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Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones.

Publication ,  Journal Article
Holl, TM; Haynes, BF; Kelsoe, G
Published in: J Immunol Methods
March 31, 2010

We describe and characterize a stromal cell independent culture system that efficiently supports pro-B cell to IgM+ B cell development with near normal levels of IgH and Igkappa diversity. Pro-B cells present in non-adherent bone marrow cells proliferate in the presence of IL-7 and subsequent to the removal of IL-7 and addition of BAFF, differentiate normally into IgM+ B cells. B cell development in vitro closely follows the patterns of development in vivo with culture-derived (CD) B cells demonstrating characteristic patterns of surface antigen expression and gene activation. IgM+ CD B cells respond to TLR stimulation by proliferation and differentiation into antibody-secreting cells. Self-reactive IgM+ B cell development is blocked in 3H9 IgH knockin mice; however, cultures of 3H9 IgH knockin pro-B cells yields high frequencies of "forbidden", autoreactive IgM+ B cells. Furthermore, serum IgG autoantibody exceeded that present in autoimmune, C4(-/-) animals following the reconstitution of RAG1(-/-) mice with IgM+ CD cells derived from BL/6 mice.

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Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

March 31, 2010

Volume

354

Issue

1-2

Start / End Page

53 / 67

Location

Netherlands

Related Subject Headings

  • Toll-Like Receptors
  • Time Factors
  • Stromal Cells
  • Precursor Cells, B-Lymphoid
  • Phenotype
  • Mice, Transgenic
  • Mice, Knockout
  • Mice, Inbred C57BL
  • Mice
  • Interleukin-7
 

Citation

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Holl, T. M., Haynes, B. F., & Kelsoe, G. (2010). Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones. J Immunol Methods, 354(1–2), 53–67. https://doi.org/10.1016/j.jim.2010.01.007
Holl, T Matt, Barton F. Haynes, and Garnett Kelsoe. “Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones.J Immunol Methods 354, no. 1–2 (March 31, 2010): 53–67. https://doi.org/10.1016/j.jim.2010.01.007.
Holl TM, Haynes BF, Kelsoe G. Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones. J Immunol Methods. 2010 Mar 31;354(1–2):53–67.
Holl, T. Matt, et al. “Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones.J Immunol Methods, vol. 354, no. 1–2, Mar. 2010, pp. 53–67. Pubmed, doi:10.1016/j.jim.2010.01.007.
Holl TM, Haynes BF, Kelsoe G. Stromal cell independent B cell development in vitro: generation and recovery of autoreactive clones. J Immunol Methods. 2010 Mar 31;354(1–2):53–67.
Journal cover image

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

March 31, 2010

Volume

354

Issue

1-2

Start / End Page

53 / 67

Location

Netherlands

Related Subject Headings

  • Toll-Like Receptors
  • Time Factors
  • Stromal Cells
  • Precursor Cells, B-Lymphoid
  • Phenotype
  • Mice, Transgenic
  • Mice, Knockout
  • Mice, Inbred C57BL
  • Mice
  • Interleukin-7