Broad neutralization coverage of HIV by multiple highly potent antibodies

Broadly neutralizing antibodies against highly variable viral pathogens are much sought after to treat or protect against global circulating viruses. Here we probed the neutralizing antibody repertoires of four human immunodeficiency virus (HIV)-infected donors with remarkably broad and potent neutralizing responses and rescued 17 new monoclonal antibodies that neutralize broadly across clades. Many of the new monoclonal antibodies are almost tenfold more potent than the recently described PG9, PG16 and VRC01 broadly neutralizing monoclonal antibodies and 100-fold more potent than the original prototype HIV broadly neutralizing monoclonal antibodies1, 2, 3. The monoclonal antibodies largely recapitulate the neutralization breadth found in the corresponding donor serum and many recognize novel epitopes on envelope (Env) glycoprotein gp120, illuminating new targets for vaccine design. Analysis of neutralization by the full complement of anti-HIV broadly neutralizing monoclonal antibodies now available reveals that certain combinations of antibodies should offer markedly more favourable coverage of the enormous diversity of global circulating viruses than others and these combinations might be sought in active or passive immunization regimes. Overall, the isolation of multiple HIV broadly neutralizing monoclonal antibodies from several donors that, in aggregate, provide broad coverage at low concentrations is a highly positive indicator for the eventual design of an effective antibody-based HIV vaccine. Subject terms:

Immunology

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Neutralization breadth at different IC50 cut-offs. Median neutralization potency against viruses neutralized with an IC50 < 50 μg ml−1. b. Figure 2: Key monoclonal antibodies fully recapitulate serum neutralization by the corresponding donor serum. respectively).1. Of note. Figure 1: Neutralization activity of the newly identified a. Serum breadth was correlated with the breadth of the broadest monoclonal antibody (mAb) for each donor (percentage of viruses neutralized at 50% neutralizing titre (NT50) > 100 or IC50 < 50 μg ml−1. PGT antibodies. 2. monoclonal antibodies isolated from donor 39 could not completely recapitulate the serum neutralization breadth. .

75–100% competition.3.000 are shown. Only glycan structures with RFU (relative fluorescent units) > 3. 126. 128 and 130 to gp120 is competed by Man9 oligodendrons but not Man4 oligodendrons. Error bars represent standard deviation. d. b12 (anti-CD4 binding site). F425/b4e8 (anti-V3). +. Man9 (314) and Man9GlcNAc2 (194) glycans directly. vaccine-achievable concentrations. ++. Binding of PGT monoclonal antibodies 125. except for the PG9 competition assay. Competition of PGT monoclonal antibodies with sCD4 (soluble CD4). Glycan microarray analysis (Consortium for Functional Glycomics (CFG). version 5. Competition assays were performed by ELISA using gp120Bal or gp120JR-FL. Boxes are coded as follows: +++. 128 and 130 contact Man8 (313). PGT-131 showed no detectable binding to the CFG glycan array but bound to Man9-oligodendrons30 (data not shown). b. a. 127. X5 (CD4-induced). −. Figure 3: Epitope mapping of PGT antibodies. which was performed on the surface of JRFLE168K or JR-CSF transfected cells. 126. Man8GlcNAc2 (193). 25–50% competition. Binding of 131 to immobilized gp120 was too low to measure any competition. quaternary) and each other. Figure 4: Certain antibodies or antibody combinations are able to cover a broad range of HIV isolates at low. 50–75% competition. Experiments were performed in duplicate. . 2G12 (anti-glycan). Error bars represent standarmean. PG9 (anti-V1/V2 and V3. c. <25% competition. 127.0) reveals that PGT monoclonal antibodies 125. and data represent an average of at least two independent experiments.

H2 and H3. PGC14. conserved functional sites on otherwise hypervariable antigens. PGV04. with minor contacts from heavy-chain complementarity-determining region 1. Percentage of viruses covered by single monoclonal antibodies (solid lines) or by at least one of the monoclonal antibodies in dual combinations of breadth (dashed black lines) dependent on individual concentrations. b12. The y-axis shows the cumulative frequency of IC50 values up to the concentration shown on the x-axis and can therefore also be interpreted as the breadth at a specific IC50 cut-off. b. which neutralizes strains from multiple subtypes of influenza A virus. PGT130–131. right Cross-neutralization of influenza A viruses mediated by a single antibody loop Immune recognition of protein antigens relies on the combined interaction of multiple antibody loops.a. Recognition of the haemagglutinin receptor-binding site is dominated by a single heavy-chain complementarity-determining region 3 loop. which provide a fairly large footprint and constrain the size and shape of protein surfaces that can be targeted. Thus. VRC01. but it is unclear whether such a mechanism is available to antibodies. The grey area in both panels is the coverage of 26 monoclonal antibodies tested on the 162-virus panel (PGT121–123. c. Here we report the isolation and characterization of an antibody called C05. Single protein loops can mediate extremely high-affinity binding. binding predominantly with a single loop can allow antibodies to target small. PGT135–137. 2F5) and depicts the theoretical maximal achievable coverage known to date. 2G12. Cumulative frequency distribution of IC50 values of broadly neutralizing monoclonal antibodies tested against a 162-virus panel. PGT141–145. PG16. and is sufficient to achieve nanomolar binding with a minimal footprint. PGT125– 128. X-ray and electron microscopy structures show that C05 recognizes conserved elements of the receptor-binding site on the haemagglutinin surface glycoprotein. . PG9. including H1. 4E10.

n = 5 mice for all studies.d. 2. 4 or 5 days post-infection (DPI) with 25× MLD50 of A/Memphis/3/2008 (c) or 33× MLD50 of A/X-31/1968 viruses (d). . group 1 is circled in blue and group 2 is circled in green. Error bars denote ± s. b. A single therapeutic dose of 15 mg kg−1 C05 IgG was delivered 1. 2. Survival and weight loss were monitored in response to varying amounts of C05 IgG administered prophylactically to mice 24 h before challenge with 25× the 50% mouse lethal dose (MLD50) of A/Memphis/3/2008 (H1N1) (a) or 33× MLD50 of A/X-31/1968 (H3N2) viruses (b). 3.1. Figure 2: C05 protects mice from lethal virus challenge. subtypes from groups 1 and 2. Figure 1: C05 neutralizes multiple influenza virus Phylogenetic tree with the two main viral lineages indicated. Strains from subtypes circled in red are bound or neutralized by C05. a. c. d.

5. Because salts are formed from neutralization reactions with equivalent concentrations of weights of acids and bases: N parts of acid will always neutralize N parts of base. 4. The neutralization of a strong acid and strong base has a pH equal to 7. Neutralization Table of Contents 1. 3. References 10.ions to generate water. Introduction When a solution is neutralized. Problems 8. Introduction 2. 7. The amount of acid needed is the amount that would give one mole of protons (H+) and the amount of base needed is the amount that would give one mole of (OH-). 2. 9. 8. . Contributors A neutralization reaction is when an acid and a base react to form water and a salt and involves the combination of H+ ions and OH. Titration 7. the resulting pH when a strong base neutralizes a weak acid will be greater than 7. Solutions 9. Weak Acid-Weak Base Neutralization 5. Determining What is a Strong Acid or Strong Base 3. 1. and conversely. 10. it means that salts are formed from equal weights of acid and base. The neutralization of a strong acid and weak base will have a pH of less than 7. Strong Acid-Strong Base Neutralization 4. pH Levels at the Equivalence Point 6. 6.3.

The solution is NaCL at the equivalence point. the resulting solution's pH will be greater than 7. the pH of the salt solution will always be 7. which means that the pH is equal to 7. Most everything elso not in this table is considered to be weak.ions forming water in a strong acid. the resulting solution's pH will be less than 7. strong base reaction: H + (aq)+OH − (aq)⇋ H 2 O(l) When a strong acid and a strong base fully neutralize then the pH is neutral. Stomach Antacids: .and H3O+. When a strong acid neutralizes a strong base. When a strong acid neutralizes a weak base. weak base reaction can be shown by the net ionic equation example: H + (aq)+NH 3 (aq)⇋ NH + 4 (aq) The equivalence point of a neutralization reaction is when both the acid and the base in the reaction have been completely consumed and neither of them are in excess. At this point of neutralization. the net ionic equation shows the H+ and OH. there are equal amounts of OH.Determining What is a Strong Acid or Strong Base This is a list of the most common strong acids and bases. There is no excess NaOH. When a strong base neutralizes a weak acid. Weak Acid-Weak Base Neutralization A weak acid. Strong Acids HCl HBr HI HCIO4 HNO3 Strong Bases LiOH NaOH KOH RbOH CsOH Ca(OH)2 Sr(OH)2 Ba(OH)2 Strong Acid-Strong Base Neutralization HCl (aq) +NaOH (aq) ⇋ NaCl (aq) +H 2 O (l) acid+base⇋salt+water When you get rid of all of the spectator ions.00 at 25 degrees Celsius.

dihydroxyaluminum sodium carbonat (NaAl(OH)2CO3). calcium carbonate (CaCO3). Som of the ingredients in antacids are: Magnesia (MgO). aluminum hydroxide gel (Al(OH)3).Antacids are supposed to decrease the amount of hydrochloric acid in the stomach by reacting with excess acid. milk of magnesia (Mg(OH)2. . They are use in the treatment of gastric hyperacidity and peptic ulcers. sodium bicarbonate (NaHCO3). Severa of these will habe top be recognized as Bronsted bases.

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