The Effect of Mumps Virus on the Resistance of Burkitt Lymphoma Cell Lines to Various Viruses1

[CANCER RESEARCH 29, 481â€”488,February 1969]

BarbaraA. Zajac,WernerHenle,2and Gertrude Henle

Virus Laboratories, The Children â€˜sHospital of Philadelphia, and the School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19146

SUMMARY EBV3 or not, yield an interferon-like inhibitor (4, 12, 13, 24, 27) which, since it may be synthesized without apparent stim Infections by certain paramyxoviruses were shown by others ulation, has been termed autogenous interferon (Zajac et a!., in to increase the susceptibility of cell cultures to various unme preparation). All attempts have failed to transmit EBV to host lated viruses. Burkitt tumor cell lines were exposed therefore systems other than cultures of cells from the hematopoietic to mumps virus in an attempt to enhance replication of the system in which again mainly low-grade persistent infections Epstein-Barr herpes group virus (EBV) present in a small pro are established4 (16). Efforts were made therefore to im portion of the cells. Persistent mumps virus infections were prove EB viral replication in existing carrier lines. Several me readily established in every line tested with nearly every cell ports have indicated that pamamyxovimuses (mumps, parainflu (>95%) synthesizing viral antigen. Average cellular growth enza, Newcastle disease viruses) may lower the resistance of mates were reduced by as little as 10% and no more than 50% cell population to various unrelated viruses (10, 17, 21-23, 31) as compared to uninfected control populations maintained in and that this effect may be due to reduced interferon synthesis parallel. After an initial peak, mumps virus titers persisted at (23) or prevention of interferon action (2, 11) or both (17, @ levels of i0@I to 50% tissue culture infective doses/ml of 18). Consequently, selected cell lines of Burkitt tumor or leu culture for at least 3 months. The mumps viral carrier state kemic origin were exposed to mumps virus in order to deter increased the susceptibility of Burkitt tumor cell cultures to mine its effect upon the resistance of the cultures to various vesicular stomatitis virus. It did not reduce autogenous or reo viruses and on the EBV viral carrier state. virus Type 3-induced interferon synthesis, but it prevented the protective action of exogenous human interferon. The supemin MATERIALS AND METHODS fection by mumps virus failed to affect the EBV carrier state. The percentage of EBV-antigen-producing cells remained con Cells. The Bumkitt cell lines selected for these studies were stant, and inoculation of various host systems with samples (a) the EB-1 line (6), which contained EBV and produced from the mumps viral carrier cultures, collected at varying autogenous interferon; (b) the EB-3 line (7), which carried the intervals after infections and mixed with a potent antiserum to virus but did not produce detectable amounts of autogenous mumps virus, provided no evidence for transmission of EBV. interferon; (c) the Raji line (26), which neither contained virus particles nor produced autogenous interferon (5) ; and (d) the INTRODUCTION Ogun line (Osunkoya and Pulvertaft, personal communica tion), which contained rare cells with EBV particles, yet pro Lines of blastoid cells derived from Burkitt's lymphomas or duced autogenous interferon. In addition, 2 EBV-free cell lines peripheral leukocytes of leukemic and other patients or of derived from peripheral leukocytes of leukemic patients were healthy donors often harbor a herpes group virus which has used for some of the experiments. These were the SK-Li line been referred to as Epstein-Barr virus after the first cell lines in (3), which produced autogenousinterferon, and the RPMI which it was discovered (7). According to recent observation, 6410 line (20), which did not. The Bumkitt cell lines were this virus is closely related to, if not identical with, the causa kindly supplied by M. A. Epstein, Bland Sutton Institute, Mid tive agent of infectious mononucleosis (15, 25). Generally dlesex Hospital, London, England. The SK-L1 line was fur only a small percentage of cells in the carrier cultures produce nished by M. B. Clarkson, Sloan-Kettering Institute, New virus particles, the majority of which lack part or all of the York, New York, and the RPMI 6410 line was furnished by J. nucleoid (7). Many of the cell lines, whether they contain

3Abbreviations used: EBV, Epstein-Barr virus; HEK, human embry onic kidney; VSV, vesicular stomatitis virus; TCD50, 50% tissue culture infective doses; ID50, 50% infectious doses; L(MCN), Earle's strain L cell cultures; NDV, Victoria strain of Newcastle disease virus; HBSS-2, â€˜This investigation was supported by Grants CA-04568 and 5T1 Hanks' balanced salt solution supplemented with 2% fetal calf serum. AI-00104 from the NIH, USPHS. 4J. S. Horoszewicz, V. C. Dunkel, and J. T. Grace, Jr. Biologica@ 2Career Award, 5-K6-AI-22,683, NIH, USPHS. Properties of a Herpes-like Virus (HLV) from Bumkitt Lymphoma Cell Received June 7, 1968; accepted October 22, 1968. Line, submitted for publication.

FEBRUARY 1969 481

T. Grace, Jr., Roswell Park Memorial Institute, Buffalo, New for detection of viral antigens: (a) human mumps hyperim York. mune @-g1obulin;(b) for EBV, the same as a but exhaustively All lines were maintained on Medium #1629 (Baltimore Bio adsorbed with mumps virus; (c) rabbit anti-VSV 7-globulin; logical Laboratories, Baltimore, Maryland), supplemented with (d) rabbit anti-NDV v-globulin; and (e) goat anti-reo VirUS 10% fetal calf serum, 2 mM glutamine, 100 units of penicillin, Type 3 7-globulin, kindly furnished by E. H. Lennette, and 100 iig streptomycin/ml. The lines were subcultured week California State Health Department, Berkeley, California. ly by seeding 1 X i0@ viable (trypan blue exclusion) cells/mi Preparation and staining of cell smears and of infected cover of fresh medium. slip cultures have been described (14). Human diploid cells (WI-38) were obtained from the Wistar Interferon Production and Assays. For autogenous inter Institute, Philadelphia, Pennsylvania, and HEK cultures from feron production, cells from various cultures were sedimented Flow Laboratories, Rockville, Maryland. Primary rabbit kid and resuspended in fresh medium to a concentration of 1 X ney cultures were kindly supplied by T. Tokumaru, of this 107/ml. The suspensions were incubated on a rotary shaker for hospital, fat head minnow cells by R. Maisberger, Lehigh Uni 24â€”72 hr as described elsewhere (Zajac et a!., in preparation). versity, Allentown, Pa., and turtle heart cell cultures by D. T. For virus-induced interferon production, from 5 X 106 to 5 X Karzon, Children's Hospital, Buffalo, New York. The above io'@ cells were resuspended in 1 ml of virus suspension, diluted cell cultures were maintained on Eagle's basal medium in to yield desired input multiplicities, and incubated at 37Â°Cfor Hanks' balanced salt solution containing 2â€”10% fetal calf se 1 hr. After centrifugation the cells were resuspended to a con rum, glutamine, penicillin, and streptomycin. The pH of the centration of 1 X 106 to 1 X 107/ml and further handled as media was adjusted to 7.4 with sodium bicarbonate. L(MCN) described. At the end of the incubation period, the cultures were maintained as described (29). were centrifuged and the supernates were dialyzed for 24 to Viruses. The lUcid strain of mumps virus was derived from a 72 hours at 4Â°C against 100 volumes of HC1-KC1 buffer at pH typical case of parotitis by amniotic inoculation of 8-day-old 2.0 and for another 24 hours against 100 volumes of phos chick embryos. Amniotic fluids of the 7â€”9th consecutive pas phate-buffered saline at pH 7.0. The samples were stored at sages were used. The stock virus preparations contained ap 4Â°Cuntil assayed. proximately 1083 ID50 for chick embryos and i0@ hemagglu Serial two-fold dilutions of interferon samples were made in tinating units/mi. HBSS-2, and 0.5-mi aliquots were added to duplicate cultures Stocks of the Indiana strain of VSV were derived from in of HEK cells. After 24 hr incubation at 37Â°C, interferon fected chick embryo fibroblast cultures. These preparations treated and control cultures were challenged with 0.2 ml of contained between 108 to iO@ TCID50/ml when assayed on VSV suspension containing 1000 TCD50. In assays of virus HEK or L(MCN) cultures respectively. induced interferons, the cultures were mefed prior to challenge NDV had been propagated for an undetermined number of with VSV. After 40 hr at 37Â°C, the controls showed total allantoic passages in embryonated chicken eggs. These prepara destruction of the cells. The endpoints of interferon activity tions contained 109.0 to i0@5 ID50 /ml for chick embryos. were taken as the highest initial dilution of sample which gave Reovirus Type 3 was obtained from S. Dales, Public Health 50% suppression of viral cytopathic effects. Reovirus-induced Research Institute of the City of New York, and passaged in interferons were diluted in HBSS-2 which contained a 1:800 HEK cultures. The stock preparation contained 108.2 dilution of monkey anti-reovimus Type 3 serum obtained from TCD50/ml when assayed in HEK cultures. the Virus Research Resources Branch of the NIH, Bethesda, Infection of Cell Cultures and Virus Assays. For infection, 5 Maryland. A dilution of 1 : 10,000 of this serum neutralized x 106cellsweresedimentedandresuspendedin1to2mlof iO@ TCD50 of reovirus when assayed in HEK cultures. virus, appropriately diluted to yield the desired input multi plicity. After incubation with intermittent shaking for 1 to 2 RESULTS hr at 37Â°C,the cells were sedimented again and resuspended in growth medium to a concentration of 2 X iO@ cells/mi. Unin Course of Mumps Virus Infection in Burkitt Tumor Cell fected control cultures were handled and maintained in paral Lines. Cells of the EB-1 , EB-3, and Raji lines were exposed to id. Samples were removed at intervals for cell counts, infectiv mumps virus at an input multiplicity of 5, and the resulting ity titrations, and preparation of cell smears for immunofluo cultures as well as uninfected controls were observed there rescence tests. after for cellular growth, percentages of cells containing Samples of cell suspensions for infectivity titrations were mumps antigen, and yields of infectious mumps virus. As seen stored in ampuls at â€”70Â°Cuntil assayed. Four tissue culture in Chart 1 , the reduction in cellular growth mate was most tubes each were inoculated with 0.2-mi volumes of given marked in the EB-3 mumps culture, ranging from 18 to 63% at 10-fold virus dilutions and incubated at 37Â°C. Mumps virus various intervals with an average of 48%. The EB-1 mumps and was assayed in HEK cultures, and the extent of infection was Raji mumps cultures showed an average reduction in cell repli determined on the 6th day by hemadsorption according to the cation of 20 and <10% respectively. The titers of mumps virus technic of Vogel and Shelokov (32). VSV was titrated in in the initial period rose at different rates and to varying peaks L(MCN) cells, and the results were calculated on the basis of depending upon the cell line, but after establishment of an vimus-specffic cytopathic effects on the third day after equilibrium the titers persisted at levels between 10@ and @ inoculation. TCID50 /ml as determined in HEK cultures. The actual Immunofluorescence Assays. The following fluorescein iso quantities of mumps virus were probably 100-fold higher, thiocyanate-conjugated antibody preparations were employed since comparative titrations of the virus in HEK cultures and

482 CANCER RESEARCH VOL.29

enhanced susceptibility to VSV was evident from a marked reduction in the cellular growth mates, a 5- to 20-fold increase in the number of cells showing VSV-specific immunoflu orescence, and a similar increase in the VSV titers obtained within 3 days after challenge. In order to establish these ob servations firmly, EB-1 cells were exposed to mumps virus as described. At various intervals thereafter, cells from this cul ture, as well as from uninfected control populations, were sedimented and challenged with VSV at an input multiplicity of 2. The resulting sublines of the EB-1 and EB-1 mumps cultures were monitored by cell counts, VSV and mumps virus specific immunofluorescence, and infectivity titers. The results (Chart 2) showed a gradually increasing susceptibility of the EB-1 mumps culture to superinfection with VSV. The en hancement, as measured by reduction in cellular growth, was @ not dramatic before the 52nd day following exposure to mumps virus. At that time and thereafter, the effect of VSV on cellular growth was rapid and pronounced. VSV-specific immunofluorescence tests revealed low degrees of enhance ment already in the first week after infection of the culture with mumps virus, and the differences between the EB-1 and EB-1 mumps cultures became more striking as time progressed. @ 30 40 n so so 40 20 30 40 At each time of challenge, the enhancement of the VSV-super 0 CONTROL DAYS AFTER INFECTION infection was most pronounced within the first 3 ensuing days. . MUMPSINFECTED Thereafter, the percentages of cells with VSV antigen declined Chart 1. Course of mumps virus infection in EB-1, EB-3, and Raji to lower levels. All the experimental cultures in this expemi cells. VP, virus particles; FA, fluorescent antibody. ment survived as triple carrier populations, harboring EBV, mumps virus, and VSV. Another EB-1 mumps carrier culture, in the chick embryo amnion generally revealed differences of 2 which had been established and maintained in the same man log10 units in favor of the latter assay system. Even with nem, succumbed to VSV when challenged on the 66th day after corresponding corrections, the titers obtained were low in view exposure to mumps virus (Chart 3). of the fact that within 5 to 10 days after exposure, and con Similar, but less extensive experiments, were carried out tinning thereafter, practically all cells contained mumps anti with EB-3 and Raji cells. The results obtained with the EB-3 gen, as evident from mumps-specific immunofluorescence. It is mumps cultures were comparable to those described for EB-1 obvious that infected cells retained their capacity to divide mumps populations. Raji mumps cultures failed to reveal sig since the cultures continued to grow at only slightly reduced nificant differences in response to VSV, as compared to the rates. Only a few of the cells contained sufficient mumps anti controls, when challenged either on the 21st or 90th day after gen to mender the whole cell intensely fluorescent. Such cells exposure to mumps virus. were seen mainly during the early incubation period. The great Effect of Mumps Virus Infection upon Interferon Synthesis. majority of cells contained well-delineated aggregates of anti Cell lines of Burkitt tumor or leukemic origin which either did gen of varying sizes (Fig. 1), often amounting to no more than or did not produce autogenous interferon were exposed to minute granules. Infection of other cell lines, i.e., the Ogun mumps virus and, after establishment of the carrier state, ex line of Burkitt tumor, and the SK-Li and RPMI-6410 lines of amined for its effect upon interferon synthesis. The results leukemic origins, yielded similar results. (Table 1) revealed that the mumps infection failed to induce The above observations beam a striking resemblance to the per se detectable interferon production and to reduce autoge persistent mumps viral infection of Chang's human conjunctiva nous interferon yields. cells as reported by Walker and Hinze (34). Later experiments The effect of mumps virus infections on production of virus (35, 36) indicated that a reduction in serum concentration in induced interferon was examined by exposure of mumps cam the culture medium increased the virus yields and the number rier and control cultures to either NDV or meovimus Type 3. of cells showing hemadsomption. Similar experiments with the The EB-1 and Ogun lines could not be stimulated by NDV to EB-1 line failed to show differences in the various parameters produce interferon in excess of autogenous inhibitor synthesis. studied, whether the cultures were fed medium with 10 or 2% The experiments with NDV (Table 2) were restricted therefore fetal calf serum immediately after exposure to mumps virus or to cell lines devoid of the capacity to yield autogenous inter after establishment of the carrier state. femon (Raji and RPM! 6410) and to the SK-Li line capable of Increased Susceptibility of Mumps Carrier Cultures to VSV. In producing interferon in excess of autogenous interferon. The preliminary experiments it was found that EB-1 mumps cul mumps viral carrier state had different effects on NDV-in tumes were more susceptible to VSV compared to uninfected duced interferon synthesis depending upon the line under test. controls and that the susceptibility to VSV appeared to in It prevented interferon production in the Raji culture, reduced crease with prolongation of the mumps viral carrier state. The synthesis 8-fold in SK-L1, but only insignificantly in RPM!

FEBRUARY 1969 483

@oo

I') 000 0

800 E

(I) -J -I 300 w C.)

-j 400

> 200

40 30 >u) (fl-i>â€”j20 @C) go

246 246 246 246 246 246 246 246 246

0 EB-I + DAYS AFTER INFECTION @ . EBlmumps VSV

Chart 2. Gradual enhancement ofsusceptibiity to VSV in an EB-1 mumps culture. VSV, vesicular stomatitis virus; FA, fluorescent antibody.

6410 cells. NDV-specific immunofluorescence tests on cell whether treated with interferon or not, the cellular growth smears prepared at the time of harvest of the interferons clear rates, VSV-specific immunofluorescence, and VSV-infectivity ly showed that the mumps carrier state did not enhance infec titers were closely similar. In contrast, EB-1 cultures free of tion by another paramyxovirus, but rather reduced it 2.5- to mumps virus and treated with interferon showed good protec 10-fold. The percentages of cells with NDV antigen in all 3 tion on the basis of all 3 parameters when compared to the mumps carrier cultures, however, were similar; yet the yields controls which were not exposed to interferon. of interferon varied from <4 to 128 units. When the same cell Effect of Mumps Virus on the Indigenous EB Virus Infec lines were exposed to reovirus Type 3 at different input multi tion. EB-1 and EB-3 cultures were tested 2, 5, 7, 14, and 50 plicities, the yields of interferon were similar in the control days after infection with mumps virus for EBV-specific im and mumps virus-infected cultures (Table 3). Test for meovirus munofluomescence, and samples taken at these intervals were specific immunofluorescence again failed to show enhance inoculated into various types of tissue cultures and newborn ment of the supeminfection by the mumps viral carrier state, and weanling animals in an effort to detect enhancement of but, in contrast to the results with NDV, there was no reduc the indigenous agent. For the first approach, cell smears from tion in the number of reovirus-infected cells. mumps-infected and control cultures were stained for mumps Effect of Mumps V@irusInfection on the Protective Action of antigens as well as EBV, using a fluorescein-labeled human Extraneous Interferon. EB-1 mumps cultures, infected 66 days mumps hypemimmune globulin before and after exhaustive previously, and controls were exposed to interferon produced adsorption with mumps virus. The adsorbed conjugate failed by SK-Li cells on stimulation by ultraviolet-inactivated NDV to detect the characteristic mumps antigen aggregates but me (Zajac et a!., in preparation). Cells were sedimented (5 X 106), tamed its capacity to stain those EB-1 and EB-3 cells which resuspended in 2.5 ml of the interferon preparation (170 harbored EBV (1â€”2% and 3â€”5% respectively). The mumps units/ml), incubated for 24 hours at 37Â°C, and then challenged virus-infected Bumkitt cell lines failed to show significant in with VSV at an input multiplicity of 2. After an adsorption creases in the percentages of EBV-positive cells at any time in period of 2 hours, the volume of the culture was adjusted to the course of the experiments. yield 2 X iO@ cells per ml. Control cultures, carried in parallel, The various samples of cell suspension, which had been pre were treated with (a) VSV only, (b) interferon only, and (c) served at â€”70Â°Cuntil assay, were exposed to sonication for 3 medium only. The experiment was monitored by cell counts, minutes (Disintegrator-Forty, Ultrasonic Industries, Inc., Plain VSV-specific immunofluomescence, and infectious VSV titers. view, New York) and then mixed with equal volumes of a 1:5 The results (Chart 4) showed that in the EB-1 mumps cultures, dilution of equine antiserum to mumps virus, kindly supplied

484 CANCER RESEARCH VOL.29

0 harvested and processed for EBV-specific immunofluor 0 escence. None of the inoculated cell cultures showed cyto -J 0 EBI CONTROL -@J pathic effects referable to EBV. In a few instances cytopathic 0 EBI+VSV effect was noted, however, which was found to be due to 6.0 unneutmalized mumps virus. Cultures without cytopathic effect 0 . EBIMUMPS were maintained by feeding and reseeding for as long as 40 . EBIMUMPS+VSV 0 days. These showed no evidence of interference at any time I- when challenged with VSV. The coverslip preparations failed 5.0 to reveal any immunofluorescence specific for EBV. w I.-. Mixtures of cell suspension and equine anti-mumps serum were inoculated also into litters of 1- and 6-day-old hamsters or mice by the intraperitoneal or intracerebral routes. The > U) 4.0 animals were checked daily for illness or death in the early > FA STAINING period and then at longer intervals for development of tumors. (DAY 3) None of the mice inoculated at birth or later developed signs 0 NONE of illness. About 12% of the hamsters infected at the newborn 600 stage became ill or were found dead after incubation periods of 10 to 17 days. The preparations involved were shown to >90% MUMPS contain unneutralized mumps virus. Passages of brain or spleen C/) homogenates from these animals into newborn animals and -J 400 -J cell cultures failed to produce illness or to show cytopathic w 2.5% VSV effects. Surviving animals of the primary and passage series 0 were kept for 18 months. No tumor formation became evi Li -J dent. 4 >90%MPS DISCUSSION > 75%VSV -Ill The persistant mumps virus infection of cell lines derived 0 5 I0 from Burkitt's lymphoma or peripheral leukocytes of leukemic patients resembles the mumps carrier state in Chang's human DAY AFTER CHALLENGE conjunctiva cell line as described by Walker and Hinze (34). Even at a high multiplicity of infection there was relatively Chart 3. Supeminfection with VSV of an EB-1 mumps culture initi little evidence of cell loss, and the cultures showed merely a ated 66 days previously. VSV, vesicular stomatitis virus; FA, fluorescent reduction in growth rates which were most marked in the EB-3 antibody; MPS, mumps. line but barely perceptible in Raji cultures. Practically all cells produced mumps antigen, but mostly in segregated, well delineated masses of variable sizes. The cells evidently were not greatly disturbed and continued to divide yielding infected 1Cell Table descendants. This type of carrier state has been termed â€œmegu mlControlMumps-infectedEB-13232Ogun2424SK-Li3232Raji<4<46410<4<4 units per lated infectionâ€• (33) or â€œendosymbiosisâ€• (9) and has been lineInterferon observed, in addition to mumps virus, with parainfluenza Type 1 (19), measles (29), rubella (28), and rabies viruses (9). The exact mechanism by which this type of carrier state is main tamed is presently unknown. Infection of cell cultures with paramyxoviruses, i.e., mumps, pamainfluenza, and Newcastle disease viruses, were shown to be Effect of mumps virus infection on the production of autogenous in accompanied by enhanced susceptibility to other, unrelated terferon. viruses (cf. 1). These observations were confirmed in that EB-1 mumps, and EB-3 mumps cultures were less resistant to VSV and also herpes simplex virus (Henle and Henle, unpublished) than the corresponding controls free of mumps virus. Raji by M. R. Hilleman, Merck Institute for Therapeutic Research, mumps cultures, however, failed to show enhanced suscepti West Point, Pa. After incubation for one hr at room tempera biity to VSV, nor did they or SK-Li mumps and RPM! 6410 ture, test tube cultures of HEK, W!-38, primary rabbit kidney, mumps cultures reveal increased susceptibility to reovirus fat head minnow, and turtle heart cells, with and without Type 3. Challenged with NDV, these cultures even showed a floating coverslips, were inoculated with 0.2 ml per tube of reduction in susceptibility which in part may be based on given mixtures. The cultures were observed for cytopathic destruction by mumps virus of cell receptors needed for ad effect and development of resistance to challenge with sorption of NDV. Obviously, enhanced susceptibility appears 100â€”1000 TCD50 of VSV. At various intervals coverslips were to be limited to certain cells as well as to certain viruses.

FEBRUARY 1969 485

Table 2

InputControlMumps-infectedNDV-specificNDV-specificCell Interferonimmunofluorescencelinemultiplicities Interferonimmunofluorescence cells)Raji of NDV (units/ml)(% + cells) (units/mi)(% +

<40.0100 <40.0 <48.650 6430.0 <414.580 25650.0 <416.0SK-L1 19653.0 80.0100 80.0 646.350 102430.0 12812.080 102430.0 12812.06410 102430.0

<40.0100 <40.0 163.250 6430.0 12811.080 25650.0 12811.0Effect 25650.0 ofNewcastleof mumps virus infection on theproduction of NDV-induced interferon.NDV, Victoria strain diseasevirus.Table

3Input

immunofluorescencelineCell multiplicities InterferonControlMumps-infectedReovirus-specificimmunofluorescence InterferonReovirus-specific cells)Raji of reovirus (units/mi)(% + cells) (units/nd)(% + 0<40.01 <40.0 <412.55 <412.1 6019.310 8016.6 24019.6SK-L1 24024.3 080.01 80.0 32032.95 32024.8 24032.110 32026.3 24029.06410 32023.4 0<40.01 <40.0 32051.05 32065.0 nt50.010 48065.0 320nt 32050.0 Effect of mumps virus infection on the production of reovirus-induced interferon. nt, not tested.

The reduction in cellular resistance has been ascribed to sup controls. Inhibition of interferon synthesis thus may be an pmession of interferon production (23) and/or prevention of occasional, but not a general, result of mumps virus infections. interferon action (2, 11, 17, 18). The data presented here have The experiments with EB-1 mumps cultures clearly indicated shown that autogenous interferon synthesis is not suppressed that the action of extraneous human interferon was prevented by a persistent mumps infection. The effects of a mumps viral by the persistent mumps infection. carrier state on virus-induced interferon production may be Despite the evidence for increased susceptibility of EB-1 influenced by the type of virus used as well as by the cultures mumps and EB-3 mumps cultures to VSV and HSV and for under test. With NDV as inducer, the results varied from com prevention of interferon action, no evidence was obtained that plete inhibition to no inhibition of interferon synthesis. This the indigenous EBV infection of these cell lines was enhanced variability was apparently unrelated to differences in adsorp in any detectable way. The percentage of cells producing EBV tion of the virus onto the cells, as akeady discussed, since antigen remained essentially stationary, except for minor fluc similar numbers of cells were infected by NDV in every in tuations also seen in controls, and efforts to transmit the virus stance. When reovirus Type 3 was used as inducer, interferon from mumps carrier cultures to other host systems failed, as synthesis was similar in mumps viral carrier cultures and their did earlier attempts with materials from stock cultures (8).

486 CANCER RESEARCH VOL.29

6. Epstein, M. A., and Barr, Y. M. Cultivation in Vitro of Human Lymphoblasts from Burkitt's Malignant Lymphoma. Lancet, 1: I200 252â€”253,1964. 7. Epstein, M. A., Barr, Y. M., and Achong, B. G. Studies with Bum I000 kitt's Lymphoma. Wistar Inst. Symp. Monograph, 4: 69â€”82,1965. I') 0 8. Epstein, M. A., Henle, G., Achong, B. G., and Barr, Y. M. Morpho logical and Biological Studies on a Virus in Cultured Lymphoblasts E 800 from Burkitt's Lymphoma. J. Exptl. Med., 121: 761â€”770, 1965. 9. Femnandes, M. V., Wiktor, T. J., and Koprowski, H. Endosymbiotic (I)-j Relationship between Animal Viruses and Host Cells. A Study of 600 Ui-j Rabies Virus in Tissue Culture. J. Exptl. Med., 120: 1099â€”1115, C.) 1964. Ui 10. Frothingham, T. E. Enhancement of Sindbis Virus Plaque Size in -J 400 Cell Cultures Treated with Mumps Virus-infected Egg Fluid. Virol 4 > ogy, 19: 583â€”586,1963. 200 11. Frothingham, T. E. Further Observations on Cell Cultures Infected Concurrently with Mumps and Sindbis Viruses. J. Immunol., 94: 521â€”529,1965. 12. Henle, G., and Henle, W. Evidence for a Persistent Viral Infection 8 in a Cell Line Derived from Burkitt's Lymphoma. J. Bacteriol., 89: 0 252â€”258,1965. 7 13. Henle, G., and Henle, W. Interference in the Detection of Viral >E (0%., 6 Carrier States. Wistar Inst. Symp. Monograph, 4: 83â€”90,1965. >0 C) 5 14. Henle, G., and Henle, W. Immunofluorescence in Cells Derived 0 C.) from Burkitt's Lymphoma. J. Bacteriol., 91: 1248â€”1256, 1966. I- 4 15. Henle, G., Henle, W., and DieM, V. Relation of Burkitt's Tumor 3 associated Herpes-type Virus to Infectious Mononucleosis. Proc. 75 Nati. Acad. Sci. U. S., 59: 94â€”101, 1968. 16. Henle, W., DieM, V., Kohn, G., zur Hausen, H., and Henle, G. C0-j 50 Herpes-type Viruses and Chromosome Marker in Normal Leuko 25 cytes after Growth with Irradiated Burkitt Cells. Science, 157: 1064â€”1065, 1967. g 3 5 7 I 3 5 7 17. Hermodsson, S. Inhibition of Interferon by an Infection with Para influenza Virus Type 3 (PIV-3). Virology, 20: 333â€”343, 1963. DAYS AFTER INFECTION 18. Hermodsson,S. Action ofParainfluenza Virus Type 3 on Synthesis of Interferon and Multiplication of Hetemologous Virus. Acta. Pathol. Microbiol. Scand., 62: 224â€”238,1964. Chart 4. Lack of protection of EB-1 mumps cultures by human inter 19. Ishida, N., Homma, M., Osata, T., Hinuma, Y., and Mijamoto, T. feron. FA, fluorescent antibody; VSV, vesicular stomatitis virus. Persistent Infection in HeLa cells with Hemadsomption Virus Type 2. Virology, 24: 670â€”672, 1964. 20. Iwakata, S., and Grace, J. T. Jr., Cultivation in vitro of Myeloblasts from Human Leukemia. N. Y. State J. Med., 64: 2279â€”2282, ACKNOWLEDGMENTS 1964. 21. Kato, N., Okada, A., and Ota, F. A Factor Capable of Enhancing The competent technical assistance of Mrs. Marjorie Cassel and Mrs. Virus Replication Appearing in Parainfluenza Virus Type 1 Marie Adams is gratefully acknowledged. (HVJ)-infected Allantoic Fluid. Virology, 26: 630â€”637,1965. 22. Kumagai, T., Shimizu, T., and Matumoto, M. Detection of Hog REFERENCES f.holera Virus by its Effect on Newcastle Disease Virus in Swine Tissue Culture. Science, 128: 336, 1958. 1. Cantell, K. Enhancement of Animal Virus Growth by Another Virus. 23. Macno, K., Yoshii, S., Nagata, I., and Matsumoto, T. Growth of In: M. Sanders and E. H. Lennette, (eds.), Medical and Applied Vi Newcastle Disease Virus in a HVJ Carrier Culture of HeLa Cells. rology, pp. 31â€”44, St. Louis, Missouri: Warren H. Green, Inc., 1968. Virology,29:255â€”263,1966. 2. Cantell, K., and Valle, M. The Ability of Sendai Virus to Overcome 24. McCombs, R. M., and Benyesh-Melnick, M. Studies on Acute Leu Cellular Resistance to Vesicular Stomatitis Virus. II. The Possible kemia and Infectious Mononucleosis of Childhood. I. Viral Inter Role of Interferon. Ann. Med. Exptl. Biol. Fennial, 43: 61â€”64, ference with Lymphoblastoid Cells of Spontaneously Transformed 1965. Bone Marrow Cultures. J. Natl. Cancer Inst., 39: 1187â€”1196, 3. Clarkson, B. D., Strife, A., and deHarven, E. Continuous Culture 1967. of Seven New Cell Lines (SK-Li to 7) from Patients with Acute 25. Niedemman,J. C., McCollum, R. W., Henle, G., and Henle, W. Infec Leukemia. Cancer, 20: 926â€”947,1967. tious Mononucleosis. Clinical Manifestations in Relation to EB Vi 4. Deinhardt, F., and Burnside, J. Spontaneous Interferon Production rus Antibodies. J. Am. Med. Assoc., 203: 205â€”209, 1968. in Cultures of a Cell Line from a Human Myeloblastic Leukemia. J. 26. Pulvertaft, R. J. V. Cytology of Burkitt's Tumor (African Lym NatI. Cancer Inst., 39: 681â€”683, 1967. phom). Lancet, 1: 238â€”240,1964. 5. Epstein, M. A., Achong, B. G., Barr, Y. M., Zajac, B., Henle, G., 27. Rabson, A. S., O'Conor, G. T., Baron, S., Whang, J. J., Legallais, and Henle, W. Morphological and Virological Investigations on Cul F. T. Morphologic, Cytogenetic and Virologic Studies in vitro of a tured Burkitt Tumor Lymphoblasts (Strain RAJI). J. Nati. Cancer Malignant Lymphoma from au African Child. Intern. J. Cancer, 1: Inst., 37: 547â€”559,1966. 89â€”106,1966.

FEBRUARY 1969 487

28. Rawls, W. F., and Melnick, J. L. Rubella Virus in Carrier Cultures 32. Vogel, J., and Shelokov, A. Adsomption-hemagglutination Test for Derived from Congenitally Infected Infants. J. Exptl. Med., 123: Influenza Virus in Monkey and Kidney Tissue Culture. Science, 795â€”816,1966. 126: 358â€”359, 1957. 29. Rodriguez, J. E., and Henle, W. Studies on Persistent Infections of 33. Walker, D. L. The Viral Carrier State in Animal Cell Cultures. Tissue Cultures. V. The Initial Stages of Infection of L(MCN) Cells Progr. Med. Virol., 6: 111â€”148,1964. by Newcastle Disease Virus. J. Exptl. Med., 119: 895â€”922, 1964. 34. Walker, D. L., and Hinze, H. C. A Carrier State of Mumps Virus in 30. Rustigian, R. Persistent Infection of Cells in Culture by Measles Human Conjunctiva Cells. I. General Characteristics. J. Exptl. [email protected] and Characteristics of HeLa Sublines Persist Med.,116: 739â€”750,1962. ently Infected with Complete Virus. J. Bactemiol., 92: 1792â€”1804, 35. Walker, D. L., and Hinze, H. C. A Carrier State ofMumps Virus in 1966. Human Conjunctiva Cells. II. Observations on Intracellular Transfer 31. Valle, M., and Cantell, K. The Ability of Sendai Vsrus to Overcome of Virus and Virus Release. J. Exptl. Med., 116: 751â€”758, 1962. Cellular Resistance to Vesicular Stomatitis Virus. I. General Char 36. Walker, D. L., Chang, P., Northrop, R. L., and Hinze, H. C. Persist actemistics of the System. Ann. Med. Exptl. Biol. Fennial, 43: ent, Noncytocidal Virus Infection: Nonsynchrony of Viral and Ccl 57â€”60,1965. lular Multiplication. J. Bacteriol., 92: 983â€”989,1966.

p @â€¢ .

â€¢@â€˜â€¢

@@ â€¢0 â€¢1 ,@ 4

E p. @ .

%. .

.@.

â€˜4. 1 & C,

Fig. 1. Mumps-specific immunofluorescence in EB-1 cells infected 15 days previously. X 1000.

488 CANCER RESEARCH VOL.29

Downloaded from cancerres.aacrjournals.org on September 25, 2021. © 1969 American Association for Cancer Research. The Effect of Mumps Virus on the Resistance of Burkitt Lymphoma Cell Lines to Various Viruses

Barbara A. Zajac, Werner Henle and Gertrude Henle

Cancer Res 1969;29:481-488.

Updated version Access the most recent version of this article at: http://cancerres.aacrjournals.org/content/29/2/481

E-mail alerts Sign up to receive free email-alerts related to this article or journal.

Reprints and To order reprints of this article or to subscribe to the journal, contact the AACR Publications Subscriptions Department at [email protected].

Permissions To request permission to re-use all or part of this article, use this link http://cancerres.aacrjournals.org/content/29/2/481. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC) Rightslink site.