Mapping of two new human B-cell epitopes on HIV-1 gp120
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Mapping of two new human B-cell epitopes on HIV-1 gp120. / Kusk, P; Holmbäck, K; Lindhardt, B O; Hulgaard, E F; Bugge, T H.
I: AIDS (Year), Bind 6, Nr. 12, 12.1992, s. 1451-6.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Mapping of two new human B-cell epitopes on HIV-1 gp120
AU - Kusk, P
AU - Holmbäck, K
AU - Lindhardt, B O
AU - Hulgaard, E F
AU - Bugge, T H
PY - 1992/12
Y1 - 1992/12
N2 - OBJECTIVE: To map epitopes on gp120 defined by human antibodies and to examine the neutralizing activity of these antibodies.DESIGN AND METHODS: Serum from HIV-1-antibody-positive individuals was used to screen a random fragment expression library representing gp120 from the HIVIIIB clone BH10. The library was based on the pUEX1 expression vector. Serum was tested for in vitro neutralizing activity using H9 cells and the HIVIIIB isolate.RESULTS: Four different epitopes defined by human antibodies were mapped on gp120. Two of these have not previously been reported and are located within amino acids (aa) 90-100 in the C1 region and aa 355-365 in the semi-conserved region between V3 and V4. The other two are located within aa 140-145 and aa 286-309. These epitopes are situated in regions that have been shown to demarcate human epitopes. Three serum samples with neutralization titres > or = 1024 were identified. None of the purified antibody fractions defining the mapped epitopes on gp120 had any neutralizing capacity against HIVIIIB.CONCLUSIONS: This study is the first demonstration of the applicability of random fragment expression libraries for the direct screening of human serum in order to map epitopes on gp120. Two new epitopes and two previously identified epitopes were mapped in this way. However, none of the linear epitopes was defined by antibody fractions neutralizing HIVIIIB, and it was not possible to map epitopes defined by neutralizing antibodies in the serum samples capable of neutralizing HIVIIIB infection of H9 cells. Thus, it appears that the neutralizing activity of serum in this study was not due to anti-gp120 antibodies defining linear epitopes.
AB - OBJECTIVE: To map epitopes on gp120 defined by human antibodies and to examine the neutralizing activity of these antibodies.DESIGN AND METHODS: Serum from HIV-1-antibody-positive individuals was used to screen a random fragment expression library representing gp120 from the HIVIIIB clone BH10. The library was based on the pUEX1 expression vector. Serum was tested for in vitro neutralizing activity using H9 cells and the HIVIIIB isolate.RESULTS: Four different epitopes defined by human antibodies were mapped on gp120. Two of these have not previously been reported and are located within amino acids (aa) 90-100 in the C1 region and aa 355-365 in the semi-conserved region between V3 and V4. The other two are located within aa 140-145 and aa 286-309. These epitopes are situated in regions that have been shown to demarcate human epitopes. Three serum samples with neutralization titres > or = 1024 were identified. None of the purified antibody fractions defining the mapped epitopes on gp120 had any neutralizing capacity against HIVIIIB.CONCLUSIONS: This study is the first demonstration of the applicability of random fragment expression libraries for the direct screening of human serum in order to map epitopes on gp120. Two new epitopes and two previously identified epitopes were mapped in this way. However, none of the linear epitopes was defined by antibody fractions neutralizing HIVIIIB, and it was not possible to map epitopes defined by neutralizing antibodies in the serum samples capable of neutralizing HIVIIIB infection of H9 cells. Thus, it appears that the neutralizing activity of serum in this study was not due to anti-gp120 antibodies defining linear epitopes.
KW - Amino Acid Sequence
KW - B-Lymphocytes/immunology
KW - Base Sequence
KW - Enzyme-Linked Immunosorbent Assay
KW - Epitopes/genetics
KW - Gene Library
KW - HIV Antibodies/immunology
KW - HIV Envelope Protein gp120/genetics
KW - HIV-1/immunology
KW - Humans
KW - Molecular Sequence Data
KW - Neutralization Tests
KW - Peptide Fragments/immunology
M3 - Journal article
C2 - 1283518
VL - 6
SP - 1451
EP - 1456
JO - AIDS
JF - AIDS
SN - 1350-2840
IS - 12
ER -
ID: 201190627