Advanced

In vitro immunization of naive human B cells yields high affinity immunuglobulin G antibodies as illustrated by phage display

Dueñas, M.; Chin, L. T.; Malmborg, A. C.; Casalvilla, R.; Ohlin, M. LU and Borrebaeck, C. A K LU (1996) In Immunology 89(1). p.1-7
Abstract

In vitro antibody responses to a synthetic immunogen, consisting of both a B cell [V3 loop of gp120 from human immunodeficiency virus type 1 (HIV-1)] and a T-helper epitope (15 amino acids of tetanus toxoid) was studied. The in vitro activation was performed by primary and secondary in vitro immunizations, using lymphocytes obtained from uninfected, seronegative donors. Analysis of the in vitro immune response demonstrated an antigen-specific isotype switch, which was dependent on the presence of antigen-specific T-helper cells, CD40 ligation and antigen. Antibody libraries were constructed from cells derived directly from the naive donors, or from primary or secondary in vitro immunized B cells. Five libraries were displayed on... (More)

In vitro antibody responses to a synthetic immunogen, consisting of both a B cell [V3 loop of gp120 from human immunodeficiency virus type 1 (HIV-1)] and a T-helper epitope (15 amino acids of tetanus toxoid) was studied. The in vitro activation was performed by primary and secondary in vitro immunizations, using lymphocytes obtained from uninfected, seronegative donors. Analysis of the in vitro immune response demonstrated an antigen-specific isotype switch, which was dependent on the presence of antigen-specific T-helper cells, CD40 ligation and antigen. Antibody libraries were constructed from cells derived directly from the naive donors, or from primary or secondary in vitro immunized B cells. Five libraries were displayed on filamentous phage and selected for anti-V3-specific Fab fragments, using a selection approach that linked recognition and phage replication. A panel of 19 recombinant antigen-specific Fab, representing different phases of the humoral in vitro immune response, were sequenced, expressed and analysed using a biosensor. Recombinant Fab fragments derived from cultures on day 12 exhibited an increase in affinity of close to two orders of magnitude compared to those obtained from cells primary immunized for 7 days. This study provides the first evidence that an antigen-specific in vitro immune response can yield high-affinity immunoglobuling G antibodies.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
in
Immunology
volume
89
issue
1
pages
7 pages
publisher
Wiley-Blackwell
external identifiers
  • Scopus:0029843718
ISSN
0019-2805
language
English
LU publication?
yes
id
e79d32b2-96e5-480e-a5dd-460e9e866a33
date added to LUP
2016-04-19 14:10:29
date last changed
2016-04-22 13:20:44
@misc{e79d32b2-96e5-480e-a5dd-460e9e866a33,
  abstract     = {<p>In vitro antibody responses to a synthetic immunogen, consisting of both a B cell [V3 loop of gp120 from human immunodeficiency virus type 1 (HIV-1)] and a T-helper epitope (15 amino acids of tetanus toxoid) was studied. The in vitro activation was performed by primary and secondary in vitro immunizations, using lymphocytes obtained from uninfected, seronegative donors. Analysis of the in vitro immune response demonstrated an antigen-specific isotype switch, which was dependent on the presence of antigen-specific T-helper cells, CD40 ligation and antigen. Antibody libraries were constructed from cells derived directly from the naive donors, or from primary or secondary in vitro immunized B cells. Five libraries were displayed on filamentous phage and selected for anti-V3-specific Fab fragments, using a selection approach that linked recognition and phage replication. A panel of 19 recombinant antigen-specific Fab, representing different phases of the humoral in vitro immune response, were sequenced, expressed and analysed using a biosensor. Recombinant Fab fragments derived from cultures on day 12 exhibited an increase in affinity of close to two orders of magnitude compared to those obtained from cells primary immunized for 7 days. This study provides the first evidence that an antigen-specific in vitro immune response can yield high-affinity immunoglobuling G antibodies.</p>},
  author       = {Dueñas, M. and Chin, L. T. and Malmborg, A. C. and Casalvilla, R. and Ohlin, M. and Borrebaeck, C. A K},
  issn         = {0019-2805},
  language     = {eng},
  number       = {1},
  pages        = {1--7},
  publisher    = {ARRAY(0x9240738)},
  series       = {Immunology},
  title        = {In vitro immunization of naive human B cells yields high affinity immunuglobulin G antibodies as illustrated by phage display},
  volume       = {89},
  year         = {1996},
}