An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus

Traggiai, Elisabetta; Becker, Stephan; Subbarao, Kanta; Kolesnikova, Larissa; Uematsu, Yasushi; Gismondo, Maria Rita; Murphy, Brian R.; Rappuoli, Rino; Lanzavecchia, Antonio

doi:10.1038/nm1080

Cited by 674 publications

(700 citation statements)

References 30 publications

Supporting

Mentioning

678

Contrasting

Unclassified

Order By: Relevance

“…PBMCs were thawed, diluted in 10 ml of IMDM (Mediatech), and centrifuged through 1 ml of heat-inactivated FBS (Invitrogen) to remove cellular debris. B cells were enriched using the Miltenyi B cell isolation kit II (Miltenyi Biotec) and then transformed as previously described (33). Titered, direct-pelleted EBV (Advanced Biotechnologies) was added to isolated B cells at 1/100 final dilution for 2 h at 37°C; then the cells were washed three times and plated.…”

Section: Memory B Cell Isolation Ebv Transformation and Culturementioning

confidence: 99%

“…B cells were isolated and transformed using EBV to stimulate proliferation and secretion (33). From our screen of Ͼ20 million primary B cells, 12 clones were recovered that met stringent specificity criteria described below.…”

Section: Single B Cell Phenotypingmentioning

confidence: 99%

“…Fully human mAbs are increasingly recognized as superior therapeutic candidates over mouse-derived products (33,34), because the mAbs have been naturally selected and therefore have high affinity, are stable, and lack off-target reactivity against human Ags. Compared with hyperimmunized mice, however, the frequency of favorable clones in the human repertoire is typically too low for practical recovery.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Potent High-Affinity Antibodies for Treatment and Prophylaxis of Respiratory Syncytial Virus Derived from B Cells of Infected Patients

Collarini

Lee

Foord

et al. 2009

The Journal of Immunology

111

View full text Add to dashboard Cite

Native human Abs represent attractive drug candidates; however, the low frequency of B cells expressing high-quality Abs has posed a barrier to discovery. Using a novel single-cell phenotyping technology, we have overcome this barrier to discover human Abs targeting the conserved but poorly immunogenic central motif of respiratory syncytial virus (RSV) G protein. For the entire cohort of 24 subjects with recent RSV infection, B cells producing Abs meeting these stringent specificity criteria were rare, <10 per million. Several of the newly cloned Abs bind to the RSV G protein central conserved motif with very high affinity (Kd 1–24 pM). Two of the Abs were characterized in detail and compared with palivizumab, a humanized mAb against the RSV F protein. Relative to palivizumab, the anti-G Abs showed improved viral neutralization potency in vitro and enhanced reduction of infectious virus in a prophylaxis mouse model. Furthermore, in a mouse model for postinfection treatment, both anti-G Abs were significantly more effective than palivizumab at reducing viral load. The combination of activity in mouse models for both prophylaxis and treatment makes these high-affinity human-derived Abs promising candidates for human clinical testing.

show abstract

Section: Memory B Cell Isolation Ebv Transformation and Culturementioning

confidence: 99%

Section: Single B Cell Phenotypingmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Potent High-Affinity Antibodies for Treatment and Prophylaxis of Respiratory Syncytial Virus Derived from B Cells of Infected Patients

Collarini

Lee

Foord

et al. 2009

The Journal of Immunology

111

View full text Add to dashboard Cite

show abstract

“…To determine whether Abs that enhance civet S entry can be detected in humans, we examined neutralizing mAbs derived from Epstein-Barr virus transformation of memory B cells from a recovered patient (15).…”

Section: Inhibition Of Entry By Human Mabs To Smentioning

confidence: 99%

Evasion of antibody neutralization in emerging severe acute respiratory syndrome coronaviruses

Yang

Werner

Kong

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

251

284

View full text Add to dashboard Cite

Molecular characterization of the severe acute respiratory syndrome coronavirus has revealed genetic diversity among isolates. The spike (S) glycoprotein, the major target for vaccine and immune therapy, shows up to 17 substitutions in its 1,255-aa sequence; however, the biologic significance of these changes is unknown. Here, the functional effects of S mutations have been determined by analyzing their affinity for a viral receptor, human angiotensin-converting enzyme 2 (hACE-2), and their sensitivity to Ab neutralization with viral pseudotypes. Although minor differences among eight strains transmitted during human outbreaks in early 2003 were found, substantial functional changes were detected in S derived from a case in late 2003 from Guangdong province [S(GD03T0013)] and from two palm civets, S(SZ3) and S(SZ16). S(GD03T0013) depended less on the hACE-2 receptor and was markedly resistant to Ab inhibition. Unexpectedly, Abs that neutralized most human S glycoproteins enhanced entry mediated by the civet virus S glycoproteins. The mechanism of enhancement involved the interaction of Abs with conformational epitopes in the hACE-2-binding domain. Finally, improved immunogens and mAbs that minimize this complication have been defined. These data show that the entry of severe acute respiratory syndrome coronaviruses can be enhanced by Abs, and they underscore the need to address the evolving diversity of this newly emerged virus for vaccines and immune therapies.angiotensin-converting enzyme 2 ͉ enhancement ͉ immunoglobulin G ͉ pseudovirus

show abstract

“…To approach this issue, we are in the process of establishing a method for making human monoclonal antibodies based on an improved method for EBV immortalisation of human memory B cells (Traggiai 2004). The advantage of EBV-immortalised B cells is to have a continuous and direct source of GAD65-specific antibody secreting and antigen presenting cells.…”

Section: Future Perspectives Human Monoclonal Gad65 Igg Antibodies Frmentioning

confidence: 99%

MS and clinically isolated syndromes: Shared specificity but diverging clonal patterns of virus‐specific IgG antibodies produced in vivo and by CSF B cells in vitro

Skorstad

Vandvik

Vartdal

et al. 2009

Euro J of Neurology

View full text Add to dashboard Cite

Background: Intrathecal synthesis of oligoclonal IgG antibodies against measles virus (MeV), varicella zoster virus (VZV) and herpes simplex virus type‐1 (HSV‐1) is a characteristic feature multiple sclerosis (MS). Methods: We have used isoelectric focusing‐immunoblot to define the clonal patterns of IgG and of IgG antibodies to MeV, VZV and HSV‐1 in supernatants of in vitro cultures of peripheral blood lymphocytes (PBL) and cerebrospinal fluid (CSF) cells and in sera and CSF from three patients with MS and three patients with clinically isolated syndromes (CIS) suspective of demyelinating disease. Results: In vitro synthesis of IgG by PBL was not detected in any patient. In contrast, in vitro synthesis by CSF cells of oligoclonal IgG and oligoclonal IgG antibodies to one or two of the three viruses tested was observed in all six patients. The clonal patterns of the in vitro synthesized IgG and virus specific IgG differed to varying extent from those synthesized intrathecally in vivo. However, in each patient, the in vitro and in vivo intrathecally produced antibodies displayed specificity for the same viruses. The addition of B cell activating factor (BAFF) had no effect on the amounts or clonal patterns of either total IgG or virus‐specific IgG produced by CSF cells in vitro. Conclusion: Virus specific B cells capable of spontaneous IgG synthesis are clonally expanded in the CSF of patients with MS. The B‐cell repertoire in CSF samples is only partially representative of the intrathecal B‐cell repertoire.

show abstract

An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus

Cited by 674 publications

References 30 publications

Potent High-Affinity Antibodies for Treatment and Prophylaxis of Respiratory Syncytial Virus Derived from B Cells of Infected Patients

Potent High-Affinity Antibodies for Treatment and Prophylaxis of Respiratory Syncytial Virus Derived from B Cells of Infected Patients

Evasion of antibody neutralization in emerging severe acute respiratory syndrome coronaviruses

MS and clinically isolated syndromes: Shared specificity but diverging clonal patterns of virus‐specific IgG antibodies produced in vivo and by CSF B cells in vitro

Contact Info

Product

Resources

About