Immunogenetic restriction on elicitation of antibodies

a technology of immunogenetic restriction and antibody elicitation, which is applied in the field of influenza neutralizing antibodies, can solve the problems of low detection concentration of sbnabs and no approach that can evaluate the ability of influenza vaccines, and achieve the effect of improving the neutralization capacity or affinity of antibodies

Inactive Publication Date: 2017-06-22
DANA FARBER CANCER INST INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about improving the ability of antibodies to neutralize influenza viruses. This is done by mutating certain amino acids in the antibodies' structure. The mutations can be made in a specific part of the antibodies' structure called the VH domain. The mutations can involve changing certain amino acids like serine, valine, isoleucine, proline, serine, arginine, valine, and others. The goal is to enhance the ability of the antibodies to bind to the influenza virus' HA protein. These mutations can be made using various techniques known in the field of molecular biology, and they can be combined to improve the antibodies' neutralization capacity. This can help in the development of better drugs or vaccines to prevent or treat influenza infections.

Problems solved by technology

However, only very low concentrations of sBnAbs are detected in the sera of seasonal influenza or H5N1 vaccines, or in commercial intravenous immunoglobulin (IVIG) preparations.
While these assays set the standard for judging the efficacy of vaccines, to date there is no approach that can evaluate the ability of influenza vaccines to induce broadly neutralizing “heterosubtypic” antibodies binding to a highly conserved hydrophobic pocket on the stem of HA (HV1-sBnAbs).

Method used

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  • Immunogenetic restriction on elicitation of antibodies
  • Immunogenetic restriction on elicitation of antibodies
  • Immunogenetic restriction on elicitation of antibodies

Examples

Experimental program
Comparison scheme
Effect test

example 1

ethods

[0248]Described herein are the general methods and assays used in the working examples.

[0249]Materials

[0250]The anti-HA antibodies F10, A66, G17, and D8 were previously described in the study of Sui et al (1) and International Application No. WO 2009 / 079259, herein incorporated by reference in their entireties. The mAbs CR6261 and CR6331 were synthesized by Genewiz, North Brunswick, N.J. Recombinant HA of H5VN04 was produced as described (1). A / California / 04 / 2009 (H1CA0409) and A / Singapore / 1 / 57 (H2 SIN 57) recombinant HAs were supplied by Biodefense and Emerging Infections Research Resources Repository (BEI Resources)

[0251]Cloning of Antibody Variants

[0252]IGHV1-69*01 germline V-segment was synthesized by Geneart (Regensburg, Germany). The germline variant of IGHV1-69 / F10 was constructed by ligating the IGHV1-69*01 gene (NcoI 5′, BssHII 3′) with F10 gene segment that included the CDR-H3+ light chain (BssHII 5′ NotI 3′) into the pET22b vector, which was digested with NcoI-NotI....

example 2

ation of Anchor Residues in the Heavy Chains of HVI-69-SBNABS

[0284]The co-crystal structures of the HV1-69-sBnAbs, F10 (1), CR6261 (3), and CR9114 (5) with H5VN04 established that binding is mediated exclusively by the IGHV1-69 heavy chains. Estimates of the binding free energy contributions for heavy chain CDR residues using ANCHOR server (17) (FIG. 1A) have identified three common anchor points: a hydrophobic residue at CDR-H2 position 53 (generally Ile / Met), a Phe at CDR-H2 position 54, and a Tyr residue at CDR-H3 position 98. Structural analysis shows (FIGS. 1B and 6) that the common aromatic pair of CDR-H2 Phe54 and CDR-H3 Tyr98 pack closely together (˜4 Å) in order to bind to adjacent pockets formed by elements of the fusion peptide. Tyr98 makes both hydrophobic interactions as well as a strong H-bond with the fusion peptide (the main chain carbonyl of Asp192), and adopts a single conformation in the 3 known structures. The side-chains of Phe54 converge in one location, packin...

example 3

nalysis of Somatic Mutations in Rearranged IGHV1-69 Genes of the SBNABS

[0286]A mean of 12.6±4.2 V-segment substitutions are found among the published HV1-69-sBnAbs, ranging from 5 in CR6331 / CR6432 to 22 in FE43 / CR6334 (FIG. 2B), which is similar to the average range for rearranged IGHV genes (21). This distribution indicates the pathways for potent HV1-69-sBnAbs formation do not necessarily require multiple maturation events, but rather the incorporation of key residues. Further examination of the V-segments of HV1-69-sBnAbs revealed common substitutions such as the hydrophobic residue at position 74 (in CDR-H4) and changes in CDR-H1 and CDR-H2 (FIG. 2A). To investigate which of these substitutions are unique for HV1-69-sBnAbs, we compared the frequency of specific somatic mutations in the HV1-69-sBnAbs V-segments with the identical mutations in a control set of unique Ab sequences derived from the IGHV1-69-51p1 germline group (IgBlast, n=287). Thirteen HV1-69-sBnAb distinctive V-se...

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Abstract

The present invention provides structural determinants important for binding to the stem domain of the HA protein of influenza virus, and methods of use thereof for production of high affinity neutralizing influenza virus antibodies based upon these determinants. The present invention further provides tools for determining the efficacy of an influenza virus vaccine. The present invention further provides a molecular signature useful for determining the efficacy of an influenza virus vaccine in a subject, or for predicting prior immunologic exposure or antigen responsiveness to vaccine or influenza virus infection.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority to, and the benefit of U.S. Ser. No. 61 / 955,678 filed on Mar. 19, 2014, and of U.S. Ser. No. 61 / 974,297 filed on Apr. 2, 2014, the contents of which are incorporated herein by reference in their entirety.GOVERNMENT INTEREST[0002]This invention was made with government support under AI074518 awarded by the National Institutes of Health and W911NF-10-1-0266 awarded by the Defense Advanced Research Projects Agency. The United States government has certain rights in the invention.FIELD OF THE INVENTION[0003]This invention relates generally to influenza neutralizing antibodies, the structural determinants of such antibodies, as well as to methods for use thereof.BACKGROUND OF THE INVENTION[0004]An influenza pandemic represents one of the greatest acute infectious threats to human health. Vaccination remains the principle means of preventing seasonal and pandemic influenza and its complications. A “universal” in...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12Q1/68G01N33/50
CPCC07K16/1018A61K47/4853G01N33/5052C12Q1/6876C07K2317/76C07K2317/565G01N2800/52C07K2317/622C07K2317/55C07K2317/54C07K2317/92C12Q2600/158C12Q2600/106C07K2317/56C07K2317/21C07K2317/567A61K47/6841A61P31/14A61K39/395A61K2300/00
Inventor MARASCO, WAYNEAVNIR, YUVAL
Owner DANA FARBER CANCER INST INC
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