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TLR2, TLR4 and the MyD88 signaling are crucial for the in vivo generation and the longevity of long-lived antibody-secreting cells.

Komegae EN, Grund LZ, Lopes-Ferreira M, Lima C - PLoS ONE (2013)

Bottom Line: TLR4 (MyD88-independent) modulated the emigration from the spleen of Bmem as well as ASC B220(pos).Terminally differentiated ASC B220(neg) required the cooperation of both signals through TLR2/TLR4 via MyD88 for longevity in peritoneum, whereas Bmem required only TLR2/MyD88 to stay in spleen, and ASC B220(pos) rested in peritoneum dependent on TLR4 signaling.Our data sustain that earlier events on memory B cells differentiation induced in secondary immune response against Natterins, after secondary lymph organs influx and egress, may be the key to determining peripheral localization of innate-like B cells and memory B cells as ASC B220(pos) and ASC B220(neg).

View Article: PubMed Central - PubMed

Affiliation: Immunoregulation Unit, Special Laboratory of Applied Toxinology, Butantan Institute and Department of Immunology, University of São Paulo, São Paulo, Brazil.

ABSTRACT
This study was undertaken to gain better insights into the role of TLRs and MyD88 in the development and differentiation of memory B cells, especially of ASC, during the Th2 polarized memory response induced by Natterins. Our in vivo findings demonstrated that the anaphylactic IgG1 production is dependent on TLR2 and MyD88 signaling, and that TLR4 acts as adjuvant accelerating the synthesis of high affinity-IgE. Also, TLR4 (MyD88-independent) modulated the migration of innate-like B cells (B1a and B2) out of the peritoneal cavity, and the emigration from the spleen of B1b and B2 cells. TLR4 (MyD88-independent) modulated the emigration from the spleen of Bmem as well as ASC B220(pos). TLR2 triggered to the egress from the peritoneum of Bmem (MyD88-dependent) and ASC B220(pos) (MyD88-independent). We showed that TLR4 regulates the degree of expansion of Bmem in the peritoneum (MyD88-dependent) and in BM (MyD88-independent) as well as of ASC B220(neg) in the spleen (MyD88-independent). TLR2 regulated the intensity of the expansion of Bmem (MyD88-independent) and ASC B220(pos) (MyD88-dependent) in BM. Finally, TLR4 signals sustained the longevity of ASC B220(pos) (MyD88-independent) and ASC B220(neg) into the peritoneum (MyD88-dependent) and TLR2 MyD88-dependent signaling supported the persistence of B2 cells in BM, Bmem in the spleen and ASC B220(neg) in peritoneum and BM. Terminally differentiated ASC B220(neg) required the cooperation of both signals through TLR2/TLR4 via MyD88 for longevity in peritoneum, whereas Bmem required only TLR2/MyD88 to stay in spleen, and ASC B220(pos) rested in peritoneum dependent on TLR4 signaling. Our data sustain that earlier events on memory B cells differentiation induced in secondary immune response against Natterins, after secondary lymph organs influx and egress, may be the key to determining peripheral localization of innate-like B cells and memory B cells as ASC B220(pos) and ASC B220(neg).

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The persistence of B1b cells in bone marrow is negatively regulated by TLR2/TLR4 and MyD88 signals.A representative dot plot of B1b cells (B220lowCD5neg) analyses is shown. Cells from peritoneum (A, D), spleen (B, E) and bone marrow (C, F) were obtained from TLR4 mutant (left) or from TLR2 KO and MyD88 KO (right) Natterins immunized mice after 48, 74 and 120 days. The bars representative of the absolute numbers of B220lowCD5neg cells were determined from total mononuclear cells by multiparametric flow cytometer using Rat IgG2ak PE-anti-mouse CD5, and Rat IgG2ak PerCP-Cy5-anti-mouse CD45R/B220. *p<0.05 compared to control mice; and #p<0.05 compared to WT mice immunized with Natterins.
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pone-0071185-g004: The persistence of B1b cells in bone marrow is negatively regulated by TLR2/TLR4 and MyD88 signals.A representative dot plot of B1b cells (B220lowCD5neg) analyses is shown. Cells from peritoneum (A, D), spleen (B, E) and bone marrow (C, F) were obtained from TLR4 mutant (left) or from TLR2 KO and MyD88 KO (right) Natterins immunized mice after 48, 74 and 120 days. The bars representative of the absolute numbers of B220lowCD5neg cells were determined from total mononuclear cells by multiparametric flow cytometer using Rat IgG2ak PE-anti-mouse CD5, and Rat IgG2ak PerCP-Cy5-anti-mouse CD45R/B220. *p<0.05 compared to control mice; and #p<0.05 compared to WT mice immunized with Natterins.

Mentions: Regarding B1b cells (B220lowCD5neg) our results showed that Natterins did not induce the expansion of B1b cells into peritoneal cavity (Fig. 4A) or in the BM (Fig. 4C) of TLR4 WT mice (white bars), but in contrast this subpopulation was expanded in the spleen only at 48 d in TLR4 WT mice (Fig. 4B). In TLR4 mutant mice (black bars) Natterins induced an increased number of B1b cells only at 120 d in peritoneal cavity (Fig. 4A), and in entire course of the response in spleen (Fig. 4B), and only in the early phase in BM (Fig. 4C), indicating that TLR4 acts as negative regulator, mainly in the transient expansion of B1b cells in spleen.


TLR2, TLR4 and the MyD88 signaling are crucial for the in vivo generation and the longevity of long-lived antibody-secreting cells.

Komegae EN, Grund LZ, Lopes-Ferreira M, Lima C - PLoS ONE (2013)

The persistence of B1b cells in bone marrow is negatively regulated by TLR2/TLR4 and MyD88 signals.A representative dot plot of B1b cells (B220lowCD5neg) analyses is shown. Cells from peritoneum (A, D), spleen (B, E) and bone marrow (C, F) were obtained from TLR4 mutant (left) or from TLR2 KO and MyD88 KO (right) Natterins immunized mice after 48, 74 and 120 days. The bars representative of the absolute numbers of B220lowCD5neg cells were determined from total mononuclear cells by multiparametric flow cytometer using Rat IgG2ak PE-anti-mouse CD5, and Rat IgG2ak PerCP-Cy5-anti-mouse CD45R/B220. *p<0.05 compared to control mice; and #p<0.05 compared to WT mice immunized with Natterins.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3733974&req=5

pone-0071185-g004: The persistence of B1b cells in bone marrow is negatively regulated by TLR2/TLR4 and MyD88 signals.A representative dot plot of B1b cells (B220lowCD5neg) analyses is shown. Cells from peritoneum (A, D), spleen (B, E) and bone marrow (C, F) were obtained from TLR4 mutant (left) or from TLR2 KO and MyD88 KO (right) Natterins immunized mice after 48, 74 and 120 days. The bars representative of the absolute numbers of B220lowCD5neg cells were determined from total mononuclear cells by multiparametric flow cytometer using Rat IgG2ak PE-anti-mouse CD5, and Rat IgG2ak PerCP-Cy5-anti-mouse CD45R/B220. *p<0.05 compared to control mice; and #p<0.05 compared to WT mice immunized with Natterins.
Mentions: Regarding B1b cells (B220lowCD5neg) our results showed that Natterins did not induce the expansion of B1b cells into peritoneal cavity (Fig. 4A) or in the BM (Fig. 4C) of TLR4 WT mice (white bars), but in contrast this subpopulation was expanded in the spleen only at 48 d in TLR4 WT mice (Fig. 4B). In TLR4 mutant mice (black bars) Natterins induced an increased number of B1b cells only at 120 d in peritoneal cavity (Fig. 4A), and in entire course of the response in spleen (Fig. 4B), and only in the early phase in BM (Fig. 4C), indicating that TLR4 acts as negative regulator, mainly in the transient expansion of B1b cells in spleen.

Bottom Line: TLR4 (MyD88-independent) modulated the emigration from the spleen of Bmem as well as ASC B220(pos).Terminally differentiated ASC B220(neg) required the cooperation of both signals through TLR2/TLR4 via MyD88 for longevity in peritoneum, whereas Bmem required only TLR2/MyD88 to stay in spleen, and ASC B220(pos) rested in peritoneum dependent on TLR4 signaling.Our data sustain that earlier events on memory B cells differentiation induced in secondary immune response against Natterins, after secondary lymph organs influx and egress, may be the key to determining peripheral localization of innate-like B cells and memory B cells as ASC B220(pos) and ASC B220(neg).

View Article: PubMed Central - PubMed

Affiliation: Immunoregulation Unit, Special Laboratory of Applied Toxinology, Butantan Institute and Department of Immunology, University of São Paulo, São Paulo, Brazil.

ABSTRACT
This study was undertaken to gain better insights into the role of TLRs and MyD88 in the development and differentiation of memory B cells, especially of ASC, during the Th2 polarized memory response induced by Natterins. Our in vivo findings demonstrated that the anaphylactic IgG1 production is dependent on TLR2 and MyD88 signaling, and that TLR4 acts as adjuvant accelerating the synthesis of high affinity-IgE. Also, TLR4 (MyD88-independent) modulated the migration of innate-like B cells (B1a and B2) out of the peritoneal cavity, and the emigration from the spleen of B1b and B2 cells. TLR4 (MyD88-independent) modulated the emigration from the spleen of Bmem as well as ASC B220(pos). TLR2 triggered to the egress from the peritoneum of Bmem (MyD88-dependent) and ASC B220(pos) (MyD88-independent). We showed that TLR4 regulates the degree of expansion of Bmem in the peritoneum (MyD88-dependent) and in BM (MyD88-independent) as well as of ASC B220(neg) in the spleen (MyD88-independent). TLR2 regulated the intensity of the expansion of Bmem (MyD88-independent) and ASC B220(pos) (MyD88-dependent) in BM. Finally, TLR4 signals sustained the longevity of ASC B220(pos) (MyD88-independent) and ASC B220(neg) into the peritoneum (MyD88-dependent) and TLR2 MyD88-dependent signaling supported the persistence of B2 cells in BM, Bmem in the spleen and ASC B220(neg) in peritoneum and BM. Terminally differentiated ASC B220(neg) required the cooperation of both signals through TLR2/TLR4 via MyD88 for longevity in peritoneum, whereas Bmem required only TLR2/MyD88 to stay in spleen, and ASC B220(pos) rested in peritoneum dependent on TLR4 signaling. Our data sustain that earlier events on memory B cells differentiation induced in secondary immune response against Natterins, after secondary lymph organs influx and egress, may be the key to determining peripheral localization of innate-like B cells and memory B cells as ASC B220(pos) and ASC B220(neg).

Show MeSH