Swine viruses

Journal oj" Biological .S'tandardization 1978 6, 173-I 79

Antibody o n s e m h .Lmans after adm s ation o f w h on" a n d vaccine prepared from two nt in u A/ "' ~ e S * VH I~t\ Z~vadov~,t V. Vonka, t E. Domor~zkov~t,t L. (ek,$ ~ . St~rek, t J. !trtdkovg¢,~ V, P l e s ~ , § V. JanoutH and i~-. Uv~lt[

O u r previous findings with s e ~ o f several ~ f f e r e n t animal species had /ndic~ated a close ~ t i g e p J c similarity ~ t w e e n t h e haemagglut/nins o f $w/nelA.,m Arbor/31 (A_~) and Sv,4ne/New J e r s e y ] ! 8 / 7 6 ( N J ) ~ru~s~. S i n ~ t h e f o y e r virus grows m u c h b e t t e r in chick e m b r y o s t h a n the N J - d e r i v e d recombirmnts (X-53 ~nd ~,IIB-3), e x p e d m e n t a l whole-vitaas vaccine f r o m A A a n d split vacc/nes f r o m AA a n d X-53 were p r e p a r e d f o r t ~ t s ~'~ h u m a n beings. G ~ u p s o f children a n d adults were ~ w i t h two ~mccine doses # y e n at a 3-week i n t e r v a l I n the case o f w h o l e - ~ f i o n vaccine b o t h d o s ~ c o n t a i n e d the A A v i m , vchJle the second d o s e o f split vaccine contained either A A o r N J . Sera o f 164 vac~=ine~s w e ~ ~d~mined in haemoggludnation-inhibition tests. I n all but two s u b j e c t , appro~d~mately t h e same a n t i b o d y titres against t h e two viruses w e r e deterrrLined in b o t h the p ~ - a n d pos tion sera. ~ i s m d i e ~ t ~ that t h e N J v i ~ c a n be s u b s t i t u t e d b y t h e A A -Hrus f o r ".~ccLn.e p r o d u c t i o n . C h i l d r e n aged 7-1S years ~ d vaccinated sue y with A A a n d N J s # i t v a c c l n ~ displayed a ly b e t t e r a n t i b o d y response t h a n those vaccinated with t w o doses o f t h e A A ~ c c i n e .

INTRODUCTION A s r-~orted in the preceding paper (Dole~alovfi, Z#.vadov~, St~rek 8,= Vord~a, 1978, previous paper), an in on o f sera from tour different species of axdmal, either infected with live or immuniz-ed with Lnactivated vi~s~es, indicated a c l o ~ antigenic rela~onship * Received f o r publication 6 J ~ e 1977 t nt o f H x p e ~ e n t a l Virology, Institute of Sera a n d V a g i n a , ~kla. :~ Institute o f H y # e n e ~ d EpldMmiology, Praglae, ~ec-hoslo-~k/a. § R e g i o n H y g i e n e C~-xt~, Ostrava, Czechoslovalda. [| D i s t d ~ H y g i e n e Centre, Olom~oue, CzechosIo-.mJkla. 0092-I1571

173 $01.00/G.'9

Prague, Czechoslo-

© 1978 The Intc~ationM Associatlo~ o f Biological Sta=tdardi~tion

173

H. Z A V A D O V A ET AL. between the .~:~53~reeombinant (possessing b o t h surface-antigefis of A]Swinc/New Jersey/18/76 virus) mud the A]Swine]Anm Arbcw/31 vir~as. Since the latter virus grows much b e ~ e r in chick embryos than the former, ~;e examined the p o ~ i b i l i t y of using the A n n Arbor (AA) virus instead of X-53 for inactivated vaccine production. T h e present paper describes o u r findings in sera of s u c c o r s i r n m u ~ z e d with either wholev i e s or split vaccine prepared front AA virus. .METHODS In A/Swlne/Ann Arbor/31 (A~P~) and X-53 viruses were the same as previously used (DolczaIov~ et aL, 1978, in this volume), tvlRC-11, ~ recombinant between the A[Po~ Chalmers]l]73 and A]PR]8134 r i n s e s , and X-47, a recombinant between A]Vietoria]3]75 mad P R 8 viruses, were kindly provided b y D r Schild (IVledieal R ~ e a r e h Council, London). Both of these recombinants po~essed the surface antigens of the respective H 3 N 2 parent viruses. (a) "Whote-vir~as vaccines, formaldehyde-inactivated, a l u m l n i u m - p h o s p ~ t e - a d s o r b e ~ vaccine Admvira (Imuna, ~. Micha/~ny, Slov~Ja). Bivaccine contalned A#= ~qrus and M R C - I I virus, m o n ne contained the AA ,4ir~ds only. T h e vaccines contained 1024 H units of each virus per I mL (b) Split vaccines were prepared b y ether disruption in our la ry as has been described (Z~vadov~, Xronka, St~rek, Domorfizkovfi, Bruj & Sk-~il, 1975). T w o monovaccines were used, one containing fi~. virus, the other X-53 ~firxls. I n both, the H ~ t i g e n content w ~ 2500 units per 1 mL

Selection of ~ e e t s and

tion

re

~2nole-virus vaccine was administered to groups of su~ects aged 18-24 (24 subjects), 37-50 (15 subjects) and 55-80 y e a ~ (20 subjects). TI~e youngest age group consisted o f healthy workem, the middle-aged group o f crippled patients living in a rehabilitation establishment ~ d the oldest group of patieras suffering from chrot~Jc di . T w o subcutaneous doses of vaccine were given to each subject at a 3-week iater-val. F o r the first dose (0-4 ml) t h e bivaccine and for the second (0.2 rnl) the monov a e d n e (A_A) was used. Split vaccine was administered to healthy subjects aged I - 3 (16 subjects living in semi-dosed children's homes), 7--!5 (47 schoolchildren) and 20--60 years (60 subjecvs, m e m b e ~ of the personnel o f a District ene Centre). ~ v o i n t r a d e ~ a I doses were injected at a 3-week interval. T h e AA virus vaccine was used for the first dose (0"2 ml), and either t h e #_A or the X-53 vinJs vaccines for t h e second dose (0-3[ ml). Clinical reactivity was followed as described o n another occasion (Z~ivado-¢~, *~ronka, Adam, D o m o ~ k o v ~ & Davenport, I972). N o u rd clinical ons were observed. T h e vaccination was c ~ r i e d out in surI~.mer and early a u t u m n 1976, at a time when there ~v~ no evidence of influenza in Czed'~oslovakia. F r o m all vaecinees blood s ~ # e s were t ~ e n immediately prior to vaccination, at administration of the second dose ~ d 3-4 after vaccination.

Hemagglutir~tion-inhibition

(HI) test

~ 1 sera were treated with R D E ( tot Dest Enzyrae, Sevac, P r a g u e ) a s recommended b y oft & M i n u s e (1964). T h e y were diluted I : 1 ~ 1 : 640 in I74

VACCINES F R O M

TV/O A / S W I N E

VIRUSES

twofold steps. T h e test was p e r f o ~ e d ,as described p r e v i o u ~ y (Tug.ova, W o n ~ & St~rek, 1968). I n calculating geometric mea~n d t r e s (G~ClT). sera negative at dilution 1 : I0 were considered positive at dilution 1 : 5. RESULTS An

to ~hoIe-virus vaccine

Sera of v a t . h a t e d subjects were tested for H I antibody both for A A and X-53 {New Jersey) vir~ases, q ~ e results arc presented i n Fig. 1. I t ca~a be seen t h a t p r i o r t o vaccination a n t i - H s w a n t i b o d i ~ were only rarely encountered in subjects born after 1925;

100 80

~05 92

S w AA/3! Sw NJ/TG

~8

i:~} 11~

32 30

~4

~0 ....

| 2~ Age Subjeets

I ~ 5 1952 - 59 24

I

| 2~

123

1925-37

15

~3 I 2 3 1~-1920 20

Fig. I . Haerrtagglutinatior~-inbJbit/ng a n t i b o d i e s agaLnst h ' f f / u ~ .A/Switte (hfsxv N z ) , A n n A r b o r (LAg.) a n d X - 5 3 p r i o r t o arid a ~ e r a d tlort o f w h o l e - v l n ~ s . re: rage o f s u b j e c t s ~ s e s s ~ g a n t i b o d y . A b s ~ s a : 1~ taken #og t o first v a ~ t e d o s e ; es ~ a t s e c o n d v a c c i n e d o s e ; 3 ~ a r n p ] e ~ ta2.~¢ax 3 ~ w e e k s a f t e r v~-eeinatiorz. ~L.'qlzitee~lu~.~.s: a n t i b o d y agaL~st #_A; b l a c k c o l u m n s : a n t i b o d y agah'mt X - 5 3 . Nua-nbers a t t o p o f e a c h ex)Iumrx ir.,d__i~te GI'v~tT.

o n the other hand, t h e r n ~ o r i t y of su born before 1920 ~sse~r,~d this type of antibody. T h e d y response to vaccination wm_~sd e a r l y age-dependent. #alter the first vaccine dose l ~ s t h a n 2 0 % of the youngest subjects developed low levels of antib o d ) : T h e second v dose b r o u g h t about a marked increase in seroconversions a n d G M T . I n subjects aged 37-50 ye;trs o n e vaccine dose was sumcient to induce a n d b o d y formation L-~ the majori H of vaceLrze~; t h e second dose Lnflu~'zce~. neither the percentage of s e r o p o s i ~ v ~ n o r the G M T . T h e results h'~ t h e oldest subjects a sirrfil~ pattern. #ffter one dose all subjects tested ~ e I-Isw antibody a n d f i e G M T had ed three- to four-fold; ~ w e v e r , t h e second ~ c ~ _ - e dose was w i t h o u t a n y p r o n o u n ~ d influence on the antibody status. Comparisort o f t h e r e s u l ~ of H I T performed with A,A or X , 5 3 virus s h o w ~ a t t h e a n t i ~ d y d i d n o t dis ate between the two ~ma~-s in the two age groups. ~¢~-zor difference:3 were only encountered ~ t h e ~ r a of t h e subjects. I t should b e added ~ a t whe~ever antibodies were detected against one b u t n o t the other vireos, the r e s p e ~ v e ar~tibody ~tre did n o t e~cceed 1 : 10. 175

!t. Z A V A D O V A E T AL. Sinee~fl~e ,vaccine contained ~ e P o r t C h a | m e r s virus @ t I P . C 1 1 ) ~ the H3 e o m p o n ~ t , it was o f interest to in te t h e anti-Victoria antibody de The resul~ are s h o ~ in Fig. 2. T h e ~ t l b o d y r~sponse was quite prono~mced, more ~ in t h e y o u n g e r than the older subjecxs,

/Lgt Sub}ee~

IZ3 1952-9 24

J93 1925-37 15

IE3 q900--20 20

Fig. 2. FIaema L'd~.i i~ agai~t influ~ X-67 (H3N2) virus prior to and after t~tion of whole-~rus . For expla.~mtion see tegend to F/g. 1. B/aek ¢~]t:~'~'~: antibody a ~ i n s t X-47.

A n t = ~ y response to split As above, sera were tc~ted for H I a n t i ~ d y against , a ~ an d X-53 . The ~sults are su ~ d in Fig. 3. T h e general pattern is clear: efficien~j increased with the age o f , I n children aged 1-3 years ~ao v a e d n e doses w e r e n e e d e d for the d nt o f low antibody levels in only a smMl ~ of eez. The ~e ~ h-~ae of ~ehoolehi t h a t the n v e ~ i o n ~ t e was p~iee as high. I n young a d d t s a c o m p e result was achieved by o n e ~ e c i n e dose~ ~ t h the d dose h a ~ n g o @ a little on th e ~ t i b o d y status. Mid d le-ag ed adults, although ng a similar pattern, at A fairly shm~p antibody was only e n c o u n t e r e d in t h e o l d ~ t age ~ o u p . O n e dose o f vaccine i n d u c e d seroc~nversion at a e n ~ and a m a r k e d in G ~ T . A further a n d ~ d y in x~s d a l t e r di e sec,o n d dose. n o f t h e results obtained v,~th t h e t w o v i ~ m e m p l o y e d no substantial d ees in ei~h er'p re- o r pos nation set'a, except for t h e oldest e ~ group ~-d-lere ~'~fibodies a~ir'mt X-53 vimas w e ~ d at m o r e f r e ~ e n t l y ~ d in sl higher t h ~ against AA ~r~as. W h i l e t h e ~ vir~as was used b r t h e first dose in all vaeeinees, either AA- or X-53derived -vacCines were used for the d dose. I n the yo age g r o u p too few s s were o ed to j u s t i ~ a n y an . I n the a d u l t groups n o appre~ a b l e d.iffe~nees were noted be those revaeclnated with o n e o r the o t h e r ~mas. 176

VACCINES FROM TWO AISWI~E

VIRUSES

Sw A A / 3 |

~00

S w NO/76

46

60

3BU

80

F t

6oi9

4oL

9

9

IO

~0

13 1

9

! 20

i23

12 3

t973-75

Age

Subiects

t6

1,23

] 23

J23

I961 - 6B

I 2 3

1946-

27

5B

iE $

~2~

{9~?-- 4!

12 3

(907~

20

20

~ 23

26

20

b~g. 3. HaemgglutLrmtion-inhibiting ~ t i b o d i ~ a g a i m t ~ f l u e n z a A / S w i n e (Haw N I ) ~ A r b o r (AA) a n d X-S3 ~ r u s c s p r i o r t o a n d after administration o f s p l l t - p r o d u ~ vaccine. F o r explanation s ~ Fig. 1. However, a marked d i f f e r e n ~ was detected i n t h e group of schoolchildren. T a b l e 1 shows that ~m~onve~sions were decidedly m o r e frequent and titres higher Jn those given t h e heterologous X-53 vivas as t h e second vaccine dose. TABLE I. Antibody response in schoolchildren ~ t e r administration of split vaccine A_A A~ group (mean age)

Vaccination s~edule

196I~9

A_A~+ AA

No. of subjects

8e~m ~arnple.P

14

(i1) 1961~9

~+X-53

13

Antibody pres~ (%)

X-53

GMT

G~clT

0 0 3 (21)

6

I

0

H

o

llI

3 (21)

I

0

~

0

o

--

o

s (61)

12

8 (61)

(Ii)

Ill

~-

Andbody presence (%)

6

12

* The fi~t ~pm was t a k e n i ~ m e d i a t e l y ~ r l o r t o v a ~ L n a ~ 0 n , t h e second at t h e . e c o n d dose ~ g n i s t r a t i o n a n d t h e t h i r d 3 weeks a f t e r i h e s e c o n d dose.

in H I

to A A

6r,~es

As h d i c a t e d above, t h e sera tested t h e same r e a c t i ~ t y against both ~ r a s e s . O n l y 2 o f t h e 164 s u b j ~ s followed were exceptional in tFds r e s p e c t . T h e fin in their sera are s h o w n in T a b l e 2, I t cart h e seen that already t h e i r ation sera were nearly s~ecifie. I n one o f these subjects t h e titre o f A A antib o d y v¢~ 16 times higher than that of X-53 aIxtibody. T h e o t h e r subject possessed a 177

~i: Z A V A D O V A E T A L . high |eveJ.of~BtiTX=53 antibody~ and l a ~ e d , anti-#~A antibody. ~raccination was without a ~marked effect on the f o y e r subject, but in the other it induced anti-#,), a n t i ~ d y fo~afion. TA~

2. H I antibody tevels L~ two s u ~ e c ~ exhihlt~mg s~M-specific reac~on to either AA or ~ 5 3 vir~s Subject V.V.

O.L.

Age* (yea~

Se~m ~amples~

;~A

X-53

~

I II III

160 160 I60

I0

54

I lI

III

10 20

< !0 20

160 160

40

320

* At the time of ble~L'~. For ~pla~tioa see Table I. Note: Both sublets we~ ~c~hated with the split vaodrve; ~ ~e ~t one the second ~ i n e d o s e e o n t ~ e d X - 5 3 v i r ~ wi~AIe Ln t h e o t h e r b o t h d o s ~ c o n t a i r t e d ~ . k ~ t a ~ .

DISCUSSION T h e n~.aLu objective of this study was to verify when-her vaccine prepared from A A v i r u s ~d be u ~ d for immmnization of huroan b e i n ~ a g a ~ s t New J e r ~ y - l l k e ~ s e s : The resulm obtained showed equal reac~vi W to both vhaases h-x pre- ~ d ~ s t - v a c c i n a t i o n sera of almost ~ o~ o f the vaccinees. T h i s indicates that the re endatiort t h a t the X-53 (New J e r ~ y ) virus should be substituted b y the better~ ng M v i e s is fully justified. T h i s e~onctusion seems to be equally valid for both whole-vires ~ d split vaccines. T h e ~ , v a r r e n ~ of v e ~ ~ r e s e ~ # v m g a strain-specific reaction is probably the c o n s e q a e n c e ~ n d l n ~ 4 ~ u M sensitlvi~ to m i n o r ~ ic d e t e ~ t s not ~ e d Sy'~*'~g~~gther virus. T h e antibody r~t~onse to~..~ wine c o m p o n e n t of the bivalent vaccine w ~ agedependenL As cx=pected, the antibody response was most p r o n o u n ~ d i~ those mdivlduals who most l ~ e l y had e x p e H e n ~ d ~ n t a c t with the influenza S v d n e - ~ e vK~ses. In a g r e ~ n e n t ~ t h some recently published data (Parkm~-h G a l ~ o , T o p & Noble, 1976), the reactivity of p e c a n s having had ~ n t a ~ with H O a~ntigen was of a higher degree t h ~ that of er subjects. I n the r n ~ o r i t y of young subjects two doses o f the~ ~-aceSne used were required for mtltibody to f o ~ . T h e p r e ~ n t s ~ d y ~,as not d ~ i ~ e d to c o m p ~ e t h e whole-virus ~ d s p g t - p r o d u ~ vaccLnes. T h e v a c ~ n ~ n ~ d differed nor only ~ the p h y s i ~ l nature of their antigen but a ] ~ b y ~mposi:tion ( b l a i n e v e r s ~ monoya ), dosage ~ d route o f Luoculalion. I n a d d i t i o n / t h e age distribution o f subjects 6 y e n w h o l e - ~ r u s or split vaccine w ~ different. For ~ a m p l e , a l t h o u # the age span o f the oldest age groups was simMar, in both ~ _ s m a n y subj'ec~ ~ o n g ~ie whole-x~rus vaccine ~r e c i # n t s were over 70, while v e ~ few among t~ose re g split ~ i n e were over 60, " ~ q e age factor was a ~ responsible for t h e ent i n d d e n c e o f a~nti-Hsw a~at~oodi~ in pre-vac~-ladon ~ = hn ~ o u p s . However, Gne s # t vac~hae, alth elidtin:g an adequate irnmunoIo#~ n ~ in elderly Lndbdduals, was clearly I ~ s effective, at fine d o ~ s u ~ d , t h ~ whMe-~¢~rus vaccine. Sim~arly, ~ in the subjects irranunized ~ t h v~ole-virus 178

V A C C I N E S F R O M T W O A/Sx,V I N E ~VIRUSES vaccine, immunological responsiveness was better in adults possessing a broad background of natural exposure to different influenza viruses than in children. T h e failure of the split vaccine to stimulate antibody formation i n small children confirms the rec~ent data o f Parkman et aL (I976). I n schoolchildren, only 40~/o o f vaccinees developed antibody after two vaccine-doses. T h e antibody were low, and definitely too l o w to prevent infection, er, it is to be exoected that in s u d l *partially' i m m u n i z e d z u ~ e c t s influenza m o ~ i d i t y m i g h t be lower and the course o f the disease milder. It would therefore seem justified to vaccinate this age group with split vaccine, should the danger of a influenza pandemde become i m m i n e n t . T h e r e is no d o u b t that t h e vaccknation effect c o u l d be e n h a n c e d by administering a larger a m o u n t of antigen ~ d ] o r using adjuvant. O n e of the present observations c o m m a n d s ~peciaI interest. T h e antibody response in t h e schoolchildren was better in those s u ~ e c t s in w h o m ~ virus administration had been ~followed b y a second dose containing X-53 "virus than in t h o ~ revaceinated with A_A "vir~. T h i s effect may have been associated with the consecutive use of two different viruses. Unfortunately, a g r o u p o f subjects receiving only X - 5 3 had n o t been included and so the possibility that t h e X-53 p ~ p used was more i m m u n o g e n i c t h a t tl~e A~A vaccine cam~ot be r~Ied out. Lu spite o f having the same H a n t i ~ n content (as d e t e r m i n e d by t h e haemagglutination test), the two preparations could have dLffered, e.g. by t h e degree of t h e subunit tion or by the amount of non-haernagglutinating antigen present. Nevertheless, it seems r e a ~ n a b t e to investigate ~a~her the possibility of increasing tile vaccination ¢qTect by dxe successive administration of p r ~ a r a t i o n s containing closely related but not identical viruses. I t d i o u l d M recalled that a sirrfilar p h e n o m e n o n was observed in some animal s ~ c i e s at an earlier stage o f our work (Dol~2alov~ et aL, 1978, in this volume). T h e m a r k e d antigenic diversity between the Port Chalmers[l[73 and Victoria/3/75 viruses is well ~ v n . It was therefore surprising to tSnd a p r o n o u n c e d increase in anti-Victoria a n t ~ o d y in Port Ch -vaccinated individuals. I t is possible that flxis effect xv~ conditioned by the extensive circulation of ~v~ictoria-|ike v i r u ~ s a few m o n t h s prior to the present vaccination. REFERENCES Davenport, F. Pd. & rvlinuse, E. (1964). ostic rer for Viral and l~tYket~n'al Diseases, 3rd ed., Ch. 13, pp. 455--469. :New York: Amerieart PgblJe HealCh Asso~ation. Dole~alov~, B., Z~vadov~. H., St~rek, N~[.& Vonka, V. (I978). Antibody response in four a n i m ~ species to tr,vo different influenza viruses, of rdigation. 6, I65-171. Park~mm, P. D., Gala~o, G. J., Top, F. H. & Noble, G. R. (I976). Frorn the National Ins~o-lte o f ,ALUergy and Infectious Diseases of the l'qafional Institutes of/-fea/th, ~ e Center for Disease Control and fhe Bureau of the Biologi~ of b e F ~ d and D r u g A d rr,~istration. S u m m a r y of ~ v . i e ~ ~ a l s of influer~a va~ines. "yournal of IM, 100-107. Tu~kovh, ~., V. & St~rek, M. (I968). Comparison of genetic properties of Lnfluen3~a virus before and after repeated in kidney cell c~-oare. Acre Virologiea 12, 316-323. Zhvadov~, H., Vonkm V., St~rek, M., "Domo , E., Braj, J. & , V. (I975). Experience with C~eh~,lo-;~k subunit anti-influenza vaccine (in Czech). Oas¢Tpis ldka~J ~esle~ch 114, I462-1466. Z~ivadov~, H., Vonka, V., Adam, E., Dora , E. & D a v e n ~ r t , F. M. (I972). reacti~t~, and ogerd~ty of ~nLn influenza vaccine. L Archiv far die gesamte u~g 36, 43-52. 179