A methodological study of E-rosette formation using AET-treated sheep red blood cells.
The influence of some of the well knwon technical variables on the E-rosette technique was examined using sheep red blood cells (SRBC) treated with 2-aminoethylisothiouronium bromide (AET). With AET treatment, E-rosette formation becomes less dependent on time and temperature and on the presence of serum. The mechanical stability of the rosettes is enhanced, and the number of SRBC attached to each rosette-forming lymphocyte (RFC) is markedly increased, leading to a sharper distinction between RFC and non-RFC. Ultimately, significantly more E-receptor carrying lymphocytes become detectable. Evidence is provided that the specificity of the E-rosette technique is unchanged after AET treatment of SRBC, in spite of the enhanced binding. A simple and reliable method for E-RFC identification is documented.
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002417:Cattle; D002498:Centrifugation; D004905:Erythrocyte Aging; D004912:Erythrocytes / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D008214:Lymphocytes / Q000276:immunology; D012397:Rosette Formation / Q000379:methods*; D012756:Sheep; D013696:Temperature; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology; D000338:beta-Aminoethyl Isothiourea / Q000494:pharmacology*
D007136:Immunoglobulins; D000338:beta-Aminoethyl Isothiourea; D014357:Trypsin