TY - JOUR

T1 - Antigenicity and immunogenicity of recombinant glutamate-rich protein of Plasmodium falciparum expressed in Escherichia coli

AU - Theisen, M

AU - Vuust, J

AU - Gottschau, A

AU - Jepsen, S

AU - Høgh, B

PY - 1995/1

Y1 - 1995/1

N2 - A recombinant Plasmodium falciparum glutamate-rich protein (GLURP) was produced in Escherichia coli as a nearly full-length protein. In order to map immunodominant regions on GLURP, the nonrepetitive amino-terminal region (R0) as well as the central repeat region (R1) and the carboxy-terminal repeat region (R2) were also produced as separate products. All four purified gene products reacted specifically with serum samples from adults living in an area of Liberia where malaria is holoendemic. It appears that the human immune response against GLURP is primarily directed against the R2 region because 94% of the serum samples reacted with this region in an immunoassay. Antibody reactivity against the R0 region was also observed in 75% of the serum samples, while the R1 region showed only weak antibody-binding activity. When the nearly full-length GLURP molecule was adsorbed to Al(OH)3 it was found to be immunogenic in mice. In these experiments, the antibody response was almost exclusively directed against the R2 region. When anti-GLURP sera were obtained from rabbits immunized with the three regions, R0, R1, and R2, respectively, they recognized in immunoprecipitation experiments authentic GLURP from P. falciparum grown in vitro. These results demonstrate that GLURP produced in E. coli can induce a humoral immune response against GLURP derived from blood-stage parasites.

AB - A recombinant Plasmodium falciparum glutamate-rich protein (GLURP) was produced in Escherichia coli as a nearly full-length protein. In order to map immunodominant regions on GLURP, the nonrepetitive amino-terminal region (R0) as well as the central repeat region (R1) and the carboxy-terminal repeat region (R2) were also produced as separate products. All four purified gene products reacted specifically with serum samples from adults living in an area of Liberia where malaria is holoendemic. It appears that the human immune response against GLURP is primarily directed against the R2 region because 94% of the serum samples reacted with this region in an immunoassay. Antibody reactivity against the R0 region was also observed in 75% of the serum samples, while the R1 region showed only weak antibody-binding activity. When the nearly full-length GLURP molecule was adsorbed to Al(OH)3 it was found to be immunogenic in mice. In these experiments, the antibody response was almost exclusively directed against the R2 region. When anti-GLURP sera were obtained from rabbits immunized with the three regions, R0, R1, and R2, respectively, they recognized in immunoprecipitation experiments authentic GLURP from P. falciparum grown in vitro. These results demonstrate that GLURP produced in E. coli can induce a humoral immune response against GLURP derived from blood-stage parasites.

KW - Amino Acid Sequence

KW - Animals

KW - Antibodies, Protozoan/biosynthesis

KW - Antigens, Protozoan/biosynthesis

KW - Base Sequence

KW - Enzyme-Linked Immunosorbent Assay

KW - Escherichia coli/genetics

KW - Female

KW - Immune Sera

KW - Immunodominant Epitopes/immunology

KW - Liberia

KW - Malaria, Falciparum/blood

KW - Mice

KW - Mice, Inbred BALB C

KW - Molecular Sequence Data

KW - Peptide Fragments/genetics

KW - Plasmodium falciparum/genetics

KW - Precipitin Tests

KW - Protozoan Proteins/biosynthesis

KW - Rabbits

KW - Recombinant Fusion Proteins/biosynthesis

KW - Repetitive Sequences, Nucleic Acid

M3 - Journal article

C2 - 7719909

VL - 2

SP - 30

EP - 34

JO - Clinical and Diagnostic Laboratory Immunology

JF - Clinical and Diagnostic Laboratory Immunology

SN - 1556-6811

IS - 1

ER -