At 22:39 -0000 8.8.2000, Lisa Whyte wrote: >We are using a 4-color mix of CD45/CD3/CD4/CD8 to determine T cell subset >ratios. However our CD4 + CD8 percentages (gated on the CD45+ CD3+ cells) >do not add up to 100%. Has anyone else had these problems or have any >suggestions? Help would be appreciated - thanks! >Lisa Whyte Dear Lisa, I don't think it is a problem (well, it depends how big the difference is). The remaining cells could be 1. TCRgamma/delta positive and 2. the "true" double negatives (I mean TCRalpha/beta positive CD4neg CD8neg). The percentage of TCRgamma/delta+ should be 1-10% of CD3+ cells in normal healthy individuals. TCRalpha/beta+ CD4-8- in peripheral blood are usually pathological and may be increased in the autoimmunity-related lymphoproliferation syndrome. They should be assessed directly (by having TCRalpha/beta and CD4 and CD8 in the same tube) not by calculating. Ondrej Ondrej Hrusak, M.D., Ph.D. Institute of Immunology Ondrej.Hrusak@lfmotol.cuni.cz fax +4202 2443 5962 http://www.lf2.cuni.cz/clip/ phone +4202 2443 5969, +4202 2443 2084 postal address: V uvalu 84 150 06 Praha 5 Czech Republic
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