Anti-mPDCA-1 – functional grade

Efficiently deplete PDCs with a functional grade antibody
Ground-breaking tool for studying PDC function in vivo
Outstanding reproducibility avoids cumbersome and expensive experiment repetitions

Overview

Anti-mPDCA-1 pure – functional grade was developed for the in vivo depletion of mouse plasmacytoid dendritic cells (PDCs) in order to study their functional role in experimental mouse models. The antibody has been used for PDC depletion in various mouse models of infection, for example, murine CMV2, cutaneous HSV3, and vaginal HSV-24 infection, where PDCs are shown to play a crucial role in antiviral protection. In addition, the functional role of PDCs after Listeria monocytogenes infection has been analyzed and revealed that the interaction between convential DCs and PDCs is essential for CpG-induced immune activation.

Details

Background information
Clone JF05-1C2.4.1 recognizes the mouse plasmacytoid dendritic cell antigen-1 (mPDCA-1). mPDCA-1 is specifically expressed on mouse PDCs — a subset of CD11c⁺ dendritic cells detected at low frequency in all lymphoid tissues, peripheral blood, and some non-lymphoid tissues.
In mouse spleen, bone marrow, and lymph nodes, mPDCA-1 is exclusively expressed on cells which are CD11c^low, Ly-6C^high, CD45R (B220)⁺, MHC Class II^low, CD8a^low/–, CD80^low/–, CD86^–, CD40^–, CD11b^–, CD90^–, CD49b (DX5)^–, CD3^–, CD19^–, i.e. on cells with the phenotype of mouse PDCs. Multi-color fluorescent staining of spleen cells clearly revealed that all CD11c^low Ly-6C^high CD45R (B220)⁺ PDCs are mPDCA-1⁺ and that mPDCA-1 expression is restricted to PDCs.

Clone	Isotype
JF05-1C2.4.1	Rat IgG2b

Figure 1

Mouse PDCs were specifically depleted in BALB/c mice by i.p. injection of 500 µg Anti-mPDCA-1 pure – functional grade. The frequency of mPDCA-1⁺ PDCs was analyzed in spleen 24 h after injection of the antibody or diluent only (control mice) by staining with Anti-mPDCA-1. To exclude blocking of Anti-mPDCA-1 staining by the in vivo applied Anti-mPDCA-1 antibody, cells were additionally stained with Anti-Ly-6C and CD45R (B220). Note that the frequency of PDCs decreases in mice which received the Anti-mPDCA-1 antibody (A) to about one fifth compared to control mice (B), independent of whether it is analyzed based on Anti-mPDCA-1 staining or on Anti-Ly-6C and CD45R (B220) staining. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence.

A) Anti-mPDCA-1 antibody

Mouse PDCs were specifically depleted in BALB/c mice by i.p. injection of 500 µg Anti-mPDCA-1 pure – functional grade. The frequency of mPDCA-1<sup>+</sup> PDCs was analyzed in spleen 24 h after injection of the antibody or diluent only (control mice) by staining with Anti-mPDCA-1. To exclude blocking of Anti-mPDCA-1 staining by the <i>in vivo</i> applied Anti-mPDCA-1 antibody, cells were additionally stained with Anti-Ly-6C and CD45R (B220). Note that the frequency of PDCs decreases in mice which received the Anti-mPDCA-1 antibody (A) to about one fifth compared to control mice (B), independent of whether it is analyzed based on Anti-mPDCA-1 staining or on Anti-Ly-6C and CD45R (B220) staining. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence.

B) Control