Expression and immunogenicity of the V3 loop from the envelope of human immunodeficiency virus type 1 in an attenuated aroA strain of Salmonella typhimurium upon genetic coupling to two Escherichia coli carrier proteins

Vaccine. 1993 Sep;11(12):1221-8. doi: 10.1016/0264-410x(93)90046-z.

Abstract

A peptide comprising residues glu293 to ser334 from the principal neutralization determinant (V3 loop) of the envelope of human immunodeficiency virus type 1 (HIV1 LAVBRU isolate) has been inserted within internal permissive sites of either LamB or MalE, two envelope proteins from Escherichia coli K12. The MalE hybrid protein (MalE133-V3 loop) was stably expressed in the periplasm of Escherichia coli K12, and the V3 loop peptide was detectable on the surface of the native protein by an anti-gp160 monoclonal antibody (mAb 110-A). The disulfide bridge between the two cysteines of the loop was formed. In contrast, genetic coupling to the outer membrane protein LamB did not allow the expression of a stable hybrid protein, and major proteolytic cleavage products of the LamB153-V3 loop were detected by mAb 110-A. The two plasmid-encoded hybrid genes were transferred to an aroA mutant of Salmonella typhimurium. Constitutive expression of the MalE133-V3 loop had no detectable effect on cell growth and on the survival in vivo of the recipient strain. The LamB153-V3 loop was not stably expressed in Salmonella, either in vitro or in vivo. Live recombinant salmonellas expressing MalE-V3 and LamB-V3 loop hybrids were used to immunize mice. The MalE-V3 loop hybrid induced anti-HIV1 envelope antibodies detectable by Western blot and ELISA, while the anti-HIV1 envelope antibodies induced by the LamB-V3 loop hybrid were only detectable by Western blot. In addition, purified MalE-V3 loop hybrid protein was able to stimulate in vitro and induce in vivo a V3 loop-specific T-cell proliferative response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Vaccines / genetics
  • AIDS Vaccines / immunology
  • AIDS Vaccines / pharmacology
  • ATP-Binding Cassette Transporters*
  • Amino Acid Sequence
  • Animals
  • Antibody Formation / drug effects
  • Antibody Formation / immunology
  • Bacterial Outer Membrane Proteins
  • Base Sequence
  • Carrier Proteins / genetics*
  • Escherichia coli / genetics*
  • Escherichia coli Proteins*
  • Gene Products, env / genetics*
  • Gene Products, env / immunology*
  • HIV Envelope Protein gp160
  • HIV-1 / genetics*
  • HIV-1 / immunology*
  • Lymphocyte Activation / drug effects
  • Lymphocyte Activation / immunology
  • Maltose-Binding Proteins
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Monosaccharide Transport Proteins*
  • Periplasmic Binding Proteins*
  • Porins
  • Protein Precursors / genetics*
  • Protein Precursors / immunology*
  • Receptors, Virus / genetics*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Salmonella typhimurium / genetics*
  • Sensitivity and Specificity
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology

Substances

  • AIDS Vaccines
  • ATP-Binding Cassette Transporters
  • Bacterial Outer Membrane Proteins
  • Carrier Proteins
  • Escherichia coli Proteins
  • Gene Products, env
  • HIV Envelope Protein gp160
  • MalE protein, E coli
  • Maltose-Binding Proteins
  • Monosaccharide Transport Proteins
  • Periplasmic Binding Proteins
  • Porins
  • Protein Precursors
  • Receptors, Virus
  • Recombinant Fusion Proteins
  • maltoporins
  • maltose transport system, E coli