Archives of Virology

Archives of Virology 57, 199--204 (1.978)

© by Springer-Verlag 1978

Stimulation of Antibodies to Epstein-Barr Virus (EBV) in Acute Viral Infections By

T. GOTLI:EB-STEMATSKY~L. I:~ANNON, A. VONSOVER, and N. VA~SA~O The Central Virological Laboratory, Ministry of Health, Tel-Aviv-Yafo, and The Tel-Aviv University, Saltier School of Medicine, Ramat-Aviv, Israel Accepted February 24, 1978 Summary Acute and convalescent sera from 77 patients with serologically confirmed influenza, measles and adenovirus infections and from 36 healthy controls were tested for the level of antibodies to Epstein-Burr (EB) virus. In the three groups of patients significantly higher titers of antibodies to EB viral capsid antigen (VCA) were found as compared to the controls. In 19 patients twofold or higher rise in antibody titers between the first and second blood sample was demonstrated. It is suggested that in patients with influenza, measles or adenovirus infections, involvement of lymphocytes leads to reactivation of EB virus and antibody formation is stimulated. Introduction

A variety of pathological conditions associated with immunosuppressed state are sometimes followed by reactivation of latent viruses, which lie dormant in the patient's cells, while under normal conditions cell mediated immunity prevents extensive multiplication of these viruses. Delayed cutaneous sensitivity indicating impairment of ceil mediated immunity is found in several viral infections. Measles morbidity is associated with anergy against tuberculin (23), and similar findings were reported also in influenza and mumps infections (t, 14, 18). Significant decrease of delayed cutaneous sensitivity to various recall antigens was demonstrated in vaecinees with attenuated rubella, virus (3). Epstein-Burr (EB) virus genome, after primary infection, becomes integrated in human lymphoeytes (25). Evidence for previous infection is obtained from the presence of antibodies to the viral capsid antigen (VCA) (15) and the virus can also be found in the oropharynx (4). In subacute sclerosing panencephalitis (SSPE) patients, we have found elevated titers of antibodies to EB virus as 14 Arch. Virol. 57/3

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T. GOTL:IEB-STElWATSKY,L. RANNOI% A, Vo~csovER, and N. V.~Rsa>-o:

compared to m a t c h e d controls (6). These findings were similar to those reported b y JOSTeAS et al. (13). Three cases of infectious mononucleosis (ISI) associated with SSPE were also reported b y F~oRI~vo et al. (2). S S P E is k n o w n to be related to chronic measles infection ( t l ) a n d i n some eases associated with impaired cell mediated i m m u n i t y (5). I t was also d e m o n s t r a t e d t h a t in acute measles infection circulating l y m p h o c y t e s become infected (17) a n d virus a n t i g e n was demonstrated in t h y m u s a n d l y m p h o i d tissues (24). I t was therefore of interest to find out. how acute viral infections affect E B virus.

Materials and Methods Acute and convalescent blood samples were obtained in the years 1975--76 from patients with meastes, infections due to influenza A and B, adenoviruses and heMthy controls. The first blood samples were obtained as follows: in the influenza patients between the second and the fifth day of illness, in the measles patients up to the seventh day after a,ppearanee of the rash and in the adenovirus group between the fourth and seventh day after beginning of illness. The interval between the first and second blood sample was seven to 40 days. Out of 77 patients, 24 were children two to 14 years oId and the rest were young adults, 15 to 24 years old. I n the control group, 13 were in the younger age group and 23 were in the older group. There was no significant difference in the age distribution of patients and controls [p (X~)>0.90]. Laboratory diagnosis was based on serologieM criteria with or without virus isolations. Antibodies to influenza A and B and the adenovirus group antigen were tested b y complement fixation (CF). Influenza antigens were prepared from A/Port Chalmers/ 1/73 and B/Victoria/98926/70 strains. Virus isolations were carried out in embryonated eggs art in primary monkey kidney cell cultures. Adenovirus CF antigen was prepared from. adenovirus type 2 grown in h u m a n kidney cell line and isolation of virus was carried out, in the same cell line. Antibodies to measles virus were determined by the hcmaggtutination inhibition (HI) test. Measles antigen was prepared from Edmonston strain grown in I-IeLa cells and treated with tween and ether (16, 19). Antibodies to EB.VCA and early antigen (EA) were determined by the indirect immunoftuorescence (IF) method (7, 10). PaHB-1 ceils were used for demonstration of EB-VCA. For detection of antibodies to EA, Raji cells superinfected with EB virus obtained from I-IR 1K cells were used. Sera were tested in twofold dilutions for VCA antibodies starting at 1 : t0 to 1 : 320. Antibodies to EA were determined in the following dilutions : 1 : 5, 1 : 10, 1 : 20. Heterophile antibodies were tested by the monotest (VV~ampole Laboratories).

Results A comparison of a n t i b o d y titers to EB-VCA in h e a l t h y individuals a n d the three groups of p a t i e n t s was carried out (Table 1). A m o n g a total of 77 patients, t e n did n o t show antibodies ot EB-VCA at 1:10 dilution in b o t h blood samples a n d a m o n g the 36 controls eight were w i t h o u t antibodies at this titcr. The distrib u t i o n of a n t i b o d y titers i n the first, a n d second blood samples of t,he p a t i e n t s as compared to the controls is presented together with the c u m u l a t i v e percentage of those possessing antibodies at 1:160 a n d above. A m o n g the p a t i e n t s the percentage r a n g e d between 12.5 to 22.5 in the first a n d 20.8 to 35.0 in the second blood sample. No antibodies were f o u n d in the control group a t this titer. The differences i n titer between the three groups of p a t i e n t s were n o t statistically significant [p (X 2) = 0 . 1 5 ] . On the other h a n d the a n t i b o d y titers in p a t i e n t s

EBV Antibodies in Acute Viral Infections

201

were significantly higher as compared to controls. [For the first blood samples p (X 2) = 0 . 0 1 5 a n d for the second p (X2) 1 : 3 2 0

Percent (%) ~t:160

Table 2. N u m b e r o/patients with and without change in antibody titer to E B - V C A /rein the acute to the convalescent stage

Group

Total tested

Without change

×2

Influenza

40

29

8

Measles

24

21

2

Adenovirus

13

8

1

Increase ×4 × >8 3 3

Percent (%) increase 27.5

1

12.5

1

38.5

I n Table 3 t e n selected p a t i e n t s with at, least fourfold rise in antibodies to E B virus between the two blood s~mples or -with a tiger 1 : 320 or abOve are presented. Details concerning clinical diagnosis, age, sex, d a y of blood sampling, a n t i b o d y tigers to influenza, measles or adenovirus together with a n t i b o d y tigers to EB-VCA a n d E A are recorded. Out of four influenza p a t i e n t s three h a d shown a fourfold rise i n antibodies to E B - V C A a n d one h a d also a rise i n antibodies to EA. F r o m p a t i e n t No. 1, herpes simplex virus was isolated together with influenza virus a n d from p a t i e n t No. 2 influenza virus was also isolated, b o t h of the A/Victoria/75 v a r i a n t . Two measles p a t i e n t s arc included, one with a rise of antibodies to VCA from t : 10 to 1 : 320 a n d to E A from less t h a n 1 : 10 to 1 : 20. The second p a t i e n t , with post measles GuillMn Barr6 syndrome, h a d shown a rise in antibodies to E A from less t h a n 1 : i 0 go 1 : 40. As to the four adenovirus patients, a d e n o v i r u s t y p e 4 was 14"

202

T. CxOTLIiEB-STE:~ATS:giY,L. II~A~'~'O~ ", A. VO~-SOVE~¢,and N. VA?aSANO:

isolated from p a t i e n t No. 7 a n d 8, P a t i e n t No, 10, a child with severe bronchop n e u m o n i a , had c o n e o m i t t a n t rise in antibodies to adenovirus a n d influenza A as well as to E B virus. Monotest was negative in all p a t i e n t s a n d no evidence for clinical infectious mononucleosis was recorded. Table 3. Antibody titers to E B virus in selected patients with influenza, measles and adenovirus in/ections Patient Clinical diagnosis

Sex/ Age

Blood Days sam- after ple onset Antigen

AntiEBV body titer VCA EA

1

Influenza

M/19

1 2

2 17

Influenza A

10 60

80 320

< 5 10

2

Influenza

M/18

1 2

4 18

Influenza A

15 60

80 320

l0 10

3

Influenza

M/18

1 2

3 17

Influenza A

I0 480

320 320

t0 10

4

Influenza

M/18

i 2

4 18

Influenza A

15 120

t0 ¢0

5 5

5

Measles

F/t1

1 2

7 35

Measles

32 128

10 320

320

i0 10

t0

Bronehopneumonia

F/3

1

2

Adenovirus Influenza A

10 15

80

< 5

2

36

Adenovirus Influenza A

40 t 20

320

5

Discussion E l e v a t i o n a n d f l u c t u a t i o n of a n t i b o d y titers to E B virus i n various chronic diseases characterized b y their i m m u n o s u p p r e s s i v e effect, as sarcoidosis or l y m p h o m a s a n d leukemia were found (8, 12). Raised a n t i b o d y titers were also described in SSPE (6, 13). The present s t u d y d e m o n s t r a t e s the effect of acute viral infections on the level of antibodies to E B virus as studied in p a t i e n t s infected with influenza, measies a n d adenovirns. A considerable percentage of p a t i e n t s had shown a m e a n i n g f u l change of a n t i b o d y titers between the first, a n d second blood samples. There was no evidence of infectious mononucleosis as d e t e r m i n e d b y the presence of heterophile antibodies or b y clinieM symptoms, Also no one of the p a t i e n t s w i t h o u t antibodies (negative a t I : 10) in their first blood samples has s h o ~ a seroconversion.

EBV Antibodies in Acute Viral Infections

203

Impaired cell-mediated immune response found during influenza and measles infections is probably related to the effect of the virus on the patient's lymphocytes. I n volunteers infected with virulent or attenuated influenza virus, suppressed T-cell function was demonstrated (14). In a similar study, reduction in the number of T-cells was evident 24 hours after infection, preceding the onset of clinical illness (20). This could explain the raised titer of antibodies to EB virus found already in the first blood sample of some patients. It was also found that circulating lymphoeytes, during acute measles are infected by the virus (9, 17). Raised antibody titers to EB virus were present already in the majority of the first blood samples of the measles patients. This could be due to the length of the incubation period of measles, in which the patient's lymphocytes become infected. Changes in interaction between subpopulation of 13~nphocytes during viral infections may lead to reactivation of the EB viral genome which is integrated in the B lymphocytes and antibody formation is stimulated even before the disease becomes overt. As for the adenovirus patients, the adenoid tissue is chronically infected by adenoviruses (21) and this m a y have a prolonged suppressing effecfG on cellmediated immune response. I t is tempting to speculate that concomittant infection of adenovirus and EB virus in the oropharynx contributes to the pathogenesis of nasopharyngeal carcinoma, in which it is still unclear how the epithelial tissue becomes infected by EB virus (22). Interactions between viruses causing acute infections in man and a virus with malignant potential should also be considered.

Acknowledgments We thank M. Modan, Department of Clinical Epidemiology, The Chaim Sheba Medical Centre, TeLHashomer, for statistical analysis of the data. The technical assistance of M. Levy and Y. Shlomo-David is greatly appreciated.

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(1976). 4. GERBER, I~., NONOYAMA, M., LUCAS, S., PERLII~, E., GOLDSTEIN, L. I." Oral excretion of Epstein-Bert virus by healthy subjects and patients with infectious mononucleosis. Lancet II, 988--989 (1972). 5. GERSON, 14[. L., I-IAsI~AM, R. li. A. : Subtle immunologic abnormalities in four boys with subacute sclerosing panencephalitis. J. Amer. reed. Ass. 216, 1201--1202 (1971). 6. GOTLIEB-STE~-VIATS!KY, T., RAI~+NON, L., VO:KSOVER, A. : Antibodies to Epstein-Barr virus in subacute sclerosing panencephalitis patients. Europ. l~eurol. 13, 418--421

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