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Joining chain-expressing and -nonexpressing B cell populations in the mouse.

Erlandsson L, Akerblad P, Vingsbo-Lundberg C, Kallberg E, Lycke N, Leanderson T - J. Exp. Med. (2001)

Bottom Line: Serum immunoglobulin (Ig)M and serum IgG were reduced six to eightfold, while serum IgA was elevated 14-fold in these mice.Analysis of single B cells stimulated by lipopolysaccharide in vitro using reverse transcription-polymerase chain reaction showed that J chain-nonexpressing B cells could be detected that had a secretory phenotype as determined by an abundance of transcript for secretory IgM.Finally, limiting dilution analysis of peripheral B cells showed that J chain expression was a clonal property already established in naive, peripheral B lymphocytes.

View Article: PubMed Central - PubMed

Affiliation: Immunology Section, Department of Cell and Molecular Biology, Lund University, S-221 84 Lund, Sweden.

ABSTRACT
The diphtheria toxin A chain (DTA) was gene targeted into the Joining chain (J chain) locus to create a mouse strain selecting against J chain-expressing cells, JDTA mice. Serum immunoglobulin (Ig)M and serum IgG were reduced six to eightfold, while serum IgA was elevated 14-fold in these mice. JDTA mice were immune competent although the serum Ig response compared with wild-type mice was reduced sixfold at day 14 but only fourfold at day 45 after immunization. Exchanging the DTA gene with a cDNA for c-myc resulted in mice with a distinct phenotype with increased Ig production and enhanced humoral immune responses. Analysis of single B cells stimulated by lipopolysaccharide in vitro using reverse transcription-polymerase chain reaction showed that J chain-nonexpressing B cells could be detected that had a secretory phenotype as determined by an abundance of transcript for secretory IgM. Finally, limiting dilution analysis of peripheral B cells showed that J chain expression was a clonal property already established in naive, peripheral B lymphocytes.

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RT-PCR analysis of J chain expression in mouse cell lines representing various stages of B cell differentiation. RT-PCR analysis was performed as described previously. The 70Z/3 cell line was analyzed both unstimulated but also after 24 and 40 h stimulation with LPS which triggers its differentiation to a surface Ig+ phenotype, as indicated.
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Figure 7: RT-PCR analysis of J chain expression in mouse cell lines representing various stages of B cell differentiation. RT-PCR analysis was performed as described previously. The 70Z/3 cell line was analyzed both unstimulated but also after 24 and 40 h stimulation with LPS which triggers its differentiation to a surface Ig+ phenotype, as indicated.

Mentions: Our data indicates that once B cells are committed to secrete Ig at a high rate, they either express J chain or not. However, it cannot be excluded that J chain expression is a clonal property decided before the B lymphocyte reaches the terminal developmental stage of high Ig secretion. Evidence in favor of early commitment to Ig secretion can be found in the observation that some human cell lines and normal cells representing early stages of B cell development have been shown to express the J chain locus 32333435. We confirmed this finding using a panel of mouse B cell lines and RT-PCR (Fig. 7). The plasmacytoma J558 and total spleen cells expressed high levels of J chain RNA as expected. However, the B cell lymphomas WEHI231, A20, and K46R also expressed detectable levels of J chain RNA, as did the Abelson transformed pre-B cell lines 18–81 and 230–238. Interestingly, J chain transcripts were undetectable in the pre-B cell line 70Z/3 both before and after stimulation with LPS.


Joining chain-expressing and -nonexpressing B cell populations in the mouse.

Erlandsson L, Akerblad P, Vingsbo-Lundberg C, Kallberg E, Lycke N, Leanderson T - J. Exp. Med. (2001)

RT-PCR analysis of J chain expression in mouse cell lines representing various stages of B cell differentiation. RT-PCR analysis was performed as described previously. The 70Z/3 cell line was analyzed both unstimulated but also after 24 and 40 h stimulation with LPS which triggers its differentiation to a surface Ig+ phenotype, as indicated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195946&req=5

Figure 7: RT-PCR analysis of J chain expression in mouse cell lines representing various stages of B cell differentiation. RT-PCR analysis was performed as described previously. The 70Z/3 cell line was analyzed both unstimulated but also after 24 and 40 h stimulation with LPS which triggers its differentiation to a surface Ig+ phenotype, as indicated.
Mentions: Our data indicates that once B cells are committed to secrete Ig at a high rate, they either express J chain or not. However, it cannot be excluded that J chain expression is a clonal property decided before the B lymphocyte reaches the terminal developmental stage of high Ig secretion. Evidence in favor of early commitment to Ig secretion can be found in the observation that some human cell lines and normal cells representing early stages of B cell development have been shown to express the J chain locus 32333435. We confirmed this finding using a panel of mouse B cell lines and RT-PCR (Fig. 7). The plasmacytoma J558 and total spleen cells expressed high levels of J chain RNA as expected. However, the B cell lymphomas WEHI231, A20, and K46R also expressed detectable levels of J chain RNA, as did the Abelson transformed pre-B cell lines 18–81 and 230–238. Interestingly, J chain transcripts were undetectable in the pre-B cell line 70Z/3 both before and after stimulation with LPS.

Bottom Line: Serum immunoglobulin (Ig)M and serum IgG were reduced six to eightfold, while serum IgA was elevated 14-fold in these mice.Analysis of single B cells stimulated by lipopolysaccharide in vitro using reverse transcription-polymerase chain reaction showed that J chain-nonexpressing B cells could be detected that had a secretory phenotype as determined by an abundance of transcript for secretory IgM.Finally, limiting dilution analysis of peripheral B cells showed that J chain expression was a clonal property already established in naive, peripheral B lymphocytes.

View Article: PubMed Central - PubMed

Affiliation: Immunology Section, Department of Cell and Molecular Biology, Lund University, S-221 84 Lund, Sweden.

ABSTRACT
The diphtheria toxin A chain (DTA) was gene targeted into the Joining chain (J chain) locus to create a mouse strain selecting against J chain-expressing cells, JDTA mice. Serum immunoglobulin (Ig)M and serum IgG were reduced six to eightfold, while serum IgA was elevated 14-fold in these mice. JDTA mice were immune competent although the serum Ig response compared with wild-type mice was reduced sixfold at day 14 but only fourfold at day 45 after immunization. Exchanging the DTA gene with a cDNA for c-myc resulted in mice with a distinct phenotype with increased Ig production and enhanced humoral immune responses. Analysis of single B cells stimulated by lipopolysaccharide in vitro using reverse transcription-polymerase chain reaction showed that J chain-nonexpressing B cells could be detected that had a secretory phenotype as determined by an abundance of transcript for secretory IgM. Finally, limiting dilution analysis of peripheral B cells showed that J chain expression was a clonal property already established in naive, peripheral B lymphocytes.

Show MeSH
Related in: MedlinePlus