[CANCER RESEARCH 40, 823-829, March 1980]

0008-5472/80/O040-0000$02.O0

Natural Feline Leukemia Virus Infection and the Immune Response of

Cats of Different Ages―
Chris K. 23 M. Essex, M. B. Gardner, and W. D. Hardy, Jr.3
Department of Microbiology, Harvard School of Public Health, Boston, Massachusetts 02 115 (C. K. G.. M. El: University of Southern California School of
Medicine, Los Angeles, California 90032 (M. B. G.j; and Sloan Kettering Institute for Cancer Research, New York, New York 1002 11W. D. H.J

ABSTRACT MCH's there is a high incidence of FeLV infection and within

such environments leukemia clusters frequently arise (4, 9, 14,
Forty-two kittens and 28 adult cats were placed as tracers in 22). Cats which remain healthy in MCH environments com
leukemia cluster environments in contact with resident cats, monly have detectable antibody to FeLV and also to virus
30% of which were persistently infected with feline leukemia induced, transformation-specific FOCMA (4, 8, 9, 16, 19, 23,
virus (FeLV). After 7 months exposure, FeLV viremia had been 37). Prospective studies have shown that FOCMA antibody is
detected in 71% of the tracer kittens, although only 55% of not detected in cats which subsequently develop tumors and
these remained persistently infected; in the same period, 11% that high titers of antibody protect cats from tumor development
of tracer adults became infected, but by 2 years the figure in most cases (4, 9, 10, 16). The humoral effector mechanism
reached 43%. Mean latent periods before detection of viremia involved appears to be CDA, since this FOCMA-CDA reactivity
were 3.4 ±1.8 (S.D.) and 13.0 ±5.9 months for kittens and is not directed at FeLV, but it does lyse FOCMA-beaning cat
adults, respectively. First detection of FeLV infection was ac tumor cells in the presence of cat complement (1 , 8, 16, 17,
companied by a sharp although transient drop in peripheral 39). In this system, we have been unable to detect antibody
white blood cell numbers, and infection onset triggered the dependent cellular cytotoxicity with FOCMA immune sera and
humomalimmune response which was comprised of separate cat effector cells.5
antibodies with virus-neutralizing and tumor lysis activities. Following virus inoculation, young kittens are most suscep
High titers of virus-neutralizing antibody appeared in transiently tible to the pathogenic effects of FeLV and the closely related
viremic cats immediately following elimination of viremia; this feline sarcoma virus (8, 16, 26). Results reported here show
antibody was rarely detected in cats that remained persistently that young cats are more rapidly infected following natural
viremic. Lytic complement-dependent antibody to feline oncor contact exposure, but that older cats become infected after a
navirus-associated cell membrane antigen appeared in most protracted exposure period. In most cats, the onset of detect
cats I to 2 weeks after FeLV infection was first detected, and able viremia was found to be directly associated with a drop in
subsequently high titers of this antibody remained in both circulating WBC numbers and, after a short delay, with the
transiently and persistently infected cats. If the rate of FeLV independent appearances of FOCMA-CDA and/or VNA. A
infection was summarized by using vinemia and/or antibody minority of cats reverted to the leukemia virus-negative state
appearance, then 95% of the kittens became infected within 1 after a transient viremia which was terminated by the appear
year and 61% of the adults within 2 years. Adult cats are, ance of VNA. While the latent period before infection became
therefore, susceptible to FeLV infection following long-term established was longer in adults than in kittens, no evidence
natural exposure, and their apparent resistance cannot be was found to suggest that this resulted from the ability of older
attributed to a protective humoral immune response that de cats to mount a protective humoral immune response shortly
veloped immediately after exposure commenced. after exposure commenced.

INTRODUCTION MATERIALS AND METHODS

FeLV4is infectious among cats, thus exemplifying a horizon Cats. The private MCH's studiedwere located in California,
tally transmitted oncomnavinusof an outbred species (5, 22, 25, Connecticut, and Massachusetts. Resident cats were predom
29, 33). Infection results after contact between individuals, and inantly random bred and of domestic short or long hair varieties;
saliva is the most likely vehicle for virus transmission (13). In most were spayed or castrated, and >20% were related.
Persistent FeLV infection was confirmed in 25, 27, and 32% of
I Supported by NCI Grants CAl 3885 and CAl

AmerIcan Cancer Society Grant DT32, and Grant 1438-C-i from the Massachu
821 6, NCI Contract CB-64001,
the resident cats in the separate MCH's used in this study. As
setts Division of the American Cancer Society. The research was also supported tracers, 42 kittens (3 to 4 months old) and 28 adults (@6
In part by the Division of Cancer Cause and Prevention, National Cancer Institute, months old) were placed in the MCH's and were allowed to
NIH, Departmentof Health, Education,and Welfare, under Contract NOl CP
91007. intermix freely. The kittens were raised in specific-pathogen
2 5@, of the American Cancer Society (Massachusetts Division). To whom free breeding colonies and screened for preexisting FeLV
requests for reprints should be addressed. infection or immunity prior to, or immediately after, MCH entry.
3 Scholar of the Leukemia Society of America.
4 The abbreviations used are: FeLV, feline leukemia virus: MCH, multiple cat
Only tracers remaining in the MCH's continuously for 12
household; FOcMA, feline oncornavirus-associated cell membraneantigen: CDA, months are reported. At regular intervals, all cats were exam
complement-dependent antibody; FOCMA-CDA, complement-dependent anti med clinically, and jugulovenous blood samples were removed
body to feline oncomavirus-associated cell membrane antigen: VNA, virus neu
tralizing antibody; GSA, group-specific antigens: FIP, feline infectious peritonitis.
Received July 19, 1979; accepted December 10, 1979. 5 J. McCartney and C. K. Grant, Manuscript in preparation.

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C. K. Grant et a!.

for testing. The mean point between 2 consecutive tests, in
which the second revealed infection or immunity, was taken as
the conversion time from commencement of exposure.
FeLV Infection. The presence of FeLV GSA in fixed blood
smears was determined by immunofluorescence (20). Positive
results from this test correlate 95% with other tests for
presence of FeLV, e.g. , infectious virus isolation and electron
microscopy (14, 20, 27). The existence of transient vinemia
was confirmed by virus isolation using the focus-forming assay
with CCC81 cells (1 1).
VNA. This procedure was modified slightly from that de
scnibed by Schaller and Olsen (38). Serum dilutions (1 :5, 1:
25, 1:125, and 1:625) were tested for their ability to neutralize
subgroup A, virus, harvested from the feline lymphoblastoid
cell line F442 (35), prior to infection of CCC81 cells (1 1).
Serum dilutions causing neutralization of 50% of the foci num
bens detected in control plates were considered positive.
Antibody to FOCMA. Serum antibody activity was titrated (at
doubling dilutions of 1:2 to 1:256) using immunofluonescence
(8) and 51Crrelease (15, 17) as previously described, except
that the 51Crrelease assay was performed in microcytotoxicity
plates using the Flow semiautomated Titertek harvesting sys
tem (Flow Labs, Inc., McLean, Va.). Target cells for both assays
were the feline FL74 lymphoblastoid suspension culture line
(40). Data from both assays correlated @98%, and this serum
reactivity was referred to as FOCMA-CDA. Geometric mean
antibody titers were calculated on all serum samples removed
from individual cats after the first sign (FeLV GSA on FOCMA
CDA) of infection was detected.
RESULTS
Prevalence of FeLV Infection and FOCMA-CDA Immunity
in MCH's. Blood samples from all resident cats in 7 MCH's
were examined for FeLV GSA and FOCMA-CDA (Table 1). One
household (Household A) was found to be free of FeLV infection
and associated humoral immunity despite a previous outbreak
of FIP; while this disease is frequently FeLV associated (4, 5,
14, 21 , 33), its manifestation in this FeLV-fnee household
demonstrates that it can appear independently. Remaining
households (Households B to G) had previously contained at
Table 1
Prevalence of FeLV GSA and FOCMA-CDA inanimalsNo.
MCH resident
of cats
forFeLVGSA
positive
andFeLV
FOCMA- FOCMA
HouseholdTotal
onlyA220 no. of catsGSA only CDA CDA
0B151 0
9C223 2 it is apparent that detectable VNA and FOCMA-CDA immune
12D183 3
9E507 2
22F564 9 19) that resisted persistent FeLV infection (i.e. , FeLV GSA
17G123 14
6Total 2
75(Households
cats1 7321 32
Bto
combined)%oftotalC12.1
G of all exposed kittens irrespective of their FeLV GSA status.
18.5
a The remaining26% of cats in HouseholdsB to G were negativefor FeLV
GSA and FOCMA-CDA.
least one cat with leukemia-lymphoma, and, when examined,
between 20 and 50% of the cats in each house were found to
be FeLV infected (FeLV GSA positive). In total, 173 resident
cats were examined, and 30.6% of these were persistently
FeLV infected, although most were otherwise healthy at time of
examination (7). Appearance of FOCMA-CDA has been shown
to be due to FeLV exposure (16, 17), and approximately 60%
of both the FeLV-free and the FeLV-infected resident cat
groups had antibody detectable in their senaat dilutions of 1:
4. In Households B to G, therefore, evidence for past onpresent
FeLV infection (as determined by FeLV GSA and/or FOCMA
CDA immunity) was found in 74% of the total resident cats.
The healthy FeLV GSA-positive cats are the source of horizon
tal FeLV infection within households, but because the majority
are FOCMA-CDA positive they are themselves protected from
developing virus-related neoplasia (9, 10, 16).
Rate of FeLV Infection In Tracer Cats. Tracers (42 kittens
and 28 adult cats), which were negative for FeLV GSA and
FOCMA-CDA, were placed in MCH's where 25 to 32% of the
resident cats were FeLV GSA positive. As controls, 20 cats
were isolated in laboratory environments free of FeLV infection;
throughout the study period, these controls remained FeLV
GSA negative and did not develop FOCMA-CDA or VNA.
The rates of FeLV infection of the tracers are shown in Chart
1. By 7 months of continuous contact with MCH residents, 30
of the 42 tracer kittens (71 %) became FeLV GSA positive,
although only 23 (55%) remained persistently vimemic until
death or experimental termination. The 7 kittens which became
FeLV GSA positive for a short period [mean, 3.2 ±1 (S.D.)
weeks] thereafter returned permanently to the FeLV GSA-neg
ative state. In future discussion, these cats are classed sepa
rately as transient vinemics, although in Chart 1 and Table 2
they are included among the FeLV GSA-positive cats.
Infection of adult tracers occurred more slowly (Chart 1).
After 7 months, only 11% of these cats were FeLV GSA
positive, but after 2 years of continuous exposure 43% of the
tracers had become infected. In this group of adult tracers,
evidence was also found for 3 transiently viremic cats, but,
inasmuch as intervals between removing samples were longer
than those used for kittens, we were unable to establish the
duration of transient viremia in the cases.
Mean intervals between commencement of exposure and
appearance of FeLV vinemia in the groups of tracers are sum
manized in Table 2.
Antibody Responses of Tracer Cats. Several classes of
response to exposure could be distinguished within the tracer
kittens according to (a) the susceptibility to FeLV infection and
(b) the subsequent appearance of humoral immunity in the form
of VNA and/or FOCMA-CDA. The number of kittens exhibiting
each type of response is shown in Table 3, and from these data
responses occurred independently. While most kittens (17 of
detected transiently onnot at all) developed VNA, this antibody
was rarely detected in cats which succumbed to persistent
viremia. On the other hand, FOCMA-CDA was detected in 69%
43.4 First detection of serum antibodies in individual tracers varied
with the onset of FeLV infection, and data from tracer kittens
are exemplified in Table 4. Neither FOCMA-CDA non VNA

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 Immunity following FeLV Infection

$11

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0
a'
.
‘.1

C.)

3 15
MONTHS CONTACT EXPOSURE
Chart 1. Rate of detection of FeLV GSA In blood smears from tracer cats placed in MCH: data from cats which became transiently infected are included. U, kittens
first exposed at 3 to 4 months of age; •,
adults first exposed at @6 months of age.

Table 2 to those seen in kittens, except that the interval between

Appearance of FeLV infection or FOCMA-CDA in tracer cats commencement of exposure and development of immunity was
exposure3-4
Age at initial relatively long. As in kittens, FOCMA-CDA appeared in adults
mos.aPosimos.6—60
soon after first detection of FeLV GSA (Table 2), and subse
quently the magnitude of the antibody responses was of the
Mean same order as that detected in young cats.
tive conversion tive conversion
(%)Mean
(mos.)71
timeb Cmos)Posi- (%) times' Ages of adult tracers and the time elapsed to infection and/
FeLV GSA detectedC ±18d ±5.9
or development of immunity are summarized in Table 5. In each
FOCMA-CDA detected 693.4 3.9 ±2.413.0 50 13.4 ±6.2 age group, the numbers of cats are small, but the data suggest
Mean ±S.D., 18.6 ±14.8 months. a trend in which cats of 12 to 17 months of age require the
@ b from initial exposure to FeLV GSA or FOCMA-CDA appearance. longest exposure period before becoming infected.
C Transiently and persistently FeLV-infected cats.
d Mean ± S.D.
Some cats developed humoral immunity without detectable
FeLV infection. If all cats which became FeLV GSA positive
appeared before FeLV GSA was detected in persistently on and/on immune are considered together, then after 1 year of
transiently infected cats, although these antibodies appeared exposure 95% of kittens had exhibited some form of infection
In a minority of exposed cats that were never vinemic (FeLV
Table 3
GSA negative) (see ‘‘Discussion―).
The mean interval sepamat
Detection
tracerkittensFeLV
of FeLV infection and/or serum antibodies in bloodfrom
ing the first appearance of FeLV GSA and FOCMA-CDA was
2.4 ±1.9 weeks. After appearance, FOCMA-CDA titers in statusNo.
. creased to peak values by 5.0 ± 2.1 weeks later, and in most

. of these cats (1 9 of 29) antibody then persisted at peak titer

VNANot GSA statusAntibody catsFOCMA-CDA of
L foratleast 6months before anydecline wasevident. Inthe detected at any time—
,@a
—

0
; remainder, antibody titers dropped after peaking to about one 4
: half
themaximum level detected,and then thetiterpersistedat 6Detected —

+
+

+2
: thelower
level. transiently (duration,—
—03.2±1.Owk)+
I VNA appeared in serum at the same time as FOCMA-CDA in —
cats which were never detectably viremic. In all transiently 2
,. vimemic
cats,
VNA was
first
detected
approximately
1week
— +

5Detected + +0

after the disappearance of FeLV GSA, although FOCMA-CDA —5>3mos.)+

persistently (duration,—
appeared and increased in titer during the transient episode of —

vimemla.In most individual cats, no correlations were apparent — + 0

+ +15 3
between FOCMA-CDA and VNA in megandto detection or to the
a Positive result signifies that, after the first appearance, antibody was sub
quantitative levels of the antibodies. sequently detected in later serum samples so that the geometric mean antibody
@ The immune responses of adult cats followed patterns similar titer of all immune samples @1 :4.

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C. K. Grant et a!.

or resistance, whereas after 2 years of exposure 61% of the VNA

FOCMA-CDA
adult tracers had responded in a similar manner. FeLV—not detected
Geometric mean titers for all antibody-positive cats are com A
3
pared in Table 6. Mean FOCMA-CDA titers were similar in the
FeLV-infected cat groups (residents and tracers) and in the
I 1@L1.11
corresponding groups of FeLV-free but immune housemates.
In contrast, almost all persistently infected cats lacked detect
2
TIf@t@@t'@Ftt
able VNA, whereas almost all exposed but immune cats had
1
high VNA titers. The mean VNA titer, in the minority of persist
ently infected cats that had detectable neutralizing antibody,
was 1 log lower than the mean titer found in transiently infected
individuals after they had eliminated their infection. B FOCMA-CDA
3 FeLVI@VNAIIIP
I@1I
Changes in Peripheral WBC Numbers Associated with E
FeLV Infection. PeripheralWBC countsvaried in the individual S
tracer kittens, but when FeLV GSA was first detected there
‘C
was a coincident decrease in hemopoietic cell numbers to 0
approximately one-half of the preinfection levels (Chart 2). In
transiently viremic cats, the duration of leukopenia was short, C.)
m
1

and cell numbers returned to normal levels by 4 to 6 weeks. In

most cats with persistent FeLV infection, the mild leukopenia VNA—not detected
C
Table 4
Appearance
meanantibody
of FeLV GSA, FOCMA-CDA, and VNA, and geometric
titers in individual
exposureTime
cats following FeLV contact

.of detection after mi- . mean titer

Geometric
tial exposure (wk)

FOCMA- FOCMA
VNAPersistent FeLV CDA VNA CDA
NDpositive
FeLV GSA 4 7 ND 1:68
ND10
8 10 ND 1 :84
ND18 10 ND 1:69
NDTransient 19 ND 1:38
F.LV gsa
detect•d
1positiveFeLV GSA 6 7 12 1:73 1:21
TIME (WEEKS)
1:16012
8 ND 12 ND
1:16014 12 16 1:21 Chart 2. Changes in WBC numbers in cats placed in MCH at 3 to 4 months of
1:30FeLV 16 18 1:160 age. Horizontal axis, weeks before and after first detection of FeLV gsa. Cats
were grouped as follows: A, noninfected (FeLV GSA not detected): B, transiently
GSA negative ND
1:92ND 12 10 1:65 FeLV infected: and C, persistently FeLV infected. Inasmuch as FeLV was never
1:121ND 20 20 1:29 detected in A, cell numbers were zeroed to first appearance of FOCMA-CDA
1:2028ND, ND 12 ND which occurred 2 weeks after FeLV GSA appearance in other groups. Horizontal
line at 2.05, WBC value for all cats prior to detection of FeLV infection; bars,
not detected.
S.D. First appearance and subsequent persistence of FOCMA-CDA, VNA, and
Table 5 FeLV GSA are indicated by horizontal bars above appropriate figures.
Age and appearance of infection or immunity in adult FeLV-exposed
catsNo.
of cats:FeLV lasted for at least 3 months.
Diseases Apparent in FeLV-exposed Tracer Cats. Twelve
in of the tracer kittens died after entry into FeLV-infected MCH's,
Age
orposure
at initial ex fected orTime to infection
(mos.)
(mos.)6-1110 ImmuneImmunity and all but one of these were persistently infected with FeLV.
—Exposed

614.8±7.9a12-176
Causes of death were nonregenenative anemia (5 cats), FIP (5
318.7±4.918-236 cats), and leukemia-lymphoma (2 cats). Adult tracer cats re
5.6>246 513.5 ± mained essentially healthy. High FOCMA-CDA titers (1:16 to
38.8±2.8a
1:64) were apparent in 3 cats with fatal FIP and one cat which
Mean ±S.D.
died from anemia, and a low titer of FOCMA-CDA (1 :4) was
Table 6 detected in sera removed both before and at death of one
catsGeometric
Geometricmean FOCMA-CDA and VNA titers of resident and tracer tracer with leukemia. No FOCMA-CDA was detected in the cat
mean
mean
titersaFeLV
VNA which developed lymphoma.
FOCMA-CDA titersCGeometric

positiveFeLV GSA DISCUSSION

FeLVFeLVGSA
GSAGSA Per
sistentbResidents1
Cat groupnegative positivenegative Transient
Laboratory studies have shown that cats inoculated with
FeLV at a young age are most susceptible to the pathological
:20Tracers1 :22 1
:23 1:191 :134 1:156 1:12 effects of the virus (8, 16, 26). Results reported here extend
a Calculatedon antibody-positivecats only. these observations and show that young cats are more sus
b Three persistently viremic cats with low but detectable levels of VNA. ceptible to naturally transmitted FeLV infection, but that many

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 Immunity following FeLV Infection
older cats eventually become persistently infected after a pro after elimination of viremia. Infection of cats occurs with FeLV
tracted exposure period. We cannot be sure that protracted A subgroup only or with a mixture of FeLV A and B subgroups
and continuous natural exposure is necessary to produce (19, 28). Theoretically, cats recently infected with virus A or
persistent viremia, because we did not remove tracers after AB might make VNA to the subgroups concerned and thereby
only short-term exposure. Nevertheless, it seems likely that eradicate their viremia (most cats), or they may respond with
long-term exposure plays some role in causing persistent in VNA to the B subgroup thereby remaining viremic with FeLV A
fection, because most pet cats that have been exposed to (cf. Ref. 30). Serologically, A and B virus subgroups sometimes
FeLV are found to be immune but not viremic, and usually cross-react in vitro (36), so that VNA directed to B subgroups
these pets are housed singly and exposure occurs through might be detected in a neutralizing test using A subgroup
short-term chance contact (10, 16, 19). indicator virus. We believe this explains the apparent paradox
The transient FeLV-associated leukopenia which was ob that a minority of viremic cats (3 of 21) had low but persistently
served confirms findings by others (7, 33) that infection causes detectable levels of VNA.
acute disturbances within hemopoietic tissue. These results The kittens which became ill following FeLV exposure were
correlate with the finding that both FOCMA-CDA and VNA almost all viremic, and the diseases were limited to FIP, anemia,
appeared in response to an episode of viremia, probably be and lymphoid tumors. FIP is caused by a coronavirus which
cause FeLV must infect and/or transform host cells before it may be activated from a subclinical state due to FeLV-associ
will present an adequate stimulus to induce humoral immunity. ated immunosuppression (4, 5, 16, 32—34).Anemia is also
It seems likely that those cats which became immune while commonly associated with FeLV infection, and, when it is not
apparently remaining FeLV GSA negative were actually tran fatal, the disease may represent a preleukemic state (4, 5, 16,
siently infected for a very short interval between tests for FeLV. 24, 33). Despite the relatively high FeLV infection rates de
In the case of FOCMA-CDA, which is directed at a tumor or tected, the appearance of only 2 cases of leukemia-lymphoma
transformation-specific antigen (4, 8, 9, 16, 17), prior virus was not surprising for 2 reasons. First, the latent period be
induced transformation of host cells would appear to be a tween detection of FeLV GSA and appearance of overt neo
prerequisite for stimulation of an antibody response. Conceiv plastic disease is protracted and may be as long as 40 months
ably, infection might sometimes occur at a localized site (e.g., in natural circumstances (12, 16). Second, many of the per
the oropharyngeal region) in which case virus would not be sistently infected cats developed FOCMA-CDA, and prospec
detected by the FeLV GSA test performed on peripheral blood tive studies have shown that most cats which develop leukemia
smears. lack FOCMA antibody at all times prior to tumor appearance;
The differing susceptibilities to FeLV infection of young and moreover, cats immunized to elicit FOCMA antibody were
old cats may be explained by the possibility that young cats subsequently resistant to progressive tumor development (4,
are less immunocompetent. If so, the difference involves mech 5, 8—1 0, 16, 19). The protective effect of FOCMA-CDA immu
anisms other than humoral immunity, since there was no evi nity is not absolute, however, since approximately 13% of
dence that most olden tracers resisted primary FeLV infection immune cats had low levels of this antibody and yet developed
by a rapid production of antibody after placement in FeLV lymphoid tumors (16—1 8). In some cases, tumor progression in
infected environments. In a separate study, we have seen 2 the face of FOCMA-CDA can be explained by severe depletion
cats become FOCMA-CDA immune and one cat become FeLV of lytic complement activity (18, 31).
GSA positive after residence in an infected MCH for 3 to 4 The possibility exists that immune mechanisms other than
years. Inasmuch as we have studied most of the adult tracer FOCMA-CDA play some role in feline leukemia resistance.
cats for approximately 2 years only, it is possible that our figure Cytotoxicity mediated by antibody-dependent cells has been
of 61% total FeLV-infected and/on immune adults (Table 5) is implicated in tumor resistance in nonfeline systems, but while
conservative at this time, since in MCH's where FeLV infection feline peripheral lymphocytes have antibody-dependent cell
has been fully established, 74% of all cats show some sign of mediated cytotoxicity effector cell activity in systems containing
infection or immunity (Table 1). Possibly the mode of virus heterologous tumor target cells, they fail to mediate lysis to
transmission plays some role in governing the latent period feline leukemia cells in the presence of FOCMA-CDA immune
between commencement of exposure and onset of infection. sera (data not shown). Two other mechanisms which may play
Virus is shed in saliva (13, 27), and it has been shown that for roles in feline leukemia immunity are natural killer cells and
infection to occur cats must be caged together rather than cytotoxic macrophages. To date, natural killer cells have been
singly in juxtaposed cages (25). Apparently, social grooming detected in unexposed and FeLV-exposed field cats but they
or sharing of feeding utensils is more important than aerosol also exist in tumor-bearing animals; cytotoxic peritoneal mac
transmission, and subjectively we have noticed that the most rophages have been detected only in cats immunized i.p. with
asocial adult cats seem to be the least readily infected. There allogeneic leukemia cells (16). Inasmuch as cats with FOCMA
seems to be no evidence for a significantly large population of antibody resist tumor progression (4, 5, 8—1 0, 16, 19), as CDA
cats which are genetically refractory to FeLV, because almost is the only cytotoxic mechanism detected in FOCMA immune
all (40 of 42) of the exposed kittens developed viremia and/or cats, and as CDA-mediated tumor cell lysis proceeds in culture
immunity, and others have shown that all cats inoculated or mixtures containing only cat components, we believe that this
otherwise exposed to virus become infected or immune as a complement-dependent lytic mechanism plays a primary role
result (5, 10, 25, 33, 37). in the immunological defense of outbred cats against lymphoid
The function of VNA seems to be to terminate FeLV infection, tumors. One factor which may underlie this emphasis on a B-
for the antibody was commonly detected in the absence of cell effector mechanism is that FeLV preferentially infects feline
virus, and in transiently vinemic cats it appeared rapidly at or T-cells and that the majority of naturally occurring lymphoid

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C. K. Grant et a!.
tumors are of T-cell origin (2, 3, 23). 5.
The concept that FOCMA-CDA reactivity is directed at a
virus-induced transformation-specific antigen and not to FeLV
per Se, (1 , 6, 16—18, 39) was supported by the finding that 6.
geometric mean titers of FOCMA-CDA were the same in both
FeLV-viremic and FeLV-free groups of immune cats (Table 6).
This finding suggests that FOCMA-CDA is not absorbed in vivo 7.
by the large quantities of cell-free FeLV or virus-replicating
cells that exist in peripherally viremic cats. Conversely, from 8.
data in Tables 3 and 4, it is apparent that cats can maintain
persistently high levels of serum VNA without such sera con 9.
taming any cytotoxic activity to virus-replicating tumor cells
(FL74). Unlike FOCMA-CDA, VNA was rarely detected in vi-@ 10.
remic cats, and it became detectable in transient viremics only
after eradication of detectable peripheral infection (Tables 4
11.
and 6). High levels of FOCMA-CDA reactivity were shown in
sera lacking antibodies to both the major FeLV antigens (a
glycoprotein and a protein with molecular weights of 70,000 12.
and 30,000, respectively) as detected by nadioimmune precip
itation (17), and recently we have lysed an established feline 13.
leukemia virus-negative tumor cell line by FOCMA-CDA.6 As 14.
such, FOCMA-CDA does lyse tumor cells which express only
FOCMA and no viral antigens, but this point is relevant to only
a minority of feline leukemias because most lymphoid tumors 15.
that arise replicate FeLV (12).
The demonstration here of transient viremia following natural
FeLV infection may be relevant to that minority of feline leu 16.
kemia virus-negative tumors (12). Recently, several lines of
evidence have emerged to associate transient FeLV infection 17.
with a subsequent development of virus-negative tumors (16),
and it is probable that in some circumstances FeLV is leuke
mogenic on a ‘‘hit
and run' ‘
basis. During transient viremia, the
opportunity exists for insertion of the onc or !euk gene into 18.
DNA of normal feline lymphoid cells, so that at a later date
transformation may result in the absence of FeLV replication.
The cats most susceptible to developing such tumors may be 19.
the minority of transient viremics that developed a protective
titer of VNA but failed to produce detectable FOCMA-CDA. 20.
ACKNOWLEDGMENTS
We thank Dr. S. Cotter for advice and help: M. Mandel, D. Hamm, and D.
Harris for excellent technical assistance: and M. Gravell and M. Rhodes for their
support and cooperation.
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Natural Feline Leukemia Virus Infection and the Immune
Response of Cats of Different Ages
Chris K. Grant, M. Essex, M. B. Gardner, et al.

Cancer Res 1980;40:823-829.

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