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Severe Developmental B Lymphopoietic Defects in Foxp3-Deficient Mice are Refractory to Adoptive Regulatory T Cell Therapy.

Riewaldt J, Düber S, Boernert M, Krey M, Dembinski M, Weiss S, Garbe AI, Kretschmer K - Front Immunol (2012)

Bottom Line: However, marginal zone B and B-1a cells were absent throughout ontogeny.Developmental B lymphopoietic defects largely correlated with defective thymopoiesis.Importantly, neonatal adoptive T(reg) cell therapy suppressed exacerbated production of inflammatory cytokines and restored thymopoiesis but was ineffective in recovering defective B lymphopoiesis, probably due to a failure to compensate production of stroma cell-derived IL-7 and CXCL12.

View Article: PubMed Central - PubMed

Affiliation: Center for Regenerative Therapies Dresden, Technical University Dresden Dresden, Germany.

ABSTRACT
The role of Foxp3-expressing regulatory T (T(reg)) cells in tolerance and autoimmunity is well-established. However, although of considerable clinical interest, the role of T(reg) cells in the regulation of hematopoietic homeostasis remains poorly understood. Thus, we analysed B and T lymphopoiesis in the scurfy (Sf) mouse model of T(reg) cell deficiency. In these experiments, the near-complete block of B lymphopoiesis in the BM of adolescent Sf mice was attributed to autoimmune T cells. We could exclude a constitutive lympho-hematopoietic defect or a B cell-intrinsic function of Foxp3. Efficient B cell development in the BM early in ontogeny and pronounced extramedullary B lymphopoietic activity resulted in a peripheral pool of mature B cells in adolescent Sf mice. However, marginal zone B and B-1a cells were absent throughout ontogeny. Developmental B lymphopoietic defects largely correlated with defective thymopoiesis. Importantly, neonatal adoptive T(reg) cell therapy suppressed exacerbated production of inflammatory cytokines and restored thymopoiesis but was ineffective in recovering defective B lymphopoiesis, probably due to a failure to compensate production of stroma cell-derived IL-7 and CXCL12. Our observations on autoimmune-mediated incapacitation of the BM environment in Foxp3-deficient mice will have direct implications for the rational design of BM transplantation protocols for patients with severe genetic deficiencies in functional Foxp3(+) T(reg) cells.

No MeSH data available.


Related in: MedlinePlus

Temporal analysis of peripheral B cell compartments during ontogeny of Sf mice. Cohorts of experimental Sf and WT mice described in Figure 3 were concomitantly employed for the analysis of peripheral B cell compartments. (A) Peripheral B cell maturation in the spleen. Representative flow cytometry of newly formed IgDlowIgMhigh and mature IgDhighIgMlow B cells (dot plots), as well as CD19+ B cells (histograms) in the spleen of 7-day-old WT (top) and Sf (bottom) mice. Graphs depict absolute numbers of respective B cell compartments from WT (blue circles) and Sf (red squares) mice, at indicated time points during ontogeny. (B) IgMhighIgDlow CD23low/−CD21high MZ B cells. Representative flow cytometry of IgMhighCD21high MZ B cells among gated IgDlowCD23low/− splenocytes of 4-week-old mice (top), and absolute numbers of MZ B cells during ontogeny of WT and Sf mice (bottom). (C) B cell subsets in the peritoneal cavity. Representative flow cytometry of IgM, IgD, and CD5 expression among PECs from 4-week-old mice, as well as absolute numbers of peritoneal B cell subsets (B-2: IgDhighIgMlow; B-1: IgDlowIgMhigh; B-1a: IgMhighIgDlowCD5+Mac-1+; B-1b: IgMhighIgDlowCD5−Mac-1+). (D) Flow cytometry of PtC-reactive B cells among gated populations of total CD19+ (left panels) and CD19+CD5+ (right panels) PECs from 3–4-week-old WT (n = 7) and Sf (n = 5) mice, as indicated. (E) Total populations of sIgM+ cells were FACS-purified from PECs of CD45.1+ WT mice and adoptively transferred i.p. into 2-week-old CD45.2+ WT (n = 3) and Sf (n = 3) recipient mice, as indicated. 14 days after cell transfer, recovered PECs were analyzed for the expression of sIgM and CD5 among gated endogenous CD45.2+ cells (left) or CD45.1+ donor cells (right). Data are representative of two independent experiments. Numbers in dot plots and histograms without parentheses indicate the percentage (A–C) and mean percentages ± SEM (D,E) of gated cells within the respective gates. Parenthesized numbers indicate mean absolute numbers (× 103 ± SEM) of PtC+ cells (D). Symbols and lines in graphs indicate individual mice and mean values, respectively.
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Figure 4: Temporal analysis of peripheral B cell compartments during ontogeny of Sf mice. Cohorts of experimental Sf and WT mice described in Figure 3 were concomitantly employed for the analysis of peripheral B cell compartments. (A) Peripheral B cell maturation in the spleen. Representative flow cytometry of newly formed IgDlowIgMhigh and mature IgDhighIgMlow B cells (dot plots), as well as CD19+ B cells (histograms) in the spleen of 7-day-old WT (top) and Sf (bottom) mice. Graphs depict absolute numbers of respective B cell compartments from WT (blue circles) and Sf (red squares) mice, at indicated time points during ontogeny. (B) IgMhighIgDlow CD23low/−CD21high MZ B cells. Representative flow cytometry of IgMhighCD21high MZ B cells among gated IgDlowCD23low/− splenocytes of 4-week-old mice (top), and absolute numbers of MZ B cells during ontogeny of WT and Sf mice (bottom). (C) B cell subsets in the peritoneal cavity. Representative flow cytometry of IgM, IgD, and CD5 expression among PECs from 4-week-old mice, as well as absolute numbers of peritoneal B cell subsets (B-2: IgDhighIgMlow; B-1: IgDlowIgMhigh; B-1a: IgMhighIgDlowCD5+Mac-1+; B-1b: IgMhighIgDlowCD5−Mac-1+). (D) Flow cytometry of PtC-reactive B cells among gated populations of total CD19+ (left panels) and CD19+CD5+ (right panels) PECs from 3–4-week-old WT (n = 7) and Sf (n = 5) mice, as indicated. (E) Total populations of sIgM+ cells were FACS-purified from PECs of CD45.1+ WT mice and adoptively transferred i.p. into 2-week-old CD45.2+ WT (n = 3) and Sf (n = 3) recipient mice, as indicated. 14 days after cell transfer, recovered PECs were analyzed for the expression of sIgM and CD5 among gated endogenous CD45.2+ cells (left) or CD45.1+ donor cells (right). Data are representative of two independent experiments. Numbers in dot plots and histograms without parentheses indicate the percentage (A–C) and mean percentages ± SEM (D,E) of gated cells within the respective gates. Parenthesized numbers indicate mean absolute numbers (× 103 ± SEM) of PtC+ cells (D). Symbols and lines in graphs indicate individual mice and mean values, respectively.

Mentions: In the spleen of Sf mice, we found numbers of newly formed IgDlowIgMhigh B cells to be reduced as early as day 7 p.p., as compared to WT littermate control mice (Figure 4A, left). While the compartment size of mature IgDhighIgMlow B cells failed to reach WT levels during ontogenetic progression, significant numbers of CD19+ B cells were maintained until Sf mice succumbed to autoimmune pathology (Figure 4A, middle and right). Thus, reduced splenic B cell maturation in Sf mice is likely to be a consequence of ceasing central B lymphopoiesis and abrogated export of immature sIgM+ B cells to the spleen.


Severe Developmental B Lymphopoietic Defects in Foxp3-Deficient Mice are Refractory to Adoptive Regulatory T Cell Therapy.

Riewaldt J, Düber S, Boernert M, Krey M, Dembinski M, Weiss S, Garbe AI, Kretschmer K - Front Immunol (2012)

Temporal analysis of peripheral B cell compartments during ontogeny of Sf mice. Cohorts of experimental Sf and WT mice described in Figure 3 were concomitantly employed for the analysis of peripheral B cell compartments. (A) Peripheral B cell maturation in the spleen. Representative flow cytometry of newly formed IgDlowIgMhigh and mature IgDhighIgMlow B cells (dot plots), as well as CD19+ B cells (histograms) in the spleen of 7-day-old WT (top) and Sf (bottom) mice. Graphs depict absolute numbers of respective B cell compartments from WT (blue circles) and Sf (red squares) mice, at indicated time points during ontogeny. (B) IgMhighIgDlow CD23low/−CD21high MZ B cells. Representative flow cytometry of IgMhighCD21high MZ B cells among gated IgDlowCD23low/− splenocytes of 4-week-old mice (top), and absolute numbers of MZ B cells during ontogeny of WT and Sf mice (bottom). (C) B cell subsets in the peritoneal cavity. Representative flow cytometry of IgM, IgD, and CD5 expression among PECs from 4-week-old mice, as well as absolute numbers of peritoneal B cell subsets (B-2: IgDhighIgMlow; B-1: IgDlowIgMhigh; B-1a: IgMhighIgDlowCD5+Mac-1+; B-1b: IgMhighIgDlowCD5−Mac-1+). (D) Flow cytometry of PtC-reactive B cells among gated populations of total CD19+ (left panels) and CD19+CD5+ (right panels) PECs from 3–4-week-old WT (n = 7) and Sf (n = 5) mice, as indicated. (E) Total populations of sIgM+ cells were FACS-purified from PECs of CD45.1+ WT mice and adoptively transferred i.p. into 2-week-old CD45.2+ WT (n = 3) and Sf (n = 3) recipient mice, as indicated. 14 days after cell transfer, recovered PECs were analyzed for the expression of sIgM and CD5 among gated endogenous CD45.2+ cells (left) or CD45.1+ donor cells (right). Data are representative of two independent experiments. Numbers in dot plots and histograms without parentheses indicate the percentage (A–C) and mean percentages ± SEM (D,E) of gated cells within the respective gates. Parenthesized numbers indicate mean absolute numbers (× 103 ± SEM) of PtC+ cells (D). Symbols and lines in graphs indicate individual mice and mean values, respectively.
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Show All Figures
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Figure 4: Temporal analysis of peripheral B cell compartments during ontogeny of Sf mice. Cohorts of experimental Sf and WT mice described in Figure 3 were concomitantly employed for the analysis of peripheral B cell compartments. (A) Peripheral B cell maturation in the spleen. Representative flow cytometry of newly formed IgDlowIgMhigh and mature IgDhighIgMlow B cells (dot plots), as well as CD19+ B cells (histograms) in the spleen of 7-day-old WT (top) and Sf (bottom) mice. Graphs depict absolute numbers of respective B cell compartments from WT (blue circles) and Sf (red squares) mice, at indicated time points during ontogeny. (B) IgMhighIgDlow CD23low/−CD21high MZ B cells. Representative flow cytometry of IgMhighCD21high MZ B cells among gated IgDlowCD23low/− splenocytes of 4-week-old mice (top), and absolute numbers of MZ B cells during ontogeny of WT and Sf mice (bottom). (C) B cell subsets in the peritoneal cavity. Representative flow cytometry of IgM, IgD, and CD5 expression among PECs from 4-week-old mice, as well as absolute numbers of peritoneal B cell subsets (B-2: IgDhighIgMlow; B-1: IgDlowIgMhigh; B-1a: IgMhighIgDlowCD5+Mac-1+; B-1b: IgMhighIgDlowCD5−Mac-1+). (D) Flow cytometry of PtC-reactive B cells among gated populations of total CD19+ (left panels) and CD19+CD5+ (right panels) PECs from 3–4-week-old WT (n = 7) and Sf (n = 5) mice, as indicated. (E) Total populations of sIgM+ cells were FACS-purified from PECs of CD45.1+ WT mice and adoptively transferred i.p. into 2-week-old CD45.2+ WT (n = 3) and Sf (n = 3) recipient mice, as indicated. 14 days after cell transfer, recovered PECs were analyzed for the expression of sIgM and CD5 among gated endogenous CD45.2+ cells (left) or CD45.1+ donor cells (right). Data are representative of two independent experiments. Numbers in dot plots and histograms without parentheses indicate the percentage (A–C) and mean percentages ± SEM (D,E) of gated cells within the respective gates. Parenthesized numbers indicate mean absolute numbers (× 103 ± SEM) of PtC+ cells (D). Symbols and lines in graphs indicate individual mice and mean values, respectively.
Mentions: In the spleen of Sf mice, we found numbers of newly formed IgDlowIgMhigh B cells to be reduced as early as day 7 p.p., as compared to WT littermate control mice (Figure 4A, left). While the compartment size of mature IgDhighIgMlow B cells failed to reach WT levels during ontogenetic progression, significant numbers of CD19+ B cells were maintained until Sf mice succumbed to autoimmune pathology (Figure 4A, middle and right). Thus, reduced splenic B cell maturation in Sf mice is likely to be a consequence of ceasing central B lymphopoiesis and abrogated export of immature sIgM+ B cells to the spleen.

Bottom Line: However, marginal zone B and B-1a cells were absent throughout ontogeny.Developmental B lymphopoietic defects largely correlated with defective thymopoiesis.Importantly, neonatal adoptive T(reg) cell therapy suppressed exacerbated production of inflammatory cytokines and restored thymopoiesis but was ineffective in recovering defective B lymphopoiesis, probably due to a failure to compensate production of stroma cell-derived IL-7 and CXCL12.

View Article: PubMed Central - PubMed

Affiliation: Center for Regenerative Therapies Dresden, Technical University Dresden Dresden, Germany.

ABSTRACT
The role of Foxp3-expressing regulatory T (T(reg)) cells in tolerance and autoimmunity is well-established. However, although of considerable clinical interest, the role of T(reg) cells in the regulation of hematopoietic homeostasis remains poorly understood. Thus, we analysed B and T lymphopoiesis in the scurfy (Sf) mouse model of T(reg) cell deficiency. In these experiments, the near-complete block of B lymphopoiesis in the BM of adolescent Sf mice was attributed to autoimmune T cells. We could exclude a constitutive lympho-hematopoietic defect or a B cell-intrinsic function of Foxp3. Efficient B cell development in the BM early in ontogeny and pronounced extramedullary B lymphopoietic activity resulted in a peripheral pool of mature B cells in adolescent Sf mice. However, marginal zone B and B-1a cells were absent throughout ontogeny. Developmental B lymphopoietic defects largely correlated with defective thymopoiesis. Importantly, neonatal adoptive T(reg) cell therapy suppressed exacerbated production of inflammatory cytokines and restored thymopoiesis but was ineffective in recovering defective B lymphopoiesis, probably due to a failure to compensate production of stroma cell-derived IL-7 and CXCL12. Our observations on autoimmune-mediated incapacitation of the BM environment in Foxp3-deficient mice will have direct implications for the rational design of BM transplantation protocols for patients with severe genetic deficiencies in functional Foxp3(+) T(reg) cells.

No MeSH data available.


Related in: MedlinePlus