- Email: [email protected]
Does migration inhibitory factor (MIF) act by promoting tubulin polymerization?
\\‘ORl~SIIOP
ON
LYhIPIIOCYTE
MEDIATORS
PI!)
licdlh. ‘l‘l~cw fractions \z crc illrubatctl wit11 10 ’ to 10 ” .lI tliisol~rq,yl l~l~o~l)lI~~fllic~ritl:ltc (1 )I 1’1;) at 37°C for 30 min. dialyzed, and thrn assayctl on their respective human intlicator cells : monucytcs ( hl Nl,) a~ltl ~~~~lynlc)~-l~l~olluclcal- Iculic~ylcs (PA1 KI,). RIIF activity \vas not diminihhc(l following trcatmcnt \vitli DIPF (10 a ill). In contrast, LIF activity \vas abolished by DIPI; (lo-” to 10-j 111) concentrations lvhich inhibit scrinc esterases. The inactivation of LIF by DIPF was dose, time, pH, and temperature dependent. In a dose-dependent fashion another esterase inhibitor, benzamidinc, also abolished LIF activity. Moreover, LIF but not MIT; preparations contained esterolytic activity for certain substrates, such as RAEE. These findings resolve the previous conflicting observations and su,,O”est that LIF, but not MIF, is an csterase. (Supported by USPHS Grant M-11729.)
Does dligvnfim I~lhibitor~l Ftrcfor (MIF) PICK, SHOSIIANA HONK, CENJAXIN University, Tel-Aviv, Irsael.
Act br GRIFFEL,
Prolrlofitcg Tulmlis PolJwzcrixtiorfB EDGAR AND ASNA GI~U~SPAN-~\VIRSI<\‘, Tel-Aviv
The following methods \vcre used to investigate the state of microtubules (AIT) in guinea pig macrophages exposed to MIF: (a) quantitation of total tubulin (TU) by [“Hlcolchicine of MT by binding; (b) assessment of the polymerized/total TU ratio; and (c) visualization immunofluorescence, using antibody against purified brain TU. It was found that: (1) MIF does not modify the amount of total TU. (2) Exposure of macrophages to MIF for 16 hr results in an increase in the amount of polymer (15~200% of the level in control cells). (3) Anti-TU antibody stains a complex array of MT. The percentage of cells with well-organized MT and active paranuclear MT generating centers was augmented by exposure to MIT;. (4) The MT stabilizing agent, deuterium oxide, inhibits macrophage migration. The enhanced TU polymerization is probably the reason for the unresponsiveness of MIF-treated macrophages to adenylate cyclase stimulators. MT disrupting drugs (colchicinc, vinblastine) prevent MIF action and also greatly enhance cyclic AMP generation in response to adenylate cyclase stimulators. We conclude that promotion of TU polymerization is an essential step in the action mechanism of IQIF. The way in which hlIF induces TU polymerization is unknown but might involve cyclic GhP. (Supported by KIH grant AI-11194 and by a grant from the U.S.-Israel Binational Science Foundation.) The Effect ROCKLIN,
of Histaurise on in Viiro A9acrophage I+zhibitovy Factor (NIF) Production. R. E. Harvard Medical School and Robert B. Brigham Hospital, Boston, Massachusetts.
There are a number of cell functions lvhich are modulated by histamine including basophil histamine release, antibody production, lysosomal enzyme release by neutrophils, and lymphocytemediated cytotoxicity. Recently, it has been shown that histamine suppresses the size of delayed hypersensitivity skin tests by 50w,O and inhibits in vitro MIF production and proliferation (Rocklin, R. E., 1. C/ix. I wz~st. 57, 1051, 1976). Studies have been carried out to analyze the inhibitory effect of histamine on antigen-induced MIF production. The histamine effect is reversible and acts at an early stage in antigen-lymphocyte interaction. H, receptor antagonists (burimamide) prevent the suppressive effects of histamine but not II1 receptor blockers (chlorpheniramine). Histamine does not prevent the macrophage response to preformed MIF, the binding of antigen to macrophages or the ability of lymphocytes to form nonspecific or antigen-specific rosettes with macrophagcs. Passage of immune lymph node cells over affinity columns containing bound histamine (prepared by Ken hlclmon) deletes a population which is responsive to histamine. The passed cells are able to make MIF in the presence of histamine. Thus, Ha receptor bearing lymphocytes may modulate hIIF production and ultimately the expression of cellular-immune reactions. (Supported by USPHS Grant AI-11729.) Inhibition of Cellular DNA Spthcsir Assoriatcd with 01 Activify. S.-C. LEE AND Z. J. LUCM, Stanford California.
a Factor Iulribitixg University Medical
DNA Polymerasc Center, Stanford,
The chemical nature of an inhibitor of DNA synthesis (IDS) in supernatants of PHAactivated human tonsil cells was studied. IDS reduces [‘Hjthymidine incorporation into PHAstimulated lymphocytes and into growing HeLa cells without being cytotoxic and is different from both a- and p-lymphotoxins. IDS, with a molecular weight of about 80,000, inhibits calf
ON
LYhIPIIOCYTE
MEDIATORS
PI!)
licdlh. ‘l‘l~cw fractions \z crc illrubatctl wit11 10 ’ to 10 ” .lI tliisol~rq,yl l~l~o~l)lI~~fllic~ritl:ltc (1 )I 1’1;) at 37°C for 30 min. dialyzed, and thrn assayctl on their respective human intlicator cells : monucytcs ( hl Nl,) a~ltl ~~~~lynlc)~-l~l~olluclcal- Iculic~ylcs (PA1 KI,). RIIF activity \vas not diminihhc(l following trcatmcnt \vitli DIPF (10 a ill). In contrast, LIF activity \vas abolished by DIPI; (lo-” to 10-j 111) concentrations lvhich inhibit scrinc esterases. The inactivation of LIF by DIPF was dose, time, pH, and temperature dependent. In a dose-dependent fashion another esterase inhibitor, benzamidinc, also abolished LIF activity. Moreover, LIF but not MIT; preparations contained esterolytic activity for certain substrates, such as RAEE. These findings resolve the previous conflicting observations and su,,O”est that LIF, but not MIF, is an csterase. (Supported by USPHS Grant M-11729.)
Does dligvnfim I~lhibitor~l Ftrcfor (MIF) PICK, SHOSIIANA HONK, CENJAXIN University, Tel-Aviv, Irsael.
Act br GRIFFEL,
Prolrlofitcg Tulmlis PolJwzcrixtiorfB EDGAR AND ASNA GI~U~SPAN-~\VIRSI<\‘, Tel-Aviv
The following methods \vcre used to investigate the state of microtubules (AIT) in guinea pig macrophages exposed to MIF: (a) quantitation of total tubulin (TU) by [“Hlcolchicine of MT by binding; (b) assessment of the polymerized/total TU ratio; and (c) visualization immunofluorescence, using antibody against purified brain TU. It was found that: (1) MIF does not modify the amount of total TU. (2) Exposure of macrophages to MIF for 16 hr results in an increase in the amount of polymer (15~200% of the level in control cells). (3) Anti-TU antibody stains a complex array of MT. The percentage of cells with well-organized MT and active paranuclear MT generating centers was augmented by exposure to MIT;. (4) The MT stabilizing agent, deuterium oxide, inhibits macrophage migration. The enhanced TU polymerization is probably the reason for the unresponsiveness of MIF-treated macrophages to adenylate cyclase stimulators. MT disrupting drugs (colchicinc, vinblastine) prevent MIF action and also greatly enhance cyclic AMP generation in response to adenylate cyclase stimulators. We conclude that promotion of TU polymerization is an essential step in the action mechanism of IQIF. The way in which hlIF induces TU polymerization is unknown but might involve cyclic GhP. (Supported by KIH grant AI-11194 and by a grant from the U.S.-Israel Binational Science Foundation.) The Effect ROCKLIN,
of Histaurise on in Viiro A9acrophage I+zhibitovy Factor (NIF) Production. R. E. Harvard Medical School and Robert B. Brigham Hospital, Boston, Massachusetts.
There are a number of cell functions lvhich are modulated by histamine including basophil histamine release, antibody production, lysosomal enzyme release by neutrophils, and lymphocytemediated cytotoxicity. Recently, it has been shown that histamine suppresses the size of delayed hypersensitivity skin tests by 50w,O and inhibits in vitro MIF production and proliferation (Rocklin, R. E., 1. C/ix. I wz~st. 57, 1051, 1976). Studies have been carried out to analyze the inhibitory effect of histamine on antigen-induced MIF production. The histamine effect is reversible and acts at an early stage in antigen-lymphocyte interaction. H, receptor antagonists (burimamide) prevent the suppressive effects of histamine but not II1 receptor blockers (chlorpheniramine). Histamine does not prevent the macrophage response to preformed MIF, the binding of antigen to macrophages or the ability of lymphocytes to form nonspecific or antigen-specific rosettes with macrophagcs. Passage of immune lymph node cells over affinity columns containing bound histamine (prepared by Ken hlclmon) deletes a population which is responsive to histamine. The passed cells are able to make MIF in the presence of histamine. Thus, Ha receptor bearing lymphocytes may modulate hIIF production and ultimately the expression of cellular-immune reactions. (Supported by USPHS Grant AI-11729.) Inhibition of Cellular DNA Spthcsir Assoriatcd with 01 Activify. S.-C. LEE AND Z. J. LUCM, Stanford California.
a Factor Iulribitixg University Medical
DNA Polymerasc Center, Stanford,
The chemical nature of an inhibitor of DNA synthesis (IDS) in supernatants of PHAactivated human tonsil cells was studied. IDS reduces [‘Hjthymidine incorporation into PHAstimulated lymphocytes and into growing HeLa cells without being cytotoxic and is different from both a- and p-lymphotoxins. IDS, with a molecular weight of about 80,000, inhibits calf