RE: double label for dendritic cells and MVE virus antigen
Leanne Sammels (lsammels@cyllene.uwa.edu.au)
Wed, 11 Jun 1997 13:23:43 +0800
>From: Snider Denis <sniderd@fhs.csu.mcmaster.ca>
>To: "'Leanne Sammels'" <lsammels@cyllene.uwa.edu.au>
>Subject: RE: double label for dendritic cells and MVE virus antigen
>Date: Tue, 10 Jun 1997 23:14:50 -0400
>
>Leanne,
>
>Is your virus in the T cell regions or in the follicles of the lymph
>nodes. As you may be aware there are two types of dendritic cells in
>lymph nodes. The interdigitating cells present antigen to T cells in
>the T cell regions, while the follicular dendritic cells trap antigen
>for stimulation of B cells in the follicles.
>
>The best antibody combinations we have used for isolation of mouse
>interdigitating dendritic cells from lymphoid or mucosal tissues are any
>good anti-class II MHC mAb (dendritic cells are bright) in combination
>with anti-CD11c antibody (also bright on these DC, but also found on B
>cells and macrophage so be careful). Try Phamingen for one source. A
>third color identification with anti-B7-1 that shows a bright stain is a
>sure identification of DC. These DC also tend to have higher forward
>and side scatter, compared to lymphocytes. You may have to use enzymes
>to get them out cleanly. My suggestions is a collagenase treatment
>(with highly purified collagenase).
>
>Follicular DC are more difficult. We have not tried this yet in flow
>but a good mAb was developed by Maria Kosco (I think she's still at the
>Basil Institute ??). The clone name was 2C11. It identifies FDC
>clearly in tissue section and I believe also in flow.
>
>Good luck.
>
>Denis Snider
>(PS. Please forward this to the flow list, in case anyone is interested.
>I can't access my address book to make the copy right now)
>
>
>
>
>
Dr Leanne Sammels
Department of Microbiology
University of Western Australia
Nedlands WA 6009
Phone: 08 9346 4872
Fax: 08 9346 2912