Summary To elucidate the mechanism of E-roscilc formation between T cells and sheep red blood cx-lls{SRBC). the effect of variousagentsaifectingihLM'unctionorcytoplasmicstruclures(microtLibules and microtilaments) and organdies (mitocliondria and rough endoplasniic reliculum) was invcstieatcd. E-rosciie formation was greath inhibited by agents that block cither the energy producmg systcnr(KCN and NaNilur the integration ot tiiicrofilaments (cytochalasin B. dihydrocytochalasin B_ ;nui cytochalasin D). On ihe other hand, theic was little or no suppression by either inhihitors ot proici'n synthesis (cytlolieximide and puromycin). agenls that block Ihe polymeti^aiion ol microtubules (coldiicine. podophyllotoxin and vinblastinc). or chemicals disconnecting surface membrane proteins from the iniracellular strLietural proloiiis (chlnrpromazine. ^dibucaine. liydroccrtlsonc. and propranolol). The calcium ionophorc A23187. which transports Cn-' into ihe cyiosol. inhibited the F-rosette formation iiUhe presence of Ca-\ but nol in its absence. I-rum these results, we concluded thai new synthesis of ATE' and the struciural inlegration o\' niicrofilaments arc indispensable for the E-rosette formation, whieh is triggered by an interaetion between the Iigand (Tl ITS) and ils corresponding receptor (CD2). A certain levdorintraeelluIarCa-^ is also involved m the E-rosctte formalion.lMRODUCTION Thymus-derived (T) lymphocytes spontaneously form E-rosctte aggregates with sheep red blood cells (SRBC). and serve as a surface marker for the earliest stage of the expression of hLiman T-Iincage cells (1-3). E-roscttes are widely used in clinical immunology lo identify or to separate T eells (4-6). although their precise intracellular mechanism is unknown.Receni studies on E-rosette formation have shown that it is dependent upon a specilic interaction between the CD2 moleeule on T eells (7-9) and the complementary structure on SRBC termed the Tl I target structure (Tl ITS: iO.ll). The TllTS has been bioehemieally characterized and shown to be a 42 kDa glycoprotcin (10.11). CD2 binds LFA-3. a 55-70 kDa moleeule expressed on human endothelial. epithelial and connective tissue cells in most tissues, as well as human T and B lymphocytes, granuloeytes. and erythrocytes (12).On the other hand, little is known about the participation of eytopiasmie structures and organelles during E-rosette formation, although several reports suggest they probably play an important role. A study of the mechanism of capping of the surface immunoglobulins(slg) on B cells by corresponding anlibodies revealed that this phenomenon was suppressed by corticosteroids maintaining fluidity of membranes. the calcium ionophore A23187 causing Ca-**" tlux. and reagents aetingon microtilaments such as eytochalasin B (13). Another stud\ of lymphocyte adhesion to eultured Feycr's patch high endothelial venule reported that the adhesion oflymphoeytes was suppressed by NaN? and cytoehalasin L) (14). Moreover, the inhibitory effe-n of eytochalasin B and NaNi on Erosette tbrmation h...