scholarly journals Broadly reactive human monoclonal antibodies targeting the pneumococcal histidine triad protein protect against fatal pneumococcal infection

2021 ◽  
Author(s):  
Jiachen Huang ◽  
Aaron D. Gingerich ◽  
Fredejah Royer ◽  
Amy V. Paschall ◽  
Alma Pena-Briseno ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Disease Prevention ◽  
Disease Incidence ◽  
Protein A ◽  
Pneumococcal Infection ◽  
Surface Protein ◽  
Human Monoclonal Antibodies ◽  
Human Mabs ◽  
Prevention And Treatment ◽  
Pneumococcal Serotype

AbstractStreptococcus pneumoniae remains a leading cause of bacterial pneumonia despite the widespread use of vaccines. While vaccines are effective at reducing the incidence of most vaccine-included serotypes, a rise in infection due to non-vaccine serotypes, and moderate efficacy against some vaccine included serotypes have contributed to high disease incidence. Additionally, numerous isolates of S. pneumoniae are antibiotic or multi-drug resistant. Several conserved pneumococcal proteins prevalent in the majority of serotypes have been examined as vaccines in preclinical and clinical trials. An additional, yet unexplored tool for disease prevention and treatment is the use of human monoclonal antibodies (mAbs) targeting conserved pneumococcal proteins. Here, we isolate the first human mAbs (PhtD3, PhtD6, PhtD7, PhtD8, PspA16) against the pneumococcal histidine triad protein (PhtD), and the pneumococcal surface protein A (PspA), two conserved and protective antigens. mAbs to PhtD target diverse epitopes on PhtD, and mAb PspA16 targets the N-terminal segment of PspA. The PhtD-specific mAbs bind to multiple serotypes, while mAb PspA16 serotype breadth is limited. mAbs PhtD3 and PhtD8 prolong the survival of mice infected with pneumococcal serotype 3. Furthermore, mAb PhtD3 prolongs the survival of mice in intranasal and intravenous infection models with pneumococcal serotype 4, and in mice infected with pneumococcal serotype 3 when administered 24 hours after pneumococcal infection. All PhtD and PspA mAbs demonstrate opsonophagocytic activity, suggesting a potential mechanism of protection. Our results provide new human mAbs for pneumococcal disease prevention and treatment, and identify epitopes on PhtD and PspA recognized by human B cells.

scholarly journals Broadly Reactive Human Monoclonal Antibodies Targeting the Pneumococcal Histidine Triad Protein Protect against Fatal Pneumococcal Infection

2021 ◽  
Vol 89 (5) ◽  
Author(s):  
Jiachen Huang ◽  
Aaron D. Gingerich ◽  
Fredejah Royer ◽  
Amy V. Paschall ◽  
Alma Pena-Briseno ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Disease Prevention ◽  
Disease Incidence ◽  
Protein A ◽  
Pneumococcal Infection ◽  
Human Monoclonal Antibodies ◽  
Human Mabs ◽  
Content Type ◽  
Prevention And Treatment ◽  
Pneumococcal Serotype

ABSTRACT Streptococcus pneumoniae remains a leading cause of bacterial pneumonia despite the widespread use of vaccines. While vaccines are effective at reducing the incidence of most serotypes included in vaccines, a rise in infection due to nonvaccine serotypes and moderate efficacy against some vaccine serotypes have contributed to high disease incidence. Additionally, numerous isolates of S. pneumoniae are antibiotic or multidrug resistant. Several conserved pneumococcal proteins prevalent in the majority of serotypes have been examined for their potential as vaccines in preclinical and clinical trials. An additional, yet-unexplored tool for disease prevention and treatment is the use of human monoclonal antibodies (MAbs) targeting conserved pneumococcal proteins. Here, we isolated the first human MAbs (PhtD3, PhtD6, PhtD7, PhtD8, and PspA16) against the pneumococcal histidine triad protein (PhtD) and the pneumococcal surface protein A (PspA), two conserved and protective antigens. MAbs to PhtD target diverse epitopes on PhtD, and MAb PspA16 targets the N-terminal segment of PspA. The PhtD-specific MAbs bind to multiple serotypes, while MAb PspA16 serotype breadth is limited. MAbs PhtD3 and PhtD8 prolong the survival of mice infected with pneumococcal serotype 3. Furthermore, MAb PhtD3 prolongs the survival of mice in intranasal and intravenous infection models with pneumococcal serotype 4 and in mice infected with pneumococcal serotype 3 when administered 24 h after pneumococcal infection. All PhtD and PspA MAbs demonstrate opsonophagocytic activity, suggesting a potential mechanism of protection. Our results identify new human MAbs for pneumococcal disease prevention and treatment and identify epitopes on PhtD and PspA recognized by human B cells.

Human and primate monoclonal antibodies for in vivo therapy.

1988 ◽  
Vol 34 (9) ◽  
pp. 1681-1688 ◽  
Author(s):  
P H Ehrlich ◽  
Z A Moustafa ◽  
J C Justice ◽  
K E Harfeldt ◽  
I K Gadi ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Western Blot ◽  
Protein A ◽  
Human Chromosomes ◽  
Human Monoclonal Antibodies ◽  
Western Blot Assay ◽  
Human Antibodies ◽  
Low Levels ◽  
High Concentrations

Abstract Human monoclonal antibodies, owing to their decreased immunogenicity, are expected to be an improvement over mouse monoclonal antibodies for in vivo therapy. Human and primate monoclonal antibodies are best produced with a human x mouse heteromyeloma. Several human chromosomes are stable in the human x (human x mouse) hybrids. Chimpanzee anti-digoxin monoclonal antibodies were prepared and characterized. Because they are structurally very similar to human antibodies, they should be well tolerated in humans. The anti-digoxin antibodies can be used for therapy of extreme overdoses or as an in vivo diagnostic tool for slight overdoses. Because the advantage of using human monoclonal antibodies is their lack of immunogenicity, preparation of the antibody must be scrupulous so as not to introduce extraneous immunogens. Analysis to ensure the purity of the preparation can be complicated by the presence of high concentrations of the antibody and the low levels of contamination that must be detected. We describe a Western blot assay for Protein A that is sensitive even in the presence of human IgG.

scholarly journals Efficacy of a Protein Vaccine and a Conjugate Vaccine Against Co-Colonization with Vaccine-Type and Non-Vaccine Type Pneumococci in Mice

Pathogens ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 278
Author(s):  
Gabriela B. C. Colichio ◽  
Giuliana S. Oliveira ◽  
Tasson C. Rodrigues ◽  
Maria Leonor S. Oliveira ◽  
Eliane N. Miyaji
Keyword(s):  
No Reduction ◽  
Protein A ◽  
Surface Protein ◽  
Protein Formulation ◽  
Protein Vaccine ◽  
Conjugate Vaccines ◽  
Serotype Replacement ◽  
Vaccine Type ◽  
Pneumococcal Serotype ◽  
Low Levels

Widespread use of pneumococcal conjugate vaccines (PCVs) has led to substitution of vaccine-type (VT) strains by non-vaccine type (NVT) strains in nasopharyngeal carriage. We compared the efficacy of PCV13 and a nasal protein formulation containing pneumococcal surface protein A (PspA) adjuvanted with the whole-cell pertussis vaccine (wP) in the protection against co-colonization challenge models in mice with VT and NVT strains expressing different PspAs. Immunized mice were challenged with two different mixtures: i. VT4 (PspA3) + NVT33 (PspA1) and ii. VT23F (PspA2) + NVT15B/C (PspA4). Results from the first mixture showed a reduction in loads of VT4 strain in the nasopharynx of mice immunized with PCV13. A statistical difference between the loads of the VT and NVT strains was observed, indicating a competitive advantage for the NVT strain in PCV13-immunized animals. In the second mixture, no reduction was observed for the VT23F strain, probably due to low levels of anti-23F polysaccharide IgG induced by PCV13. Interestingly, a combination of the PspA formulation containing wP with PCV13 led to a reduction in colonization with both strains of the two mixtures tested, similar to the groups immunized nasally with wP or PspA plus wP. These results indicate that a combination of vaccines may be a useful strategy to overcome pneumococcal serotype replacement.

scholarly journals Functional cooperativity mediated by rationally selected combinations of human monoclonal antibodies targeting the henipavirus receptor binding protein

2021 ◽  
Author(s):  
Michael P. Doyle ◽  
Nurgun Kose ◽  
Viktoriya Borisevich ◽  
Elad Binshtein ◽  
Moushimi Amaya ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Receptor Binding ◽  
Binding Protein ◽  
Severe Disease ◽  
Hendra Virus ◽  
Human Monoclonal Antibodies ◽  
Human Mabs ◽  
Viral Attachment ◽  
Antigenic Sites ◽  
Receptor Binding Protein

AbstractHendra virus (HeV) and Nipah virus (NiV), the prototypic members of the Henipavirus (HNV) genus, are emerging, zoonotic paramyxoviruses known to cause severe disease across six mammalian orders, including humans (Eaton et al., 2006). While several research groups have made strides in developing candidate vaccines and therapeutics against henipaviruses, such countermeasures have not been licensed for human use, and significant gaps in knowledge about the human immune response to these viruses exist. To address these gaps, we isolated a large panel of human monoclonal antibodies (mAbs) from the B cells of an individual with prior occupation-related exposure to the equine HeV vaccine (Equivac® HeV). Competition-binding and hydrogen-deuterium exchange mass spectrometry (HDX-MS) studies identified at least six distinct antigenic sites on the HeV/NiV receptor binding protein (RBP) that are recognized by human mAbs. Antibodies recognizing multiple antigenic sites potently neutralized NiV and/or HeV isolates in vitro. The most potent class of cross-reactive antibodies achieved neutralization by blocking viral attachment to the host cell receptors ephrin-B2 and ephrin-B3. Antibodies from this class mimic receptor binding by inducing a receptor-bound conformation to the HeV-RBP protein tetramer, exposing an epitope that appears to lie hidden in the interface between protomers within the HeV-RBP tetramer. Antibodies that recognize this cryptic epitope potently neutralized HeV and NiV. Flow cytometric studies using cell-surface-displayed HeV-RBP protein showed that cross-reactive, neutralizing mAbs from each of these classes cooperate for binding. In a highly stringent hamster model of NiVB infection, antibodies from both classes reduced morbidity and mortality and achieved synergistic protection in combination and provided therapeutic benefit when combined into two bispecific platforms. These studies identified multiple candidate mAbs that might be suitable for use in a cocktail therapeutic approach to achieve synergistic antiviral potency and reduce the risk of virus escape during treatment.

scholarly journals Comparison of Two Strategies to Generate Antigen-Specific Human Monoclonal Antibodies: Which Method to Choose for Which Purpose?

2021 ◽  
Vol 12 ◽  
Author(s):  
Anna M. Ehlers ◽  
Constance F. den Hartog Jager ◽  
Tineke Kardol-Hoefnagel ◽  
Miriam M.D. Katsburg ◽  
André C. Knulst ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Heterologous Expression ◽  
Single Cell ◽  
Antigen Binding ◽  
Human Monoclonal Antibodies ◽  
Human Mabs ◽  
Single Cell Sequencing ◽  
2S Albumins ◽  
Functional Features ◽  
Set Up

Human monoclonal antibodies (mAbs) are valuable tools to link genetic information with functional features and to provide a platform for conformational epitope mapping. Additionally, combined data on genetic and functional features provide a valuable mosaic for systems immunology approaches. Strategies to generate human mAbs from peripheral blood have been described and used in several studies including single cell sequencing of antigen-binding B cells and the establishment of antigen-specific monoclonal Epstein-Barr Virus (EBV) immortalized lymphoblastoid cell lines (LCLs). However, direct comparisons of these two strategies are scarce. Hence, we sought to set up these two strategies in our laboratory using peanut 2S albumins (allergens) and the autoantigen anti-Rho guanosine diphosphate dissociation inhibitor 2 (RhoGDI2, alternatively ‘ARHGDIB’) as antigen targets to directly compare these strategies regarding costs, time expenditure, recovery, throughput and complexity. Regarding single cell sequencing, up to 50% of corresponding V(D)J gene transcripts were successfully amplified of which 54% were successfully cloned into expression vectors used for heterologous expression. Seventy-five percent of heterologously expressed mAbs showed specific binding to peanut 2S albumins resulting in an overall recovery of 20.3%, which may be increased to around 29% by ordering gene sequences commercially for antibody cloning. In comparison, the establishment of monoclonal EBV-LCLs showed a lower overall recovery of around 17.6%. Heterologous expression of a mAb carrying the same variable region as its native counterpart showed comparable concentration-dependent binding abilities. By directly comparing those two strategies, single cell sequencing allows a broad examination of antigen-binding mAbs in a moderate-throughput manner, while the establishment of monoclonal EBV-LCLs is a powerful tool to select a small number of highly reactive mAbs restricted to certain B cell subpopulations. Overall, both strategies, initially set-up for peanut 2S albumins, are suitable to obtain human mAbs and they are easily transferrable to other target antigens as shown for ARHGDIB.

scholarly journals Genetic Immunization with the Region Encoding the α-Helical Domain of PspA Elicits Protective Immunity againstStreptococcus pneumoniae

2001 ◽  
Vol 69 (9) ◽  
pp. 5456-5463 ◽  
Author(s):  
Joseph R. Bosarge ◽  
James M. Watt ◽  
D. Olga McDaniel ◽  
Edwin Swiatlo ◽  
Larry S. McDaniel
Keyword(s):  
Protein A ◽  
Survival Rates ◽  
Serum Antibody ◽  
Pneumococcal Infection ◽  
Surface Protein ◽  
Eukaryotic Expression ◽  
Dependent Manner ◽  
Genetic Immunization ◽  
Helical Domain ◽  
Dose Dependent

ABSTRACT Pneumococcal surface protein A (PspA) is a pneumococcal virulence factor capable of eliciting protection against pneumococcal infection in mice. Previous studies have demonstrated that the protection is antibody mediated. Here we examined the ability ofpspA to elicit a protective immune response following genetic immunization of mice. Mice were immunized by intramuscular injections with a eukaryotic expression vector encoding the α-helical domain of PspA/Rx1. Immunization induced a PspA-specific serum antibody response, and immunized mice survived pneumococcal challenge. Survival and antibody responses occurred in a dose-dependent manner, the highest survival rates being seen with doses of 10 μg or greater. The ability of genetic immunization to elicit cross-protection was demonstrated by the survival of immunized mice challenged with pneumococcal strains differing in capsule and PspA types. Also, immunized mice were protected from intravenous and intratracheal challenges with pneumococci. Similar to the results seen with immunization with PspA, the survival of mice genetically immunized with pspA was antibody mediated. There was no decline in the level of protection 7 months after immunization. These results support the use of genetic immunization to elicit protective immune responses against extracellular pathogens.

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