Neutralizing GP41 antibodies and their use
Inventors
Connors, Mark • Huang, Jinghe • Laub, Leo • Mascola, John • Longo, Nancy • Doria-Rose, Nicole
Assignees
US Department of Health and Human Services
Publication Number
US-10047147-B2
Publication Date
2018-08-14
Expiration Date
2032-11-07
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Abstract
Monoclonal neutralizing antibodies are disclosed that specifically bind to the HIV-1 gp41 membrane-proximal external region (MPER). Also disclosed are compositions including the disclosed antibodies that specifically bind gp41, nucleic acids encoding these antibodies, expression vectors including the nucleic acids, and isolated host cells that express the nucleic acids. The antibodies and compositions disclosed herein can be used for detecting the presence of HIV-1 in a biological sample, or detecting an HIV-1 infection or diagnosing AIDS in a subject. In additional, the broad neutralization breadth of the disclosed antibodies makes them ideal for treating a subject with an HIV infection. Thus, disclosed are methods of treating and/or preventing HIV infection.
Core Innovation
The invention provides isolated human monoclonal neutralizing antibodies that specifically bind to the HIV-1 gp41 membrane-proximal external region (MPER). These antibodies, including 10E8 and 10E8-like antibodies, exhibit broad neutralization breadth and potency against diverse HIV-1 isolates. The invention also discloses compositions containing such antibodies, nucleic acids encoding them, expression vectors, and host cells expressing these nucleic acids. The antibodies bind a previously uncharacterized epitope on gp41, designated the 10E8 epitope, enabling them to detect HIV-1 presence in samples, diagnose infection or AIDS, and treat or prevent HIV infection.
The problem being addressed is the inability of current HIV-1 vaccines to induce potent and broadly reactive neutralizing antibodies (NAbs) effective against most circulating HIV-1 strains. Existing neutralizing monoclonal antibodies that target the gp41 MPER, such as 2F5, 4E10, and Z13E1, are limited in either strain cross-reactivity or neutralization potency and exhibit autoreactivity, reducing their therapeutic viability. There is a need for methods to prepare monoclonal broadly neutralizing antibodies that provide effective protection from HIV infection.
The invention solves this problem by identifying human monoclonal antibodies that bind the gp41 MPER at the 10E8 epitope characterized structurally and functionally. These antibodies are broadly neutralizing, do not exhibit autoreactivity or phospholipid binding, and thus overcome prior limitations. The crystal structure of antibody 10E8 complexed with gp41 reveals the atomic-level interface and identifies critical conserved residues for neutralization, providing insight for vaccine design and therapeutic application. Methods for treatment or prevention of HIV infection by administering these antibodies or their encoding nucleic acids are also disclosed.
Claims Coverage
The patent claims comprise a single main inventive method with multiple defined steps concerning the production of antibodies that specifically bind to a target antigen from memory B cells.
Isolation and expansion of memory B cells with defined phenotype
Sorting a population of memory B cells isolated from a subject where at least 90% of the cells are CD19+IgA−IgD−IgM− B cells, into wells of a microtiter plate, wherein the cells are not immortalized.
Incubation with growth medium containing IL-21, IL-2, and mouse CD40L
Incubating the memory B cells with an effective amount of IMDM growth medium comprising IL-21, IL-2, and mouse CD40L for 10-15 days to induce expansion and antibody secretion.
Screening for target antigen binding or neutralization activity
Following incubation, screening supernatant from the wells for specific binding activity for the target antigen, neutralization of the target antigen, or both.
Selection and variable region amplification of memory B cells producing target-binding antibodies
Selecting expanded cells from wells with specific binding activity for the target antigen, amplifying nucleic acids encoding heavy and light chain variable regions from those cells.
Expression of heavy and light chain variable regions to produce target antigen-specific antibody
Expressing the amplified heavy and light chain variable regions to produce an antibody comprising these regions that specifically binds the target antigen.
The claimed method centers on sorting specific memory B cells, expanding them with cytokines and CD40L, screening secreted antibodies for target antigen activity, and molecularly cloning and expressing variable regions to produce specific monoclonal antibodies.
Stated Advantages
The antibodies exhibit broad and potent neutralization of diverse HIV-1 isolates, neutralizing about 98% of tested viruses with high potency.
The antibodies do not bind phospholipids or autoantigens, thus do not exhibit autoreactivity, overcoming a major drawback associated with earlier MPER antibodies.
They have improved access to the epitope on the viral envelope spike compared to prior MPER antibodies like 2F5 and 4E10.
The structure-based understanding of the antibody-epitope interaction supports vaccine design efforts to elicit broadly neutralizing MPER-specific antibodies.
Documented Applications
Detecting the presence of HIV-1 in a biological sample by specifically binding gp41 using the disclosed antibodies or fragments thereof.
Diagnosing HIV-1 infection or AIDS in a subject through increased binding of the antibodies to a sample relative to a control.
Treating or preventing HIV-1 infection in a subject by administering therapeutically effective amounts of monoclonal neutralizing antibodies that specifically bind gp41.
Use in post-exposure prophylaxis or to reduce or eliminate viral reservoirs in combination with anti-viral therapy.
Use of antibody nucleic acids or expression constructs to elicit in vivo antibody production.
Screening a vaccine or immunogen for proper epitope conformation by detecting binding of the antibodies.
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