In article <ferbas.5.00122E4A at cyto.medsch.ucla.edu>, ferbas at cyto.medsch.ucla.edu (John Ferbas) writes:
>I have used an anti-MHC class I antibody (W632) in an attempt to demonstrate
>MHC class I restriction in 2 CD8+ CTL clones, but have had no success. I know
>that the antibody is saturating the targets by FACS analysis. Also, the
>antibody is added before the assay and present throughout. Is there any
>alternative way to demonstrate class I restriction? Your comments are greatly
>appreciated.
>===========================================================================================
>John Ferbas Ph.D. Ferbas at cyto.medsch.ucla.edu>UCLA MED/Cellular Immunology & Cytometry Voice: (310) 825 6340
>12-236 Factor Bldg. FAX: (310) 794 2145
>10833 Le Conte Ave.
>Los Angeles CA 90024-1745
Dear John,
I am a graduate student working on HLA class I structure. Is it
possibe that W6/32 is not blocking your CTL epitope? Is your CTL response
Allo or syngeneic? It is possible that an allo CTL is responding to
another class I epitope unrelated to the W6/32 epitope and may not be
sterically hindered. Why not try some other monomorphic determinant
monoclonals such as TP 25.99 or some of the B2m specific monoclonals such
as BBM 1 or L368.
hope this was helpful.
CPT
