CHARACTERIZATION OF A RECOMBINANT EIMERIA-ACERVULINA ANTIGEN EXPRESSED IN SPOROZOITE AND MEROZOITE DEVELOPMENTAL STAGES

被引:17
作者
CASTLE, MD [1 ]
JENKINS, MC [1 ]
DANFORTH, HD [1 ]
LILLEHOJ, HS [1 ]
机构
[1] USDA ARS, BELTSVILLE AGR RES CTR,PROTOZOAN DIS LAB, BLDG 1180,BARC E, BELTSVILLE, MD 20705 USA
关键词
D O I
10.2307/3283125
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
A cDNA from Eimeria acervulina encoding an immunogenic region of antigens shared between sporozoites and merozoites was cloned and expressed in Escherichia coli. Immunofluorescence staining of sporozoites, merozoites, and coccidial-infected intestinal tissue with monoclonal antibody used to detect the recombinant clone indicated that the epitope was present on internal parasite proteins. Immunostaining of nitrocellulose paper containing protein from both asexual stages revealed numerous sporozoite antigens (18-120 kDa) and only 1 merozoite antigen (150 kDa). Northern blot hybridization assays using this cDNA clone for probing sporozoite and merozoite RNA showed that distinct transcripts were present in both asexual stages. Similar to the immunofluorescence studies, many homologous RNAs were observed in sporozoites (3.7 - 13 kb) and only 1 prominent hybridizing species was found in merozoites (3.3 kb). The recombinant coccidial antigen, designated MA16, is a 125- kDa beta-galactosidase fusion protein, representing about 10 kDa of parasite protein. In blastogenesis assays, purified recombinant MA16 antigen is capable of activating T lymphocytes obtained from E. acervulina-immune inbred chickens. DNA sequencing of MA16 clone and analysis of the predicted amino acid sequence indicated several putative T cell epitopes that may be responsible for the observed in vitro blastogenic response.
引用
收藏
页码:384 / 390
页数:7
相关论文
共 36 条
[1]  
Abu Ali M., 1976, Veterinary Parasitology, V1, P309, DOI 10.1016/0304-4017(76)90033-9
[2]   EIMERIA-FALCIFORMIS - EFFECTS OF CO-60 IRRADIATION ON INFECTIVITY AND IMMUNOGENICITY OF SPORULATED OOCYSTS [J].
COX, AB ;
DUNCAN, S ;
LEVY, CK .
JOURNAL OF PARASITOLOGY, 1977, 63 (05) :927-929
[3]   SPECIFICITY AND CROSSREACTIVITY OF IMMUNE SERUM AND HYBRIDOMA ANTIBODIES TO VARIOUS SPECIES OF AVIAN COCCIDIA [J].
DANFORTH, HD ;
AUGUSTINE, PC .
POULTRY SCIENCE, 1983, 62 (11) :2145-2151
[4]   GENETICALLY ENGINEERED ANTIGEN CONFERS PARTIAL PROTECTION AGAINST AVIAN COCCIDIAL PARASITES [J].
DANFORTH, HD ;
AUGUSTINE, PC ;
RUFF, MD ;
MCCANDLISS, R ;
STRAUSBERG, RL ;
LIKEL, M .
POULTRY SCIENCE, 1989, 68 (12) :1643-1652
[5]  
DAVIS LG, 1986, BASIC METHODS MOL BI, P143
[6]   PRODUCTION OF MONOCLONAL-ANTIBODIES - STRATEGY AND TACTICS [J].
DESTGROTH, SF ;
SCHEIDEGGER, D .
JOURNAL OF IMMUNOLOGICAL METHODS, 1980, 35 (1-2) :1-21
[7]   RAPID MINIPREPARATIONS OF BACTERIOPHAGE-LAMBDA DNA [J].
DUMANSKI, JP ;
RUTTLEDGE, M ;
DATTA, S .
NUCLEIC ACIDS RESEARCH, 1988, 16 (18) :9044-9044
[8]   STUDIES ON TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS [J].
HANAHAN, D .
JOURNAL OF MOLECULAR BIOLOGY, 1983, 166 (04) :557-580
[9]   EIMERIA-TENELLA - IMMUNOGENICITY OF ARRESTED SPOROZOITES IN CHICKENS [J].
JEFFERS, TK ;
LONG, PL .
EXPERIMENTAL PARASITOLOGY, 1985, 60 (02) :175-180
[10]   IDENTIFICATION OF IMMUNODOMINANT SURFACE-ANTIGENS OF EIMERIA-ACERVULINA SPOROZOITES AND MEROZOITES [J].
JENKINS, MC ;
DAME, JB .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1987, 25 (02) :155-164