Limits...
Alleviation of skin inflammation after Lin(-) cell transplantation correlates with their differentiation into myeloid-derived suppressor cells.

Jeong Ryu S, Ju JM, Kim W, Bum Kim M, Hee Oh K, Sup Lee D, Lee H, Eun Oh J, Soo Park K, Young Choi E - Sci Rep (2015)

Bottom Line: Longitudinal in vivo imaging and flow cytometry analyses revealed that Lin(-) cells home directly to inflamed skin within 6 h, where they undergo extensive expansion with the peak on day 14 post-transplantation, and preferential differentiation into CD11b(+)Ly6G(int)Ly6C(+) cells by day 7.Cells with phenotypic profiles of neutrophils, macrophages, and DCs appeared in inflamed skin on day 14.Progenies of transplanted Lin(-) cells showed similar kinetics of expansion and myeloid differentiation in BM.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.

ABSTRACT
To understand the cellular mechanism underlying the therapeutic effects exerted by hematopoietic stem cell transplantation in the repair of tissue damage, we investigated the in vivo dynamics of bone marrow (BM) lineage-negative (Lin(-)) cells transplanted into mice with hyper sensitivity dermatitis. Longitudinal in vivo imaging and flow cytometry analyses revealed that Lin(-) cells home directly to inflamed skin within 6 h, where they undergo extensive expansion with the peak on day 14 post-transplantation, and preferential differentiation into CD11b(+)Ly6G(int)Ly6C(+) cells by day 7. Cells with phenotypic profiles of neutrophils, macrophages, and DCs appeared in inflamed skin on day 14. Progenies of transplanted Lin(-) cells showed similar kinetics of expansion and myeloid differentiation in BM. However, differentiation into CD11b(+)Ly6G(int)Ly6C(+) cells in the inflamed skin on day 7 was more skewed toward CD115(+) cells (≥60%) with immune suppressive function and higher expression levels of iNOS, arginase, and IL-10, compared with those in the BM. Transplantation of Lin(-) cells reduced the levels of Cd3 transcript and CD4(+)/CD8(+) cells in inflamed skin. These results demonstrate differentiation of transplanted Lin(-) cells into myeloid-derived suppressor cells in inflamed skin to be the basis of the alleviation of skin inflammation after Lin(-) cell transplantation.

No MeSH data available.


Related in: MedlinePlus

Differentiation of Lin− cells into myeloid and granulocytic cells at the site of inflammation.(a) Schematic overview of treatment with anti-Gr-1 depleting antibody and transplantation of Lin− or Lin+ cells (5 × 105). Dermatitis mice were injected i.p. with an anti-Gr-1 antibody prior to and 3 days after transplantation with Luc-Tg Lin− cells; rat-IgG was treated as a control. (b) BLI images were collected on days 1, 3, and 7 post-transplantation. Images shown are representative of two independent experiments (n = 3 mice/group/experiment).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4594128&req=5

f5: Differentiation of Lin− cells into myeloid and granulocytic cells at the site of inflammation.(a) Schematic overview of treatment with anti-Gr-1 depleting antibody and transplantation of Lin− or Lin+ cells (5 × 105). Dermatitis mice were injected i.p. with an anti-Gr-1 antibody prior to and 3 days after transplantation with Luc-Tg Lin− cells; rat-IgG was treated as a control. (b) BLI images were collected on days 1, 3, and 7 post-transplantation. Images shown are representative of two independent experiments (n = 3 mice/group/experiment).

Mentions: Having established the source of cells expanded at the site of inflammation, we next investigated differentiation of Lin− progenies at the site of inflammation, as these processes were presumed to play a role in their therapeutic effects. To determine the lineage of the differentiating cells, we selectively depleted each cell lineage by treating the recipients with antibodies against lineage markers (Gr-1, CD4, CD8, or B220) prior to and 3 days after transplantation of Luc-Tg Lin− cells (Fig. 5a). Then, differentiation into a specific lineage was determined by specific signal abrogation in the recipients with the corresponding depletion treatment in BLI analyses. Treatment with a Gr-1-depleting antibody abrogated the luminescence signals at the site of inflammation when the recipients of Luc-Tg Lin− cells were analyzed on day 7, but not on days 1 or 3 post-transplantation (Fig. 5b), as did clodronate-treatment which depletes macrophage/monocytes27 (data now shown). This indicated depletion of Gr-1-expressing cells originated from transplanted Luc-Tg Lin− cells in the inflamed skin on day 7. Prior flow cytometric analysis had verified depletion of Gr-1hi granulocytes in the inflamed skin of dermatitis mice with one time treatment of anti-Gr-1 antibody (data not shown). Other treatments including those targeting CD8, CD4, and B220 had no effect on signal detection (data not shown). These results indicated that, despite their multi-lineage potency, Lin− cells preferentially differentiated into myeloid and granulocytic lineage cells within the first 7 days, in consistent with the promoted myeloid/granulocytic differentiation in the presence of inflammation, relative to steady-state hematopoiesis528.


Alleviation of skin inflammation after Lin(-) cell transplantation correlates with their differentiation into myeloid-derived suppressor cells.

Jeong Ryu S, Ju JM, Kim W, Bum Kim M, Hee Oh K, Sup Lee D, Lee H, Eun Oh J, Soo Park K, Young Choi E - Sci Rep (2015)

Differentiation of Lin− cells into myeloid and granulocytic cells at the site of inflammation.(a) Schematic overview of treatment with anti-Gr-1 depleting antibody and transplantation of Lin− or Lin+ cells (5 × 105). Dermatitis mice were injected i.p. with an anti-Gr-1 antibody prior to and 3 days after transplantation with Luc-Tg Lin− cells; rat-IgG was treated as a control. (b) BLI images were collected on days 1, 3, and 7 post-transplantation. Images shown are representative of two independent experiments (n = 3 mice/group/experiment).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4594128&req=5

f5: Differentiation of Lin− cells into myeloid and granulocytic cells at the site of inflammation.(a) Schematic overview of treatment with anti-Gr-1 depleting antibody and transplantation of Lin− or Lin+ cells (5 × 105). Dermatitis mice were injected i.p. with an anti-Gr-1 antibody prior to and 3 days after transplantation with Luc-Tg Lin− cells; rat-IgG was treated as a control. (b) BLI images were collected on days 1, 3, and 7 post-transplantation. Images shown are representative of two independent experiments (n = 3 mice/group/experiment).
Mentions: Having established the source of cells expanded at the site of inflammation, we next investigated differentiation of Lin− progenies at the site of inflammation, as these processes were presumed to play a role in their therapeutic effects. To determine the lineage of the differentiating cells, we selectively depleted each cell lineage by treating the recipients with antibodies against lineage markers (Gr-1, CD4, CD8, or B220) prior to and 3 days after transplantation of Luc-Tg Lin− cells (Fig. 5a). Then, differentiation into a specific lineage was determined by specific signal abrogation in the recipients with the corresponding depletion treatment in BLI analyses. Treatment with a Gr-1-depleting antibody abrogated the luminescence signals at the site of inflammation when the recipients of Luc-Tg Lin− cells were analyzed on day 7, but not on days 1 or 3 post-transplantation (Fig. 5b), as did clodronate-treatment which depletes macrophage/monocytes27 (data now shown). This indicated depletion of Gr-1-expressing cells originated from transplanted Luc-Tg Lin− cells in the inflamed skin on day 7. Prior flow cytometric analysis had verified depletion of Gr-1hi granulocytes in the inflamed skin of dermatitis mice with one time treatment of anti-Gr-1 antibody (data not shown). Other treatments including those targeting CD8, CD4, and B220 had no effect on signal detection (data not shown). These results indicated that, despite their multi-lineage potency, Lin− cells preferentially differentiated into myeloid and granulocytic lineage cells within the first 7 days, in consistent with the promoted myeloid/granulocytic differentiation in the presence of inflammation, relative to steady-state hematopoiesis528.

Bottom Line: Longitudinal in vivo imaging and flow cytometry analyses revealed that Lin(-) cells home directly to inflamed skin within 6 h, where they undergo extensive expansion with the peak on day 14 post-transplantation, and preferential differentiation into CD11b(+)Ly6G(int)Ly6C(+) cells by day 7.Cells with phenotypic profiles of neutrophils, macrophages, and DCs appeared in inflamed skin on day 14.Progenies of transplanted Lin(-) cells showed similar kinetics of expansion and myeloid differentiation in BM.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.

ABSTRACT
To understand the cellular mechanism underlying the therapeutic effects exerted by hematopoietic stem cell transplantation in the repair of tissue damage, we investigated the in vivo dynamics of bone marrow (BM) lineage-negative (Lin(-)) cells transplanted into mice with hyper sensitivity dermatitis. Longitudinal in vivo imaging and flow cytometry analyses revealed that Lin(-) cells home directly to inflamed skin within 6 h, where they undergo extensive expansion with the peak on day 14 post-transplantation, and preferential differentiation into CD11b(+)Ly6G(int)Ly6C(+) cells by day 7. Cells with phenotypic profiles of neutrophils, macrophages, and DCs appeared in inflamed skin on day 14. Progenies of transplanted Lin(-) cells showed similar kinetics of expansion and myeloid differentiation in BM. However, differentiation into CD11b(+)Ly6G(int)Ly6C(+) cells in the inflamed skin on day 7 was more skewed toward CD115(+) cells (≥60%) with immune suppressive function and higher expression levels of iNOS, arginase, and IL-10, compared with those in the BM. Transplantation of Lin(-) cells reduced the levels of Cd3 transcript and CD4(+)/CD8(+) cells in inflamed skin. These results demonstrate differentiation of transplanted Lin(-) cells into myeloid-derived suppressor cells in inflamed skin to be the basis of the alleviation of skin inflammation after Lin(-) cell transplantation.

No MeSH data available.


Related in: MedlinePlus