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Continuous Culture of Mouse Primary B Lymphocytes by Forced Expression of Bach2.

Publication ,  Journal Article
Finney, J; Kelsoe, G
Published in: J Immunol
September 1, 2021

Stable, long-term culture of primary B lymphocytes has many potential scientific and medical applications, but remains an elusive feat. A major obstacle to long-term culture is that in vitro mitogens quickly drive B cells to differentiate into short-lived plasma cells (PCs). PC differentiation is governed by opposing teams of transcription factors: Pax5, Bach2, and Bcl6 suppress PC commitment, whereas IFN regulatory factor 4 and Blimp1 promote it. To determine whether transcriptional programming could prolong B cell culture by blocking PC commitment, we generated mouse primary B cells harboring gain- or loss-of-function in the key transcription factors, continuously stimulated these cells with CD154 and IL-21, and determined growth potential and phenotypes in vitro. We found that transgenic expression of Bach2 prohibits PC commitment and endows B cells with extraordinary growth potential in response to external proliferation and survival cues. Long-term Bach2-transgenic B cell lines have genetically stable BCRs [i.e., do not acquire V(D)J mutations], express high levels of MHC class II and molecules for costimulation of T cells, and transduce intracellular signals when incubated with BCR ligands. Silencing the Bach2 transgene in an established transgenic cell line causes the cells to secrete large quantities of Ig. This system has potential applications in mAb production, BCR signaling studies, Ag presentation to T cells, and ex vivo clonal expansion for adoptive cell transfer. Additionally, our results provide insight into molecular control over activated B cell fate and suggest that forced Bach2 expression in vivo may augment germinal center B cell or memory B cell differentiation at the expense of PC commitment.

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Published In

J Immunol

DOI

EISSN

1550-6606

Publication Date

September 1, 2021

Volume

207

Issue

5

Start / End Page

1478 / 1492

Location

United States

Related Subject Headings

  • Receptors, Antigen, B-Cell
  • Proto-Oncogene Proteins c-bcl-6
  • Primary Cell Culture
  • PAX5 Transcription Factor
  • Mutation
  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice
  • Immunology
  • Immunologic Memory
 

Citation

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Finney, J., & Kelsoe, G. (2021). Continuous Culture of Mouse Primary B Lymphocytes by Forced Expression of Bach2. J Immunol, 207(5), 1478–1492. https://doi.org/10.4049/jimmunol.2100172
Finney, Joel, and Garnett Kelsoe. “Continuous Culture of Mouse Primary B Lymphocytes by Forced Expression of Bach2.J Immunol 207, no. 5 (September 1, 2021): 1478–92. https://doi.org/10.4049/jimmunol.2100172.
Finney J, Kelsoe G. Continuous Culture of Mouse Primary B Lymphocytes by Forced Expression of Bach2. J Immunol. 2021 Sep 1;207(5):1478–92.
Finney, Joel, and Garnett Kelsoe. “Continuous Culture of Mouse Primary B Lymphocytes by Forced Expression of Bach2.J Immunol, vol. 207, no. 5, Sept. 2021, pp. 1478–92. Pubmed, doi:10.4049/jimmunol.2100172.
Finney J, Kelsoe G. Continuous Culture of Mouse Primary B Lymphocytes by Forced Expression of Bach2. J Immunol. 2021 Sep 1;207(5):1478–1492.

Published In

J Immunol

DOI

EISSN

1550-6606

Publication Date

September 1, 2021

Volume

207

Issue

5

Start / End Page

1478 / 1492

Location

United States

Related Subject Headings

  • Receptors, Antigen, B-Cell
  • Proto-Oncogene Proteins c-bcl-6
  • Primary Cell Culture
  • PAX5 Transcription Factor
  • Mutation
  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice
  • Immunology
  • Immunologic Memory