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Modulation of macrophage (MØ) activities by tuftsin and two formylated analogs
W S 1 6 - 6 *s ACTIVATION OF PLASMA MEMBRANE PHOSPHOLIPID METABOLISM IS A NESESSARY EARLY SIGNAL FOR INTERLEUKIN-2 PRODUCTION IN HUMAN LYMPHOCYTES K.Resch, B.Krebs,M.Kracht,R.Kurrle*,B.Rehermann, M.Golombek, M.SchmidL and M.Szamel Institute of Molecular Pharmacology, Department of Pharmacology and Toxicology, Medical School, Hannover, *Behringwerke Ag,Marburg ,FRG Activation of human peripheral blood lymphocytes by anti-CD3 or anti- TCR-CD3 monoclonal antibodies led to interleukin-2 (IL-2) production, receptor expression and proliferation. Similarly, cells stimulated with the phorbol ester (TPA) and ionomycin produced IL-2 and expressed high affinity receptors. In contrast lymphocytes that were activated by dioctanoylglycerol (diCS) and ionomycin expressed high-affinity IL-2 receptors, however were not able to synthetise IL2. Addition of polyunsaturated fatty acids, like linoleic acid to diCS-ionomycin treated cells led to IL-2 production and proliferation of lymphocytes. Phospholipase A2 was activated tree fold by the monoclonal antibodies and by TPA, activation of the enzyme was measurable up to 4 hours. In contrast,in diC8 treated cells a maximal activation of phospholipase A2 was achieved after 30 min. Lysophosphatid acyltransferase, the enzyme specifically incorporating polyunsaturated fatty acids into plasma membrane phospholipids was also activated by the monoclonal antibodies and TPA, activation was detectable even after 4 hours of stimulation. These results suggest that long Lerm activation of the phospholipid metabolising enzymes and thus elevated amounts of polyunsaturated fatty acids in plasma membrane phospholipids are necessary early signals for IL-2 production and proliferation in human lymphocytes.
W S 1 6 - 7 *7 I. FLOR[NTIN (1), V. CHUNG (2), d. MNTTINEZ (2), J. HkRAL (3), H. TiS~T (4), JP. 8ROUD (4), 0. HATIIE (I) (1) I.C.l.O., UA CNRS 1163, H~ttal Paul Brousse, VlZZejuK (2) Centre de ~ ~ l o g t e , Montp~nwr, (z) ~ d ~ ~,d~, ~p~al d~ ~ S J k ~ r * , PARm, (4) UA CM~S 595, ~ t ~ l Cochin, PkRIS, FRANCE. Tuflsin (71v"-Lus-Pro-Jv'g) ~; a natural I ~ aofivator w h ~ aots throu~ binding to s ~ receptors. Studu~ the lmmunologfc ¢onsequerces ofa ~ ) e i.v. ~.k,ctk~n of tuft.tin to mice we demote'cared tl~t It mod~wd all I ~ funotim~ examk'wd. It amplified p ~ t o s i s - i n d u o e d dweMlumineso~noe, oVto~ztio acttv~, 'mterleuld~l pro4uctton and slimubted Pt~E2release. Tlwse effects peakedon day 7 and paralleled an inhibition of efftotor and r~)ui,~or V funofims of 19mp~c~tes. Formg¼t~l mabgs : f ~ a.d (f--I~t~)-'oJfl~ vwe sunthet~z~. T I ~ s~ht~u d ~ c ~ l t ~ l tuflsin ~ ~ t th~ ,~lUred some affinit 9 for receptors to for'rn~jl~tedpeptkJes. ~ the~" effeots to tuftsin on I'~, we o~J not observed nefther Qualitathre nor great quint#afire d~t"erenoes e x i t on PGE2 proekJet~ which ~ r n ~ bs$ affected b9 the analogs. This fir o0,-,o~-~,it~ntwith ~n abseece of depre-ss~,eeffeo~s on ~mpho~*es. When t u f t ~ or anato~s were admin~ered t w ~ 7 d. apart, PI~ dea¢~vatio~ occurred w~JHn24 h follov~g the 2 d. ~njeetJon.Th~ vas shorn on the ohem~mines~,n~ respeese vhie~ thereafter deveiope4 as after • single ~n.k,~t~. In tut~-~tre~ed mk~e POE2 pro4uetion v ~ also abre~ated and th~ parageled nm-n~HzaHon of IU.mphoovte fue~'ff~. The de~efiv4~on phenomef~e~did aot oootrred v ~ h 2 ~jeotiens 14 d . . , ~ aHhough I'~ were still st~mubted ~ the time of the 2rid injection.
WS16-8
*8
PITUITARY EXTRACTS CONTAIN A FACTOR WHICH INDUCES THYMIC EPITHELIAL CELL PROLIFERATION AND SECRETION J.W. Hadden, A. Galy, H. Chen, C. Dinarello, M. Jolivet, and E.M. Hadden Program of Immunopharmacology, University of South Florida Medical College, Tampa, Florida, U.S.A. A close link between pituitary and thymus has been suggested by many studies. In search of a putative "thymotrophic" hormone we examined the effect of bovine pituitary extracts (BPE) on human thymic epithelial (HUTE) cells in vitro. BPE (5-50 ug/ml) induced thymidine incorporation and nuclear labelling of the majority of Keratin-positive, esterasenegative HUTE cells. The thymidine incorporation and degree of nuclear labelling with BPE compared to the effects of i-I0 ng/ml of fibroblast and epidermal growth factors (FGF and EGF). Purified human growth hormone, prolactin, ACTH, TSH, LH, FSH, insulin, testosterone, estradiol, and MSA II were inactive to stimulate proliferation of HUTE cells. The effect of BPE was not influenced by hydrocortisone, indomethacin, or endotoxin. In a preliminary experiment BPE stimulated ~ interleukin I (ILI~) production by these cells. Preliminary characterization indicates the factor is > i0,000 daltons, preciptitates between 50-80% ammonium sulphate, and is labile < pH 4.5. Chromatographic separations are in progress. It remains to be determined whether the pituitary factor represents FGF or a new hormone which may link the pituitary and thymus in an endocrine feedback network comparable to the adrenal.
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W S 1 6 - 7 *7 I. FLOR[NTIN (1), V. CHUNG (2), d. MNTTINEZ (2), J. HkRAL (3), H. TiS~T (4), JP. 8ROUD (4), 0. HATIIE (I) (1) I.C.l.O., UA CNRS 1163, H~ttal Paul Brousse, VlZZejuK (2) Centre de ~ ~ l o g t e , Montp~nwr, (z) ~ d ~ ~,d~, ~p~al d~ ~ S J k ~ r * , PARm, (4) UA CM~S 595, ~ t ~ l Cochin, PkRIS, FRANCE. Tuflsin (71v"-Lus-Pro-Jv'g) ~; a natural I ~ aofivator w h ~ aots throu~ binding to s ~ receptors. Studu~ the lmmunologfc ¢onsequerces ofa ~ ) e i.v. ~.k,ctk~n of tuft.tin to mice we demote'cared tl~t It mod~wd all I ~ funotim~ examk'wd. It amplified p ~ t o s i s - i n d u o e d dweMlumineso~noe, oVto~ztio acttv~, 'mterleuld~l pro4uctton and slimubted Pt~E2release. Tlwse effects peakedon day 7 and paralleled an inhibition of efftotor and r~)ui,~or V funofims of 19mp~c~tes. Formg¼t~l mabgs : f ~ a.d (f--I~t~)-'oJfl~ vwe sunthet~z~. T I ~ s~ht~u d ~ c ~ l t ~ l tuflsin ~ ~ t th~ ,~lUred some affinit 9 for receptors to for'rn~jl~tedpeptkJes. ~ the~" effeots to tuftsin on I'~, we o~J not observed nefther Qualitathre nor great quint#afire d~t"erenoes e x i t on PGE2 proekJet~ which ~ r n ~ bs$ affected b9 the analogs. This fir o0,-,o~-~,it~ntwith ~n abseece of depre-ss~,eeffeo~s on ~mpho~*es. When t u f t ~ or anato~s were admin~ered t w ~ 7 d. apart, PI~ dea¢~vatio~ occurred w~JHn24 h follov~g the 2 d. ~njeetJon.Th~ vas shorn on the ohem~mines~,n~ respeese vhie~ thereafter deveiope4 as after • single ~n.k,~t~. In tut~-~tre~ed mk~e POE2 pro4uetion v ~ also abre~ated and th~ parageled nm-n~HzaHon of IU.mphoovte fue~'ff~. The de~efiv4~on phenomef~e~did aot oootrred v ~ h 2 ~jeotiens 14 d . . , ~ aHhough I'~ were still st~mubted ~ the time of the 2rid injection.
WS16-8
*8
PITUITARY EXTRACTS CONTAIN A FACTOR WHICH INDUCES THYMIC EPITHELIAL CELL PROLIFERATION AND SECRETION J.W. Hadden, A. Galy, H. Chen, C. Dinarello, M. Jolivet, and E.M. Hadden Program of Immunopharmacology, University of South Florida Medical College, Tampa, Florida, U.S.A. A close link between pituitary and thymus has been suggested by many studies. In search of a putative "thymotrophic" hormone we examined the effect of bovine pituitary extracts (BPE) on human thymic epithelial (HUTE) cells in vitro. BPE (5-50 ug/ml) induced thymidine incorporation and nuclear labelling of the majority of Keratin-positive, esterasenegative HUTE cells. The thymidine incorporation and degree of nuclear labelling with BPE compared to the effects of i-I0 ng/ml of fibroblast and epidermal growth factors (FGF and EGF). Purified human growth hormone, prolactin, ACTH, TSH, LH, FSH, insulin, testosterone, estradiol, and MSA II were inactive to stimulate proliferation of HUTE cells. The effect of BPE was not influenced by hydrocortisone, indomethacin, or endotoxin. In a preliminary experiment BPE stimulated ~ interleukin I (ILI~) production by these cells. Preliminary characterization indicates the factor is > i0,000 daltons, preciptitates between 50-80% ammonium sulphate, and is labile < pH 4.5. Chromatographic separations are in progress. It remains to be determined whether the pituitary factor represents FGF or a new hormone which may link the pituitary and thymus in an endocrine feedback network comparable to the adrenal.
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