Harvard College

Tun-Hou Lee

Myron E Essex

Xiaofang Yu

Chun-Nan Lee

Jonathan S Allan

Woan-Ruoh Lee

Selective deglycosylation of HIV-1 envelope proteins enhances their ability to elicit a protective immune response in people. Glycosylation can reduce or prevent immunological recognition of envelope protein domains. Selective deglycosylation exposes these domains and improves the opportunity for a protective immune response. Deglycosylation which produces substantial conformational changes (as determined by loss of infectivity) should be avoided. Recombinant HIV-1 envelope glycoproteins are generated which have primary amino acid sequence mutation(s) in consensus sequence(s) for N-linked glycosylation (sugar attachment), so as to prevent glycosylation at that site(s). The position of such genetic deglycosylation is important and should be between the C terminus of gp120 and the Cys at the N-terminal side of the cysteine loop containing the hyper-variable region 3 (V3) (this Cys is generally positioned about at residue 296, counting from the N-terminus of gp120). The mutant glycoprotein should be deglycosylated such that the total molecular mass of the mutant gp120 component is less than 90% (more preferably less than 75%) of the corresponding fully glycosylated wild type gp120 component to maximize a useful immune response.

Selectively deglycosylated human immunodeficiency virus type 1 envelope

Nucleic acid constructs encoding mutated human immunodeficiency virus gp41 polypeptides are described. The mutated polypeptides are effective to disrupt viral replication of HIV or disrupt the assembly of viral Env proteins in an HIV infected cell.

HIV gp41 mutants

A first glycoprotein having a molecular weight of approximately 120,000 daltons in the H9/HTLV-III cell line, of which approximately 90,000 daltons is the unglycosylated moiety, is obtained from cells infected with human T-cell leukemia virus, type III. A second glycoprotein having a molecular weight of approximately 160,000 daltons is also obtained from such cells, of which approximately 90,000 daltons is the unglycosylated moiety and is substantially identical to the unglycosylated moiety of the first glycoprotein. The presence, in a biological specimen, of either of these unglycosylated or of the unglycosylated moiety is indicative of the presence of cells infected by human T-cell leukemia virus. An assay for the glycoprotein or its unglycosylated moiety is a useful diagnostic procedure for determining such infection in biological specimens.

Assay for detecting infection by HTLV-III

Nucleic acid constructs encoding mutated human immunodeficiency virus matrix proteins are described. The mutated proteins lower the incorporation of envelope polypeptides in viral particles, disrupt viral assembly or disrupt viral entry into uninfected cells.

Nucleic acids encoding mutated human immunodeficiency virus matrix

Cells infected with HTLV-III yield a protein (p27) having an apparent molecular weight of about 27,000 daltons.

T-cell lymphotrophic virus protein and assay

A first glycoprotein having a molecular weight of approximately 61,000-68,000 daltons in the MJ, C5-MJ, C91 PL or HUT-102 cell lines, of which 46,000 to 48,000 is the unglycosylated moiety, is obtained from cells infected with human T cell leukemia virus. A second glycoprotein having a molecular weight of approximately 45,000-52,000 daltons is also obtained from such cells and is in large part identical to the NH.sub.2 -terminal end of the first glycoprotein. The presence, in a biological specimen, of antibody to the antigenic determinant of either of these proteins is indicative of the presence of cells infected by human T cell leukemia virus. An assay for the antibody is a useful diagnostic procedure for determining such infection in biological specimens.

Method and products for detection of human T cell leukemia virus

A first glycoprotein having a molecular weight of approximately 61,000-68,000 daltons in the MJ, C5-MJ, C91 PL or HUT-102 cell lines, of which 46,000 to 48,000 is the unglycosylated moiety, is obtained from cells infected with human T cell leukemia virus. A second glycoprotein having a molecular weight of approximately 45,000-52,000 daltons is also obtained from such cells and is in large part identical to the NH.sub.2 -terminal end of the first glycoprotein. The presence, in a biological specimen, of antibody to the antigenic determinant of either of these proteins is indicative of the presence of cells infected by human T cell leukemia virus. An assay for the antibody is a useful diagnostic procedure for determining such infection in biologial specimens.

A first glycoprotein having a molecular weight of approximately 120,000 daltons in the H9/HTLV-III cell line, of which approximately 90,000 daltons is the unglycosylated moiety, is obtained from cells infected with human T-cell leukemia virus, type III. A second glycoprotein having a molecular weight of aproximately 160,000 daltons is also obtained from such cells, of which approximately 90,000 daltons is the unglycosylated moiety and is substantially identical to the unglycosylated moiety of the first glycoprotein. The presence, in a biological specimen, of either of these glycoproteins or of the unglycosylated moiety is indicative of the presence of cells infected by human T-cell leukemia virus. An assay for the glycoprotein or its unglycosylated moiety is a useful diagnostic procedure for determining such infection in biological specimens.

Growing community of inventors

Newton, MA, United States of America

Tun-Hou Lee