Neutralizing and non-neutralizing monoclonal antibodies against dengue virus E protein derived from a naturally infected patient.
BACKGROUND
Antibodies produced in response to infection with any of the four serotypes of dengue virus generally provide homotypic immunity. However, prior infection or circulating maternal antibodies can also mediate a non-protective antibody response that can enhance the course of disease in a subsequent heterotypic infection. Naturally occurring human monoclonal antibodies can help us understand the protective and pathogenic roles of the humoral immune system in dengue virus infection.


RESULTS
Epstein-Barr Virus (EBV) transformation of B cells isolated from the peripheral blood of a human subject with previous dengue infection was performed. B cell cultures were screened by ELISA for antibodies to dengue (DENV) envelope (E) protein. ELISA positive cultures were cloned by limiting dilution. Three IgG1 human monoclonal antibodies (HMAbs) were purified and their binding specificity to E protein was verified by ELISA and biolayer interferometry. Neutralization and enhancement assays were conducted in epithelial and macrophage-like cell lines, respectively. All three HMAbs bound to E from at least two of the four DENV serotypes, one of the HMAbs was neutralizing, and all were able to enhance DENV infection.


CONCLUSIONS
HMAbs against DENV can be successfully generated by EBV transformation of B cells from patients at least two years after naturally acquired DENV infections. These antibodies show different patterns of cross-reactivity, neutralizing, and enhancement activity.
Section of Pediatric Infectious Disease, Department of Pediatrics, Tulane University School of Medicine, New Orleans, LA, USA. jschieff@tulane.edu
John S
JS
Schieffelin
Joshua M
JM
Costin
Cindo O
CO
Nicholson
Nicole M
NM
Orgeron
Krystal A
KA
Fontaine
Sharon
S
Isern
Scott F
SF
Michael
James E
JE
Robinson
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D057134:Antibodies, Neutralizing / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D019067:Antibody-Dependent Enhancement; D001402:B-Lymphocytes / Q000276:immunology / Q000821:virology; D002460:Cell Line; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D003716:Dengue Virus / Q000276:immunology*; D004847:Epithelial Cells / Q000821:virology; D004854:Herpesvirus 4, Human / Q000254:growth & development; D006801:Humans; D008264:Macrophages / Q000821:virology; D011485:Protein Binding; D014481:United States; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D014759:Viral Envelope Proteins