospiras do sorovar Hardjo inativadas, bem como, com proteínas de membrana externa (PME) extraidas desse sorovar. As galinhas imunizadas com uma suspensão de leptospiras inativadas ou com PME de Leptospira interrogans do sorovar Hardjo produziram anticorpos reativos a PME Hardjo detectáves por teste ELISA.

The aim was to determine whether hens immunized with an inactivated suspension of Leptospira and a solution of outer membrane proteins extracted from the serovar Hardjo, could produce specific polyclonal antibodies to Leptospira, detected in ELISA assay. Eight hens White Leghorn race with 25-weeks-old were immunized, three with an inactivated suspension of Leptospira, three with a solution of outer membrane proteins (OMP) extracted from the serovar Hardjo and two controls immunized with saline. Blood samples were collected fortnightly and eggs daily. The IgY was purified from the egg yolk using the method for the delipidation of dilution with water acidic and ammonium sulfate precipitation. The ELISA assay was performed to verify the specificity of the IgY, these was possible to observe the production of specific antibody to Leptospira both in serum and purified egg yolk. The specific antibody titers peaked in the fifth week post immunization. The production of polyclonal IgY was effective for producing high titers of specific antibodies.

Production and evaluation of chicken antibodies against a synthetic peptide from glial growth factor / Producción y evaluación de anticuerpos de gallinas contra un péptido sintético del factor de crecimiento glial

Neuregulins (NRG) are proteins that belong to the family of epidermal growth factors. It is well established that these factors are essential for the development and maintenance of the nervous system. Due to the difficulty of purifying enough quantities of these factors and the lack of specificity from commercially available antibodies, the aim of this work was to produce antibodies against a synthetic peptide capable to detect and identify neuregulin GGFb isoforms. To accomplish this goal, polyclonal antibodies were raised in hens against a synthetic peptide designed from the GGFb1 extracellular sequence. The sequence analysis was made using different epitope-predicting programs. Our results showed that the peptide sequence selected was immunogenic because it was capable of inducing a specific type B immune response in the experimental animal model. These antibodies were also capable of recognizing a recombinant GGF protein and GGF isoforms present in different samples. Our results suggest that the development of immunoglobulin Y (IgY) using synthetic peptides represents, a valuable tool for neuroscience research.

Las Neuregulinas (NRG) son proteínas que pertenecen a la familia de los factores de crecimiento epidermal. Se ha demostrado que estos factores son esenciales para el desarrollo y mantenimiento de la funcionalidad del sistema nervioso. Debido a la dificultad para purificar estas proteínas y la falta de especificidad de los anticuerpos disponibles comercialmente, el objetivo de este trabajo fue producir anticuerpos contra un péptido sintético capaz de detectar e identificar una isoforma de la Neuregulina (GGFb). Para lograr este objetivo, se desarrollaron anticuerpos en gallinas (IgY) contra un péptido sintético diseñado a partir de la secuencia aminoacídica de la región extracelular de GGFb, utilizando programas de predicción de epítopes. Los resultados demuestran que el péptido seleccionado fue immunogénico debido a que estimuló una respuesta inmune específica tipo B en el modelo utilizado. Estos anticuerpos fueron también capaces de reconocer una proteína recombinante e isoformas de GGF presentes en diferentes muestras biológicas. Nuestros resultados demuestran el potencial valor de las inmunoglobulinas Y (IgY) contra péptidos sintéticos como una herramienta de aplicación para la investigación en neurociencia.

Production of immunoglobulin Y (IgY) against synthetic peptide analogs of the immunogenic epitopes of the hepatitis B surface antigen

INTRODUCTION: Several studies have been conducted on the use of Immunoglobulin Y (IgY) technology in the fields of diagnostics and therapeutics. IgY is the avian counterpart of the mammalian immunoglobulin G (IgG) which is exclusively transferred from the hen to the yolk thus conferring passive immunization to the growing embryo. However, despite the advantages it offers over the use of mammalian immunoglobulin, IgY technology has remained underutilized. OBJECTIVE:The objective of this study is to produce an IgY with activity against synthetic peptide analogs of known immunogenic epitopes of the Hepatitis B Surface Antigen (HBsAg) - a molecular marker of Hepatitis B infection. METHODS: Chickens were immunized with synthetic peptide analogs of previously reported immunogenic epitopes of the S and the pre-S1 regions of the Hepatitis B surface antigen (HBsAg). IgY specific for the synthetic peptides was isolated by delipidation and salt precipitation and was further purified by affinity chromatography. Purity and molecular weights of the whole IgY molecule and its subunits were assessed and determined by SDS-PAGE. Anti-peptide activity and specificity were determined by indirect ELISA. The study was approved by the Ethical Review Board (ERB) and Technical Review Board of the Research Implementation and Development Office (RIDO), University of the Philippines Manila. RESULTS AND CONCLUSION: The IgY that was purified in this study had an approximate molecular weight of 165 kilodaltons. The heavy and light chains are 60 and 28 kilodaltons, respectively. The affinity purified IgY demonstrated anti-peptide antibody activity against synthetic peptide analogs of known immunogenic epitopes of the HBsAg. Specific binding against a battery of synthetic peptides also revealed that the affinity purified IgY specifically binds to the known immunogenic epitope of the HBsAg.

Anti-Helicobacter pylori effects of IgY from egg york of immunized hens / 한국실험동물학회지

Effects of egg york containing IgY specific for Helicobacter pylori on the bacterial growth and intragastric infection were investigated in comparison with a proton-pump inhibitor pantoprazole. For in vitro anti-bacterial activity test, H. pylori (1x108 CFU/mL) was incubated with a serially diluted IgY for 3 days. As a result, IgY fully inhibited the bacterial growth at 16 mg/mL, which was determined to a minimal inhibitory concentration. In vivo elimination study, male C57BL/6 mice were infected with the bacteria by intragastric inoculation (1x108 CFU/mouse) 3 times at 2-day intervals, and 2 weeks later, orally treated twice a day with 50, 100, 200 or 500 mg/kg IgY for 18 days. After the final administration, biopsy sample of the gastric mucosa was assayed for the bacterial identification via urease, oxidase, catalase, nitrate reduction and H2S tests in addition to microscopic examination for mucosal inflammation. In CLO kit test, 75, 50, 12.5 and 12.5% of the animals revealed positive reaction following treatment with 50, 100, 200 and 500 mg/kg IgY, respectively, resulting in a superior efficacy at 200 mg/kg than 30 mg/kg pantoprazole that displayed 75% elimination. The CLO test results were confirmed by bacterial identification. Microscopic examination revealed that H. pylori infection caused severe gastric mucosal inflammation, which were not observed in the CLO-negative mice following treatment with IgY or pantoprazole. Taken together, IgY inhibited the growth of H. pylori, and improved gastritis and villi injuries by eliminating the bacteria from the stomach. The results indicate that IgY could be a good candidate overcoming tolerance of antibiotics for the treatment of H. pylori-mediated gastric ulcers.

Use of hen egg derived immunoglobulin against scolopendra (Scolopendra gigantea) venom

Las picaduras de escolopendra (Scolopendra gigantea) en seres humanos y animales domésticos representan un accidente agudo y muy doloroso. La necrosis y otros daños ocasionados por este veneno pueden ser prevenidos, si se inyecta un antiveneno. Este estudio propone producir anticuerpos policlonales en gallinas hiper-inmunizadas contra el veneno de escolopendra (Scolopendra gigantea Linneaus 1758). Un grupo de gallinas fue inyectado subcutánea e intramuscularmente con diluciones de venenos, de acuerdo con tres rutinas diferentes. Los huevos fueron recogidos diariamente y los anticuerpos en la yema fueron purificados con un método modificado de polietilen-glicol y cloroformo. Los niveles de anticuerpos en yema fueron calculados con prueba de precipitación de gel de agar y pruebas de protección (ED50). Los huevos cosechados 15 días post-inyección tenían los títulos más altos de anticuerpos. Después de seis meses, los anticuerpos liofilizados y guardados a 5°C mantenían su actividad. Ratones envenenados, inyectados posteriormente con anticuerpos purificados, tuvieron un 100% de supervivencia al compararse con los controles. La limpieza, la eficacia, y la sencillez de producir los antivenenos en gallina, y la incapacidad de estos anticuerpos (IgY) para fijarse al complemento humano, formulan una alternativa interesante a otros antivenenos producidos en mamíferos. Este estudio puntualiza que los anticuerpos de huevo de gallina pueden ser provechosos como un instrumento terapéutico para tratar escolopendrismo en seres humanos y animales domésticos. Además, abre un campo terapéutico para la fabricación de otros antivenenos contra el espectro amplio de las toxinas y como probable herramienta de diagnóstico.

A scolopendra sting in humans and domestic animals is an acute and highly painful accident. The present study was an attempt to raise specific hyper-immune polyclonal antibodies against scolopendra centipede (Scolopendra gigantea Linneaus 1758) venom. A group of hens were injected with venoms subcutaneously and intramuscularly according to three different routines. This protocol was found to be effective for hyperimmunization. Eggs were gathered daily and antibodies were purified from yolk with a polyethylene-glycol and chloroform modified method. Titers of antibodies in yolk were estimated with an agar gel precipitation test, and a serum protection (ED50) test. Eggs harvested at 15 days post-injection had maximum antibody titers. After six months, antibodies lyophilized and stored at 5°C still maintained their activity. Envenomed mice were injected with purified antibodies, which induced 100% recovery as compared to those not treated with the antibodies. The cleanliness, effectiveness, and simplicity of producing antibodies against scolopendra venom in avian egg yolk, and their incapability to attach human complement, formulates an interesting option to equine and other mammalian antivenoms. This study infers that avian egg yolk antibodies may be useful as a therapeutic tool in treating scolopendrism in humans and domestic animals. It also opens a field for the production of other antivenoms against the wide spectrum of toxins as well as the use of these antibodies as a diagnostic tool.

Passive immunization using purified IgYs against infectious bursal disease of chickens in Pakistan

Infectious bursal disease (IBD) is an acute and highly contagious disease of young chickens caused by Birnavirus. Mortality of infected birds can be best prevented if injected with antibodies. The present study was an attempt to raise specific hyper-immune polyclonal antibodies against IBD virus in Pakistan. Commercial layers divided into four groups were injected with IBD vaccine subcutaneously according to four different treatment regimens. Eggs were collected daily and antibodies were purified from yolk with dextran sulphate. Titers of antibodies in serum and yolk were evaluated with enzyme linked immunosorbant assay and agar gel precipitation test. Antibody titers were significantly higher in yolk than serum. Eggs collected at 28 days post-vaccination had maximum antibody titers. Of treatment regimens, T3 was found to be most effective for hyperimmunization. Lyophilized antibodies stored at 4oC did not lose their activity till the end of experiment. IBD virus infected birds were injected with purified antibodies which induced 92% recovery as compared to control birds. The study implicates that the purified antibodies may be useful as a therapeutic agent to cure IBD infected birds.

Therapeutic Effect of Anti-Rotavirus Chicken Egg Yolk Immunoglobulin (IgY) on Diarrhea by Infection of Rotavirus / 소아과

PURPOSE: Rotavirus is an enteric pathogen that affects millions of children globally each year. But no specific therapy is available for the management of rotavirus diarrhea. Due to the clear need to define improved modality for treatment of rotavirus diarrhea, we evaluated the efficacy of anti- rotavirus IgY in the treatment of infants and children with gastroenteritis. METHODS: First, the amount of viral particle in the stools of thirteen patients (seven were given IgY, 6 placebo) infected by rotavirus were evaluated for 3 days with the quantitative RT-PCR method. Second, 36 children with known rotavirus infection identified by ELISA or semi-quantitative RT- PCR were evaluated. We gave 5 g anti-rotavirus egg yolk daily in two equally divided doses for 3 days to two groups (an 18 IgY group and an 18 placebo group), respectively after parenteral consent. Daily vomiting frequency, stool frequency, oral intake and urine output were monitored for 3 days, and electrolyte and blood chemistry were checked at the first and third days. RESULTS: First, in the placebo group, the amount of virus particles increased daily, but in the IgY group it decreased daily. Second, when IgY and placebos were given to children infected with rotavirus, diarrhea on the third day decreased significantly in the IgY group, compared with the placebo group. CONCLUSION: Treatment with antirotavirus immunoglobulin from immunized chicken's egg resulted in a decrease in the amount of viral particles in stools and diarrhea frequency in children. These results suggest that anti-rotavirus IgY is effective in the treatment of rotavirus gastroenteritis.

Effects of anti-lipopolysaccharide egg yolk immunoglobulin on LPS in vitro / 第三军医大学学报

Objective To observe the effects of anti lipopolysaccharide (LPS) immunoglobulin Y (IgY) on LPS in vitro . Methods The direct in vitro neutralization of different concentrations of IgY and LPS was conducted for the detection of the remained amount of LPS to observe the dose effect relationship. The primary cultured human umbilical vein endothelial cells (HUVECs) were cultured for 48-72 h after addition of certain concentrations of fluorescein isothiocyanate LPS (FITC LPS) and IgY. Changes of the fluorescent intensity in HUVECs were observed at different time points. After addition of certain concentrations of LPS and IgY for incubation with HUVECs for 4, 8, and 12 h, the changes of lactate dehydrogenase (LDH) activity were measured. Results The dose effect relationship was satisfactory when the ratio was 16︰1. At 4, 8, and 12 h, fluorescent intensity was somewhat strong in FITC LPS group, but weak in IgY group, especially at 8 and 12 h. The LDH activity in medium increased obviously after incubation for 8 h in LPS group ( P

Preparation of IgY against Actinobacillus actinomycetemcomitans and growth suppression of actinobacillus actinomycetemcomitans and capnocytophaga gingivalis by specific IgY against Actinobacillus actinomycetemcomitans / 实用口腔医学杂志

Objective:To prepare specific IgY production using Actinobacillus actinomycetemcomitans(A.a) immunizing hen, and then to investigate anti-Actinobacillus actinomycetemcomitans IgY inhibiting growth of A.a and Capnocytophaga gingivalis(C.g). Methods:Using immunization method, water-dilution method, two-step ammonium sulfate precipitation, inhibiting bacteria growth test in liquid anaerobic culture, and ELISA, IgY were induced, extracted, purified, and inhibiting growth of A.a and C.g by the IgY was roundly evaluated. Results:The IgY purity reached to 85.6%~90.3% through 550 g/L and 330 g/L ammonium sulfate precipitation, and efficacy value was 1∶32 000. The IgY efficacy value of anti- A.a was 1∶8 000 against C.g in cross-reactivity.When IgY concentrations of anti-A.a were in the 5.0,1.0,0.1 g/L and concentration of A.a was in the 5?108 CFU/L, the suppression rate of A.a growth were 31.60%(P=0.004),10.24%(P=0.024),-3.30% respectively during 24 h culture and were 64.20%(P=0.004),53.21%(P=0.002),11.20% respectively in 72 h culture. When the concentration of A.a was in the 1?108 CFU/L, the suppression rate of A.a growth were 35.71%(P=0.004),30.95% (P=0.012),11.11% respectively during 24 h culture, and were 65.11%(P=0.005),54.04%(P=0.002),16.17% respectively during 72 h culture. When 5.0 g/L IgY of anti-A.a was cultured with 1?108 CFU/L C.g for 24 h, the suppression rate of C.g growth was 41.61%(P=0.005), and for 72 h it was 86.99%(P=0.014). Conclusion:The hen is able to be induced to produce high efficacy value IgY of anti-A.a by A.a. The specific IgY of anti-A.a is capable of inhibiting A.a and C.g growth. There are common antigens and cross immunizing reactivity between A.a and C.g.

Preparation of immunoglobulin Y against avain influenza virus(H_5N_1)and studies on the inhibitory effect to influenza virus A (FM_1) in vitro / 中国免疫学杂志

Objective:To prepare the immunoglobulin Y(IgY) against Avain Influenza Virus (H5N1) and to investgate its inhibitory activity to Influenza virus A (FM1).Methods:Iimmunized by Avain Influenza vaccine,IgY against Avain Influenza Virus was purified from the yolk and then the antibody was enriched by the means:caprylate acid-two-step salt precipitation-gel chromatograph.TD0 of IgY and the inhibory effectiveness were showed by nhibiting cytopathic effect (CPE) thst in MDCK infected with Influenza virus A(FM1).Results:IgY against Avain Influenza Virus was obtained.The TD0 value of IgY against Avain Influenza to MDCK cell line was 1.764 mg/ml,while at the dosage as low as 0.082 8 mg/ml the lnfluenza virus A(FM1) was still inhibited in vitro.Connchusion:Caprylate acid-two-step salt precipitation-gel chromatograph can prepare IgY successfully and the inhibitory effectiveness of IgY is good for inhibition of Influenza virus A(FM1).