1. 1 A n al ysis of I F N - γ pr o d ucti o n b y P B M C. T he n u m ber of I F N -γ pr o d uci n g cells will be e val uate d usi n g E LI S P O T assa ys as des cri be d earlier 3 9. Bri efl y, 2 x 1 0 5 m o n o n uclear cells o btai ne d fr o m t he peri p her al bl o o d of patie nts will be plate d i n q ua dr u plicates i n 9 6 -well m ultiscree n H A filtrati o n plates ( Milli p ore, Be df or d, M A) pre -c o ate d wit h m o use a nti -h u m a n I F N  m o n ocl o nal a nti b o d y ( M A B 2 8 5, R & D S yste m, Mi n nea p olis, M N) a n d i nc u bate d f or 3 6 hr at 3 7 o C wit h A L V A C -p 5 3, or A L V A C -c a n d 2 n g/ ml I L -2. T otal v ol u me is 2 0 0 l. A fter i nc u bati o n, t he wells will be was he d a n d t he n i nc u bate d o ver ni g ht at 4 o C wit h a bi oti n ylate d g oat a nti -h u ma n I F N -γ a nti b o d y ( B A F 2 8 5, R & D S yste m), f oll o we d b y i nc u bati o n wit h a vi di n -al kali ne p h os p hatase ( Si g ma). T he wells will t he n be was he d a n d s p ots will be vis ualize d wit h B CI P/ N B T s u bstrate ( Si g ma). T he plates will be air -drie d a n d t he c ol ore d s p ots c o u nte d usi n g C. T.