Induction of a protein-targeted catalytic response in autoimmune prone mice: antibody-mediated cleavage of HIV-1 glycoprotein GP120.
We have induced a polyclonal IgG that degrades the HIV-1 surface antigen, glycoprotein gp120, by taking advantage of the susceptibility of SJL mice to a peptide-induced autoimmune disorder, experimental autoimmune encephalomyelitis (EAE). Specific pathogen-free SJL mice were immunized with structural fragments of gp120, fused in-frame with encephalitogenic peptide MBP(85-101). It has resulted in a pronounced disease-associated immune response against antigens. A dramatic increase of gp120 degradation level by purified polyclonal IgG from immunized versus nonimmunized mice has been demonstrated by a newly developed fluorescence-based assay. This activity was inhibited by anti-mouse immunoglobulin antibodies as well as by Ser- and His-reactive covalent inhibitors. A dominant proteolysis site in recombinant gp120 incubated with purified polyclonal IgG from immunized mice was shown by SDS-PAGE. The SELDI-based mass spectrometry revealed that these antibodies exhibited significant specificity toward the Pro484-Leu485 peptide bond. The sequence surrounding this site is present in nearly half of the HIV-I variants. This novel strategy can be generalized for creating a catalytic vaccine against viral pathogens.
Ponomarenko
Natalia A
NA
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, RAS, 16/10, Miklukho-Maklaya str., Moscow 117871, Russia.
Vorobiev
Ivan I
II
Alexandrova
Elena S
ES
Reshetnyak
Andrew V
AV
Telegin
Georgy B
GB
Khaidukov
Sergey V
SV
Avalle
Bérangère
B
Karavanov
Alexander
A
Morse
Herbert C
HC
Thomas
Daniel
D
Friboulet
Alain
A
Gabibov
Alexander G
AG
D000818:Animals; D017151:Antibodies, Catalytic / Q000378:metabolism*; D000915:Antibody Affinity; D001327:Autoimmune Diseases / Q000139:chemically induced / Q000276:immunology; D002384:Catalysis; D004591:Electrophoresis, Polyacrylamide Gel; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology*; D005456:Fluorescent Dyes / Q000737:chemistry; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1 / Q000737:chemistry*; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D007930:Leucine / Q000737:chemistry; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D011392:Proline / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014765:Viral Vaccines / Q000276:immunology
D017151:Antibodies, Catalytic; D005456:Fluorescent Dyes; D015699:HIV Envelope Protein gp120; D007074:Immunoglobulin G; D010446:Peptide Fragments; D014765:Viral Vaccines; D011392:Proline; D007930:Leucine