Specific bacterial inhibition of lymphocyte transformation

259

SPECIFIC BACTERIAL INHIBITION OF LYMPHOCYTE TRANSFORMATION. J.C. Houck and L.M. Part, Virginia Mason Research Center, I000 Seneca St., Seattle, WA 98101

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A number of years ago we reported that Pseudomonas putida and subsequently, a patient-derived Pseudomonas aeruginosa , contained a macromolecule which apparently specifically inhibited lymphocyte transformation in vitro. This inhibitor was effective against both T-cells and B-cells, but not against bone marrow or non-lymphoid cell systems. All the inhibition was concentrated in a molecular weight area on Ultrogel ACA34 at an elutlon point appropriate to a molecular weight of between 80 to 70,000 daltons. Considerable purification of this inhibitor was possible by a) smashing the bacteria with glass beads, and b) extracting the soluble components which were partially clarified by streptomycin sulfate precipitation; c) further purification with ethanol fractlonatlon 30-60% Fraction and d) Ion-exchange c o l ~ chromatography (DEAESephacel) was performed with the final purification step being elution from Ultrogel. The inhibitor, although not terrlbly stable and partlally destroyed by lymphilization, was active at a concentration of approximately i0 ng/ml in inhibiting two-way MLC of histolncompa~ible mouse splenic lymphocyges in vitro. The still impure, but very active ractlon is not slgnlflcangly toxic when injected in vivo in mice, but will cause a prolongatlon of the survival of histoincompatlble mouse skin grafts in vivo. This project was supported by a grant from NIAID from NIH. IMMUNOMODULATION INDUCED BY GLYCOPROTEIN FRACTIONS ISOLATED FROM KLEBSIELLA PNEUMONIAE C. GRISCELLI, A. DORANDY, J.L. VIRELIZIER, P. SMETS, C. MARCHIANI, R. ZALISZ, B. FOURNET Groupe d'I-,,unologie et Rhumatologie INSERM U 132, Paris and ROUSSEL-UCLAF, Paris

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A glycoproteic extract of Klebsiella pneumoniae serotype 2 (GEK2) and a main glycoproteic soluble fraction (FI) were studied. Beside effacts of GEK2 and FI given orally in animal models on antibody formation, cellular immunity, polymorphonuclear and macrophage functions, GEK2 given orally tu humans was shown to enhance delayed hypersensitivity to antigens and to increase the in ui;~ao proliferative responses to tetanos toxold of leukocytes from previously sensitized subjects. F1 added (O.O1 to I mg) to human leukocyte cultures provoked a decrease of proliferative responses initiated with antigens or allogenic cells (MLR) but induced generation of plasma cells, when used alone, and enhancement of plas ma cell generation induced by Pokeweed mltogen. F1 was also shown to enhance antibody production of IgG but not of IgM classes in an i~ v ~ O system using a polysacharid antigen of Candida albicans. Furthermore, FI increased the specific cytotoxic activity generated during the MLR and the natural killer (NK) activity. First trials of therapy using GEK2 and FI will be presented. CLONED T CELLS MONOCYTOGENES: S.H.E.

Kaufmann

WITH SPECIFICITY TO THE INTRACELLULAR BACTERIUM B I O L O G I C A L F U N C T I O N S IN V I T R O A N D IN V I V O . and

H.

LISTERIA

Hahn

Institute of Medical Microbiology, Free University, Basel Institute for Immunology, Basel, Switzerland

Berlin,

F.R.G.

and

P e r i t o n e a l e x u d a t e T l y m p h o c y t e s f r o m L. m o n o c y t o g e n e s - i m m u n i z e d m i c e were c l o n e d in d o u b l e l a y e r s o f t a g a r in t h e p r e s e n c e of a c c e s s o r y c e l l s a n d listerial antigen. Colony-derlved T c e l l s w e r e p i c k e d a n d e x p a n d e d in m e dium containing TCGF, accessory cells and antigen. These T cell lines exe r t e d L. m o n o c y t o g e n e s - s p e c i f i c proliferative responses, interleukin ind u c t i o n a s w e l l a s b y - s t a n d e r h e l p f o r B c e l l s in v i t r o . In v i v o t h e T c e l l l i n e s w e r e c a p a b l e of c o n f e r r i n g d e l a y e d - t y p e h y p e r s e n s i t i v i t y to sol u b l e l i s t e r l a l a n t i g e n a n d p r o t e c t i o n t o l l v e L. m o n o c y t o g e n e s upon syngeneic recipient mice. The biological activities were restricted by the H - 2 I A - l o c u s o f t h e M H C . L. m o n o c y t o g e n e s - s p e c i f i c T cells, which had been r e c l o n e d b y l i m i t i n g d i l u t i o n o r in d o u b l e l a y e r s o f t a g a r e x e r t e d an i d e n t i c a l f u n c t i o n a l s p e c t r u m . It is c o n c l u d e d t h a t a s i n g l e T c e l l p o p u l a t i o n is c a p a b l e o f m e d i a t i n g d i f f e r e n t b i o l o g i c a l a c t i v i t i e s o f a c q u i r e d cellular immunity. S u p p o r t e d in p a r t b y D F G g r a n t Ka 5 7 3 - I / I

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