Molecular characterization of a Leishmania donovani cDNA clone with similarity to human 20S proteasome a-type subunit

TY - JOUR

T1 - Molecular characterization of a Leishmania donovani cDNA clone with similarity to human 20S proteasome a-type subunit

AU - Christensen, C B

AU - Jørgensen, L

AU - Jensen, A T

AU - Gasim, S

AU - Chen, M

AU - Kharazmi, A

AU - Theander, T G

AU - Andresen, K

N1 - Keywords: Animals; Antibodies, Protozoan; Base Sequence; Blotting, Northern; Blotting, Western; Cloning, Molecular; Cysteine Endopeptidases; DNA, Complementary; Enzyme-Linked Immunosorbent Assay; Humans; Leishmania donovani; Leishmaniasis, Visceral; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Multienzyme Complexes; Proteasome Endopeptidase Complex; Recombinant Fusion Proteins; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Vaccines, DNA

PY - 2000

Y1 - 2000

N2 - Using plasma from patients infected or previously infected with Leishmania donovanii, we isolated a L. donovanii cDNA clone with similarity to the proteasome a-type subunit from humans and other eukaryotes. The cDNA clone, designated LePa, was DNA sequenced and Northern blot analysis of L. donovanii poly(A(+))mRNA indicated the isolation of a full length cDNA clone with a transcript size of 1.9 kb. The expressed recombinant LePa fusion protein induced proliferation of peripheral blood mononuclear cells in one out of seven patients who had suffered from visceral leishmaniasis. Plasma from 16 out of 25 patients with visceral leishmaniasis and four out of 18 patients with cutaneous leishmaniasis contained IgG antibodies which reacted with the purified LePa fusion protein as evaluated in an ELISA. The LePa DNA sequence was inserted into an eukaryotic expression vector and Balb/c mice were vaccinated. DNA vaccination of Balb/c mice with LePa generated an initial significant reduction in lesion size after challenge.

AB - Using plasma from patients infected or previously infected with Leishmania donovanii, we isolated a L. donovanii cDNA clone with similarity to the proteasome a-type subunit from humans and other eukaryotes. The cDNA clone, designated LePa, was DNA sequenced and Northern blot analysis of L. donovanii poly(A(+))mRNA indicated the isolation of a full length cDNA clone with a transcript size of 1.9 kb. The expressed recombinant LePa fusion protein induced proliferation of peripheral blood mononuclear cells in one out of seven patients who had suffered from visceral leishmaniasis. Plasma from 16 out of 25 patients with visceral leishmaniasis and four out of 18 patients with cutaneous leishmaniasis contained IgG antibodies which reacted with the purified LePa fusion protein as evaluated in an ELISA. The LePa DNA sequence was inserted into an eukaryotic expression vector and Balb/c mice were vaccinated. DNA vaccination of Balb/c mice with LePa generated an initial significant reduction in lesion size after challenge.

M3 - Journal article

C2 - 10564720

SN - 0304-4165

VL - 1500

SP - 77

EP - 87

JO - B B A - General Subjects

JF - B B A - General Subjects

IS - 1

ER -