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Incunabular immunological events in prion trafficking.

Michel B, Meyerett-Reid C, Johnson T, Ferguson A, Wyckoff C, Pulford B, Bender H, Avery A, Telling G, Dow S, Zabel MD - Sci Rep (2012)

Bottom Line: Monocytes and dendritic cells (DCs) required Complement for optimal prion delivery to lymph nodes hours later in a second wave of prion trafficking.B cells constituted the majority of prion-bearing cells in the mediastinal lymph node by six hours, indicating intranodal prion reception from resident DCs or subcapsulary sinus macrophages or directly from follicular conduits.These data reveal novel, cell autonomous prion lymphotropism, and a prominent role for B cells in intranodal prion movement.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine and Biomedical Sciences, Department of Microbiology, Immunology and Pathology, Prion Research Program at Colorado State University, Fort Collins, Colorado 80521, USA.

ABSTRACT
While prions probably interact with the innate immune system immediately following infection, little is known about this initial confrontation. Here we investigated incunabular events in lymphotropic and intranodal prion trafficking by following highly enriched, fluorescent prions from infection sites to draining lymph nodes. We detected biphasic lymphotropic transport of prions from the initial entry site upon peripheral prion inoculation. Prions arrived in draining lymph nodes cell autonomously within two hours of intraperitoneal administration. Monocytes and dendritic cells (DCs) required Complement for optimal prion delivery to lymph nodes hours later in a second wave of prion trafficking. B cells constituted the majority of prion-bearing cells in the mediastinal lymph node by six hours, indicating intranodal prion reception from resident DCs or subcapsulary sinus macrophages or directly from follicular conduits. These data reveal novel, cell autonomous prion lymphotropism, and a prominent role for B cells in intranodal prion movement.

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Flow cytometric analysis of passive and active transport of prion rods.Prion rods were injected IP into donor mice and two hours later immune cells harvested from the PC and mediastinal lymph nodes. Peritoneal cells were washed to remove unbould prion rods and transferred into the PC of recipient mice. MedLN from donor mice 2 HPI (panels a–e), and recipient mice 2 and 16 HPI (panels f–k), were analyzed for prion-bearing resident and migratory immune cells, respectively. Total cells from the MedLN (grey dots in panels a, f and g) are plotted to show relative size (forward scatter, linear scale), granularity (side scatter, log scale) and proportion of total live cells that bear prions (red dots). Resident MedLN cells were analyzed for SSChiCD11b+CD169+ SCS MΦs (b and d) and SSChiCD11c+CD8α+ DCs (c and e) bearing prion rods. Prion-loaded cells from donor mice were injected into the PC of recipient mice and lymph nodes harvested 2 and 16 hours later. Migratory immune cells were also analyzed for monocytes (depicted as red peaks (h and i) and dots (Jjand k)), DCs (green) and MΦs (purple) using the same phenotypic markers used in Figure 3. These cell subsets were compared to cells in MedLN from PBS inoculated control mice (blue). Cell counts in graphs d, e, j and k are shown as log10 per 105 total cells.
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f5: Flow cytometric analysis of passive and active transport of prion rods.Prion rods were injected IP into donor mice and two hours later immune cells harvested from the PC and mediastinal lymph nodes. Peritoneal cells were washed to remove unbould prion rods and transferred into the PC of recipient mice. MedLN from donor mice 2 HPI (panels a–e), and recipient mice 2 and 16 HPI (panels f–k), were analyzed for prion-bearing resident and migratory immune cells, respectively. Total cells from the MedLN (grey dots in panels a, f and g) are plotted to show relative size (forward scatter, linear scale), granularity (side scatter, log scale) and proportion of total live cells that bear prions (red dots). Resident MedLN cells were analyzed for SSChiCD11b+CD169+ SCS MΦs (b and d) and SSChiCD11c+CD8α+ DCs (c and e) bearing prion rods. Prion-loaded cells from donor mice were injected into the PC of recipient mice and lymph nodes harvested 2 and 16 hours later. Migratory immune cells were also analyzed for monocytes (depicted as red peaks (h and i) and dots (Jjand k)), DCs (green) and MΦs (purple) using the same phenotypic markers used in Figure 3. These cell subsets were compared to cells in MedLN from PBS inoculated control mice (blue). Cell counts in graphs d, e, j and k are shown as log10 per 105 total cells.

Mentions: SSChiCD11b+CD169+ subcapsulary sinus MΦs (SCS MΦs, Figure 5a) isolated from donor mice show a high propensity for prion uptake 2 HPI (Figure 5b (red cloud) and 5d (red dots), median = 94; IQR = 27 to 148 per 100,000 cells, n = 17 mice, p<0.01) compared to mice inoculated with PBS (Figure 5b (blue cloud) and 5d (blue squares, median = 6; IQR 2 to 7, n = 3 mice). SSChiCD11c+CD8α+ resident DCs also show a significant ability to take up prions (Figure 5c and 5e (red cloud and dots) median = 8; IQR 4 to 11, n = 17, p<0.05) compared to PBS controls (blue cloud and dots, median = 2; IQR 1 to 5, n = 3).


Incunabular immunological events in prion trafficking.

Michel B, Meyerett-Reid C, Johnson T, Ferguson A, Wyckoff C, Pulford B, Bender H, Avery A, Telling G, Dow S, Zabel MD - Sci Rep (2012)

Flow cytometric analysis of passive and active transport of prion rods.Prion rods were injected IP into donor mice and two hours later immune cells harvested from the PC and mediastinal lymph nodes. Peritoneal cells were washed to remove unbould prion rods and transferred into the PC of recipient mice. MedLN from donor mice 2 HPI (panels a–e), and recipient mice 2 and 16 HPI (panels f–k), were analyzed for prion-bearing resident and migratory immune cells, respectively. Total cells from the MedLN (grey dots in panels a, f and g) are plotted to show relative size (forward scatter, linear scale), granularity (side scatter, log scale) and proportion of total live cells that bear prions (red dots). Resident MedLN cells were analyzed for SSChiCD11b+CD169+ SCS MΦs (b and d) and SSChiCD11c+CD8α+ DCs (c and e) bearing prion rods. Prion-loaded cells from donor mice were injected into the PC of recipient mice and lymph nodes harvested 2 and 16 hours later. Migratory immune cells were also analyzed for monocytes (depicted as red peaks (h and i) and dots (Jjand k)), DCs (green) and MΦs (purple) using the same phenotypic markers used in Figure 3. These cell subsets were compared to cells in MedLN from PBS inoculated control mice (blue). Cell counts in graphs d, e, j and k are shown as log10 per 105 total cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3368226&req=5

f5: Flow cytometric analysis of passive and active transport of prion rods.Prion rods were injected IP into donor mice and two hours later immune cells harvested from the PC and mediastinal lymph nodes. Peritoneal cells were washed to remove unbould prion rods and transferred into the PC of recipient mice. MedLN from donor mice 2 HPI (panels a–e), and recipient mice 2 and 16 HPI (panels f–k), were analyzed for prion-bearing resident and migratory immune cells, respectively. Total cells from the MedLN (grey dots in panels a, f and g) are plotted to show relative size (forward scatter, linear scale), granularity (side scatter, log scale) and proportion of total live cells that bear prions (red dots). Resident MedLN cells were analyzed for SSChiCD11b+CD169+ SCS MΦs (b and d) and SSChiCD11c+CD8α+ DCs (c and e) bearing prion rods. Prion-loaded cells from donor mice were injected into the PC of recipient mice and lymph nodes harvested 2 and 16 hours later. Migratory immune cells were also analyzed for monocytes (depicted as red peaks (h and i) and dots (Jjand k)), DCs (green) and MΦs (purple) using the same phenotypic markers used in Figure 3. These cell subsets were compared to cells in MedLN from PBS inoculated control mice (blue). Cell counts in graphs d, e, j and k are shown as log10 per 105 total cells.
Mentions: SSChiCD11b+CD169+ subcapsulary sinus MΦs (SCS MΦs, Figure 5a) isolated from donor mice show a high propensity for prion uptake 2 HPI (Figure 5b (red cloud) and 5d (red dots), median = 94; IQR = 27 to 148 per 100,000 cells, n = 17 mice, p<0.01) compared to mice inoculated with PBS (Figure 5b (blue cloud) and 5d (blue squares, median = 6; IQR 2 to 7, n = 3 mice). SSChiCD11c+CD8α+ resident DCs also show a significant ability to take up prions (Figure 5c and 5e (red cloud and dots) median = 8; IQR 4 to 11, n = 17, p<0.05) compared to PBS controls (blue cloud and dots, median = 2; IQR 1 to 5, n = 3).

Bottom Line: Monocytes and dendritic cells (DCs) required Complement for optimal prion delivery to lymph nodes hours later in a second wave of prion trafficking.B cells constituted the majority of prion-bearing cells in the mediastinal lymph node by six hours, indicating intranodal prion reception from resident DCs or subcapsulary sinus macrophages or directly from follicular conduits.These data reveal novel, cell autonomous prion lymphotropism, and a prominent role for B cells in intranodal prion movement.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine and Biomedical Sciences, Department of Microbiology, Immunology and Pathology, Prion Research Program at Colorado State University, Fort Collins, Colorado 80521, USA.

ABSTRACT
While prions probably interact with the innate immune system immediately following infection, little is known about this initial confrontation. Here we investigated incunabular events in lymphotropic and intranodal prion trafficking by following highly enriched, fluorescent prions from infection sites to draining lymph nodes. We detected biphasic lymphotropic transport of prions from the initial entry site upon peripheral prion inoculation. Prions arrived in draining lymph nodes cell autonomously within two hours of intraperitoneal administration. Monocytes and dendritic cells (DCs) required Complement for optimal prion delivery to lymph nodes hours later in a second wave of prion trafficking. B cells constituted the majority of prion-bearing cells in the mediastinal lymph node by six hours, indicating intranodal prion reception from resident DCs or subcapsulary sinus macrophages or directly from follicular conduits. These data reveal novel, cell autonomous prion lymphotropism, and a prominent role for B cells in intranodal prion movement.

Show MeSH
Related in: MedlinePlus