Int. J . Cancer: 24, 819-825 (1979)

ACTIVATION OF MOUSE MACROPHAGES BY PYRAN COPOLYMER A N D ROLE IN AUGMENTATION OF NATURAL KILLER ACTIVITY Paolo PUCCETTI l x 3 Angela SANTONI I, Carlo RICCARDI ', Howard T. HOLDENz, and Ronald B. HERBERMAN Pharmacology, University of Perugia, 06/00 Perugia, Italy; and Laboratory of Immunodiagnosis, National Cancer Institute, Bethesda, Md. 20205, USA

' Institute of

Inoculation of mice with pyran copolymer resulted in activation o f natural k i l l e r (NK) cells as well as macrophages. Conditions optimal for the boosting of N K activity seemed t o differ from those optimal for macrophage activation as assessed by cytostasis of tumor target cells. Peak levels of macrophage cytostatic reactivity were found at about 7 days after drug injection and were only achieved by the highest doses of pyran tested. Macrophage activation was consistently higher i n the peritoneal cavity than i n the spleen, regardless of route of administration, in contrast t o the failure of i.v. pyran t o induce high N K reactivity i n peritoneal exudate cells. A t 2-3days after pyran treatment of older mice, N K augmentation reached peak levels, but only minimal macrophage activation was found. Despite these differences, macrophages played a role i n regulating N K activity i n pyran-treated mice. Functional macrophages appeared t o be required for augmentation of N K activity by pyran, since boosting was impaired by prior in vivo inoculation of silica. Macrophagesalsoappeared ableto inhibit NKactivity. I n younger mice that exhibited high spontaneous levels of N K activity, pyran treatment produced a substantial reduction in N K activity t o levels below those of untreated mice. This depression coincided w i t h the t i m e of peak levels of macrophage cytostasis. Furthermore, removal of adherent cells f r o m the spleen cells of these pyran-treated mice resulted in levels of N K activity almost as high as those of untreated mice. The possibility that the depression of N K activity i n young mice by pyran copolymer is due t o suppressor cells i s discussed.

In the accompanying paper (Santoni e f al., 1979), we provided evidence that an early response of older mice to treatment with pyran copolymer was an augmentation of the cell-mediated cytotoxicity against tumor cells exerted by natural killer (NK) lymphocytes (Herberman et al., 1975; Kiessling et al., 1975). Boosting of the cytotolytic activity varied among strains of mice and was demonstrable in the spleen as well as in the peritoneal cavity. The relative extent of the augmentation at either site was dependent on the route of drug administration. Within the range of doses tested, similar levels of augmentation were seen within 2-3 days of pyran treatment. Pyran copolymer is one of the most powerful stimulants of the reticuloendothelial system (Puccetti and Giampietri, 1978) and also induces the production of interfiron (Merigan, 1967). Interferon production and macrophage activation appear to be closely linked events. Interferon has been shown to augment the cytotoxic reactivity of macrophages (Schultz et al., 19776), and also plays a central role in boosting of N K activity in mice (Djeu eta/., 1979a, Gidlund eta/., 19781, rats (Oehler e t a / . ,

1978), and man (Herberman etal., 1979). In addition, macrophages have been shown to be required for interferon production and boosting of NK activity by poly I:C (Djeu era[., 19796). With the evidence for multiple effects of pyran, it has become important to determine the possible interrelationships among these effects and their roles in mediating the anti-tumor resistance induced by pyran (Harme1 and Zbar, 1975; Morahan and Kaplan, 1976). Jn the present study, an attempt has been made to assess the role that macrophages might play in modulating NK activity, by comparing the levels of macrophage and NK-cell activation at various times after pyran administration and after administration of the agent by various routes. The data suggest a role for macrophages in both boosting and depressing NK activity. MATERIAL AND METHODS

Mice Inbred CBA and BIO.A(2R) and hybrid [BALB/c >i DBA/2]F, (CDZF,) and [C57BL/6 x DBA}2]FI (BD2FJ mice were obtained from the Mammalian Gctietics Section, National Cancer Institute, Bethesda, Md., and tested at 8-16 weeks of age. Drugs Pyran copolymer (NCS 4601 5 ) was obtained from Hercules Research Center, Wilmington, Del. Pyran was dissolved in sterile 0.9% NaCl solution and the PH was adjusted to 7.0 using I .O N NaOH. The drug was administered intraperitoneally (i.p.) or intravenously (i.v.) to the mice in an injection volume equal to 1 "/, of their body weight. Silica (Santocel, grade 68, average agglomerated size 3 ,um, Monsanto Industrial Chemical Co., St. Louis, Mo.) was suspended at a concentration of 12 pg/ml in 0.9 % NaCl and injected in a volume of 0.5 ml/mouse. Preparation of effector cells Effector cells from various lymphoid organs were prepared as previously described (Ortiz de Landazuri and Herberman, 1972). Removal of adherent cells was performed by passage through a nylon column according to the method of Julius e t a l . (1973). Phagocytic cells were identified after an 18-h incubation at 37" C with 2-pm latex spheres; after two To whom reprint requests should be addressed. Received: June 1, 1979 and in revised form August 29,

a

I979.

820

PUCCETTI ET AL.

washes to remove residual latex, the cells were scored microscopically for uptake of latex particles.

100 r

slCr release cytotoxicity assay

Effector cells at various concentrations were incubated with 1 x lo4 51Cr-labelledtarget cells for 4 h at 37" C in round-bottomed 96-well plastic microtiter plates. After incubation, the plates were centrifuged at 800 g for 10 min and the radioactivity in 0.1 ml of supernatant was measured in a gamma scintillation counter. All combinations were performed in quadruplicate. A tissue culture cell line of YAC, a Moloney virus-induced lymphoma of A/Sn origin, YAC-1 (Cikes et al., 1973) was used as the target for NK activity. Baseline 51Cr release was determined by the use of unlabelled autologous target cells in place of lymphoid cells. This baseline was 10-18% of the total 51Cr incorporated into the target cells. The percentage of specific lysis was calculated as follows: test cum autologous control cpm "/, Cytotoxicity = x 100 total cpm autologous control cpm where test cpm is the mean cpm released in the presence of effector cells, autologous control cpm is the mean cpm released by targets incubated with autologous cclls, and total cpm is the total amount of "Cr incorporated into target cells. Growth inhibition assay ( C I A ) The human myeloid cell line K-562 (Lozzio and Lozzio, 1975), which is poorly sensitive to killing by murine NK cells (Haller et al., 1977; Nunn and

80

1

70 60 50 -

40 -

30 20 10 O-t

I 3 5 1 7 Day After wran Administration

FIGURE 1 - CD2FI mice treated with pyran (75 mg/kg) at different times (days 1 , 3, 5 , 7) served as a source of effector cells in a GIA. 0-0, spleen cells (A/T ratio 20:l); B-B, PEC (A/T ratio 5 : l ) ; 0 , B, untreated controls.

' 0'O P 2.5

8.3

25

75

225

Pyran (rnglkg) FIGURE 2 - BD2F, mice treated with different doses of pyran copolymer were a source of effector cells in a CIA performed 7 days after drug treatment. W-B, sp!een cells (A/T ratio 20:l); 0-0, PEC (A/T ratio 5 : l ) . 0 , B, untreated controls. Hcrbzrman, 1979), was used as the target for the macrophage-mediated cytostasis. These cells were maintained as stationary suspension cultures and were subcultured three times weekly. K-562 cells at 2 x 104/0.2 ml were incubated as quadruplicate samples either alone or in the presence of effector cells at different ratios in round-bottomed wells of tissue culture treated plates in RPMI-1640 (supplemented with 10% fetal calf serum, 25 mM Hepes buffer and 1 % gentamycin sulphate) in a 5 % CO, incubator at 37" c. Eighteen hours before harvesting, 0.1 pCi/well of 5-[1251]-2'-deo~y~ridine ([lT]dUrd, spec. act. 5 Ci/mg, Radiochemical Centre, Amersham, England) together with 0.01 pg/well of 5-fluoro-2'-deoxyuridine (FldUrd) were added. The test was terminated at 48 h by harvesting of the cells on a multiple suction-filtration apparatus (Mash TI, Microbiological Ass., Inc., Bethesda, Md.) by means of a fiberglass filter paper (Grade 934 AH, Reeve Angel Co., Clifton, N.J.). Radioactivity of paper discs containing the [1251]dUrd-labellednucleic acid was read in a scintillation counter (Packard mod. 5142). Targets incubated without effector cells always incorporated 75% of the added radioactivity and served as a baseline control. Results of experimental groups are expressed as percentage [lZ51]dUrduptake inhibition according to the formula: mean of counts % growth in the group x 100 inhibition = 100 mean of counts in baseline control Standard errors, usually less than 5 %, have not been reported.

82 1

AUGMENTATION OF N K ACTIVITY BY PYRAN TABLE I

MACROPHAGE ACTIVATION BY P Y R A N COPOLYMER: COMPARISON BETWEEN DIFFERENT ROUTES O F ADMINISTRATION ”/, growth inhibition

Treatment

Effector cells

2O:l

40:l

None Pyran 25 nig/kg i.p., day 7 Pyran 25 mg/kg i.v., day 7

‘ A/T ratios.

Spleen cells PEC Spleen cells PEC Spleen cells PEC

20.8

9.5

93.1

69.2

95.0

78.2

10:l

5:l

I .2 12.5 27.0 98.2 35.0 98.0

2S:l

I .2

3.8

96.5

77.5

81.3

29.0

All vdues were significantly higher than those detectable with untreated controls ( p . 0.01)

RESULTS

The efect of pyran treatment on macrophage-merliuted cytostasis Macrophage activation, following pyran treatment, has been shown to reach peak levels of cytotoxic activity within 5-7 days of drug injection (Kaplan et a/., 1974). In order to better define the possible relationship of pyran-induced macrophage activation with NK-cell activity, we systematically analyzed various parameters affecting pyran-mediated changes in macrophage activity. Macrophage activation was evaluated in terms of cytostatic effects against cultured K-562 target cells. Spleen and peritoneal exudate cells from 10-weekold pyran-treated (75 mg/kg) CDZF, mice were tested at various times after i.p. injection of pyran (Fig. 1). The macrophage-mediated cytostatic activity gradually developed over the 7-day observation period. PEC at an A/T ratio of 5:1 were consistently more efficient than spleen cells at an A/T ratio of 20:l. Significant levels of activity could be detected no earlier than day 3 in PEC and day 5 in the spleen. A dose-response study was then performed on day 7 after treatment. Sixteen-week-old BD2F, mice were given a wide range of drug doses (2.5-225 mg/ kg) and PEC, as well as spleen cells, were assayed (Fig. 2). A clear dose-dependency was found in that doses lower than 25 mg/kg failed to result in marked macrophage activation. Doses above 25 mg/kg, however, produced similar levels of cytostasis, when measured either at the A/T ratios shown in Figure 2 or at lower A/T ratios (data not shown). At this point we investigated whether different routes of administration would result in differences in the activation of macrophages by pyran. Spleen cells and PEC of 14-week-old BlO.A(2R) mice, given pyran 25 mg/kg 7 days earlier either intraperitoneally or intravenously, were tested in a GIA (Table I). A significant difference was found between the two routes of administration only at the lower A/T ratios, suggesting that the i.p. inoculation caused more substantial activation of PEC than i.v. inoculation. K-562 tumor cells are known to be poorly sensitive to murine NK effectors expressing spontaneous

levels of activity (Haller et al., 1977; Nunn and Herberman, 1979). However, since some mouse NK activity can be detected against K-562, it was important to assess the adherent nature of pyraninduced effectors operating in the GIA. Table I1 shows that spleen cells from 9-week-old CDZF, mice treated with pyran 75 mg/kg 7 days earlier lost most of their activity when passed twice through a nylon column. TABLE I1 EFFECT OF REMOVAL OF ADHERENT CELLS O N CYTOSTATIC ACTIVITY OF SPLEEN CELLS FROM PYRAN-TREATED MICE ~~

~~~~~

Treatment

depletion

:4 growth inhibition 4O:l

2O:l

None None

9.8 82.6

t 1 .O

Nylon-passed twice

19.5

9.7

~

None Pyran 75 mg/kg i.p., day 7 Pyran 75 mg/kg i.p., day 7

52.1

’ AIT ratios The cfik-t of silica particles in vivo on N K activity and on its boosting by pyran copdymer Silica, which is sclzctivc;ly toxic to macrophages (Allison, 1976), has been shown to sharply diminish the spontaneous levels of NK activity in rats and mice and the boosting effect of polyinosinic-polycytidylic acid (Djeu et a / . , 1979b; Oehler and Herberman, 1978). To determine the effects of silica in vivo on boosting of NK activity by pyran copolymer in mice, 16-week-old BD2F, animals were given silica particles (6 mg, i.p.) 1 day before pyran (25 mg/kg i.v.) and 4 days before testing of spleen cells for NK activity in a 4-h CRA against YAC-1 targets (Table 111). As previously observed (Djeu et a/., 1979b), the spontaneous levels of NK activity were significantly decreased by silica when compared to those of untreated controls. In a similar way, silica treatment also prevented maximal boosting of NK activity by pyran.

822

PUCCETTI ET AL. TABLE III

EFFECT OF SILICA ON PYRAN-INDUCED BOOSTING OF NATURAL KILLER ACTIVITY IN THE SPLEENS OF 16-WEEK-OLD BDZF, MICE

% cytotoxicity Treatment

1OO:l

None Silica 6 mg/niouse i.p., day 4 Pyran 25 mg/kg i.v., day 3 Pyran 25 mg/kg i.v., day 3, silica 6 mglmouse i.p., day 4

'

18.7 11.9 28.4 22.8

5O:l

25:l

13.9 9.9 20.9 14.8

10.5 5.8 15.9 10.7

AIT ratios.

The eject of pyran treatment on natural cytotoxicity of younger animals as measured during the development of macrophage activation As indicated above and in the accompanying paper (Santoni et al., 1979) the spontaneous levels of spleen NK activity in older, low-reactive mice, were boosted by i.p. treatment with pyran. Augmentation of reactivity, however, was a n early response, with decline by 5 days after administration. No such augmentation of NK levels could be shown in the spleen of younger mice after pyran treatment, but the existence of high levels of reactivity in untreated mice allowed a more prolonged follow-up on the effect of pyran on NK expression in these mice. Younger (7-week-old) CD2F1 mice were, therefore, given pyran 75 mg/kg i.p. and, at various times, their spleen cells were tzsted for N K a n d macrophage-mediated cytostasis (Fig. 3). The levels of NK activity, in addition to not being boosted on day 3, were found to be substantially lower than untreated controls by day 7, when macrophage-mediated cytostasis was reaching peak levels of reactivity. Dzpres-

a 0

40

c

sion in NK activity also correlated with the dose range of pyran needed to induce macrophage-mediated cytostasis. Pyran doses, which boosted NK activity at day 3 but were unable to activate macrophages at day 7, were also found to be incapable of decreasing the expression of NK, as shown in Table IV. Removal of nylon-adherent cells from the spleens of pyran-treated animals One possible explanation for the above results was that the activated macrophages were inhibiting or suppressing NK activity. Therefore, spleen cells froni younger (8-week-old) CBA mice treated with pyran (75 mg/kg, i.p.) 7 days earlier were passed twice through a nylon column and the eluted cells were tested for NK and for the presence of phagocytic

TABLE IV EFFECT OF VARIOUS DOSES OF PYRAN ON NK ACTIVITY A N D MACROPHAGE-MEDIATED CYTOSTASIS 7 DAYS AFTER TREATMENT Treatment

None Pyran

8.3 rng/kg 25.0 75.0 225.0

cytolysis

'

9.9 10.6 5.2 5.6 4.4

%growth inhibition

8.0 14.2 76.6 83.3 94.5

' A / T ratio in the CRA, 5O:I. - *AjT ratio in the GIA, 20:l. Source ofeffectors were spleens from 16-week-old BDZF, inice treated with various doses of pyran i.p. I days earlier.

cells by the uptake of latex particles. Table V shows that in association with a decrease in the number of phagocytic cells after column passage, the N K activity was increased and was almost as high as in the cells from untreated mice. DISCUSSION

3

5

7

Days After b r a n Administration

FIGURE3 - Spleen cells from CDZF, mice treated with pyran 75 mg/kg i.p. at different times (days 3,5,7) or untreated (day 0) were tested for NK against YAC-1 targets (A/T ratio 100:1), as well as for macrophagemediated cytostasis of K-562 tumor cells (A/T ratio 20:l). o--o, %cytolysis; ~ - - - m , % cytostasis; 0 , m, untreated controls.

The present study represents an approach to the problem of whether any functional relationship can be found between the levels of NK cell and macrophage activity following treatment of mice with the synthetic polyanion pyran copolymer. RES-stimulating agents can boost the spontaneous levels of NK activity (Herberman e f al., 1977) and recent evidence indicates that augmentation of NK expression is mediated largely through production or release of interferon (Djeu et al., 1979a; Gidlund et al., 1978; Herberman et al., 1979; Oehler et al., 1978). Pyran copolymer is both a strong RES-stimulator and interferon-inducer (reviewed by Puccetti and Giampietri, 1978), and in the preceding paper we provided evidence that pyran can indeed boost the spontaneous levels of NK activity (Santoni et a/., 1979). Such augmentation, most impressive in older mice, was a n early response t o drug treatment and was largely independent of the doses being employed. The level of boosting at a particular site varied with different routes of administration.

823

AUGMENTATION OF N K ACTIVITY BY P Y R A N

Our first approach to the role of macrophage in boosting of NK activity was represented by experiments in which we explored the effect of in vivo silica treatment on NK boosting by pyran. Intraperitoneal silica injections were found to exert a depressive effect on pyran-induced boosting of spleen activity as well as on the spontaneous levels of NK. The reduction of the spontaneous activity was indeed greater than that of the boosted activity. This was in line with a recent report by Oehler et al. (1978), who observcd that silica had a more depressive effect on the cytotoxicity of spleen cells from normal rats than on the cytotoxicity of poly 1:C-treated animals. A depressive effect of silica on mouse NK activity in vivo has also been observed (Djeu et al., 19796; Kiessling et al., 1977). Silica is considered to be selectively toxic to mononuclear phagocytes (Allison, 1976), whereas there is general agreement that N K cells are non-adherent and non-phagocytic (reviewed

mor cells and cytotoxicity is usually assessed as cytostasis or cytolysis of target cells (Harmel and Zbar, 1975; Schultz et al., 1976, 1977~).We decided, therefore, to monitor the state of macrophage activation after pyran treatment in terms of cytostatic activity against K-562 cultured target cells. Our main goals were to establish the kinetics of macrophage activation as well as to compare the conditions suitable for NK boosting and for macrophage activation. As expected, peak levels of cytostatic activity were found to occur at about 7 days, both in the spleen and in the peritoneal cavity, and the effector cells were nylon-adherent. Contrary to what had been found for N K boosling (Santoni et al., 1979) macrophage activation exhibited considerable dose-dependency but inoculation by the intravenous or intraperitoneal route gave similar results. These findings suggested that NK boosting and macrophage activation might be linked by a com-

TABLE V

REMOVAL OF ADHERENT CELLS O N NK ACTIVITY OF SPLEEN CELLS FROM PYRAN-TREATED 8-WEEK-OLD CBA MICE

Treatment

None Pyran 75 mg/kg i.p., day 7

Cell depletion

None Nylon-passed once None Nylon-passed once Nylon-passed twice

%cell recovery

% !atexpositive cells

100 27

5.0 0.9 14.1 1.2 0.7

100

32 13

% cytolysis

60.1 65.7 34.3 54.3

NT

50:l

25:l

46.8 59.2 31.5 44.9 46.2

36.5 50.1 24.7 31.8 34.3

~

Not tested.

by Herberman et al., 1977). It is likely, therefore, that silica may not act on NK cells themselves, but on phagocytic cells, which maintain or regulate the levels of NK activity in vivo. It has recently been shown that macrophages are required for production of interferon and boosting of NK activity by another agent, poly I :C (Djeu et al., 19796). To explore further the relationship between macrophages and NK cells, it was important to measure separately the effects of pyran on macrophages. Much available information concerning this point indicated that i.p. treatment of mice with pyran (usually 25-100 mg/kg) produces macrophage activation which is maximal at 5-7 days (Harmel and Zbar, 1975; Kaplan et al., 1974; Munson et al., 1970). In conjunction with the appearance of high levels of interferon and possibly as a consquence of this (Schultz et al., 19776), the resting macrophage changes into a metabolically hyperactive cell, exhibiting enhanced attachment and spreading on glass (Rabinovitch et al., 1977) and higher responsiveness to chemotactic stimuli (Meltzer and Oppenheim, 1977). This is accompanied by an increase in phagocytic activity (Kapila et al., 1971) and in the ability to produce lymphocyte-activating factor (Meltzer and Oppenheim, 1977). Pyran-activated macrophages are also selectively cytotoxic in vitro for tu-

mon initial event-possibly release of interferonbut might, thereafter, develop independently. Further experiments, however, again raised the question of a possible role for macrophages in regulating the levels of N K activity. The spontaneously high levels of NK reactivity exhibited by younger mice were found to be markedly decreased by 7 days following pyran treatment, when macrophage activation was maximal. Only the doses of pyran which activated macrophages led to a decrease in spleen NK activity. Passage of spleen cells through a nylon column partially restored the activity to the level of the untreated controls. A mere dilution of NK effectors in the spleen must be considered as one possible explanation for the depressed NK activity at 7 days after pyran. However, it seems unlikely that the increase in spleen cellularity which we observed can alone account for all of the loss in N K reactivity. The spleens of normal and pyran-treated CBA mice contained 380 and 108 total lytic units, respectively (a lytic unit being defined as the number of cells needed to produce a cytotoxicity 30% above the baseline). Furthermore, preliminary results (data not shown) from mixture experiments indicated that, 7 days after pyran treatment, the spleen cells contained adherent suppressor cells that could inhibit the activity of normal spleen cells. Recently, it has

824

PUCCETTI ET AL.

been shown that C. parvum (Savary and Lotzova, 1978) and other treatments (Cudkowicz and Hochman, 1979) can induce suppressor cells for NK activity. Further studies are in progress to determine whether the decreased NK activity in spleens 7 days after pyran treatment can be accounted for entirely by suppressor cells or whether other inhibitory or inactivating mechanisms are involved. In contrast to most studies in which pyran copolymer appeared to act as an anti-tumor agent, there have been a few reports regarding enhanced tumorigenesis following pyran administration (Gazdar et al., 1972; Kripke and Borsos, 1974). Recently Mohr et al. (1976) observed enhancement of a tumor allograft in hybrid F1 mice and no adequate explanation has been so far provided. General considerations o n the possible in vivo role for NK cells in

immunosurveillance (Herberman and Holden, 1978). along with these findings, may lead to a better understanding of the mechanisms by which pyran treatment augmented oncogcnesis or tumor allograft development. On the whole, the data presented in this paper suggest that the state of macrophage activation and NK expression, two major functions influenced by pyran copolymer lreatment, may be closely associated, and this supports tho hypothesis of a role for macrophages in regulating the levels on NK activity. ACKNOWLEDGEMENTS

This work was supported by Italy-USA cooporative program contract 78.01871.65 C.N.R., Italy.

L’ACTIVATION DES MACROPHAGES DE SOURIS PAR LE COPOLYMERE PYRANNIQUE ET SON ROLE DANS L’AUGMENTATION D E L’ACTIVITE N K L’inoculation de copolymere pyrannique a declenche, chez des souris, une activation des cellules N K et des macrophages. Les conditions optimales pour cette activation ne semblent pas &re les mimes pour les cellules N K et pour les macrophages, comme le montre la cytostase des cellules cibles tumorales. On a observe des pics d’activite cytostatique des macrophages 7 jours environ apres I’injection d u produit et seulement avec les plus elevees des doses testees. L’activation des macrophages etait regulierement plus forte dans la cavite peritonkale que dans la rate, quelle que soit la voie d’inoculation, alors que le pyranne en i.v. n’induisait pas de forte reaction NK dans les cellules d’exsudat peritoneal. Deux ou trois jours apres le traitement de souris Lgees, on observait un pic d’activite NK alors que I’activation des macrophagx etait minime. Malgre ces differences, les macrophages jouaient un rBle dans la regulation de I’activite NK chez les souris traitees au pyranne. Des macrophages fonctionnels semblaient necessaires pour que l’activite N K soit accrue par le pyranne, puisque I’activation etait freinee par inoculation prealable in vivo de silice. Les macrophages semblaient aussi pouvoir inhiber I’activit8 NK. Chez les jeunes souris, qui avaient des niveaux naturellement eleves d’activite NK, le traitement au pyranne produisait une diminution substantielk de cette activite, jusqu’a des niveaux inferieurs a ceux des souris non traitees. Cette depression colncidait avec le pic de I’effet cytostatique des macrophages. En outre, si I’on separait les cellules adherentes des cellules spleniques des souris traitees, le niveau de I’activite NK devenait presque aussi eleve que chez les souris non traitbes. La p-ssibilite que la depression de I’activite N K causee chez les jeunes souris par le copolymere pyrannique soit due A des cellules suppressives est discutee.

REFERENCES

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AUGMENTATION OF NK ACTIVITY BY PYRAN

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Activation of mouse macrophages by pyran copolymer and role in augmentation of natural killer activity.

Int. J . Cancer: 24, 819-825 (1979) ACTIVATION OF MOUSE MACROPHAGES BY PYRAN COPOLYMER A N D ROLE IN AUGMENTATION OF NATURAL KILLER ACTIVITY Paolo PU...
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