Characterization of variable immunodominant antigens of Cowdria ruminantium by ELISA and immunoblots

Cross‐immunization experiments have revealed a significant antigenic diversity of the isolate of Cowdria ruminantium which needs to be characterized for the development of vaccines. We identified polymorphic immunodominant antigens by ELISA and immunoblot. Using serum from a goat immune to the Gardel stock of Cowdria (isolated in Guadeloupe) adsorbed on antigen of the Senegal stock of this pathogen, distinct serogroups were revealed by ELISA among six isolates from different geographical origins. Furthermore, a goat serum directed against the Senegal stock and adsorbed on Gardel antigens was shown to be specific for the Senegal stock, thus confirming the existence of serotypes in Cowdria. The Major Antigenic Protein 1 (MAP1) of Cowdria was shown to have variable antigenic determinants. Also in a group of variable proteins ranging from 23 to 29 kDa, one antigen of 26–27 kDa had a determinant specific for the Gardel isolate. These polymorphic antigens may be relevant components of Cowdria ruminantium for a vaccine as the sera revealing these antigens originated from a goat surviving a lethal challenge. However, the presence of T‐cell epitopes and the ability of these antigens to confer protection to ruminants remain to be investigated. The production of a rabbit antiserum against this group of polypeptides will be of great use for their purification and for the screening of expression libraries.

[1]  D. McKeever,et al.  The Use of CD4+ T‐cell Lines to Screen for Immunogenic Proteins of Cowdria ruminantium a , 1998, Annals of the New York Academy of Sciences.

[2]  F. Jongejan,et al.  Tropical Veterinary Medicine: Molecular Epidemiology, Hemoparasites and Their Vectors, and General Topics. Proceedings of the 4th biennial meeting of the Society for Tropical Veterinary Medicine. Montpellier, France, May 5-9, 1997. , 1998, Annals of the New York Academy of Sciences.

[3]  D. Martinez,et al.  Detection of genomic polymorphisms among isolates of the intracellular bacterium Cowdria ruminantium by random amplified polymorphic DNA and Southern blotting. , 1997, FEMS microbiology letters.

[4]  E. Peterson,et al.  Immunization with an acellular vaccine consisting of the outer membrane complex of Chlamydia trachomatis induces protection against a genital challenge , 1997, Infection and immunity.

[5]  D. McKeever,et al.  Analysis of T-cell responses in cattle immunized against heartwater by vaccination with killed elementary bodies of Cowdria ruminantium , 1997, Infection and immunity.

[6]  D. Martinez,et al.  Comparative efficacy of Freund's and Montanide ISA50 adjuvants for the immunisation of goats against heartwater with inactivated Cowdria ruminantium. , 1996, Veterinary parasitology.

[7]  G. Reddy,et al.  Sequence heterogeneity of the major antigenic protein 1 genes from Cowdria ruminantium isolates from different geographical areas , 1996, Clinical and diagnostic laboratory immunology.

[8]  D. Martinez,et al.  Use of a specific immunogenic region on the Cowdria ruminantium MAP1 protein in a serological assay , 1995, Journal of clinical microbiology.

[9]  K. Sumption,et al.  The Cowdria ruminantium groE operon. , 1995, Microbiology.

[10]  A. Barbet,et al.  Immunisation of sheep against heartwater with inactivated Cowdria ruminantium. , 1995, Research in veterinary science.

[11]  F. Jongejan,et al.  Size variation of the major immunodominant protein of Cowdria ruminantium. , 1994, Clinical and diagnostic laboratory immunology.

[12]  F. Jongejan,et al.  Molecular cloning of a gene encoding the immunogenic 21 kDa protein of Cowdria ruminantium. , 1994, Microbiology.

[13]  D. Martinez,et al.  Protection of goats against heartwater acquired by immunisation with inactivated elementary bodies of Cowdria ruminantium. , 1994, Veterinary immunology and immunopathology.

[14]  F. Jongejan,et al.  Molecular cloning, sequence analysis, and expression of the gene encoding the immunodominant 32-kilodalton protein of Cowdria ruminantium , 1994, Infection and immunity.

[15]  C. Nyathi,et al.  An immunoblotting diagnostic assay for heartwater based on the immunodominant 32-kilodalton protein of Cowdria ruminantium detects false positives in field sera , 1993, Journal of clinical microbiology.

[16]  D. Martinez,et al.  Protection of goats against heartwater acquired by immunization with inactivated elementary bodies of Cowdria ruminantium , 1993 .

[17]  D. Martinez,et al.  Detection of antibodies to Cowdria ruminantium in the serum of domestic ruminants by indirect ELISA. , 1993, Revue d'elevage et de medecine veterinaire des pays tropicaux.

[18]  J. D. du Plessis An in vitro test to demonstrate the inhibitory effect of homologous immune serum on the infectivity of Cowdria ruminantium. , 1993, Onderstepoort Journal of Veterinary Research.

[19]  A. Barbet,et al.  The development of antibody to Cowdria ruminantium in mice and its role in heartwater disease. , 1993, Revue d'Elevage et de Médecine Vétérinaire des Pays Tropicaux.

[20]  A. Neitz,et al.  Isolation and characterization of antigenic proteins of Cowdria ruminantium. , 1993, Revue d'elevage et de medecine veterinaire des pays tropicaux.

[21]  P. Berche,et al.  T cell-mediated immunity to Cowdria ruminantium in mice: the protective role of Lyt-2+ T cells. , 1991, Onderstepoort Journal of Veterinary Research.

[22]  E. Oaks,et al.  Molecular cloning and sequence analysis of a Rickettsia tsutsugamushi 22 kDa antigen containing B- and T-cell epitopes. , 1991, Microbial pathogenesis.

[23]  F. Jongejan,et al.  Antigenic diversity of Cowdria ruminantium isolates determined by cross-immunity. , 1991, Research in veterinary science.

[24]  M. de Groot,et al.  Competitive enzyme-linked immunosorbent assay for heartwater using monoclonal antibodies to a Cowdria ruminantium-specific 32-kilodalton protein. , 1991, Veterinary microbiology.

[25]  G. E. Jones,et al.  Protection of sheep against Chlamydia psittaci infection with a subcellular vaccine containing the major outer membrane protein , 1990, Infection and immunity.

[26]  D. Martinez,et al.  Comparaison de trois antigènes pour le sérodiagnostic de la cowdriose par immunofluorescence indirecte , 1990 .

[27]  G. Uilenberg Present and future possibilities for the control of cowdriosis and anaplasmosis. , 1990, The Veterinary quarterly.

[28]  D. Martinez,et al.  [Comparison between 3 antigens for the serodiagnosis of heartwater disease by indirect immunofluorescence]. , 1990, Revue d'Elevage et de Médecine Vétérinaire des Pays Tropicaux.

[29]  M. P. Jongejan,et al.  Theories on Channeling; Reviewed and Compared to Experiments , 1990 .

[30]  F. Jongejan,et al.  Serotypes in Cowdria ruminantium and their relationship with Ehrlichia phagocytophila determined by immunofluorescence. , 1989, Veterinary microbiology.

[31]  F. Jongejan,et al.  Identification of an immunodominant antigenically conserved 32-kilodalton protein from Cowdria ruminantium , 1989, Infection and immunity.

[32]  J. Olivier,et al.  The heterogenicity of Cowdria ruminantium stocks: cross-immunity and serology in sheep and pathogenicity to mice. , 1989, The Onderstepoort journal of veterinary research.

[33]  F. Jongejan,et al.  Antigenic differences between stocks of Cowdria ruminantium. , 1988, Research in veterinary science.

[34]  Z. Kowalski,et al.  The pathogenesis of heartwater. , 1987, The Onderstepoort journal of veterinary research.

[35]  G. Hui,et al.  Merozoite surface coat precursor protein completely protects Aotus monkeys against Plasmodium falciparum malaria. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[36]  J. Plessis A METHOD FOR DETERMINING THE COWDRIA RUMINANTIUM INFECTION RATE OF AMBLYOMMA HEBRAEUM: EFFECTS IN MICE INJECTED WITH TICK HOMOGENATES , 1985 .

[37]  N. Barré,et al.  Quelques observations sur une souche de Cowdria ruminantium isolée en Guadeloupe (Antilles françaises) , 1985 .

[38]  A. ter Huurne,et al.  Cross immunity between strains of Cowdria ruminantium. , 1983, Research in veterinary science.

[39]  A. Holder,et al.  Immunization against blood-stage rodent malaria using purified parasite antigens , 1981, Nature.

[40]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[41]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[42]  M. Kindruk,et al.  On a method for determining the , 1973 .

[43]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[44]  J. D. du Plessis Pathogenesis of heartwater. I. Cowdria ruminantium in the lymph nodes of domestic ruminants. , 1970, The Onderstepoort journal of veterinary research.