Tandem synthetic HIV-1 peptides

• US 5,759,769 A
• Filed: 06/02/1995
• Issued: 06/02/1998
• Est. Priority Date: 06/09/1993
• Status: Expired due to Term

• Current Assignee: Connaught Laboratories Limited
• Original Assignee: Connaught Laboratories Limited
• Inventors: Chong, Pele, Klein, Michel H., Sia, Charles D. Y.
• Primary Examiner(s): Smith, Lynette F.
• Application Number:
  US08/460,602
• Time in Patent Office: 1,096 Days
• Field of Search: 435/5, 435/7.1, 424/208.1
• US Class Current: 435/5
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 2740/16022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2740/16034 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2740/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2740/16222 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: G01N 33/56988 HIV or HTLV
• CPC Class Codes: Y10S 977/802 Virus-based particle
• CPC Class Codes: Y10S 977/917 Vaccine
• CPC Class Codes: Y10S 977/92 Detection of biochemical

Abstract

Novel synthetic peptides are provided which are candidate vaccines against HIV-1 and which are useful in diagnostic application. The peptides comprise an amino acid sequence of a T-cell epitope of the gag protein of HIV-1, specifically p24E linked directly to an amino acid sequence of a B-cell epitope of the V3 loop protein of an HIV-1 isolate and containing the sequence GPGR, and/or the gp41 containing the sequence ELKDWA. Multimeric forms of the tandem synthetic peptides are provided.

Claims

1. A diagnostic kit useful for detecting HIV specific antibodies in a test sample, the kit comprising:

(a) a surface;

(b) at least one peptide immobilized on the surface and having an amino acid sequence epitopically specific for the HIV-specific antibodies, wherein said peptide is selected from the group consisting of;

(i) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate and is selected from the group consisting of P24N, P24L, P24M and P24H having the respective amino acid sequences OMREPRGSDIAGTTSTL (SEQ ID NO;

70), EEMMTACOGVGGPGHK (SEQ ID NO;

73), GHKARVLAEAMSQVT (SEQ ID NO;

76) and PIVQNIQGQMVHQAI (SEQ ID NO;

79) linked at the C-terminal end of said T-cell epitope, to at least one amino acid sequence which is a B-cell epitope of the V3 loop of the envelope protein of an HIV isolate,

(ii) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof to at least one amino acid sequence which contains a B-cell epitope comprising a hybrid V3 loop sequence from at least two HIV-1 isolates;

(iii) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof to at least one amino acid sequence which contains a B-cell epitope comprising a consensus sequence of the V3 loop of at least two HIV-1 primary isolates;

(iv) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolates linked at the C-terminal end thereof to at least two amino acid sequences each containing a B-cell epitope, said B-cell epitope containing amino acid sequences each comprising a V3 loop sequence from a different HIV-1 isolate or HIV-isolate consensus sequences;

(v) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof, to at least one amino acid sequence which contains a B-cell epitope of the gp41 protein of an HIV isolate comprising the amino acid sequence X₁ LKDWX₂ wherein X₁ is E, A, G or Q and X₂ is A or T or an amino acid sequence capable of eliciting an HIV-specific antiserum and recognizing the amino acid sequence X₁ LKDWX₂ ; and

(vi) a synthetic peptide, which comprises a plurality of individual synthetic peptides linked to form a multimeric molecule, each said individual synthetic peptide comprising an amino acid sequence which contains a T-cell epitope of a gag or envelope protein of a human immunodeficiency virus (HIV) isolate linked to an amino acid sequence which contains a B-cell epitope of a gag or envelope protein of an HIV isolate; and

(c) reagent for detecting a complex formed between HIV specific antibodies in the test sample and the at least one immobilized peptide.

2. A diagnostic kit for detecting HIV antigens in a test sample, the kit comprising:

(a) a surface;

(b) an antibody immobilized on the surface and epitopically specific and noncross-reactive for distinct epitopes of the HIV antigen and raised to a peptide which is selected from the group consisting of;

(i) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate selected from the group consisting of P24N, P24L, P24M and P24H having the respective amino acid sequences OMREPRGSDIAGTTSTL (SEQ ID NO;

70), EEMMTACOGVGGPGHK (SEQ ID NO;

73), GHKARVLAEAMSQVT (SEQ ID NO;

76) and PIVONIOGOMVHOAI (SEQ ID NO;

79) linked at the C-terminal end of said T-cell epitope, to at least one amino acid sequence which contains a B-cell epitope of the V3 loop of the envelope protein of an HIV isolate,

(ii) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof to at least one amino acid sequence which contains a B-cell epitope comprising a hybrid V3 loop sequence from at least two HIV-1 isolates;

(iii) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof to at least one amino acid sequence comprising a B-cell epitope which contains a consensus sequence of the V3 loop of at least two HIV-1 primary isolates;

(iv) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolates linked at the C-terminal end thereof to at least two amino acid sequences each containing a B-cell epitope, said B-cell epitope containing amino acid sequence each comprising a V3 loop sequence from a different HIV-1 isolate or HIV-isolate consensus sequence;

(v) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof, to at least one amino acid sequence which contains a B-cell epitope of the gp41 protein of an HIV isolate comprising the amino acid sequence X₁ LKDWX₂ wherein X₁ is E, A, G or Q and X₂ is A or T or an amino acid sequence capable of eliciting an HIV-specific antiserum and recognizing the amino acid sequence X₁ LKDWX₂ ; and

(vi) a synthetic peptide, which comprises a plurality of individual synthetic peptides linked to form a multimeric molecule, each said individual synthetic peptide comprising an amino acid sequence which contains a T-cell epitope of a gag or envelope protein of a human immunodeficiency virus (HIV) isolate linked to an amino acid sequence which contains a B-cell epitope of a gag or envelope protein of an HIV isolate; and

(c) reagent for detecting a complex formed between HIV antigens in the test sample and the at least one immobilized antibody.

3. An antibody specific for a peptide which is selected from the group consisting of:

(i) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate selected from the group consisting of P24N, P24L, P24M and P24H having the respective amino acid sequences OMREPRGSDIAGTTSTL (SEQ ID NO;

70) EEMMTACOGVGGPGHK (SEQ ID NO;

73) GHKARVLAEAMSQVT (SEQ ID NO;

76) and PIVONIOGOMVHOAI (SEQ ID NO;

79) linked at the C-terminal end of said T-cell epitope, to at least one amino acid sequence which contains a B-cell epitope of the V3 loop of the envelope protein of an HIV isolate,

(ii) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof to at least one amino acid sequence which contains a B-cell epitope comprising a hybrid V3 loop sequence from at least two HIV-1 isolates;

(iii) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof to at least one amino acid sequence which contains a B-cell epitope comprising a consensus sequence of the V3 loop of at least two HIV-1 primary isolates;

(iv) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolates linked at the C-terminal end thereof to at least two amino acid sequences each containing a B-cell epitope, said B-cell epitope containing amino acid sequences each comprising a V3 loop sequence from a different HIV-1 isolate or HIV-isolate consensus sequence;

(v) a synthetic peptide, which comprises at least one amino acid sequence which contains a T-cell epitope of the gag protein of a human immunodeficiency virus (HIV) isolate linked at the C-terminal end thereof, to at least one amino acid sequence which contains a B-cell epitope of the gp41 protein of an HIV isolate comprising the amino acid sequence X₁ LKDWX₂ wherein X₁ is E, A, G or Q and X₂ is A or T or an amino acid sequence capable of eliciting an HIV-specific antiserum and recognizing the amino acid sequence X₁ LKDWX₂ ; and

(vi) a synthetic peptide, which comprises a plurality of individual synthetic peptides linked to form a multimeric molecule, each said individual synthetic peptide comprising an amino acid sequence which contains a T-cell epitope of a gag or envelope protein of a human immunodeficiency virus (HIV) isolate linked to an amino acid sequence which contains a B-cell epitope of a gag or envelope protein of an HIV isolate.