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A member of the dendritic cell family that enters B cell follicles and stimulates primary antibody responses identified by a mannose receptor fusion protein.

Berney C, Herren S, Power CA, Gordon S, Martinez-Pomares L, Kosco-Vilbois MH - J. Exp. Med. (1999)

Bottom Line: Dendritic cells (DCs) are known to activate naive T cells to become effective helper cells.Furthermore, they produced significantly more of the lymphocyte chemoattractant, macrophage inflammatory protein (MIP)-1alpha, than isolated interdigitating cells.Taken together, these results provide evidence that a subset of DCs enters primary follicles, armed with the capacity to attract and provide antigenic stimulation for T and B lymphocytes.

View Article: PubMed Central - PubMed

Affiliation: Serono Pharmaceutical Research Institute, CH-1228 Geneva, Switzerland.

ABSTRACT
Dendritic cells (DCs) are known to activate naive T cells to become effective helper cells. In addition, recent evidence suggests that DCs may influence naive B cells during the initial priming of antibody responses. In this study, using three-color confocal microscopy and three-dimensional immunohistograms, we have observed that in the first few days after a primary immunization, cells with dendritic morphology progressively localize within primary B cell follicles. These cells were identified by their ability to bind a fusion protein consisting of the terminal cysteine-rich portion of the mouse mannose receptor and the Fc portion of human immunoglobulin (Ig)G1 (CR-Fc). In situ, these CR-Fc binding cells express major histocompatibility complex class II, sialoadhesin, and CD11c and are negative for other markers identifying the myeloid DC lineage, such as (CD11b), macrophages (F4/80), follicular DCs (FDC-M2), B cells (B220), and T cells (CD4). Using CR-Fc binding capacity and flow cytometry, the cells were purified from the draining lymph nodes of mice 24 h after immunization. When injected into naive mice, these cells were able to prime T cells as well as induce production of antigen-specific IgM and IgG1. Furthermore, they produced significantly more of the lymphocyte chemoattractant, macrophage inflammatory protein (MIP)-1alpha, than isolated interdigitating cells. Taken together, these results provide evidence that a subset of DCs enters primary follicles, armed with the capacity to attract and provide antigenic stimulation for T and B lymphocytes.

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Localization of CR-Fc binding cells in lymph nodes with time after a primary immunization. Three-color confocal analysis was used in order to determine when and where CR-Fc binding cells appeared in the tissue. Cryostat sections prepared from murine lymph nodes obtained at days 0–4 (A–E) of a primary response were processed for IgM expression (red), N418 labeling (green), and CR-Fc binding (blue). The images were then digitally transformed using a computer program in order to map the MFI (in arbitrary units) for either single (CR-Fc+; N418+) or double (CR-Fc+/ N418+) positive cells. The resulting 3D immunohistograms revealed that CR-Fc binding cells accumulated with time in the outer part of the B cell follicles. Many of these cells were also N418+ (see the CR-Fc+/N418+ column). The scale 0–250 represents the MFI, with the colors yellow, blue, green, and red representing values in the 100–150, 150–200, 200–250, and >250 range, respectively.
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Figure 1: Localization of CR-Fc binding cells in lymph nodes with time after a primary immunization. Three-color confocal analysis was used in order to determine when and where CR-Fc binding cells appeared in the tissue. Cryostat sections prepared from murine lymph nodes obtained at days 0–4 (A–E) of a primary response were processed for IgM expression (red), N418 labeling (green), and CR-Fc binding (blue). The images were then digitally transformed using a computer program in order to map the MFI (in arbitrary units) for either single (CR-Fc+; N418+) or double (CR-Fc+/ N418+) positive cells. The resulting 3D immunohistograms revealed that CR-Fc binding cells accumulated with time in the outer part of the B cell follicles. Many of these cells were also N418+ (see the CR-Fc+/N418+ column). The scale 0–250 represents the MFI, with the colors yellow, blue, green, and red representing values in the 100–150, 150–200, 200–250, and >250 range, respectively.

Mentions: During an immune response, it has been previously shown that within lymph nodes, cells with dendritic morphology bind the CR-Fc fusion protein 12. To further investigate their nature, we developed a system involving immunohistochemistry and three-color microscopy. As shown in Fig. 1, the B cell follicles were visualized using an anti-IgM antibody conjugated to Texas red. CR-Fc binding cells were labeled with Cy5-conjugated reagents. A third marker associated with FITC was then used to determine either the phenotype of the CR-Fc binding cells (double positive = cyan) or the proximity of these cells to other cell types. 3D histograms were then created in order to accurately assess the number and location of single or double positive cells.


A member of the dendritic cell family that enters B cell follicles and stimulates primary antibody responses identified by a mannose receptor fusion protein.

Berney C, Herren S, Power CA, Gordon S, Martinez-Pomares L, Kosco-Vilbois MH - J. Exp. Med. (1999)

Localization of CR-Fc binding cells in lymph nodes with time after a primary immunization. Three-color confocal analysis was used in order to determine when and where CR-Fc binding cells appeared in the tissue. Cryostat sections prepared from murine lymph nodes obtained at days 0–4 (A–E) of a primary response were processed for IgM expression (red), N418 labeling (green), and CR-Fc binding (blue). The images were then digitally transformed using a computer program in order to map the MFI (in arbitrary units) for either single (CR-Fc+; N418+) or double (CR-Fc+/ N418+) positive cells. The resulting 3D immunohistograms revealed that CR-Fc binding cells accumulated with time in the outer part of the B cell follicles. Many of these cells were also N418+ (see the CR-Fc+/N418+ column). The scale 0–250 represents the MFI, with the colors yellow, blue, green, and red representing values in the 100–150, 150–200, 200–250, and >250 range, respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195630&req=5

Figure 1: Localization of CR-Fc binding cells in lymph nodes with time after a primary immunization. Three-color confocal analysis was used in order to determine when and where CR-Fc binding cells appeared in the tissue. Cryostat sections prepared from murine lymph nodes obtained at days 0–4 (A–E) of a primary response were processed for IgM expression (red), N418 labeling (green), and CR-Fc binding (blue). The images were then digitally transformed using a computer program in order to map the MFI (in arbitrary units) for either single (CR-Fc+; N418+) or double (CR-Fc+/ N418+) positive cells. The resulting 3D immunohistograms revealed that CR-Fc binding cells accumulated with time in the outer part of the B cell follicles. Many of these cells were also N418+ (see the CR-Fc+/N418+ column). The scale 0–250 represents the MFI, with the colors yellow, blue, green, and red representing values in the 100–150, 150–200, 200–250, and >250 range, respectively.
Mentions: During an immune response, it has been previously shown that within lymph nodes, cells with dendritic morphology bind the CR-Fc fusion protein 12. To further investigate their nature, we developed a system involving immunohistochemistry and three-color microscopy. As shown in Fig. 1, the B cell follicles were visualized using an anti-IgM antibody conjugated to Texas red. CR-Fc binding cells were labeled with Cy5-conjugated reagents. A third marker associated with FITC was then used to determine either the phenotype of the CR-Fc binding cells (double positive = cyan) or the proximity of these cells to other cell types. 3D histograms were then created in order to accurately assess the number and location of single or double positive cells.

Bottom Line: Dendritic cells (DCs) are known to activate naive T cells to become effective helper cells.Furthermore, they produced significantly more of the lymphocyte chemoattractant, macrophage inflammatory protein (MIP)-1alpha, than isolated interdigitating cells.Taken together, these results provide evidence that a subset of DCs enters primary follicles, armed with the capacity to attract and provide antigenic stimulation for T and B lymphocytes.

View Article: PubMed Central - PubMed

Affiliation: Serono Pharmaceutical Research Institute, CH-1228 Geneva, Switzerland.

ABSTRACT
Dendritic cells (DCs) are known to activate naive T cells to become effective helper cells. In addition, recent evidence suggests that DCs may influence naive B cells during the initial priming of antibody responses. In this study, using three-color confocal microscopy and three-dimensional immunohistograms, we have observed that in the first few days after a primary immunization, cells with dendritic morphology progressively localize within primary B cell follicles. These cells were identified by their ability to bind a fusion protein consisting of the terminal cysteine-rich portion of the mouse mannose receptor and the Fc portion of human immunoglobulin (Ig)G1 (CR-Fc). In situ, these CR-Fc binding cells express major histocompatibility complex class II, sialoadhesin, and CD11c and are negative for other markers identifying the myeloid DC lineage, such as (CD11b), macrophages (F4/80), follicular DCs (FDC-M2), B cells (B220), and T cells (CD4). Using CR-Fc binding capacity and flow cytometry, the cells were purified from the draining lymph nodes of mice 24 h after immunization. When injected into naive mice, these cells were able to prime T cells as well as induce production of antigen-specific IgM and IgG1. Furthermore, they produced significantly more of the lymphocyte chemoattractant, macrophage inflammatory protein (MIP)-1alpha, than isolated interdigitating cells. Taken together, these results provide evidence that a subset of DCs enters primary follicles, armed with the capacity to attract and provide antigenic stimulation for T and B lymphocytes.

Show MeSH
Related in: MedlinePlus