INFECTION AND IMMUNITY, Apr. 1970, p. 327-333 Vol. 1, No.

4
Copyright © 1970 American Society for Microbiology Printed in U.S.A.

Function of the Reticuloendothelial System

IV. Evidence for Two Types of Particle-Induced Reticuloendothelial Paralysis'
SIGURD J. NORMANN
Departmenzt of Pathology, University of Florida, Gainesville, Florida 32601
Received for publication 28 November 1969

Reticuloendothelial system (RES) phagocytosis has been quantitated after

Downloaded from http://iai.asm.org/ on April 10, 2021 by guest

intravenous injection of two different sets of particles by determining the clearance
rate of subsequently injected identical or nonidentical particles. Injection of carbon
produced a biphasic RES paralysis consisting of an early transient phase followed
by a delayed sustained phase. The two phases were separated by a distinct interval
of greatly augmented clearance rates. The injection of aggregated albumin was
followed only by a single period of depressed clearance, which corresponded to the
first phase of carbon-induced inhibition. This first phase, designated immediate RES
paralysis, was initiated by particle injection and its duration was related to the rate
of particle removal, to the dose of particles injected, and to the presence of the
particles in the circulation. The second phase, designated delayed RES paralysis,
began sometime after the particles had been engulfed by the cells, was independent
of the rate of particle removal, and persisted without the presence of measurable
particles in the circulation. The evidence indicates that the immediate paralysis
arises from a competition between the particles in the circulation, whereas the
delayed paralysis arises from a cellular derangement inhibitory to further phago-
cytosis. In contrast to the usual description of RES blockade as a single sustained
period of depression, the present experiments indicate that the phenomenon has two
phases which can be dissociated in time and mechanism.

The uptake of certain colloids by phagocytic
cells in vivo can be inhibited by the prior injec-
tion of the same or a different colloid. Frequently,
inert particles are used to induce this depressed
state known as "reticuloendothelial (RES)
blockade." Because such particles cannot be
eliminated from the cell, it was thought that the
decreased clearance rates represented cellular
saturation and that the onset of the "blockade"
dated from the progressive accumulation within
the cells of the first set of particles injected (2, 3).
Recently, Normann, Lagunoff, and Benditt (20)
measured simultaneously the vascular clearance
of two dissimilar colloids (carbon and aggre-
gated albumin), after their sequential injection
into the circulation of a rat. The clearance rate
of both particles was inhibited when both were
present in the circulation. Their findings could
not be explained on the basis of cellular satiation
and suggested that one phase of RES paralysis
is associated with the immediate events arising
from the presence of particles in the circulation.
On the other hand, Parker and Finney (22) ob-
1 A preliminary report of the material presented here was made
at the 51st annual meeting of the Federations of American
Societies for Experimental Biology, 1967.
327
served in mice that a period of normal clearance
may follow particle injection and exist prior to
the onset of RES paralysis. This latter observation
suggests that a second period of RES depression
may follow particle iniection and represent some
form of cellular derangement induced by colloid
ingestion. Therefore. it appears probable that the
usual description of "RES blockade" as a single
sustained period of depression, originating with
particle injection, may not be correct; rather, the
phenomenon may be composed of component
parts which may be dissociated in time and
mechanism.
The present report examines the possibility
that different phases of RES paralysis exist by
investigating the behavior of the system immedi-
ately after particle exposure in contrast to the
behavior evident 24 hr later. Two dissimilar col-
loids were studied because the response of the
RES to inert colloids such as carbon may be
different from the response to metabolizable
colloids such as aggregated albumin. The experi-
ments demonstrate that two periods of reduced
clearance rates are associated with "RES block-
ade" and that the immediate paralysis initiated
by particle injection is a decidedly different event
328 NORMANN
than the delayed paralysis manifested several
hours later.
MATERIALS AND METHODS
Male Sprague-Dawley rats weighing between 150
and 250 g were used after anesthesia with intraperi-
toneal pentobarbital sodium. The carbon suspension
manufactured by Gunther-Wagner lot C-11-1431a
was obtained from John Henschel and Co., New
York, and used as supplied. Gelatin powder derived
from swine skin was obtained from Sigma Chemical
Co. (St. Louis, Mo.). Gelatin for injection was pre-
pared as a 3% solution in 0.15 M saline. In some ex-
periments, gelatin was added to the carbon prepara-
tion prior to its injection into animals by the method of
Biozzi, Benacerraf, and Halpern (2). However, the
total amount of gelatin added to carbon was held con-
stant so that only 5 mg of gelatin per 100 g of body
weight was injected into any animal irrespective of the
carbon dose. Aggregated albumin was prepared by
controlled heating of bovine serum albumin and was
labeled with the fluorescent reagent dimethylamino-
napthelene suphonyl chloride (DNS-Cl from Sigma
Chemical Co.) as described previously (20).
The vascular clearance of carbon or DNS-labeled
aggregated albumin was determined by injecting in-
travenously a standard dose of either 10 mg of carbon
or 4 mg of aggregated albumin per 100 g. After injec-
tion, blood samples were obtained from the femoral
vein at regular time intervals. Clearance was measured
over a 15-min period except in those instances of ac-
celerated clearance. Blood was obtained in heparin-
washed syringes with particular care taken not to in-
troduce the anticoagulant into the circulation. The
carbon concentration in each sample was determined
by optical density measurement (20), where as the
clearance of the aggregated albumin was measured by
the decrease in plasma fluorescence (20). The plot of
the logarithm of the particle concentration versus time
yielded a straight line for both particle suspensions;
the slope of this line was computed by linear regression
and designated the clearance rate constant or K. Com-
parisons between the clearance rates obtained in
different experiments was performed using Student's t
test for comparison between group means with un-
equal population sizes (21).
In the studies to be described, the initial particle
injection was followed by a subsequent injection of
eithercarbonorlabeled aggregated albumin. The time
interval between the two injections was varied so that
the effect induced by the initial injection could be
evaluated at different times. Multiple determinations
were made at each interval, and the value recorded
represents an average determination on two or more
animals.
RESULTS
Immediate RES paralysis. A typical curve rep-
resenting the different rates of carbon clearance
observed after injection of various doses of ag-
gregated albumin is shown in Fig. 1. When the
carbon was administered shortly after the albu-
min, a marked reduction in carbon clearance
INFEC. IMMUN.
0
co 30
, .080-
10
< 060-
0
u
, .040-
Normal0
0 ----------
z .020-
J
2 4 6 8 !0 12 14 16
Downloaded from http://iai.asm.org/ on April 10, 2021 by guest
Aggregated T i lE IN HOURS
Albu mi n
FIG. 1. Effect of aggregated albumin injection on
the clearance rate of subsequently administered carbon.
The dose of aggregated albumin injected is indicated at
the top of each carbon line. The 95% confidenice limit
for the normal rate of carbon removal was K = 0.033
to K = 0.039.
rate was observed for each dose. However, the
rate of carbon removal was relatively constant
and independent of the amount of albumin in-
jected (K = 0.015 compared to a control of 0.036
with a standard deviation of 0.009 determined on
50 animals). A more effective paralysis of RES
function as measured by carbon was not achieved
by increasing the dose of aggregated albumin. On
the other hand, the duration of the paralysis was
dependent on the albumin dose and lengthened as
the dose of the particle injected increased. For
aggregated albumin doses greater than 4 mg/100
g of body weight, the paralysis terminated in a
state of accelerated clearance (K greater than
0.060). Induction of the accelerated clearance
state was not as effective nor was the duration sus-
tained when the dose was reduced to 1 mg/100 g
and was barely apparent at a dose of 0.5 mg/100
g. Moreover, the accelerated clearance state in-
duced by a 10-mg dose had disappeared by 24 hr
(Table 1), whereas that induced with a 30-mg
dose produced significant elevation in clearance
rate at that time. Thus, it appeared that the in-
duction and the duration of the increased carbon
removal rate depended upon the dose of aggre-
gated albumin injected.
In the next series of experiments, the sequence
of particle injection was reversed and the effect
of an initial injection of carbon was evaluated by
the clearance of a subsequent injection of labeled
aggregated albumin (Fig. 2). Three different in-
jections of carbon were made of 10, 20, and 30
mg/100 g. When the aggregated albumin was in-
jected immediately after the carbon and essentially
during its removal, the rate of albumin clearance
(K = 0.004) was significantly less than the rate
observed in the absence of carbon (control albu-
min clearance K = 0.008 with standard deviation
VOL. 1, 1970 FUNCTION OF RETICULOENDOTHELIAL SYSTEM. IV 329
TABLE 1. Effect of carbonz or aggregated albumin inijectioni onl the clearalnce rate of a
subsequentt injectiont of carbon (10 mg/100 g)
4Dose initial Avg clearance rate K of a subsequent carbon injectiona
Initial injection injection
(mg/lO00 g) 6 24

Aggregated albumin 10 0.017 + 0.007c 0.078 ± 0.020 0.040 i 0.018

30 0.017 i 0.005c 0.017 4 0 007c 0.060 i 0.013
Carbon 10 0.020 i 0.009c 0.054 i 0.020
0.036 ± 0.007
30 0.017 + 0.004c 0.020 ± 0.009c
0.083 ± 0.030
a Control: K = 0.036 ± 0.009 (50 determinations). Each value recorded represents the mean ± 1 stand-

Downloaded from http://iai.asm.org/ on April 10, 2021 by guest

ard deviation determined on 10 animals. Two periods of clearance depression are evident for large carbon
doses, but only a single period for small carbon doses. Aggregated albumin regardless of dose produces
only one period of depression.
b Hours after initial injection.
c Value significantly less than control (P < 0.01).

.090-

080-

070 -

C 060-

z 050~
60 75
Carbon TINIE IN rMINUTES
FIG. 2. Effect of carboni injectionz onl the clearance c 020-
040-
Nor ma
rate of subsequenitly injected aggregated albumin. The
95% confidence limit for the normal rate of aggregated
albumin removal was K = 0.0074 to K = 0.0087. ,020
, 030-

of 0.001 determined on 10 animals). Eventually,

the rate of albumin clearance returned to normal
and, in some instances, actually exceeded the
normal clearance value. A longer period of de-
pressed albumin clearance was observed with the
larger carbon doses.
This depressant effect of carbon on RES phago-
cytosis was also evaluated by the clearance of a
subsequent injection of the same particle. This
sequence of particles was chosen because carbon
is frequently used to induce "RES blockade," and
the extent of the paralysis induced by carbon (and
other particles) is often evaluated by a subsequent
carbon injection. However, very early time in-
tervals could not be evaluated due to residual
carbon in the circulation. Accordingly, the earliest
time at which the second carbon injection was
made depended upon the removal rate of the
first carbon dose and was given when 10% or less
of the initial carbon remained in the circulation
(generally 0.5 hr for 10 mg and 1 hr for 30 mg/100
.010 -j o-O 10mg Carbon / 100gm
A^^ 30mg Carbon 100gm
-I
2 3 4 5 6
Carbon TI tE IN HOURS AFTER INJECTION
FIG. 3. Effect of carbon injection on the clearance
rate of a subsequent injection of a standardized dose of
carbon. The 95% confidence limit for the normal rate
of carbonz removal was K = 0.033 to K = 0.039.
g). Under these conditions, a second carbon in-
jection was cleared at a rate slower than that ob-
served when the same dose of carbon was given
alone (Fig. 3). At 2 hr after the initial carbon in-
jection, the rate of clearance after a 10-mg dose
averaged 0.040, a value greater than the control
(K = 0.036), although not significantly different.
330 NORMANN
At the same time period, the clearance rate after
an initial injection of 30 mg of carbon per 100 g
was still significantly retarded (P < 0.05), averag-
ing K = 0.030. However, the clearance rate was
now recovering rapidly for, at 3 hr, greatly aug-
mented clearance rates were observed. This ele-
vated clearance was found after injection of 30
mg of carbon but not 10 mg of carbon per 100 g.
Thus depressed clearance rates, as measured by
carbon, did follow prior carbon injection, and the
duration of this depression appeared to be longer
with larger carbon doses.
The above experiments showed that an initial
injection of either carbon or aggregated albumin
delays the vascular clearance of a subsequent
colloid injection, provided the second colloid is
injected shortly after the first and essentially
during its clearance from the circulation. This
latter fact can be deduced from the observations
that closely spaced injections of similar and dis-
similar colloids produce inhibition, from the
calculated clearances of each colloid (dc/dt =
-2.3 Kt where c is concentration and t is time),
and from the observation that inhibition induced
by carbon lasts but a short time beyond the point
of detectable carbon in the circulation.
Delayed RES paralysis. Table 1 presents data
comparing the immediate and delayed responses
of the RES to injection of either carbon or ag-
gregated albumin. Both a large (30 mg/100 g) and
a small (10 mg/100 g) dose of each colloid were
examined. Whereas the aggregated albumin pro-
duced only a single period of depressed clearance,
two periods of inhibited clearance clearly existed
for the larger carbon dose. The first period oc-
curred early, coincident with particle injection,
and lasted only a short time. The second period
occurred late and was evident at 24 hr. The two
periods of retarded clearance were clearly sepa-
rated by an interval of accelerated clearance, as
evident by the values reported for 6 hr. In con-
trast, only the first period of depression was found
TABLE 2. Effect of gelatin on the clearance rate of subsequently injected carbon
Interval Avg rate of clearance of a 2nd injection of 10 mg/100 g of carbon, after a 1st injection per 100 g ofa
between
1st and
2nd
injection 5 mg of gelatin 30 mg of carbon
hr
2 0.020 i 0.006 0.030 :1: 0.007
4 0.019 i 0.007 0.085 i 0.022
8 0.018 i 0.006 0.080 ±L 0.029
24 0.023 ± 0.006 0.020 ±i0.008
aControl: K = 0.036 ±t 0.009 (50 determinations). Each value recorded represents the mean i 1 stand-
ard deviation determined on five animals. Gelatin confers a continuous sustained clearance depression
on subsequently injected carbon.
INFEC. IMMUN.
for the smaller carbon dose, as there was no de-
pression of clearance rate at 24 hr; in fact, the
clearance rate was actually greater than normal.
Thus, a second period of clearance rate depression
can occur after large doses of carbon, and the on-
set of the depression begins some time after the
particles from the initial injection have been
cleared from the circulation.
Effect of gelatin injection: a frequently used
particle-stabilizing agent. Whereas aggregated
albumin needs no stabilizing agent, carbon must
be stabilized to prevent its flocculation in blood.
Downloaded from http://iai.asm.org/ on April 10, 2021 by guest
Although the carbon preparation supplied by
Gunther-Wagner is stabilized with partially hy-
drolyzed fish gelatin, many investigators-in-
cluding Biozzi, Benacerraf, and Halpern (2),
who developed the carbon clearance method of
evaluating RES function-increase the carrier
colloid presumably to render the suspension more
stable in blood. It appears probable that the con-
tent of gelatin and its source might be important
variables. Accordingly, gelatin alone and gelatin
addition to the carbon preparation were ex-
amined for their effect on carbon removal and
induction of "RES blockade."
When gelatin was added to the carbon prepara-
tion in vitro and prior to its injection into animals,
the carbon clearance rate decreased. This effect
has been previously observed (10, 17). In the
present experiments, the same effect could also be
achieved by the injection of gelatin alone. Table 2
compares the effect upon carbon removal rate of
a prior injection of gelatin alone (5 mg/100 g),
of carbon alone without supplemental gelatin,
and of carbon with supplemental gelatin. The
following features emerged. First, the gelatin
alone was sufficient to produce a prolonged de-
pression in the clearance rate of subsequently in-
jected carbon. This effect occurred coincident to
the gelatin injection and persisted for up to 24 hr.
Secondly, if gelatin were added to the carbon
preparation in vitro and prior to its injection into
30 mg of carbon + 10 mg of carbon +
5 mg of gelatin 5 mg of gelatin
0.017 ± 0.005 0.019 i 0.004
0.014 i 0.004 0.024 ± 0.005
0.017 i 0.006 0.022 4 0.007
0.022 i 0.004 0.026 i 0.006
VOL. 1, 1970 FUNCTION OF RETICULOENDOTHELIAL SYSTEM. IV
animals, a continuous and sustained period of de-
pressed clearance resulted irrespective of the dose
of carbon injected. Thus, gelatin addition elimi-
nated the period of normal or accelerated clear-
ance observed between the two periods of block-
ade produced by an injection of 30 mg of carbon
per 100 g. Further, gelatin addition to either a
small or a large carbon dose produced an effect
equivalent to an injection of gelatin alone (Fig.
4). Injection of the supernatant fluid of the origi-
nal carbon preparation obtained after centrifuga-
tion at 45,000 X g for 2 hr had no effect upon
331
essential differences between the two forms of
paralysis.
Whereas two phases of RES paralysis were
demonstrated for carbon, the injection of a differ-
ent type of particle, aggregated albumin, was fol-
lowed by a single period of depressed clearance,
which corresponded to the first phase of carbon
induced inhibition. It would appear that the
nature of the particles is an important factor in
the production of the second phase of paralysis.
Since delayed paralysis is initiated after particles
have been cleared from the circulation, the pri-

Downloaded from http://iai.asm.org/ on April 10, 2021 by guest

carbon removal rate.
DISCUSSION
Phagocytic inhibition produced by colloid in-
jection (RES blockade) is thought to be a single
period of RES paralysis due either to a depletion
of blood opsonins (8, 16, 18) or to a cellular
limitation to further phagocytosis (1-3). In the
present experiments, carbon injection produced
a biphasic RES paralysis consisting of an early
transient phase followed by a delayed sustained
phase. These experiments provide evidence that
RES blockade may be composed of at least two
phases and each phase may involve a different
limiting mechanism in phagocytosis. The phases
have been designated as immediate or delayed
paralysis to reflect their onset relative to the first
set of particles injected. Table 3 summarizes the
mary effect of the particles presumably is on the
cell. The ability of the cell to catabolize the par-
ticle may be an important factor because a me-
tabolizable particle, such as aggregated albumin,
would be expected to have different cellular
effects than inert particles such as carbon and
thorotrast or toxic particles, such as silicon diox-
ide (23), certain lipid emulsions (26, 27), or endo-
toxin (7, 14). Indeed, a prolonged sustained
period of RES depression has been associated
with injection of each of the latter type of par-
ticles.
Injection of either carbon or aggregated al-
bumin was followed by an immediate period of
RES paralysis. The duration of this effect was re-
lated to the rate of particle removal, to the
amount of particles injected, and to the presence
of the particles in the circulation. Therefore, in
immediate RES paralysis, there appears to be

z
,-
0
co
NcJ
0

;-
tY
J

o-o 10mg Carbon / lOOgm

-^ ^30mg Carbon I lOOgm
5mg Gelatin Alone or
K\'\\Iq 1 1mg Carbon + 5mg Gelatin I 100gm

t TIME IN HOURS AFTER INJECTION

FIG. 4. Composite graph showing the effect of carbon, gelatin, and gelatin-carbon injection on the subsequent
clearance of a standard carbon dose. The different preparations were injected only once, at time zero, and the
effect on the reticuloendothelial system determined in the time periods thereafter. Note that the larger carbon dose.
without gelatin produces two distinct periods of clearance depression, whereas gelatin alone or gelatin with carbon
produces only a single sustained period ofdepressed clearance.
332 NORMANN INFEC. IMMUN.
TABLE 3. Summary of two types of particle-induced reticuloendothelial (RES) paralysis in rats
Characteristic Immediate RES paralysis Delayed RES paralysis

Onset Immediate, coincident with particle Delayed, develops sometime (hours)

injection after particle injection
Particle types Metabolizable or inert Inert or toxic
Examples of particles Carbon, aggregated albumin, gelatin Carbon, thorotrast, endotoxin, cer-
tain lipid emulsions
Duration Usually short (minutes to hours) but Prolonged (hours to days); duration
may be prolonged; duration de- not dependent upon dose or pres-
pendent upon particle dose, rate of ence of circulating particles
removal, and presence of particles

Downloaded from http://iai.asm.org/ on April 10, 2021 by guest

Particle specificity
Possible mechanism
in circulation
Yes Unknown
Two-particle interaction in circula- Cellular derangement as (i) tempor-
tion as (i) adsorption onto particle ary failure to synthesize opsonin
surface, or (ii) competition for or replace cell constituents, or (ii)
phagocytic sites or opsonin sustained failure requiring cell re-
placement for recovery.

some form of competition between the particles
in the circulation, whereas, in delayed RES
paralysis, there appears to be a primary cellular
derangement.
The demonstration of two phases of RES
paralysis depended upon producing a sufficiently
short period of immediate paralysis so that a re-
covery period of normal or augmented clearance
existed betwen the two phases of paralysis. Such
a short period of clearance inhibition might easily
be overlooked, since some particles of the first
injection will be present in the circulation dur-
ing this period. Adequate demonstration depends
upon an observed change in the clearance rate
of a second particle injected during clearance
of the first. Thus, the clearance of DNS-labeled
aggregated albumin was inhibited when injected
during carbon clearance and, conversely, the clear-
ance of carbon was inhibited when injected during
aggregated albumin removal. Furthermore, the
simultaneous determination of the clearance rates
of both particles showed that there is an inhibition
in the clearance rates of both particles (20). How-
ever, not all particles would possess the necessary
properties to compete with each other and a cer-
tain degree of specificity in the phenomenon
would be expected; indeed, such specificity has
been described (13, 16, 25, 28).
Since the duration of immediate paralysis is
dependent upon the numbers of particles injected
as well as their rates of clearance, it should be
possible to extend the period of immediate
paralysis sufficiently so that an overlapping of the
two periods of retarded clearance occurred. Gela-
tin addition to the carbon suspension prior to in-
jection of the suspension into animals slows the
rate of carbon removal. In addition, the presence
of gelatin, which is very slowly removed, can
itself retard the clearance of certain particles in-
jected subsequently (13). Thus, the prolonged
slow removal of carbon observed after gelatin
addition would be an example of the immediate
type of RES paralysis sustained by the slow re-
moval of the first particle injected (gelatin). Be-
cause gelatin can sustain a depression in carbon
clearance rate, this fact alone, or the large carbon
doses used, could account for the observations of
Biozzi et al. of only a single period of depression
after carbon injection (2).
Recently, Jeunet and Good (9) suggested that
RES depression involves both cellular and hu-
moral factors which could be dissociated in an
isolated perfused liver system. Although their
conclusions are substantiated by the present
study, an important distinction between the two
model systems should be pointed out. Whereas
repeated additions of aggregated albumin to an
isolated perfused liver system produced an im-
paired clearance at the cellular level, repeated in-
jections of either aggregated albumin (19) or
carbon (4) in the intact animal results in faster
and faster rates of clearance. That the system can
actively increase its appetite for inert particles on
each successive exposure in vivo indicates that,
for all practical purposes, there is no limit to the
capacity of the system to engulf particles. This fact
renders untenable the hypothesis that the cause
of delayed RES paralysis could be due to any
form of cellular saturation (2), despite the his-
toric acceptance of this theory. Therefore, a
different form of cellular derangement must be
involved. One possibility is a temporary failure
of the cell to produce blood factors essential for
phagocytosis. Pisano et al. (24) reported that
puromycin pretreatment prevents recovery of
clearance rates depressed by injection of a test
VOL. 1, 1970 FUNCTION OF RETICULOENDOTHELIAL SYSTEM. IV
lipid emulsion and concluded that new synthesis
of opsonic protein may be involved in recovery.
Alternatively, there could be a need for synthesis
of new cell membrane or other cellular constitu-
ents consumed in the process of phagocytosis.
Finally with more extensive cellular derangement,
there may need to be a replacement of cells to
effect recovery. Kelly and co-workers (11, 12)
showed that ingestion of particles by cells results
in a marked increase in incorporation of tritiated
thymidine into deoxyribonucleic acid of Kupffer
cells, and it has been reported that nitrogen mus-
tard, by blocking cell replication, can prolong
RES blockade (1).
The immediate RES paralysis that occurs when
two particles are in the circulation involves some
form of interaction between the particles. How-
ever, it remains unclear how the presence of the
particles actually interferes with phagocytosis, al-
though at least three possibilities suggest them-
selves. (i) There is a direct interaction between the
particles, with one particle being adsorbed onto
the surface of the other, changing the surface
properties of either or both; (ii) there is a compe-
tition between the particles at the phagocytic
site on the cell (6); or (iii) there is a competition
for, with resulting depletion of, available serum
factors essential to optimal phagocytosis (5, 15,
18).
ACKNOWLEDGMMENTS
This investigation was supported under contract DADA
17-68c-8125 with the United States Army Medical Research and
Development Command.
LITERATURE CITED
1. Benacerraf, B., B. N. Halpern, G. Biozzi, and S. A. Benos.
1954. Quantitative study of the granulopectic activity of
the reticuloendothelial system. III. The effect of cortisone
and nitrogen mustard on the regenerative capacity of the
RES after saturation with carbon. Brit. J. Exp. Pathol.
35:98-106.
2. Biozzi, G., B. Benacerraf, and B. Halpern. 1953. Quantitative
study of the granulopectic activity of the reticuloendothelial
system. II. A study of the kinetics of the granulopectic
activity of the RES in relation to the dose of carbon in-
jected. Relationship between the weight of the organs and
their activity. Brit. J. Exp. Pathol. 34:441-457.
3. Biozzi, G., C. Stiffel, B. Halpern, and D. Mouton. 1963.
Lack of action of serum opsonins in phagocytosis of inert
particles by cells of reticuloendothelial system. Proc. Soc.
Exp. Biol. Med. 112:1017-1020.
4. Dobson, E. L., L. S. Kelly, and-C. R. Finney. 1967. Kinetics
of the phagocytosis of repeated injection of colloidal
carbon: Blockade, a latent period or stimulation? A ques-
tion of timing and dose, p. 63-73. In N. R. DiLuzio and R.
Paoletti (ed.), The reticuloendothelial system and atherio-
sclerosis. Plenum Press, New York.
5. Filkins, J. P., and J. J. Smith. 1965. Plasma factor influencing
carbon phagocytosis in the isolated perfused rat liver. Proc.
Soc. Exp. Biol. Med. 119:1181-1184.
6. Fred, R. K., and M. L. Shore. 1967. Application of a mathe-
matical model to the study of RES phagocytosis in mice,
333
1-17. In N. R. DiLuzio and R. Paoletti (ed.), The reticulo-
p.
endothelial system and artheriosclerosis. Plenum Press,
New York.
7. Heilman, D. 1965. The selective toxicity of endotoxin for
phagocytic cells of the reticuloendothelial system. Int.
Arch. Allergy 26:63-79.
8. Jenkins, C. R., and D. Rowley. 1961. The role of opsonins in
the clearance of living and inert particles by cells of the
reticuloendothelial system. J. Exp. Med. 114:363-374.
9. Jeunet, F. S., and R. A. Good. 1969. Recticuloendothelial
function in the isolated perfused liver. II. Phagocytosis of
heat aggregated bovine serum albumin. Demonstration of
two components in the blockade of the reticuloendothelial
system. Res. J. Reticuloendothel. Soc. 6:94-107.
Downloaded from http://iai.asm.org/ on April 10, 2021 by guest
10. Kampschmidt, R., H. Upchurch, and A. Park. 1966. Factors
in the suspending media which alter the carbon clearance
rate. Res. J. Reticuloendothel. Soc. 3:214-221.
11. Kelly, L. S., B. A. Brown, and E. L. Dobson. 1962. Cell
division and phagocytic activity in liver reticulo-endothelial
cells. Proc. Soc. Exp. Biol. Med. 110:555-559.
12. Kelly, L. S., E. L. Dobson, C. R. Finney, and J. D. Hirsch.
1960. Proliferation of the reticuloendothelial system in the
liver. Amer. J. Physiol. 198:1134-1138.
13. Koenig, M. G., R. M. Heyssel, M. S. Melly, and D. E. Rogers.
1965. The dynamics of reticuloendothelial blockade. J.
Exp. Med. 122:117-142.
14. Levy, E., and B. H. Ruebner. 1968. Hepatic changes induced
by a single dose of endotoxin in germfree mice. Amer. J.
Pathol. 52:97-110.
15. Murray, I. M. 1963. Clearance rate in relation to agglutinins
for gelatin-stabilized colloid in the rat. Amer. J. Physiol.
204:655-659.
16. Murray, I. M., 1963. The mechanism of blockade of reticulo-
endothelial system. J. Exp. Med. 117:139-147.
17. Normann, S. J., and E. P. Benditt. 1965. Function of the
reticuloendothelial system. I. A study of the phenomenon
of carbon clearance inhibition. J. Exp. Med. 122:693-707.
18. Normann, S. J., and E. P. Benditt. 1965. Function of the
reticuloendothelial system. II. Participation of a serum
factor in carbon clearance. J. Exp. Med. 122:709-719.
19. Normann, S. J., and E. P. Benditt. 1965. Reticuloendothelial
blockade: The importance of the sequence and dose of
administered particles. Res. J. Reticuloendothel. Soc. 2:
345.
20. Normann, S. J., D. Lagunoff, and E. P. Benditt. 1968. Func-
tion of the reticuloendothelial system. III. Simultaneous
measurement of two particles during clearance inhibition.
Lab. Invest. 19:353-357.
21. Snedecor, G. W. 1961. Statistical methods, p. 90-92. Iowa
State University Press, Ames, Iowa.
22. Parker, H. G., and C. R. Finney. 1968. Latent period in the
induction of reticuloendothelial blockade. Amer. J. Physiol.
198:916-920.
23. Pearsal, N. N., and R. S. Weiser. 1968. The macrophage in
allograft immunity. I. Effects of silica as a specific macro-
phage toxin. Res. J. Reticuloendothel. Soc. 5:107-120.
24. Pisano, J. C., J. T. Patterson, and N. R. DiLuzio. 1968.
Reticuloendothelial blockade: Effect of puromycin on
opsonin-dependent recovery. Science 162:565-567.
25. Schapiro, R. L., W. J. Maclntyre, and D. I. Schapiro. 1966.
The effect of homologous and heterologous carrier on the
clearance of colloidal material by the reticuloendothelial
system. J. Lab. Clin. Med. 68:286-299.
26. Stuart, A. E. 1962. Experimental necrosis of spleen. J. Pathol.
Bacteriol. 84:193-200.
27. Stuart, A. E., C. Biozzi, C. Stiffel, B. N. Halpern, and D.
Mouton. 1960. The stimulation and depression of reticulo-
endothelial pathocytic function by simple lipids. Brit. J.
Exp. Pathol. 41:599-604.
28. Wagner, H. N., Jr., and M. io. Studies of the reticuloendo-
thelial system (RES). III. Blockade of the RES in man.
J. Clin. Invest. 42:1525-1532.