Immunization with antigen-pulsed dendritic cells significantly improves the immune response to weak self-antigens

Pablo Vargas, Claudio Cortés, Leonardo Vargas, Mario Rosemblatt, María Rosa Bono

Resultado de la investigación: Article

5 Citas (Scopus)

Resumen

Dendritic cells (DCs) are the only professional antigen-presenting cells endowed with the ability to stimulate naïve T cells and initiate a primary immune response. For this reason, DC-based immunization has been shown to be highly effective in eliciting CTL responses to viruses and tumor-associated antigens. Here we report on the use of DC immunization to enhance the B cell-mediated humoral immune response to highly conserved proteins and the application of this approach to the generation of monoclonal antibodies (mAbs) against these proteins. To illustrate the technique we describe the production of mAbs to class II transactivator (CIITA), the major histocompatibility complex (MHC) CIITA, a difficult immunogen owing to its high degree of identity among species. We show that mice immunized with a combination of an intravenous injection of DCs pulsed with recombinant fragments of CIITA followed by intraperitoneal injection of the antigen in incomplete Freund's adjuvant induced a detectable antibody response against CIITA, while sera from mice immunized using the traditional method (i.e. intraperitoneal immunization with 50 μg of protein in complete Freund's adjuvant) gave an almost undetectable response. Furthermore, a total of four fusion experiments demonstrate that immunization with Ag-pulsed DCs is necessary for the efficient generation of hybridomas and a good yield of mAbs specific for the recombinant and the native endogenous CIITA protein. Conversely, four independent fusions carried out with splenocytes from mice immunized using the traditional method failed to produce anti-CIITA hybridomas. We propose that immunization with antigen-loaded DCs should be the method of preference when attempting to raise mAbs against weak self-immunogens.

Idioma originalEnglish
Páginas (desde-hasta)29-36
Número de páginas8
PublicaciónImmunobiology
Volumen211
N.º1-2
DOI
EstadoPublished - 22 feb 2006

Huella dactilar

Autoantigens
Dendritic Cells
Immunization
Antigens
Monoclonal Antibodies
Hybridomas
Proteins
Freund's Adjuvant
Immunoglobulin Isotypes
Neoplasm Antigens
Antigen-Presenting Cells
Humoral Immunity
Major Histocompatibility Complex
Intraperitoneal Injections
Intravenous Injections
Antibody Formation
MHC class II transactivator protein
B-Lymphocytes
Viruses
T-Lymphocytes

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Hematology

Citar esto

Vargas, Pablo ; Cortés, Claudio ; Vargas, Leonardo ; Rosemblatt, Mario ; Bono, María Rosa. / Immunization with antigen-pulsed dendritic cells significantly improves the immune response to weak self-antigens. En: Immunobiology. 2006 ; Vol. 211, N.º 1-2. pp. 29-36.
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title = "Immunization with antigen-pulsed dendritic cells significantly improves the immune response to weak self-antigens",
abstract = "Dendritic cells (DCs) are the only professional antigen-presenting cells endowed with the ability to stimulate na{\"i}ve T cells and initiate a primary immune response. For this reason, DC-based immunization has been shown to be highly effective in eliciting CTL responses to viruses and tumor-associated antigens. Here we report on the use of DC immunization to enhance the B cell-mediated humoral immune response to highly conserved proteins and the application of this approach to the generation of monoclonal antibodies (mAbs) against these proteins. To illustrate the technique we describe the production of mAbs to class II transactivator (CIITA), the major histocompatibility complex (MHC) CIITA, a difficult immunogen owing to its high degree of identity among species. We show that mice immunized with a combination of an intravenous injection of DCs pulsed with recombinant fragments of CIITA followed by intraperitoneal injection of the antigen in incomplete Freund's adjuvant induced a detectable antibody response against CIITA, while sera from mice immunized using the traditional method (i.e. intraperitoneal immunization with 50 μg of protein in complete Freund's adjuvant) gave an almost undetectable response. Furthermore, a total of four fusion experiments demonstrate that immunization with Ag-pulsed DCs is necessary for the efficient generation of hybridomas and a good yield of mAbs specific for the recombinant and the native endogenous CIITA protein. Conversely, four independent fusions carried out with splenocytes from mice immunized using the traditional method failed to produce anti-CIITA hybridomas. We propose that immunization with antigen-loaded DCs should be the method of preference when attempting to raise mAbs against weak self-immunogens.",
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Immunization with antigen-pulsed dendritic cells significantly improves the immune response to weak self-antigens. / Vargas, Pablo; Cortés, Claudio; Vargas, Leonardo; Rosemblatt, Mario; Bono, María Rosa.

En: Immunobiology, Vol. 211, N.º 1-2, 22.02.2006, p. 29-36.

Resultado de la investigación: Article

TY - JOUR

T1 - Immunization with antigen-pulsed dendritic cells significantly improves the immune response to weak self-antigens

AU - Vargas, Pablo

AU - Cortés, Claudio

AU - Vargas, Leonardo

AU - Rosemblatt, Mario

AU - Bono, María Rosa

PY - 2006/2/22

Y1 - 2006/2/22

N2 - Dendritic cells (DCs) are the only professional antigen-presenting cells endowed with the ability to stimulate naïve T cells and initiate a primary immune response. For this reason, DC-based immunization has been shown to be highly effective in eliciting CTL responses to viruses and tumor-associated antigens. Here we report on the use of DC immunization to enhance the B cell-mediated humoral immune response to highly conserved proteins and the application of this approach to the generation of monoclonal antibodies (mAbs) against these proteins. To illustrate the technique we describe the production of mAbs to class II transactivator (CIITA), the major histocompatibility complex (MHC) CIITA, a difficult immunogen owing to its high degree of identity among species. We show that mice immunized with a combination of an intravenous injection of DCs pulsed with recombinant fragments of CIITA followed by intraperitoneal injection of the antigen in incomplete Freund's adjuvant induced a detectable antibody response against CIITA, while sera from mice immunized using the traditional method (i.e. intraperitoneal immunization with 50 μg of protein in complete Freund's adjuvant) gave an almost undetectable response. Furthermore, a total of four fusion experiments demonstrate that immunization with Ag-pulsed DCs is necessary for the efficient generation of hybridomas and a good yield of mAbs specific for the recombinant and the native endogenous CIITA protein. Conversely, four independent fusions carried out with splenocytes from mice immunized using the traditional method failed to produce anti-CIITA hybridomas. We propose that immunization with antigen-loaded DCs should be the method of preference when attempting to raise mAbs against weak self-immunogens.

AB - Dendritic cells (DCs) are the only professional antigen-presenting cells endowed with the ability to stimulate naïve T cells and initiate a primary immune response. For this reason, DC-based immunization has been shown to be highly effective in eliciting CTL responses to viruses and tumor-associated antigens. Here we report on the use of DC immunization to enhance the B cell-mediated humoral immune response to highly conserved proteins and the application of this approach to the generation of monoclonal antibodies (mAbs) against these proteins. To illustrate the technique we describe the production of mAbs to class II transactivator (CIITA), the major histocompatibility complex (MHC) CIITA, a difficult immunogen owing to its high degree of identity among species. We show that mice immunized with a combination of an intravenous injection of DCs pulsed with recombinant fragments of CIITA followed by intraperitoneal injection of the antigen in incomplete Freund's adjuvant induced a detectable antibody response against CIITA, while sera from mice immunized using the traditional method (i.e. intraperitoneal immunization with 50 μg of protein in complete Freund's adjuvant) gave an almost undetectable response. Furthermore, a total of four fusion experiments demonstrate that immunization with Ag-pulsed DCs is necessary for the efficient generation of hybridomas and a good yield of mAbs specific for the recombinant and the native endogenous CIITA protein. Conversely, four independent fusions carried out with splenocytes from mice immunized using the traditional method failed to produce anti-CIITA hybridomas. We propose that immunization with antigen-loaded DCs should be the method of preference when attempting to raise mAbs against weak self-immunogens.

KW - Class II transactivator (CIITA)

KW - Dendritic cells

KW - Immune response

KW - Immunization

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M3 - Article

C2 - 16446168

AN - SCOPUS:31444449825

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JF - Immunobiology

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