Evaluation of Hyperimmune Hen Egg Yolk Derived Anti-Human Rotavirus Antibodies (Anti-HRVIgY) against Rotavirus Infection

Manika Buragohain, Ganesh S. Dhale, Govind R. Ghalsasi, Shobha D. Chitambar

Enteric Viruses Department, National Institute of Virology, Pune, India.
SPF Eggs Division, Venky’s (India) Ltd., Pune, India.

DOI: 10.4236/wjv.2012.22010                

Abstract

Oral delivery of specific IgY has been reported to be beneficial against rotavirus infection. However, the production of IgYs against globally prevalent human rotavirus (HRV) serotypes and their evaluation detailing the influence on the virological/histopathological consequences have not been reported to date. In the present study, anti-HRVIgY was generated in the eggs of specific pathogen free hens immunized with HRV serotypes G1 - G4 and G9 independently. Purified anti-HRVIgY preparations were tested to determine the ELISA and neutralizing antibody titers respectively in an indirect ELISA and cell culture based neutralization assay. Efficacy of pre and post infection treatment of anti-HRV-3IgY was assessed in an infant BALB/c mouse model of human rotavirus infection by monitoring percent diarrhea, severity and duration of diarrhea, intestinal viral load and histopathology. High (1:64000 - 1:512000) titered anti-HRVIgYs were obtained from the egg yolk of immunized hens with peak titer value (1:256000/1:512000) at 40 - 60 day of immunization. In-vitro, each of the anti-HRVIgY preparations showed the presence of multiserotypic neutralizing activity with high (1:1600 - ≥1:6400) homologous and low (≤1:50 - 1:800) heterologous titers. However, anti-HRV-3IgY neutralized all of the serotypes tested in the study indicating broader in-vitro neutralizing activity. In mice, post exposure treatment with anti-HRV-3IgY significantly reduced the extent of diarrhea and intestinal virus load and inhibited histopathological changes whereas pre exposure anti-HRV-3IgY treatment imparted immediate protection from development of rotavirus gastroenteritis. Thus, the anti-HRVIgY administered orally decreased morbidity and disease incidences in mice suggesting its potential implication in prophylactic and therapeutic usage in human to achieve reduction in rotavirus disease burden.

Keywords

IgY; Rotavirus; Mouse Model; Oral Immunotherapy

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	U. D. Parashar, C. J. Gibson, J. S. Bresee and R. I. Glass, “Rotavirus and Severe Childhood Diarrhea,” Emerging Infectious Disease, Vol. 12, No. 2, 2006, pp. 304-306. doi:10.3201/eid1202.050006
[2]	C. M. Zimmerman, J. S. Bresee, U. D. Parashar, T. L. Riggs, R. C. Holman and R. I. Glass, “Cost of DiarrheaAssociated Hospitalizations and Outpatient Visits in an Insured Population of Young Children in the United States,” Pediatric Infectious Disease Journal, Vol. 20, No. 1, 2001, pp. 14-19. doi:10.1097/00006454-200101000-00004
[3]	Y. L. Chun, L. L. Tsai, H. F. Yin, Y. W. Chung, H. H. Yu, L. H. Che, Y. C. Luan, Y. L. Chin and M. H. Li, “Disease Burden and Related Medical Costs of Rotavirus Infection in Taiwan,” BMC Infectious Diseases, Vol. 6, 2006, pp. 176-184. doi:10.1186/1471-2334-6-176
[4]	J. E. Tate, S. D. Chitambar, D. H. Esposito, R. Sarkar, B. Gladstone, S. Ramani, M. V. Raghava, T. V. Sowmyanarayanan, S. Gandhe, R. Arora, U. D Parashar and G. Kang, “Disease and Economic Burden of Rotavirus Diarrhoea in India,” Vaccine, Vol. 27, No. S5, 2009, pp. F18-F24. doi:10.1016/j.vaccine.2009.08.098
[5]	M. Pathak and K. Haque, “Oral Immunoglobulin for the Prevention of Rotavirus Infection in Low Birth Weight Infants,” Cochrane Database System of Review, Vol. 3, 2003, Article ID: CD003740. doi:10.1002/14651858.CD003740
[6]	A. Z. Kapikian, L. Simonsen, T. Vesikari, Y. Hoshino, D. M. Morens, R. M. Chanock, J. R. La Montagne and B. R. Murphy, “A Hexavalent Human Rotavirus-Bovine Rotavirus (UK) Reassortant Vaccine Designed for use in Developing Countries and Delivered in a Schedule with the Potential to Eliminate the Risk of Intussusceptions,” Journal of Infectious Diseases, Vol. 192, No. S1, 2005, pp. S22-S29. doi:10.1086/431510
[7]	N. Pant, A. Hultberg, Y. Zhao, L. Svensson, Q. PanHammarstrom, K. Johansen, P. H. Pouwels, F. M. Ruggeri, P. Hermans, L. Frenken, T. Boren, H. Marcotte and L. Hammarstrom, “Lactobacilli Expressing Variable Domain of Llama Heavy Chain Antibody Fragments (Lactobodies) Confer Protection against Rotavirus Induced Diarrhea,” Journal of Infectious Diseases, Vol. 194, No. 11, 2006, pp. 1580-1588. doi:10.1086/508747
[8]	P. H. Dennehy, “Rotavirus Vaccines: An Overview,” Clinical Microbiology Review, Vol. 21, No. 1, 2008, pp. 198-208. doi:10.1128/CMR.00029-07
[9]	G. E. Armah, S. O. Sow, R. F. Breiman, M. J. Dallas, M. D. Tapia, D. R. Feikin, F. N. Binka, A. D. Steele, K. F. Laserson, N. A. Ansah, M. M. Levine, K. Lewis, M. L. Coia, M. Attah-Poku, J. Ojwando, S. B. Rivers, J. C. Victor, G. Nyambane, A. Hodgson, F. Sch?del, M. Ciarlet and K. M. Neuzil, “Efficacy of Pentavalent Rotavirus Vaccine against Severe Rotavirus Gastroenteritis in Infants in Developing Countries in Sub-Saharan Africa: A Randomised, Double-Blind, Placebo-Controlled Trial,” Lancet, Vol. 376, No.
[10]	K. Zaman, D. A. Dang, J. C. Victor, S. Shin, M. Yunus, M. J. Dallas, G. Podder, D. T. Vu, T. P. Le, S. P. Luby, H. T. Le, M. L. Coia, K. Lewis, S. B. Rivers, D. A. Sack, F. Sch?del, A. D. Steele, K. M. Neuzil and M. Ciarlet, “Efficacy of Pentavalent Rotavirus Vaccine against Severe Rotavirus Gastroenteritis in Infants in Developing Countries in Asia: A Randomised, Double-Blind, PlaceboControlled Trial,” Lancet, Vol. 376, No. 9741, 2010, pp. 615-623. doi:10.1016/S0140-6736(10)60755-6
[11]	D. H. Esposito, J. E. Tate, G. Kang and U. D. Parashar, “Projected Impact and Cost-Effectiveness of a Rotavirus Vaccination Program in India, 2008,” Clinical Infectious Diseases, Vol. 52, No. 2, 2011, pp. 171-177. doi:10.1093/cid/ciq094
[12]	K. Y. Green and A. Z. Kapikian, “Identification of VP7 Epitopes Associated with Protection against Human Rotavirus Illness or Shedding in Volunteers,” Journal of Virology, Vol. 66, No. 1, 1992, pp. 548-553.
[13]	S. M. Matsui, P. A. Offit, P. T. Vo, E. R. Mackow, D. A. Benfield, R. D. Shaw, L. Padilla-Noriega and H. B. Greenberg, “Passive Protection against Rotavirus-Induced Diarrhea by Monoclonal Antibodies to the Heterotypic Neutralization Domain of VP7 and the VP8 Fragment of VP4,” Journal of Clinical Microbiology, Vol. 27, No. 4, 1989, pp. 780-782.
[14]	P. A. Offit, H. F. Clark, G. Blavat and H. B. Greenberg, “Reassortant Rotaviruses Containing Structural Proteins VP3 and VP7 from Different Parents Induce Antibodies Protective against Each Parental Serotype,” Journal of Virology, Vol. 60, No. 2, 1986, pp. 491-496.
[15]	P. A. Offit, R. D. Shaw and H. B. Greenberg, “Passive Protection against Rotavirus-Induced Diarrhea by Monoclonal Antibodies to Surface Proteins of VP3 and VP7,” Journal of Virology, Vol. 58, No. 2, 1986, pp. 700-703.
[16]	F. M. Ruggeri, K. Johansen, G. Basile, J. P. Kraehenbuhl and L. Svensson, “Antirotavirus Immunoglobulin a Neutralizes Virus in Vitro after Transcytosis through Epithelial Cells and Protects Infant Mice from Diarrhea,” Journal of Virology, Vol. 72, No. 4, 1998, pp. 2708-2714.
[17]	L. E. Westerman, H. M. McClure, B. Jiang, J. W. Almond and R. I. Glass, “Serum IgG Mediates Mucosal Immunity against Rotavirus Infection,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 102, No. 20, 2005, pp. 7268-7273. doi:10.1073/pnas.0502437102
[18]	L. Hammarstrom, “Passive Immunity against Rotavirus in Infants,” Acta Paediatrica, Vol. 88, No. 430, 1999, pp. 127-132.
[19]	G. W. Warr, K. E. Magor and D. A. Higgins, “IgY: Clues to the Origins of Modern Antibodies,” Immunology Today, Vol. 16, No. 8, 1995, pp. 392-398. doi:10.1016/0167-5699(95)80008-5
[20]	A. Larsson and J. Sjoquist, “Chicken IgY: Utilizing the Evolutionary Difference,” Comparative Immunology Microbiology Infectious Diseases, Vol. 13, No. 4, 1990, pp. 199-201. doi:10.1016/0147-9571(90)90088-B
[21]	M. Kuroki, Y. Ikemori, H. Yokoyama, R. C. Peralta, F. C. Icatlo Jr. and Y. Kodama, “Passive Protection against Bovine Rotavirus-Induced Diarrhea in Murine Model by Specific Immunoglobulins from Chicken Egg Yolk,” Veterinary Microbiololgy, Vol. 37, 1993, pp. 135-146. doi:10.1016/0378-1135(93)90188-D
[22]	T. Ebina, “Prophylaxis of Rotavirus Gastroenteritis Using Immunoglobulin,” Archives of Virology, Vol. 12, 1996, pp. 217-223.
[23]	S. A. Sarker, N. Pant, L. R. Juneja and L. Hammarstrom, “Successful Treatment of Rotavirus-Induced Diarrhoea in Suckling Mice with Egg Yolk Immunoglobulin,” Journal of Health and Population Nutrition, Vol. 25, No. 4, 2007, pp. 465-468.
[24]	S. D. Kelkar, V. S. Bhide, S. S. Ranshing and S. S. Bedekar, “Rapid ELISA for the Diagnosis of Rotavirus,” Indian Journal of Medical .Research, Vol. 119, No. 2, 2004, pp. 60-65.
[25]	S. D. Kelkar, P. G. Ray and S. S. Bedekar, “Assay of Neutralizing Antibodies to Animal Rotavirus Strains and Human Rotavirus Serotype G8 by Modified Method in the Resident of Pune, India,” Journal of Diarrhoeal Disease Research, Vol. 14, No. 2, 1996, pp. 101-106.
[26]	M. S. Poruchynsky and P. H. Atkinson, “Rotavirus Protein Rearrangements in Purified Membrane-Enveloped Intermediate Particles,” Journal of Virology, Vol. 65, No. 9, 1991, pp. 4720-4727.
[27]	O. H. Lowry, N. J. Rosebrough, A. L. Farr and R. J. Randall, “Protein Measurement with the Folin Phenol Reagent,” The Journal of Biological Chemistry, Vol. 193, No. 1, 1951, pp. 265-275.
[28]	A. Polson, T. Coetzer, J. Kruger, E. von Maltzahn and K. J. van der Merwe, “Improvements in the Isolation of IgY from the Yolks of Eggs Laid by Immunized Hens,” Immunological Investigations, Vol. 14, No. 4, 1985, pp. 323-327.
[29]	R. H.Yolken, F. Leister, S. B. Wee, R. Miskuff and S. Vonderfecht, “Antibodies to Rotavirusesin Chicken’s Eggs: A Potential Source of Antiviral Immunoglobulins Suitable for Human Consumption,” Pediatrics, Vol. 81, No. 2, 1988, pp. 291-295.
[30]	P. G. Ray and S. D. Kelkar, “Prevalence of Neutralizing Antibodies against Different Rotavirus Serotypes in Children with Severe Rotavirus-Induced Diarrhea and Their Mothers,” Clinical and Diagnostic Laboratory Immunology, Vol. 11, No. 1, 2004, pp. 186-194. doi:10.1128/CDLI.11.1.186-194.2004
[31]	D. Katz, E. Freeman, M. Davidson, M. Abramson and P. Fuchs, “Detection of Anti-Rabies Neutralizing Antibodies in Humans and Cattle by a Combined Tissue Culture and Enzyme Linked Immunoassay,” Israel Journal of Veterinary Medicine, Vol. 53, No. 4, 1998, pp. 132-142.
[32]	M. Buragohain, G. S. Dhale, C. G. Raut, G. Kang and S. D. Chitambar, “Analyses of Clinical, Pathological and Virological Features of Human Rotavirus Strain, YO Induced Gastroenteritis in Infant BALB/c Mice,” Microbes and Infection, Vol. 13, No. 4, 2011, pp. 331-338. doi:10.1016/j.micinf.2010.12.001
[33]	K. Takahashi, K. Ohashi, Y. Abe, S. Mori, K. Taniguchi, T. Ebina, O. Nakagomi, M . Terada and S. Shigeta, “Protective Efficacy of a Sulfated Sialyl Lipid (NMSO3) against Human Rotavirus-Induced Diarrhea in a Mouse Model,” Antimicrobial Agents and Chemotherapy, Vol. 46, No. 2, 2002, pp. 420-424. doi:10.1128/AAC.46.2.420-424.2002
[34]	G. Kang, M. Iturriza-Gomara, J. G. Wheeler, P. Crystal, B. Monica, S. Ramani, B. Primrose, P. D. Moses, C. I. Gallimore, D. W. Brown and J. Gray, “Quantitation of Group A Rotavirus by Real-Time Reverse-TranscriptionPolymerase Chain Reaction: Correlation with Clinical Severity in Children in South India,” Journal of Medical Virology, Vol. 73, No. 1, 2004, pp. 118-122. doi:10.1002/jmv.20053
[35]	N. Pant, H. Marcotte, H. Brüssow, L. Svensson and L. Hammarstrom, “Effective Prophylaxis against Rotavirus Diarrhea Using a Combination of Lactobacillus rhamnosus GG and Antibodies,” BMC Microbiology, Vol. 7, 2007, p. 86. doi:10.1186/1471-2180-7-86
[36]	G. Phillips, B. Lopman, C. Tam, M. Iturriza-Gomara, D. Brown and J. Gray, “Diagnosing Rotavirus a Associated IID: Using ELISA to Identify a Cut-Off for Real Time RT-PCR,” Journal of Clinical Virology, Vol. 44, No. 3, 2009, pp. 242-245. doi:10.1016/j.jcv.2008.12.001
[37]	L. G. Luna, “Manual of Histological Staining Methods of the Armed Forces Institute of Pathology,” 3rd Edition, McGraw Hill, New York, 1968.
[38]	R. A. Fisher, “Statistical Methods for Research Workers,” Oliver and Boyd, Edinburgh, 1954.
[39]	Kruskal and Wallis, “Use of Ranks in One-Criterion Variance Analysis,” Journal of the American Statistical Association, Vol. 47, No. 260, 1952, pp. 583-621. doi:10.2307/2280779
[40]	F. Wilcoxon, “Individual Comparisons by Ranking Methods,” Biometrics Bulletin, Vol. 1, No. 6, 1945, pp. 80-83. doi:10.2307/3001968
[41]	J. A. Boshuizen, J. H. Reimerink, A. M. Korteland-van Male, V. J. van Ham, M. P. Koopmans, H. A. Büller, J. Dekker and A. W. Einerhand, “Changes in Small Intestinal Homeostasis, Morphology, and Gene Expression during Rotavirus Infection of Infant Mice,” Journal of Virology, Vol. 77, No. 24, 2003, pp. 13005-13016. doi:10.1128/JVI.77.24.13005-13016.2003
[42]	J. Kovacs Nolan and Y. Mine, “Avian Egg Antibodies: Basic and Potential Applications,” Avian and Poultry Biology Reviews, Vol. 15, No. 1, 2004, pp. 25-46. doi:10.1128/JVI.77.24.13005-13016.2003
[43]	R. Schade and A. Hlinak, “Egg Yolk Antibodies, State of the Art and Future Prospects,” Alternatives to Animal Experimentation, Vol. 13, No. 5, 1996, pp. 5-9.
[44]	R. Chalghoumi, Y. Beckers, D. Portetelle and A. Théwis, “Hen Egg Yolk Antibodies (IgY), Production and Use for Passive Immunization against Bacterial Enteric Infections in Chicken: A Review,” Biotechnology, Agronomy, Society and Environment, Vol. 13, No. 2, 2009, pp. 295-308.
[45]	P. S. Paul and Y. S. Lyoo, “Immunogens of Rotaviruses,” Veterinary Microbiology, Vol. 37, No. 3-4, 1993, pp. 299-317. doi:10.1016/0378-1135(93)90031-2
[46]	M. E. Cook and D. L. Trott, “IgY—Immune Component of Eggs as a Source of Passive Immunity for Animals and Humans,” World’s Poultry Science Journal, Vol. 66, No. 2, 2010, pp. 215-226. doi:10.1017/S0043933910000279
[47]	T. Ebina, K. Tsukada, K. Umezu, M. Nose, K. Tsuda, H. Hatta, M. Kim and T. Yamamoto, “Gastroenteritis in Suckling Mice Caused by Human Rotavirus Can Be Prevented with Egg Yolk Immunoglobulin (IgY) and Treated with a Protein-Bound Polysaccharide Preparation (PSK),” Microbiology and Immunology, Vol. 34, No. 7, 1990, pp. 617-629.
[48]	R. R. Marquardt, L. Z. Jin, J. W. Kim, L. Fang, A. A. Frohlich and S. K. Baidoo, “Passive Protective Effect of Egg-Yolk Antibodies against Enterotoxigenic Escherichia coli K88+ Infection in Neonatal and Early-Weaned Piglets,” FEMS Immunology and Medical Microbioogyl, Vol. 23, No. 4, 1999, pp. 283-288.
[49]	S. Rahimi, Z. M. Shiraz, T. Z. Salehi, M. A. K. Torshizi and J. L. Grimes, “Prevention of Salmonella Infection in Poultry by Specific Egg-Derived Antibody,” International Journal of Poultry Science, Vol. 6, No. 4, 2007, pp. 230-235. doi:10.3923/ijps.2007.230.235
[50]	P. A. Offit and H. F. Clark, “Maternal Antibody-Mediated Protection against Gastroenteritis Due to Rotavirus in Newborn Mice Is Dependent on Both Serotype and Titer of Antibody,” Journal of Infectious Diseases, Vol. 152, No. 6, 1985, pp. 1152-1158. doi:10.1093/infdis/152.6.1152
[51]	R. J. Gorrell and R. F. Bishop, “Homotypic and Heterotypic Serum Neutralizing Antibody Response to Rotavirus Proteins Following Natural Primary Infection and Reinfection in Children,” Journal of Medical Virology, Vol. 57, No. 2, 1999, pp. 204-211. doi:10.1002/(SICI)1096-9071(199902)57:2<204::AID-JMV20>3.0.CO;2-Y
[52]	J. G. Lecce, H. L. Leary Jr., D. A. Clarke and R. P. Batema, “Protection of Agammaglobulinemic Piglets from Porcine Rotavirus Infection by Antibody against Simian Rotavirus SA-11,” Journal of Clinical Microbiology, Vol. 29, 1991, pp. 1382-1386.
[53]	G. Menchaca, L. P. Noriega, M. M. Toss, J. F. Contreras, F. I. Puerto, H. Guiscafré, F. Mota, I. Herrera, R. Cedillo, O. Munoz, R. Ward, Y. Hoshino, S. López and C. F. Arias, “Serotype Specificity of the Neutralizing-Antibody Response Induced by the Individual Surface Proteins of Rotavirus in Natural Infections of Young Children,” Clinical and Diagnostic Laboratory Immunology, Vol. 5, No. 3, 1998, pp. 328-334.
[54]	S. E. Crawford, M. K. Estes, M. Ciarlet, C. Barone, C. M. O’Neal, J. Cohen and M. E. Conner, “Heterotypic Protection and Induction of a Broad Heterotypic Neutralization Response by Rotavirus-Like Particles,” Journal of Virology, Vol. 73, No. 6, 1999, pp. 4813-4822.
[55]	D. Y. Graham, G. R. Dufour and M. K. Estes, “Minimal Infective Dose of Rotavirus,” Archives of Virology, Vol. 92, No. 3-4, 1987, pp. 261-271. doi:10.1007/BF01317483
[56]	L. M. Bell, H. F. Clark, E. A. O’Brien, M. J. Kornstein, S. A. Plotkin and P. A. Offit, “Gastroenteritis Caused by Human Rotaviruses (Serotype Three) in a Suckling Mouse Model,” Proceedings of the Society for Experimental Biology and Medicine, Vol. 184, No. 1, 1987, pp. 27-132.