>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
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