journal of biotechnology lecture

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J. gen. ViroL 1981), 57, 233-237. P r i n t e d i n G r e a t B r i t a i n

Key words: I F N c t su b t y p e s / a n t i v i r a l a c t i v i ty / c l o n e d

233

Co m parison o f the Ant iv ira l Act iv it ie s o f Var ious Cloned H um an

Interferon a Subtypes in Mam m alian Cel l Cultures

A c c e p t e d 1 5 J u l y 1 9 8 1 )

SUMMARY

Five human in t e r f e ron -a ( l eukocy t e ) sub types de r ived f rom genes c loned i n

E s c h e r i c h i a c o i l have been compared fo r t he i r ab i l i t y t o i nduce an t i v i r a l a c t i v i t y

aga ins t ves icu la r s tom at i t i s v irus infec t ion of var ious m am m al ia n cel l cu l tures . These

in te r fe rons , des igna ted L eIF -A ( IFN-ct2) , -B, -C, -D ( IFN-a~) an d Le IF- F, s how

di f fe r en t r e la t ive ac t i vi ti e s whe n a s sayed on hum an ; bov ine , hams te r , mouse , r abb i t

and m on key cell lines . As w i th a na tu r a l hum an bu f fy - coa t i n t e r f e ron -a p r epa ra t i on ,

t h r ee sub types (Le IF -B , -C a nd -D) showed cons ide r ab l e ac t i v it y on R K-13 r abb i t

cel ls , bu t two (Le IF -D and -F ) a lso showed some ac t i v it y on m ouse L -929 cel ls . Of

the f i ve i n t e r f e ron sub types examined , Le IF -F demons t r a t ed t he h ighes t deg ree o f

species sp ecif ici ty.

P repa ra t i ons o f human in t e r f e ron -a ( l eukocy t e ) have shown an t i v i r a l a c t i v i t y i n va r i ous

ma mm al i an ce l l l ine s and t h i s ha s been pos tu l a t ed t o a r is e f rom the p r e sence o f d is t i nc t

mo lecu l a r sub types o f i n te r f e ron (Gre s se r

e t a l . ,

1974 ; S t ewar t & Desmy te r , 1975 ; De sm yte r

& Stewar t , 1976; Rubins te in

e t a l . ,

1981) Human in t e r f e ron -a p r epa ra t i ons gene ra l l y show

some molecu l a r he t e rogene i t y w i th mo s t a c t i v i t y a t a m o l . w t. o f abou t 18 000 and some a t a

mo l . w t . o f 22 000 , wh i l e a ll a c t i v i ty was foun d t o have a mo l . w t. o f 18 000 a f t e r t r e a tm en t w i th

glycos idases (Br idgen

e t a l . ,

1977; Otto

e t a l . ,

1980). T hus , g lycosy la t ion , a l tho ugh poss ib le

fo r a f r ac t i on o f t he na tu r a l ma t e r i a ls , wou ld no t s eem to be e s sen ti a l f o r an t i v i ra l a c t iv i t y o f

hum an in t e r f e ron -a and t he p ro t e in mo ie ty wou ld s eem to be e s sen ti a ll y hom ogen eous w i th

r ega rd t o mo l . w t .

C lon ing o f in t e r f e rons i n bac t e r i a by r ecom binan t D N A t echn iques ha s r evea l ed t ha t t he

hum an in t e r f e rons - a a re a f ami ly o f re l a t ed p ro t e in s o f 165 o r 166 am ino ac id r e s idues

(Goedde l

e t a l . ,

1980, 1981; Nagata

e t a l . ,

1980a , b ; Yelver ton

e t a l . ,

1981). In the present

s t udy w e have com pared t he i nduc t i on o f an t i v ir a l a c t iv i t y i n va r ious m am ma l i an ce ll cu l tu r e s

by f ive d if f e ren t hum an in t e r f e ron -a sub types ob t a ined by c lon ing p la smids con t a in ing hum an

in ter fe ron-a genes in E s c h e r i c h i a c o l i . Th e am ino ac id sequences of the f ive in te r fe rons , a l l

de r ived by c lon ing f rom m R N A , d i f f e r i n 18 t o 32 pos i t i ons and a r e t e rmed Le lF -A , -B, -C ,

-D , -F . Le lF -E i s a s imi l a r i n t e r f e ron gene , a l t hough i t c anno t be exp re s sed because i t

con t a in s s eve ra l s t op codons (Goedde l

e t a l . ,

1981) . The Le lF -A an d -D o f Goed de l

e t a l .

(1981) co r r e spo nd t o I FN -a 2 and IFN-a~ r e spec ti ve ly , o f Nag a t a

e t a l .

(1980b) . The resu l t s

r epo r t ed he r e i nd i ca t e t ha t t he f ive mo lecu l a r sub types o f hum an in t e r f e ron -a s t ud ied have

dis t inc t ly d i f fe rent capabi l i t i es of inducing an t iv i ra l ac t iv i t i es in var ious mammal ian ce l l

cu l tures .

Co n t inuou s cel l cu l t u re s o f hum an am nion cells (W ISH) , m ouse f i b rob l a s ts (L -929 ) and

bov ine k idney ce l l s (MDBK) were ob t a ined f rom the Amer i can Type Cu l tu r e Co l l ec t i on ,

Rockv i l le , Md . H eL a ce ll s, m onk ey k idney ce l ls (Ve ro) , hams t e r c e lls (BH K-21) and r abb i t

k idney ce l l s (RK-13) we re pu rchased f rom F low Labora to r i e s . P r imary r abb i t k idney ce l l s

we re ob t a ined f rom M A Biop roduc t s , W a lke rsv i ll e , Md . , U .S .A . A l l ce lls w e re ma in t a ined i n

Eag l e ' s m in ima l e s sen t ia l med ium and 5 % foe t a l c a l f s e rum f rom G ran d I s l and B io log i ca l s,

0022-1317/81/0000-4729 02.00 © 1981 SGM



234 S h o r t c o m m u n i c a t i o n s

Table 1. R e l a t i v e a n t i v i r a l a c t i v it ie s o f c l o n e d a n d n a t u r a l h u m a n i n te r f e ro n s o n d i ff e r e n t

ce l l s

In t e r f e ron - ct

•

C e l l s A B C D F B u f f y - c o a t

W I S H 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0

H e L a 3 3 2 2 5 6 3 1 6 5 0

M D B K 3 1 6 1 0 0 0 3 9 8 6 3 0 0 2 0 0 1 5 85

B H K 2 0 . 3 5 0 6 3 0 . 3 3 2

L - 9 2 9 0 . 5 1 - 0 0 . 8 5 5 0 . 2

P r i m a r y r a b b i t k i d n e y 0 . 3 5 . 0 8 . 0 6 3 - 8 0

R K - 1 3 2 1 5 8 1 0 0 1 5 8 0 . 1 1 5 8

V e r o 1 6 6 3 6 3 1 0 0 0 . 8 2 5 1

* D a t a a r e n o r m a l i z e d t o t h e a n t i v i r a l a c ti v i ty o f th e i n t e r f e ro n p r e p a r a t i o n s o n W I S H c e ll s ch a l l e n g e d w i th

V S V . T h e l og ~0 d i lu t io n g i v in g 5 0 p l a q u e r e d u c t i o n o n W I S H c e ll s w a s : L e I F - A , 6 . 0 ; - B , 4 . 5 ; - C , 4 . 6 ; - D , 5 . 2 ;

F , 4 - 5 ; b u f f y - c o a t , 5 . 8 .

G r a n d I s l a n d , N e w Y o r k , U . S . A . V e s i c u l a r s t o ma t i t i s v i r u s ( V S V ) I n d i a n a s t r a i n , w a s

p u r c h a s e d f r o m t h e A me r i c a n T y p e C u l t u r e C o l l e c t i o n a n d p a s s a g e d i n B H K - 2 1 c e l l s . T h e

c o n s t r u c t i o n o f b a c t e r i a l p l a s mi d s c o n t a i n i n g t h e v a r i o u s h u ma n l e u k o c y t e i n t e r f e r o n g e n e s

a n d p r e p a r a t i o n o f i n t e r f e r o n e x t r a c t s f r o m b a c t e r i a l c e l l s h a v e b e e n d e s c r i b e d p r e v i o u s l y

( G o e d d e l e t a l . , 1980 , 1981) . The superna tan t s genera ted by th i s ex t rac t ion p rocedure were

c o l l e c t e d a n d u s e d i n p l a q u e r e d u c t i o n a s s a y s ( Y e l v e r t o n e t a l . , 1981) o r cy topa th ic e f fec t

i n h i b i t i o n a s s a y s ( C a mp b e l l e t a l . , 1 97 5) u s i n g t h e N a t i o n a l I n s t i tu t e s o f H e a l t h l e u k o c y t e

i n t er f e ro n s t a n d a r d G - 0 2 3 - 9 0 1 - 5 2 7 . T h e s e p r e p a r a t i o n s o f in t e r fe r o n s d e ri v e d f r o m b a c t e r i a

had spec if ic ac t iv i ti e s in a r ange f rom 3 .2 x 104 to 1 .6 x 105 U /m g pro te in and ex t rac t s o f

bac te r ia l ce ll s con ta in ing p lasm ids l ack ing in te r fe ron gene sequen ces had no an t iv i ra l ac t iv i ty .

A l l s a mp l e s f r o m p H 2 t r e a t me n t s , 5 6 ° C t r e a t me n t s a n d a n t i b o d y n e u t r a l i z a ti o n e x p e r i me n t s

w e r e me a s u r e d f o r r e s i d u a l i n t e r f e r o n a c t i v i t y b y c y t o p a t h i c e f f e c t i n h i b i t i o n u s i n g WI S H

c el ls a n d V S V . A n t i b o d y n e u t r a l iz a t i o n o f t h e i n t e r fe r o n p r e p a r a t i o n s w a s d e t e r m i n e d w i t h

p o l y v a l e n t r a b b i t a n t i - L e l F a n d a n t i - F I F a n t i b o d i e s k i n d l y s u p p l i e d b y t h e A n t i v i r a l

S u b s t a n c e s P r o g r a m m e o f t h e N a t i o n a l I n s t it u t e s o f A l le r g y a n d I n f e c t i o u s D i s e a s e s

( N I A I D ) . T h e s e a n t i - F I F a n d a n t i - L e I F a n t i b o d y p r e p a r a t i o n s h a d t i tr e s o f 10 24 a n d

1 0 0 0 0 / ml a f t e r i n c u b a t i o n w i t h 2 0 u n i ts o f th e i r r es p e c ti v e i n te r f e ro n s t a n d a r d s . A n a t u r a l

h u m a n b u f f y - c o a t i n te r fe r o n p r e p a ra t io n w a s o b t a i n e d f r o m D r N o r w o o d H i ll , W a d l e y

Ins t i tu te fo r Molecu la r Medic ine , Da l la s , Texas , U .S .A . Th is na tu ra l ma te r ia l had a spec i f i c

ac t iv i ty o f 1 x 106 U /m g pro te in .

Di f fe ren t cel ls l ines hav e s how n w ide va r ia t ions in re sponse to the a n t iv i ra l e f fec t s o f na tu ra l

in te r fe rons whe n te s ted e i the r by p laqu e redu c t ion o r inh ib i t ion o f cy to pa th ic e ffec t s (S tewar t ,

1979 ; Laza r e t a l . , 1980 ; S tew ar t Hav e l l , 1980). W e exam ined the i n v i t r o a c t i v i ty o f

va r ious c loned in te r fe ron-a sub types in e igh t d i f fe ren t mammal ian ce l l cu l tu res . The resu l t s

p resen ted in Tab le 1 showed tha t a l l the c loned in te r fe ron sub types had h igher ac t iv i ty on

b o v i n e M D B K c el ls t h a n o n h u m a n c ell s a n d t h a t t w o ( L e I F - D a n d - F ) s h o w e d s o m e a c t iv i t y

on mouse L-929 ce l l s . As has been repor ted fo r na tu ra l human- f l ( f ib rob las t ) and -a ( l euko-

c y t e ) i n t e rf e r o n p r e p a r a t i o n s ( D e s m y t e r

e t a l . ,

1970 ; McGi l l

e t a l2 ,

1976) , four o f the c lo ned

in te r fe rons demons t ra ted re la t ive ly h igh an t iv i ra l ac t iv i ty on a con t inuous rabb i t ce l l l ine

R K - 1 3 . I n c o n t r a s t , o n l y L e I F - D a n d t h e n a t u r a l l e u k o c y t e i n t e rf e r o n p r e p a r a t i o n c a u s e d

p r o n o u n c e d p r o t e c t i o n o f p r i ma r y r a b b i t k i d n e y c el ls a g a i n s t V S V i n f e c t i o n . T h e c l o n e d

l e u k o c y t e i n t e r f e r o n s u b t y p e L e I F - F d e mo n s t r a t e d t h e h i g h e s t d e g r e e o f s p e c i e s s p e c i f i c i t y

of a l l the in te r fe rons t e s ted . Thus , i t appea rs tha t the c loned human in te r fe ron sub types d i f fe r



S h o r t c o m m u n i c a t io n s

2 3 5

T ab l e 2 .

I n h i b i ti o n o f E M C v i r u s o r V S V p la q u e f o r m a t i o n i n V e ro c e ll s f o l l o w i n g t r e a t m e n t

with in ter feron-a

Interferon-c~ EMC virus (U/m l) VSV (U/m l)

LelF-A 31 620 316 230

LelF-B 15 850 3162 0

LelF-C 31 620 31620

LelF-D 50120 316 230

LelF -F 100 000 100

Buffy coat 50120 1000000

* Vero c ell monolayers we re treated for 18 to 20 h wit h half-step log~0 dilutions of interferon pr ior to

inoculation with 40 to 50 p.f.u, of the indicated virus. The data are presented as the reciprocal of the dilution

of the interferon preparation causing 50 reduction of virus plaque formation.

s i g n i f i c an t l y f r o m o n e t o an o t h e r i n t h e i r ab i l i t y t o co n f e r p r o t ec t i o n ag a i n s t V S V i n f ec t i o n o f

dif fere nt ce l l l ines .

T h e s e r e s u l t s r a i s ed th e p o s s i b i l it y t h a t v a r i a t i o n i n t h e an t i v i ra l a c t i v i t y o f n a t u r a l h u m an

l e u k o c y t e i n t e r f e r o n p r e p a r a t i o n s a g a i n s t d i f f e r e n t v i r u s e s m a y b e d u e t o d i f f e r e n c e s i n t h e

p r o p e r t i e s o f t h e co n s t i t u en t s u b t y p es . W e , t h e r e f o r e , co m p a r ed t h e an t i v i r a l a c ti v i t y o f t h e

c l o n ed i n t e r f e r o n s u b t y p es a g a i n s t V S V o r E M C v i r u s in f ec t i o n o f V e r o ce l ls . T h e r e s u l t s

d em o n s t r a t ed t h a t t h e v a r i o u s c l o n ed i n t e r f e r o n s u b t y p es h av e d i f f e r en t r e l a t i v e ac t i v i t i e s

ag a i n s t V S V an d E M C v i r u s i n a m o n k e y ce l l l in e ( T ab l e 2 ) . T h e d i f fe r en ces i n an t i v i r a l

a c t iv i t y a g a i n s t E M C v i ru s o f t h e v a r i o u s c l o n e d i n t e rf e r o n s in t h e se e x p e r i m e n t s d i d n o t v a r y

g r e a t l y e x c e p t f o r L e I F - F , f o r w h i c h h i g h e r a c t i v i t y w a s o b s e r v e d . T h e v a r i a t i o n s i n t h e

r e l a ti v e ac t i v i t y i n d i f fe r en t c e l ls i n d i ca t e t h a t t h e i n t e r f e r o n s u b t y p es m ay d i f f e ren t i a ll y a f f ec t

b i o c h e m i c a l p a t h w a y s i n d u c e d b y i n t e r fe r o n , s u c h a s p h o s p h o r y l a t i o n o f e I F - 2 o r a c t i v a t i o n

o f R N a s e F ( B a g li o n i, 1 9 7 9 ; D . A . E p s t e in , C . W . C z a r n i e c k i , H . J a c o b s e n & R . M .

F r i ed m an , u n p u b l i s h ed r e s u lt s ). T h a t d i f fe r en t v i r u s e s r e s p o n d t o o n e o r o t h e r o f t h e

i n t e rf e r o n - in d u c e d b i o c h e m i c a l p a t h w a y s i s i n d i c a te d b y s tu d i e s w it h n a t u r a l , u n f r a c t i o n a t e d

m o u s e i n t e r f e r o n p r e p a r a t i o n s ( C z a r n i e c k i

et a l . ,

1 9 8 1 ; D . A . E p s t e i n

et al. ,

u n p u b l i s h e d

resul ts ) .

S i n ce t h e v a r i o u s c l o n ed g en es f o r t h e i n t e r f e r o n s u b t y p es w e r e p r o d u ced i n

E. eo l i

f r o m

es s en t i a ll y t h e s a m e s e r i e s o f g en e t ic m an i p u l a t i o n s an d s i n ce t h e i n t e r fe r o n s w e r e ex t r ac t ed i n

t h e s a m e w a y f r o m s i m i la r ly g r o w n c u l tu r e s , o n e m i g h t e x p e c t t h e ir y ie l d in t e r m s o f a m o u n t s

o f p r o t e i n t o b e e s s en t i a l ly i d en t ic a l . T h e u s e o f a c e l l -f l e e s y s t e m an d b ac t e r i a l m i n i ce l ls h a s

d e m o n s t r a t e d t h a t s i m i l a r a m o u n t s o f i n t e rf e r o n s u b t y p e s a r e m a d e f r o m t h e r e c o m b i n a n t

p l a s m i d D N A s , t h e g r ea t e s t d i f f e r en ce i n y i e l d b e i n g n o m o r e t h a n f i v e f o l d i n m i n i ce l ls

(S t reu l i

et a l . ,

1 9 8 1 ) an d t w o f o l d i n t h e ce l l -f l e e s y s t em ( P . K . W e ek , S . A p p e r s o n , N . S t eb b i n g ,

P . W . G r a y , D . L e u n g , M . S h e p a r d & D . V . G o e d d e l , u n p u b l i s h e d r e s u l t s ) . T h u s t h e

d i f f e ren ces i n a c t i v i t y o f t h e v a r i o u s i n t e r f e r o n s u b t y p es a r e p r i m a r i l y d u e t o i n h e r en t

d i f f e renc es in spec i f ic ac t iv i ty .

T h e an t i g en i c s p ec i fi c it ie s o f t h e c l o n ed i n t e r f e ro n s - c t w a s d e t e r m i n ed b y an t i b o d y

n e u t r a li z a t io n t e s t s. T h e N I A I D a n t i - L e l F a n t i b o d y p r e p a r a t i o n n e u t r a l i z e d 2 0 u n it s o f t h e

c l o n e d i n t e rf e r o n s u b t y p e s a t d i lu t io n s o f 1 : 6 4 0 f o r L e I F - A a n d : B , 1 : 2 4 0 f o r L e I F - C a n d - F ,

a n d 1 : 6 0 f o r L e I F - D . T h e a n t i - F I F a n t i b o d y h a d n o e f f e ct o n a n t iv i r al a c t iv i t y o f a n y o f th e

i n t e rf e r o n -c t s u b t y p es . A l l t h e i n t e r f e r o n s u b t y p es s h o w ed s i m i l a r st ab i l it y t o p r o l o n g ed

i n cu b a t i o n a t p H 2 . O n l y o n e o f t h e c l o n ed i n t e r f e r o n s ( L e I F - D ) l o s t a p o r t i o n o f i ts an t i v ir a l

a c t i v i t y an d t h e lo s s w a s o n l y 2 5 a f t e r 2 4 h, t h e s am e a s t h a t o b s e r v ed f o r t h e n a t u r a l

l e u k o c y t e p r e p a r a t i o n . H e a t t r e a t m e n t o f t h e v a r i o u s i n t e rf e r o n s u b t y p e s i n d ic a t e d t h a t

L e I F - D w a s t h e m o s t s t a b l e o f t h e c l o n e d i n t e rf e r o n s, a n d t h a t L e I F - A , -B , - C a n d - F l o st a t

l e a s t 9 0 o f t h e ir a n ti v ir a l a c t i v it y f o ll o w i n g 3 h in c u b a t i o n a t 56 ° C . L e I F - D l o s t 7 5



236

S h o r t c o m m u n i c a t i o n s

ac t iv i ty a f t e r 3 h hea t ing and 90 a f te r 6 h . Never the less , the p roper t i e s o f the c loned

l e u k o c y t e i n t e rf e r o n s d i d n o t d i ff e r g re a t l y f r o m t h e k n o w n c h a r a c t e r i s ti c s o f th e n a t u r a l

m a te r ia l in t e rms o f hea t o r ac id l ab i li ty (S tewar t , 1979).

T h e v a r i o u s i n t e r f e r o n - a s u b t y p e s in v e s t i g a te d in t h e p r e s e n t s t u d y a p p e a r t o h a v e d i s t i n c t

p roper t i e s in te rm s o f the i r ab i l ity to indu ce an t iv i ra l ac t iv i ty in va r ious m am m al ia n cel l

cu l tu res . Th is conc lus ion i s suppor ted by ea r l i e r obse rva t ions showing tha t an t iv i ra l ac t iv i ty

of na tu ra l in te r fe ron-a p repa ra t ions on rabb i t ce l l s i s a s soc ia ted wi th a pa r t i cu la r

e lec t rophore t i c f rac t ion (S tewar t & De sm yte r , 1975), a nd th a t an t iv i ra l ac t iv i ty on ca t ce l ls i s

n o t a f f e c t e d b y S D S t r e a t me n t s u n d e r r e d u c i n g c o n d i t i o n s , b u t a c t i v i t y o n h u ma n c e l l s i s

d e s t r o y e d b y s u c h t r e a t m e n t ( D e s m y t e r & S t e w a r t , 1 9 76 ). H o w e v e r , t h e re l a t io n s h i p b e t w e e n

t h e c l o n e d s u b t y p e s a n d t h e mo l e c u l a r h e t e r o g e n e i t y o f n a t u r a l l e u k o c y t e i n t e r f e r o n

prep ara t ions and the i r an t iv i ra l p roper t i e s i s no t c lea r . To da te , an t iv i ra l ac t iv i ty o f na tu ra l

l euko cy te in te r fe rons has b een com par ed in va r ious ce ll l ines us ing on ly impure p rep ara t ion s

t h a t a r e p r e s u ma b l y mi x t u r e s o f t h e v a r i o u s l e u k o c y t e i n t e r f e r o n s . A n t i v i r a l a c t i v i t y o f

n a t u r a l h u m a n i n t e r f e r o n - a p r e p a r a t io n s o n m o u s e c ells i s o n l y a f r a c t i o n o f a p e r c e n t o f t h a t

o n W I S H c ells , b u t t w o o f t h e c l o n e d s u b t y p e s ( L e I F - D a n d - F ) s h o w a b o u t 5 c r o ss - sp e c ie s

ac t iv i ty on L-929 ce ll s. T he l ack o f ac t iv i ty in mou se cel ls o f the na tu ra l l euko cy te

p r e p a r a t i o n c o u l d b e d u e t o a n t a g o n i s t i c i n t e r a c t i o n s b e t w e e n t h e i n t e r f e r o n s u b t y p e s a n d

s t u d i e s w i t h t w o o f t h e c l o n e d i n t e r f e r o n s u b t y p e s ( L e I F - A a n d - D ) h a v e a l r e a d y d e mo n -

s t ra ted tha t the re a re in te rac t ive e ffec t s (S tebb ing e t a l . 1981).

T h e f r e q u e n c y o f t h e v a r i o u s c l o n e d i n t e rf e r o n s u b t y p e s i n n a t u r a l l e u k o c y t e in t e r f e r o n

p r e p a r a t i o n s i s u n c e r t a i n . H o w e v e r , a s s u mi n g t h a t mR N A f r e q u e n c y i n i n d u c e d c e l l s i s

p r o p o r ti o n a l to t h e a m o u n t o f th e t r a n s la t i o n p ro d u c t s , L e I F - A a n d L e I F - D w o u l d a p p e a r t o

b e th e mo s t f r e q u e n t su b t y p e s , c o r r e s p o n d i n g t o a b o u t 7 0 o f t h e t o t a l i n t~ r f e ro n s p r e s e n t i n

b u f f y - c o a t o r K G - 1 c ell li n e p r e p a r a t i o n s ( N a g a t a

e t a t .

1980 b ; Go edde l

e t a l .

1981) . On the

b a si s o f m R N A f r e q u e n c y , L e I F - D s h o u l d b e th e p r e d o m i n a n t in t e rf e ro n s u b t y p e in n a t u r a l

l e u k o c y t e i n t e r fe r o n p r e p a r a t i o n s a n d t h i s is s u p p o r t e d b y t h e o b s e r v a t i o n t h a t t h e s p e c t r u m

o f a c t i v it y o f L e I F - D o n a r a n g e o f m a m m a l i a n c e lls c o r r e s p o n d s m o s t c l o s el y t o t h a t o f a

buf fy -coa t in te r fe ron p repara t ion (Tab le 1 ).

The an t iv i ra l ac t iv i ty o f the d i f fe ren t c loned in te r fe ron sub types in ce l l cu l tu res may be

mo d i f i e d

i n v i v o

w h e r e c e l l- me d i a t e d e f f ec t s m a y b e o f p a r a m o u n t i mp o r t a n c e ( G i d l u n d

e t a l .

1 9 78 ; H e r b e r m a n e t a l . 1978; Stebbing e t a l . 1978). T ha t in te r fe ron p repara t ion s inac t ive in

ce l l cu l tu res can be ac t ive i n v i v o h a s b e e n a l r e a d y d e mo n s t r a t e d ( S c h e l l e k e n s e t a l . 1979).

M oreov er , Le IF-A , which has re la t ive ly low ac t iv i ty in rabb i t ce ll s (Tab le 1 ) has p rove d to be

a s e f f ec t iv e as a h u m a n b u f f y - c o a t i n te r f e r o n p r e p a r a t i o n a g a i n s t H S V - 1 i n f e c ti o n o f t h e

r a b b i t e y e ( S mo l i n e t a l . 1981). Ho we ver , the p resen t r e su l t s ind ica te tha t the va r ious

l e u k o c y t e i n te r f e r o n s u b t y p e s h a v e d i s t in c t b i o lo g i c a l p r o p e r ti e s a n d m a y p r o v e o p t i ma l l y

use fu l aga ins t d i f fe ren t v i rus in fec t ions . Such poss ib i l i t i e s a re under inves t iga t ion us ing the

i n t e rf e r o n s d i s c u s se d h e r e , a s w e ll a s a n e x a m i n a t i o n o f i n d u c t io n o f v a ri o u s b i o c h e m i c a l

p a t h w a y s f o l l o w i n g t r e a t m e n t w i t h t h e v a r i o u s c l o n e d i n t e r f e r o n s u b t y p e s .

S e c t i o n o f V ir o lo g y G e n e n t e c h I n c .

4 6 0 P o i n t S a n B r u n o B o u l e v a r d

S o u t h S a n F r a n c is c o C a l i f o rn i a 9 4 0 8 0

U . S . A .

PHILLIP K . W E C K *

S H A R O N A P P E R S O N

L A U R I E M A Y

N O W E L L S T E B B I N G

R E F E R E N C E S

BAGLION1, C . ( 1 9 7 9 ). I n t e r f e r o n - in d u c e d e n z y m a t i c a c t iv i t i e s a n d th e i r r o le in th e a n t iv i r a l s t a t e .

Cell

1 7 , 2 5 5 - 2 6 4 .

BRIDGEN, P . J. , ANFINSEN, C. B., CORLEY, L. , BOSE, S . , ZOON, K. C. RUEGG , U. T. (1977) . H um an lym ph ob las to i d

i n t e r f e r o n : l a rg e s c a l e p r o d u c t i o n a n d p a r t i a l p u r i f i c a ti o n . Journal o f iological Chem istry 2 5 2, 6 5 8 5 - 6 5 8 7 .



Shor t communica t ions 2 3 7

CAMPBELL, J. B., GRUNBERGER, T., KOCHMAN, M. A. WHITE S. L. ( 1 9 7 5 ). A m i c r o p l a q u e r e d u c t i o n a s s a y f o r

h u m a n a n d m o u s e i n t er fe r o n. Canadian Journal of Microbiology 2 1 , 1 2 4 7 - 1 2 5 3 .

CZARNIECKI, C. W., SREEVALSAN, T., FRIEDMAN, R. M. PANET, A. ( 1 9 8 1 ) . D i s s o c i a t i o n o f i n t e r f e r o n e f f e c t s o n

m u r i n e l e u k e m i a v i r u s a n d e n c e p h a l o m y o c a r d i t i s v i r u s r e p l i c a t i o n i n m o u s e c e l l s . Journal of Virology 37 ,

8 2 7 - 8 3 1 .

D E SM Y TE R, J . S T EW A R T , W . E . ( 1 9 7 6 ) . M o l e c u l a r m o d i f i c a t i o n o f i n t e r f e r o n : a t t a i n m e n t o f h u m a n i n t e r f e r o n i n a

c o n f o r m a t i o n a c t i v e o n c a t c e ll s h u t i n a c ti v e o n h u m a n c e ll s. Virology7 0 , 4 5 1 - 4 5 8 .

D ESM Y TER , J . , RA W LS, W . C . M ELN IC K , J . L . ( 1970) . Sens i t i v i t y o f r abb i t c e l l s t o p r im a te i n t e r f e r ons . Annals of the

New York Academy of Sciences

1 7 3 , 4 9 2 - 5 0 4 .

G ID LUN D , M . , O m ~ , a . , W IG ZELL, n . , SEN IK , A . G R ESSER , I . ( 1978) . En ha nc ed N K ce l l a c t i v i t y i n m ice i n j ec t ed

w i t h i n t e r fe r o n a n d i n t e r f e r o n i n d u c e r s . Nature, London 2 7 3 , 7 5 9 - 7 6 1 .

GOEDDEL, D. V., YELVERTON, E., ULLRICH, A., HEYNEKER, H. L., MIOZZARI, G., HOLMES, W., SEEBURG, P. H., DULL,

T., MAY, L., STEBBING~ N., CREA, R., MAEDA, S., MCCANDLISS, R., SLOMA, A., TABOR, J. M., GROSS, M.,

FAMILLETTI, P. C. FESTK A, S. (1980). H u m a n l e u k o cy t e i nt e rf e ro n p r o d u c e d b y E. coli s b io log i ca l l y ac t i ve .

Nature, London 2 8 7 , 4 1 1 - 4 1 6 .

GOEDDEL, D. V., LEUNG, D. W., DULL, T. J., GROSS, M., LAWN, R. M., MCCANDLISS, R, SEEBURG, P. H., ULLRICH, A.,

V EL VE RT ON , E . G R A Y , P . W . ( 1 9 8 1 ) . T h e s t r u c t u r e o f e i g h t d i s t in c t c l o n e d h u m a n l e u k o c y t e i n t e r f e r o n

c D N A s . Nature, London 2 9 0 , 2 0 - 2 6 .

G R ESSER , I ., B A N D U, M . -T ., B R OU TY-BO Y E, D . TO V EV , M . (1974 ) . P r on ou nc ed an t i v i r a l a c t i v i t y o f h u m an in t e r f e ro n

o n b o v i n e a n d p o r c i n e c e l l s . Nature, London 2 5 1 , 5 4 3 - 5 4 5 .

HERBERMAN, R. B., DJEU, J. V., ORTALDO, J. R., HOLDEN, H. T., WEST, W. H. BONNARD, G. D. ( 1 9 7 8 ) . R o l e o f

i n t er f e ro n i n a u g m e n t a t i o n o f n a t u r a l a n d a n t i b o d y d e p e n d e n t c e l l -m e d i a t e d -c y t o t o x ic i t y . Cancer Treatment

Reports

6 2 ,

1893-1896.

LA ZA R , R ., B RErNIN G M . K ., A R M STR O NG , J . A . H O , M . (1980 ) . R es po ns e o f c l on ed p rog en y o f c l i n i ca l i so l a t e s o f

h e r p e s s i m p l e x v i r u s to h u m a n l e u k o c y t e i n t e r fe r o n . Infection and Immunity 2 8 , 7 0 8 - 7 1 2 .

MCGILL, J. I., COLLINS, P., CANTELL, K., JONES, B. a. FINTER, N. B. ( 1 9 7 6) . O p t i m a l s c h e d u l e s f o r u s e o f i n t er f e r o n

i n t h e c o r n e a s o f r a b b i t s w i t h h e r p e s s i m p l e x k e r a t i t is . Journal of Infectious Diseases 1 3 3 , 1 3 - 1 9 .

NAGATA, S., TAIRA, H., HALL, A., JOHNSRUD, L., STREULI, M., ECSODI, J., BOLL, W., CANTELL, K. WEISSMANN, C.

( 1 9 8 0 a ) . S y n t h e s i s in E. coli o f a p o l y p e p t id e w i t h h u m a n l e u k o c y t e i n t er f e r o n a c t iv i ty . Nature, London 2 8 4 ,

3 1 6 - 3 2 0 .

N A G AT A , S ., M A N T El, N . W E IS SM A NN , C . ( 1 9 8 0 b ) . T h e s t r u c t u r e o f o n e o f t h e e i g h t o r m o r e d i s t i n c t c h r o m o s o m a l

g e n e s f o r h u m a n i n t e r f e r o n - tt . Nature, London 2 8 7 , 4 0 1 4 0 8 .

OTTO, M. J., SEDMAK, J. J. GROSSBERG, S. E.

( 1 9 8 0 ) . E n z y m a t i c m o d i f i c a t io n s o f h u m a n f i b r o b la s t a n d l e u k o c y t e

i n t e r f e r o n s . Journal of Virology3 5 , 3 9 0 - 3 9 9 .

RUBINSTEIN, M., LEVY, W. P., MOSCHERA, J. A. , LAI, C.-Y., HERSHBERG, R. D., BARTLETT, R. T. PESTKA, S. ( 1 9 8 1 ) .

H u m a n l e u k o c y t e i n t e r fe r o n : i s o l a ti o n a n d c h a r a c t e r i z a t i o n o f s e v e ra l m o l e c u l a r f o r m s .

Archives of

Biochemistry and Biophysics in pres s ) .

SC HELLEK EN S, rL , W EIM A R , W . , C A N TE ta , K . STITZ, L . ( 1979 ) . A n t iv i r a l e f f ec t o f i n t e r f e ron in vivo m a y b e

m e d i a t e d b y t h e h o s t . Nature, London 2 7 8 , 7 4 2 .

SMOLIN, G., STEBBING, N., FRIEDLAENDER, M., FRIEDLAENDER, R. OKUMOTO, M. ( 1 9 8 1 ) . A n t i v i r a l e f f e c t s o f

n a t u r a l a n d c l o n e d h u m a n l e u k o c y t e i n t e r f e r o n s i n h e r p e s v i r u s i n f ec t i o n s o f t h e r a b b i t e y e . Archives of

Ophthalmology in pres s ) .

STEBBING, N., DAWSON, K. M. LINDLEY, I. J. D. (1978 ). R e q u i r e m e n t f o r m a c r o p h a g e s f o r i n t e rf e r o n t o b e e f f e ct iv e

a g a i n s t e n c e p h a l o m y o c a r d i t i s v i r u s i n f e c ti o n o f m i c e . Infection and Immunity 1 9 , 5 - 1 1 .

STEBBING, N. , WECK, P. K ., FENNO, J . T., APPERSON, S. LEE , S. r~ . (198 1) . C om pa r i so n o f the biol ogic al p rop er t i es

o f n a tu r a l a n d r e c o m b i n a n t D N A d e ri v ed h u m a n i n t er f er o n s. The Biology of the Interferon System.

A m s t e r d a m : E l s e v i e r / N o r t h - H o l l a n d ( i n p r e s s ).

STEWART, W. E. H . (19 79) . The Interferon System. ~ N ew Y o r k : S p r i n g e r - V e r l a g .

S TE W A RT , W . E . D E SM Y T ER , J . ( 1 9 7 5 ) . M o l e c u l a r h e t e r o g e n e i t y o f h u m a n l e u k o c y t e i n t e r f e r o n : t w o p o p u l a t i o n s

d i f f e ri n g i n m o l e c u l a r w e i g h t s , r e q u i r e m e n t s f o r r e n a t u r a t i o n , a n d c r o s s - s p e c i e s a n t iv i r a l a c t iv i t y .

Virology

67 ,

6 8 - 7 3 .

S TE W A RT , W . E . H A V EL L , E . A . ( 1 9 8 0 ) . C h a r a c t e r i z a t i o n o f a s u b s p e c i e s o f m o u s e i n t e r f e r o n c r o s s - r e a c t i v e o n

h u m a n c e l ls a n d a n t i g en i c a ll y r e la t e d t o h u m a n l e u k o c y t e i n t e r fe r o n . Virology 1 0 1 , 3 1 5 - 3 1 8 .

STREULI, M., HALL, A., BOLL, W., STEWART, W. E., NAGATA, S. WEISSMANN, C. ( 1 9 8 1 ) . T a r g e t c e l l s p e c i fi c i t y o f t w o

s p e c i e s o f h u m a n i n t e r f e r o n - a p r o d u c e d i n Escherichia coli a n d o f h y b r i d m o l e c u l e s d er i v e d f r o m t h e m .

Proceedings of the National Aeademy of Sciences of the United States of America 7 8 , 2 8 4 8 - 2 8 5 2 .

Y ELV ER TO N , E . , LEU Y G , D . , W EC K , P . , G ~ Y , P . W . G O ED D EL, D . V . (1981) . B ac t e r i a l s yn the s i s o f a nove l hu m an

l e u k o c y t e i n t e r f e r o n . Nucleic Acids Research 9 , 7 3 1 - 7 4 1 .

Received 12 May 1981)

journal of biotechnology lecture

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