Polyclonal Broadly Neutralizing Antibody Activity Characterized by CD4 Binding Site and V3-Glycan Antibodies in a Subset of HIV-1 Virus Controllers

Nyanhete, Tinashe; Edwards, Robert J.; LaBranche, Celia C.; Mansouri, Katayoun; Eaton, Amanda; Dennison, S. Moses; Saunders, Kevin O.; Goodman, Derrick; Janowska, Katarzyna; Spreng, Rachel L.; Zhang, Lu; Mudrak, Sarah V.; Hope, Thomas J.; Hora, Bhavna; Bradley, Todd; Georgiev, Ivelin S.; Montefiori, David C.; Acharya, Priyamvada; Tomaras, Georgia D.

doi:10.3389/fimmu.2021.670561

Cited by 3 publications

(10 citation statements)

References 64 publications

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“…The kinetics of the biomolecular interactions were examined by immobilizing the ligand on an appropriate sensor surface and keeping the analyte in solution ( Figure 1A ). Antigens were loaded onto Streptavidin (SA) or Aminopropylsilane (APS) or Amine reactive (AR2G) biosensors as detailed previously for testing the binding antibodies ( 8 , 11 – 13 , 22 ). For this method, the form of the antibody will influence the kinetics of binding to the immobilized antigen as demonstrated by differences in the kinetics of epitope-matched Fab and IgG1 (Fab form, Figure 1B ; IgG1 form, Figure 1C ).…”

Section: Methodsmentioning

confidence: 99%

“…Sample analyses were performed with approval from the Duke Medicine Institutional Review Board for Clinical Investigations (Protocol Pro00074497, Pro00104803 and Pro00009701). The efficacy and/or immunological evaluations for these studies were reported earlier ( 8 , 11 , 13 , 22 , 29 – 31 ). All study participants had previously provided consent for future use of samples for research, and all samples were de-identified.…”

Section: Materials and Equipmentmentioning

confidence: 99%

“…While avidity is classically measured by monitoring binding under chaotropic conditions using ELISA and other methods ( 10 ), kinetics based methods for measuring avidity of pAbs using surface plasmon resonance (SPR) and biolayer interferometry (BLI) techniques have been widely employed ( 8 , 11 – 22 ). Both SPR and BLI are label-free techniques for studying kinetics of biomolecular interactions where one of the binding partners is immobilized (ligand) on an appropriate sensor chip or surface and the other binding partner, termed as analyte, is kept in solution to monitor interaction.…”

Section: Introductionmentioning

confidence: 99%

“…All the above manifest often in complex time-course profiles. Currently, the standard practice in SPR and BLI analysis of pAbs binding to a given antigen is to measure the antigen specific binding response as a direct readout and estimate the k d of the interaction using a Langmuir dissociation model assuming a 1:1 interaction ( 8 , 11 – 22 ),…”

Section: Introductionmentioning

confidence: 99%

“…The inadequacy of the Langmuir model to describe the data will be conspicuous in the residuals plot when the goodness of fit of pAbs dissociation time-courses is judged. Yet the pAbs dissociation time-courses are continued to be analyzed using Langmuir dissociation model ( 8 , 11 – 22 ) or dissected into fast and slow phases manually ( 25 ) as convenient tools are lacking. Thus, models that account for the contribution of various antibodies differing in k d to the observed dissociation time-courses of pAbs-antigen interaction would be appropriate for better understanding of the heterogeneity in avidity of pAbs.…”

Section: Introductionmentioning

confidence: 99%

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A tool for evaluating heterogeneity in avidity of polyclonal antibodies

Dodds²,

Spreng³

et al. 2023

Front. Immunol.

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Diversity in specificity of polyclonal antibody (pAb) responses is extensively investigated in vaccine efficacy or immunological evaluations, but the heterogeneity in antibody avidity is rarely probed as convenient tools are lacking. Here we have developed a polyclonal antibodies avidity resolution tool (PAART) for use with label-free techniques, such as surface plasmon resonance and biolayer interferometry, that can monitor pAb-antigen interactions in real time to measure dissociation rate constant (kd) for defining avidity. PAART utilizes a sum of exponentials model to fit the dissociation time-courses of pAb-antigens interactions and resolve multiple kd contributing to the overall dissociation. Each kd value of pAb dissociation resolved by PAART corresponds to a group of antibodies with similar avidity. PAART is designed to identify the minimum number of exponentials required to explain the dissociation course and guards against overfitting of data by parsimony selection of best model using Akaike information criterion. Validation of PAART was performed using binary mixtures of monoclonal antibodies of same specificity but differing in kd of the interaction with their epitope. We applied PAART to examine the heterogeneity in avidities of pAb from malaria and typhoid vaccinees, and individuals living with HIV-1 that naturally control the viral load. In many cases, two to three kd were dissected indicating the heterogeneity of pAb avidities. We showcase examples of affinity maturation of vaccine induced pAb responses at component level and enhanced resolution of heterogeneity in avidity when antigen-binding fragments (Fab) are used instead of polyclonal IgG antibodies. The utility of PAART can be manifold in examining circulating pAb characteristics and could inform vaccine strategies aimed to guide the host humoral immune response.

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Section: Methodsmentioning

confidence: 99%

Section: Materials and Equipmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A tool for evaluating heterogeneity in avidity of polyclonal antibodies

Dodds²,

Spreng³

et al. 2023

Front. Immunol.

Self Cite

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Characterization of antibody‐dependent cellular phagocytosis in patients infected with hepatitis C virus with different clinical outcomes

Adhikari,

Abayasingam,

Brasher

et al. 2024

Journal of Medical Virology

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Early neutralizing antibodies against hepatitis C virus (HCV) and CD8 + T cell effector responses can lead to viral clearance. However, these functions alone are not sufficient to protect patients against HCV infection, thus undefined additional antiviral immune mechanisms are required. In recent years, Fc‐receptor‐dependent antibody effector functions, particularly, antibody‐dependent cellular phagocytosis (ADCP) were shown to offer immune protection against several RNA viruses. However, its development and clinical role in patients with HCV infection remain unknown. In this study, we found that patients with chronic GT1a or GT3a HCV infection had significantly higher concentrations of anti‐envelope 2 (E2) antibodies, predominantly IgG1 subclass, than patients that cleared the viruses while the latter had antibodies with higher affinities. 97% of the patients with HCV had measurable ADCP of whom patients with chronic disease showed significantly higher ADCP than those who naturally cleared the virus. Epitope mapping studies showed that patients with antibodies that target antigenic domains on the HCV E2 protein that are known to associate with neutralization function are also strongly associated with ADCP, suggesting antibodies with overlapping/dual functions. Correlation studies showed that ADCP significantly correlated with plasma anti‐E2 antibody levels and neutralization function regardless of clinical outcome and genotype of infecting virus, while a significant correlation between ADCP and affinity was only evident in patients that cleared the virus. These results suggest ADCP was mostly driven by antibody titer in patients with chronic disease while maintained in clearers due to the quality (affinity) of their anti‐E2 antibodies despite having lower antibody titers.

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Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis

Basu,

Gohain,

Kim

et al. 2023

Cells

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Developing a preventative vaccine for HIV-1 has been a global priority. The elicitation of broadly neutralizing antibodies (bNAbs) against a broad range of HIV-1 envelopes (Envs) from various strains appears to be a critical requirement for an efficacious HIV-1 vaccine. To understand their ability to neutralize HIV-1, it is important to characterize the binding characteristics of bNAbs. Our work is the first to utilize microscale thermophoresis (MST), a rapid, economical, and flexible in-solution temperature gradient method to quantitatively determine the binding affinities of bNAbs and non-neutralizing monoclonal antibodies (mAbs) to HIV-1 recombinant envelope monomer and trimer proteins of different subtypes, pseudoviruses (PVs), infectious molecular clones (IMCs), and cells expressing the trimer. Our results demonstrate that the binding affinities were subtype-dependent. The bNAbs exhibited a higher affinity to IMCs compared to PVs and recombinant proteins. The bNAbs and mAbs bound with high affinity to native-like gp160 trimers expressed on the surface of CEM cells compared to soluble recombinant proteins. Interesting differences were seen with V2-specific mAbs. Although they recognize linear epitopes, one of the antibodies also bound to the Envs on PVs, IMCs, and a recombinant trimer protein, suggesting that the epitope was not occluded. The identification of epitopes on the envelope surface that can bind to high affinity mAbs could be useful for designing HIV-1 vaccines and for down-selecting vaccine candidates that can induce high affinity antibodies to the HIV-1 envelope in their native conformation.

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Polyclonal Broadly Neutralizing Antibody Activity Characterized by CD4 Binding Site and V3-Glycan Antibodies in a Subset of HIV-1 Virus Controllers

Cited by 3 publications

References 64 publications

A tool for evaluating heterogeneity in avidity of polyclonal antibodies

A tool for evaluating heterogeneity in avidity of polyclonal antibodies

Characterization of antibody‐dependent cellular phagocytosis in patients infected with hepatitis C virus with different clinical outcomes

Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis

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