Influence of amino acid substitutions on antigenicity of immunodominant regions of the HTLV type I envelope surface gylcoprotein: a study using monoclonal antibodies raised against relevant peptides.
By the use of sera of human T cell leukemia virus type I (HTVL-I)-infected individuals it was shown that amino acid substitutions at positions 192 (proline to serine) and 250 (serine to proline) in major immunodominant regions (175-199 and 239-261) of the surface envelope glycoprotein (gp46) of the virus may influence the humoral response. Since human sera are polyclonal in nature, one cannot readily discriminate between an immunoglobulin-specific recognition and multiple bindings of diverse antibodies. To overcome this difficulty we generated murine monoclonal antibodies to synthetic peptides mimicking all or portions of these gp46 regions. The reactivity of some of these antibodies to synthetic peptides harboring (or not harboring) the preceding amino acid substitutions at position 192 or 250, to denatured gp46 by Western blotting, and to live (variously substituted) HTLV-I-infected cells, combined with blocking experiments with various peptides, allow us to conclude that the major epitopes (positions 183-191, 190-197, 190-199, and 246-252) in the two immunodominant regions may elicit different antibody responses according to their sequences. It is worth noting that in a reporter gene inhibition assay, it was found that a neutralizing monoclonal antibody (MF1), the epitope for which is located between residues 190 and 197, had a high level of activity when cells (2060) harboring a gp46 with proline at position 192 were used and had no activity toward cells (1010) with a serine at this position. Therefore our results establish that certain amino acid substitutions of gp46 may drastically affect the antigenicity of the molecule and the biological activity of the antibodies elicited.
Londos-Gagliardi
D
D
INSERM, U328, Structures et Fonctions des Rétrovirus Humains, Institute Bergonié, France.
Jauvin
V
V
Armengaut
M H
MH
Astier-Gin
T
T
Goetz
M
M
Huet
S
S
Guillemain
B J
BJ
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D015480:Deltaretrovirus Antibodies / Q000096:biosynthesis / Q000276:immunology*; D015485:Deltaretrovirus Antigens / Q000737:chemistry / Q000276:immunology*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D019084:Fluorescent Antibody Technique, Indirect; D015686:Gene Products, env / Q000138:chemical synthesis / Q000276:immunology*; D015368:Human T-lymphotropic virus 1 / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000276:immunology; D008099:Liver / Q000166:cytology / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology*; D015682:Retroviridae Proteins, Oncogenic / Q000138:chemical synthesis / Q000276:immunology*; D014709:Vero Cells
D000911:Antibodies, Monoclonal; D015480:Deltaretrovirus Antibodies; D015485:Deltaretrovirus Antigens; D018985:Epitopes, B-Lymphocyte; D015686:Gene Products, env; D016056:Immunodominant Epitopes; D007132:Immunoglobulin Isotypes; D010455:Peptides; D015682:Retroviridae Proteins, Oncogenic; C055266:gp46 protein, Human T-cell leukemia virus type I