title: Synergy in monoclonal antibody neutralization of HIV-1 pseudoviruses and infectious molecular clones.
abstract: BACKGROUND
  Early events in HIV infection are still poorly understood; virus derived from acute infections, the transmitted/founders IMCs, could provide more reliable information as they represent strains that established HIV infection in vivo, and therefore are investigated to elucidate potentially shared biological features.
  
  
  METHODS
  This study examined synergy in neutralization by six monoclonal antibodies targeting different domains in gp120 and gp41 and assayed in pairwise combination against 11 HIV-1 clade B strains, either Env pseudoviruses (PV, n = 5) or transmitted/founder infectious molecular clones (T/F IMCs, n = 6). Three of the early-infection env tested as PV were juxtaposed with T/F viruses derived from the same three patients, respectively.
  
  
  RESULTS
  All antibodies reaching IC50 were assayed pairwise (n = 50). T/F IMCs showed overall lower sensitivity to neutralization by single antibodies than PV, including within the three patient-matched pairs. Remarkably, combination index (CI) calculated using the Chow and Talalay method indicated synergy (CI < 0.9) in 42 data sets, and occurred in T/F IMC at similar proportions (15 of 17 antibody-T/F IMC combinations tested) as in pseudoviruses (27 of 33). CI values indicative of additivity and low-level antagonism were seen in 5 and 3 cases, respectively. Most pairs showed comparable synergic neutralizing effects on both virus groups, with the 4E10 + PG16 pair achieving the best synergic effects. Variability in neutralization was mostly observed on pseudovirus isolates, suggesting that factors other than virus isolation technology, such as env conformation, epitope accessibility and antibody concentration, are likely to affect polyclonal neutralization.
  
  
  CONCLUSIONS
  The findings from this study suggest that inhibitory activity of bNAbs can be further augmented through appropriate combination, even against viruses representing circulating strains, which are likely to exhibit a less sensitive Tier 2 neutralization phenotype. This notion has important implications for the design and development of anti-Env bNAb-inducing vaccines and polyclonal sera for passive immunization.
authors.affiliation: Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy. migliettariccardo@gmail.com.|Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA. migliettariccardo@gmail.com.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Miglietta
authors.affiliation: Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy. pastori.claudia@hsr.it.
authors.forename: Claudia
authors.initials: C
authors.lastname: Pastori
authors.affiliation: Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy. Assuassuntavenuti@gmail.com.
authors.forename: Assunta
authors.initials: A
authors.lastname: Venuti
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA. christinaochsenbauer@uabmc.edu.|CFAR, University of Alabama at Birmingham, Birmingham, AL, USA. christinaochsenbauer@uabmc.edu.
authors.forename: Christina
authors.initials: C
authors.lastname: Ochsenbauer
authors.affiliation: Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy. lopalco.lucia@hsr.it.
authors.forename: Lucia
authors.initials: L
authors.lastname: Lopalco
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D057809:HEK293 Cells; D015497:HIV-1 / Q000276:immunology* / Q000472:pathogenicity; D006801:Humans; D009500:Neutralization Tests
chemical_list: D000911:Antibodies, Monoclonal