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AssigneeBenjamin Rovinski et al
InventorsBenjamin Rovinski, Joel Haynes, Shi Xian Cao, Michel Henri Klein
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This invention is directed toward diagnostic kits and antigens useful for the detection of viral antigens in a test sample. The kit contains antigens obtained from self-assembled, non-infectious, non-replicating, immunogenic retrovirus-like particles comprising modified HIV-1 genomes devoid of long terminal repeats and containing nucleotide sequences encoding chimeric envelope glycoproteins. One preferred embodiment discloses the engineering of a series of expression vectors in which a synthetic oligomer encoding gp120 residues 306 to 328 (amino acids YNKRKRIHIGP GRAFYTTKNIIG) from the V3 loop of the MN viral isolate was inserted at various positions within the endogenous HIV-1.sub.LAI env gene. Expression studies revealed that insertion of the heterologous V3(MN) loop segment resulted in the secretion of fully assembled HIV-like particles containing chimeric LAI/MN envelope glycoproteins. Both V3 loop epitopes were recognized by loop-specific neutralizing antibodies. Immunization with HIV-like particles containing chimeric envelope proteins induced specific antibody responses against both the autologous and heterologous V3 loop epitopes, including cross-neutralizing antibodies against the HIV-1.sub.LAI and HIV-1.sub.MN isolates.
An immunoassay diagnostic kit for the detection of antibodies in a test sample comprising an antigen consisting of a self-assembled, non-infectious, non-replicating, immunogenic, retrovirus-like particle encoded by a modified HIV genome devoid of long terminal repeats containing a nucleotide sequence coding for a chimeric envelope glycoprotein, said chimeric envelope glycoprotein having a first retroviral envelope amino acid sequence and a second retroviral envelope amino acid sequence, wherein said first amino acid sequence contains the HIV-1 gp120 conserved region 2 and said second amino acid sequence contains a retroviral envelope amino acid sequence of a heterologous strain of HIV-1, HIV-2, HTLV-I, or HTLV-II inserted into said first retroviral envelope amino acid sequence by inserting a nucleotide sequence encoding said second retroviral envelope amino acid sequence into a nucleotide sequence encoding said first retroviral envelope amino acid sequence at an endogenous conserved region 2 restriction site selected from the group consisting of BglII and StuI.
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