Handbook of Human Stress and Immunity

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 CONTRIBUTORS

Numbers in parentheses indicate the pages on which the authors' contributions begin.

Robert Ader (1), Department of Psychiatry, The Center for Psychoneuroim-

munology, University of Rochester Medical Center, Rochester, New York
14642
Julie A. Anderson (183), Department of Psychiatry, TGH-University Psychi­
atry Center, University of South Florida College of Medicine, Tampa,
Florida 33613
Michael Antoni (267), Departments of Psychology and Psychiatry, Univer­
sity of Miami, Coral Gables, Florida 33124
Jacqueline A. Bartlett (217), Department of Psychiatry, University of Medi­
cine and Dentistry, New Jersey Medical School, Newark, New Jersey
07103
Donna Benton (341), Department of Psychiatry and Biobehavioral Sciences,
University of California at Los Angeles, Los Angeles, California 90024

xvii
xviîî Contributors

Robert H. Bonneau (125), Department of Microbiology and Immunology,

The Pennsylvania State University College of Medicine, Hershey Medical
Center, Hershey, Pennsylvania 17033
Stephen M. Breneman (1), Department of Neurobiology and Anatomy, The
Center for Psychoneuroimmunology, University of Rochester Medical
Center, Rochester, New York 14642
Gary J. Brenner (1), Department of Microbiology and Immunology, The
Center for Psychoneuroimmunology, University of Rochester Medical
Center, Rochester, New York 14642
John T. Cacioppo (321), Department of Psychology, The Ohio State Univer­
sity, Columbus, Ohio 43210
Robert Cocke (1), Department of Psychiatry, The Center for Psycho­
neuroimmunology, University of Rochester Medical Center, Rochester,
New York 14642
Nicholas Cohen (1), Departments of Psychiatry and Microbiology and Im­
munology, The Center for Psychoneuroimmunology, University of
Rochester Medical Center, Rochester, New York 14642
Sheldon Cohen (301), Department of Psychology, Carnegie-Mellon Univer­
sity, Pittsburgh, Pennsylvania 15213
Sharon O. Cummings (77), Department of Anatomy, Physiology and Cell
Biology, University of California at Davis, Davis, California 95616
Cathleen M. Dobbs (101), Department of Medical Microbiology and Im­
munology, College of Medicine, The Ohio State University, Columbus,
Ohio 43210
Joel M. Dopp (1), Department of Neurobiology and Anatomy, The Center
for Psychoneuroimmunology, University of Rochester Medical Center,
Rochester, New York 14642
Brian Esterling (267), Department of Psychology, University of Miami, Cor­
al Gables, Florida 33124
Fawzy I. Fawzy (365), Department of Psychiatry, University of California at
Los Angeles School of Medicine, Los Angeles, California 90024
Nancy W. Fawzy (365), Department of Psychiatry, University of California
at Los Angeles, School of Medicine, Los Angeles, California 90024, and
John Wayne Cancer Institute, St. John's Hospital and Health Center, Los
Angeles, California 90404
Suzanne Y. Feiten (1), Departments of Psychiatry and Neurobiology and
Anatomy, The Center for Psychoneuroimmunology, University of
Rochester Medical Center, Rochester, New York 14642
Monika Fleshner (161), Department of Psychology, University of Colorado
at Boulder, Boulder, Colorado 80309
 Contributors xix

Mary Ann Fletcher (267), Department of Medicine, University of Miami,

Coral Gables, Florida 33124, and The Miami Veterans Administration
Medical Center, Miami, Florida 33136
Ronald Glaser (321), Department of Medical Microbiology and Immunol­
ogy, The Ohio State University College of Medicine, Columbus, Ohio
43210
Ann C. Griffin (77), Department of Pathology, Dartmouth Medical School,
Dartmouth University, Lebanen, New Hampshire 03756
Lee J. Grota (1), Department of Psychiatry, The Center for Psychoneuroim-
munology, University of Rochester Medical Center, Rochester, New York
14642
Nicholas R. S. Hall (183), Department of Psychiatry and Behavioral Medi­
cine, TGH-University Psychiatry Center, University of South Florida Col­
lege of Medicine, Tampa, Florida 33613
Gail Ironson (267), Departments of Psychology and Psychiatry, University
of Miami, Coral Gables, Florida 33124
Jonathan D. Karp (1), Department of Psychiatry, The Center for Psycho-
neuroimmunology, University of Rochester Medical Center, Rochester,
New York 14642
Steven E. Keller (217), Department of Psychiatry, University of Medicine
and Dentistry, New Jersey Medical School, Newark, New Jersey 07103
Margaret E. Kemeny (245), Department of Psychiatry and Biobehavioral
Sciences, University of California at Los Angeles School of Medicine, Los
Angeles, California 90024
Janice K. Kiecolt-Glaser (321), Department of Psychiatry, The Ohio State
University College of Medicine, Columbus, Ohio 43210
Nancy Klimas (267), Department of Medicine, University of Miami, Coral
Gables, Florida 33124, and The Miami Veterans Administration Medical
Center, Miami, Florida 33136
Mahendra Kumar (267), Department of Psychiatry, University of Miami,
Coral Gables, Florida 33124
Alex Kusnecov (23), The Brain, Behavior, and Immunity Center, and the
Department of Pathology, The University of Pittsburgh School of Medi­
cine, Pittsburgh, Pennsylvania 15213
Arthur LaPerriere (267), Departments of Psychology and Psychiatry, Uni­
versity of Miami, Coral Gables, Florida 33124
Mark L. Laudenslager (161), Department of Psychiatry, University of Colo­
rado Health Sciences Center, Denver, Colorado 80220
William B. Malarkey (321), Department of Medicine, The Ohio State Uni­
versity College of Medicine, Columbus, Ohio 43210
xx Contributors

Jan A. Moynihan (1), Department of Psychiatry and Microbiology and

Immunology, The Center for Psychoneuroimmunology, University of
Rochester Medical Center, Rochester, New York 14642
Maureen P. O'Grady (183), Department of Psychiatry, TGH-University Psy­
chiatry Center, University of South Florida College of Medicine, Tampa,
Florida 33613
Bruce S. Rabin (23), The Brain, Behavior, and Immunity Center, and the
Department of Pathology, The University of Pittsburgh School of Medi­
cine, Pittsburgh, Pennsylvania 15213
Steffanie Rasnick (23), The Brain, Behavior, and Immunity Center, and the
Department of Psychiatry, University of Pittsburgh School of Medicine,
Pittsburgh, Pennsylvania 15213
Steven J. Schleifer (217), Department of Psychiatry, University of Medicine
and Dentistry, New Jersey Medical School, Newark, New Jersey 07103
Neil Schneiderman (267), Departments of Psychology, Psychiatry, and Med­
icine, University of Miami, Coral Gables, Florida 33124
John F. Sheridan (101), Department of Medical Microbiology and Immu­
nology, Section of Oral Biology, Colleges of Dentistry and Medicine, The
Ohio State University, Columbus, Ohio 43210
Samuel C. Shiflett (217), Department of Psychiatry, University of Medicine
and Dentistry, New Jersey Medical School, Newark, New Jersey 07103
Michael Shurin (23), The Brain, Behavior, and Immunity Center, and the
Department of Pathology, The University of Pittsburgh School of Medi­
cine, Pittsburgh, Pennsylvania 15213
George F. Solomon (341), Geriatric Research, Education and Clinical Cen­
ters, Sepulveda Veterans Affairs Administration Medical Center, Sep-
ulveda, California 91343
Caroline C. Whitacre (77), Department of Medical Microbiology and Im­
munology, College of Medicine, The Ohio State University, Columbus,
Ohio 43210
Daohong Zhou (23), The Brain, Behavior, and Immunity Center, and the
Department of Pathology, The University of Pittsburgh School of Medi­
cine, Pittsburgh, Pennsylvania 15213
Bruce S. Zwilling (53), Department of Microbiology, College of Biological
Sciences, The Ohio State University, Columbus, Ohio 43210
 PREFACE

There is increasing evidence that the central nervous system (CNS) can
influence the immune response, the body's defense against infectious and
malignant diseases. This book focuses on human studies, as well as relevant
animal models for human infectious and autoimmune illness. We chose to
highlight these particular animal models because they demonstrate the di­
rect impact of various Stressors on the immune system and the subsequent
modulation of infectious and autoimmune diseases. In addition, these ani­
mal studies help to demonstrate the range of physiological changes associ­
ated with Stressors. We believe that the chapters in this book will provide
the reader with a current summary of the field of psychoneuroimmunology
as it pertains to stress and illness outcomes.
Psychological distress can lead to adverse immunological changes pro­
viding one physiological pathway through which major and minor life
changes might result in an increased incidence of infectious and malignant
disease. However, most individuals who experience major life changes do

xxi
xxH Preface

not become ill, or they only experience short illness episodes. Actual organ­
ically based episodes of illness are a function of differential exposure to
pathogens and/or carcinogens, genetics, as well as the prior health of the
individual, particularly regarding immune function. Thus, in individuals
with equal exposure to an infectious agent (e.g., a virus), the probability of
clinical illness and the intensity and duration of the illness are, in part, the
product of the prior status of the individual's immune system. Following
this line of reasoning, individuals who are presumably most likely to show
health changes in response to Stressors are those whose immune function is
already compromised to some extent, particularly the elderly, since aging is
associated with impaired immune function.
The adrenal cortex produces glucocorticoids in response to Stressors
through the hypothalemic-pituitary axis. Individuals with an adreno-
cortical insufficiency such as Addison's Disease have physiological difficulty
handling stress. In addition, an overabundance of glucocorticoids during
stress also can have major physiological consequences. Research on the
relationships among the CNS, immune system and endocrine systems pro­
vides evidence of physiological pathways, including the glucocorticoids, and
other 'stress hormones" such as the catecholamines, prolactin, and growth
hormone, through which distress may modulate immune function.
There also are other pathways through which the CNS may influence
immune function, such as direct innervation of both primary and secondary
lymphoid organs. Our knowledge about the interactions among the CNS,
the immune system, and the endocrine system in regard to mechanisms of
action is still limited. However, there is an active and growing literature
describing such relationships, and their health-related consequences. These
and other issues are covered in the chapter in this book by well-known
experts in their respective areas.

Ronald Glaser
Janice K. Kiecolt-Glaser
 D
Stress-Induced Modulation of Immune
Function in Mice
Jon A. Moynihan, GoryJ. Brenner, Robert Cocke, Jonothon 0. Korp,
Stephen M. Brenemon, JoelM. Oopp, Robert Ader, Nicholos Cohen,
Lee J. Groto, and Suzanne l Feiten

I. INTRODUCTION

Stress, which is broadly defined as the response of an organism to

stimulation or change (Selye, 1950), is characterized by activation of both
the autonomie nervous system and the hypothalamo-pituitary-adrenal
(HPA) axis. The resulting neurochemical changes have been demonstrated
to affect immune function both directly and indirectly. Direct effects are
possible because lymphocytes bear receptors on their surfaces for many
neurohormones and transmitters, and lymphocytes are exposed to neuro-
chemicals in lymphoid organs and in peripheral blood (Carr &c Blalock,
1991; Roszman & Carlson, 1991; Feiten &c Feiten, 1991). More indirect
mechanisms for neural-immune interactions may involve changes in lympho­
cyte trafficking resulting from changes in sympathetic vascular tone (Ottaay,
1991).
The elucidation of pathways of communication between the central

Handbook of Human Stress and Immunity -

Copyright © 1994 by Academic Press, Inc. All rights of reproduction in any form reserved. I
2 Jan A. Moynihan et al.

nervous system and the immune system has advanced, to a large extent,
from studies utilizing Stressor administration in animal models. Application
of a Stressor in animals results in diverse changes in both specific humoral
and cell-mediated immune parameters, and in nonspecific natural killer
(NK) cell number and/or activity (reviewed in Kiecolt-Glaser &c Glaser,
1991; Shavit, 1991; Moynihan &c Cohen, 1992; Ader & Cohen, 1993).
Many of the studies that have addressed stress-induced changes in disease
outcome and/or immune function also illustrate the complexities of the
interactions between the central nervous and immune systems. It is certainly
apparent that: ( 1 ) not all Stressors produce the same pattern of neurochemi-
cal responses; (2) not all strains, species, or sex of animals respond to a
given Stressor in the same behavioral, neurochemical or immunological
manner; and (3) not all immune effector functions are equally affected by
Stressor application. Thus, in response to Stressors, immune effector func­
tions are suppressed, enhanced, or unchanged. As outlined in Table 1, sev­
eral variables have been demonstrated to be important determinants of the
immunological consequences of Stressor administration. These variables re­
flect the complexity of both the neuroendocrine responses to a Stressor,
and the interaction of these responses with the cascade of molecular and
cellular immune responses to an antigen. This neuroendocrine complexity
can be highlighted by pointing out that, although in some studies, stressor-
associated immunomodulation correlates with altered levels of glucocor-
ticoids (Blecha, Kelley, & Satterlee, 1982; Keller, Weiss, Schleifer, Miller, &
Stein, 1983; Okimura, Ogawa, Yamauchi, &c Sasaki, 1986; Coe, Rosenberg,

TABLE 1 Interacting Variables in Stress-Induced Immune Modulation

A. Experimental subject
1. Species of subject
2. Strain of animal
3. Age and sex of animal
4. Previous history (e.g., other Stressors)
5. Orcadian and other rhythms
B. Stressor
1. Nature of the Stressor (e.g., physical versus psychosocial)
2. Duration of exposure
3. Intensity of Stressor
4. Timing of Stressor administration in relation to immune challenge
C. Immune measure
1. Choice of antigen and concentration of antigen
2. Lymphoid compartment examined
3. In vitro versus in vivo immunological assay
4. Type of immune response measured (e.g., humoral versus cell-
mediated)
 1 ■ Stress-Induced Modulation of Immune Function in Mice 3

Fisher, &c Levine, 1987) or opioids (Shavit, Lewis, Terman, Gale, &C Lie­
beskind, 1984; Shavit, 1991), altered levels of these glucocorticoids and/or
opioids have not been found in all studies that report effects of Stressors on
immune responses (Cunnick, Lysle, Armfield, & Rabin, 1988; Ben-Eliyahu,
Yirmiya, Shavit, &c Liebeskind, 1990; Rabin, Cunnick, & Lysle, 1990;
Moynihan & Cohen, 1992; Ader & Cohen, 1993). Further, at least 17
neuroendocrine peptides have immunomodulatory potential (Khansari,
Murgo, &c Faith, 1990), and many of these hormones or peptides are al­
tered by Stressors. Finally, hypothalamic hormones such as corticotropin-
releasing hormone (CRH) may exert immunomodulatory effects via both
the pituitary-adrenal axis (Munck &c Guyre, 1991) and the sympathetic and
parasympathetic nervous system release of norepinephrine and other cate-
cholamines (Sundar, Cierpial, Kilts, Ritchie, &c Weiss, 1990; Irwin, Hauger,
Jones, Provencio, & Britton, 1990; Irwin, Vale, & Rivier, 1990). Thus,
multiple neuroendocrine pathways are undoubtedly involved in stress-
induced alteration of immune function.
Although it is critical to study stress-induced neurochemical changes
from the perspective of the immunologist it is also necessary to understand
what events in the immune cascade that are triggered by antigen or patho­
gen are changed as a consequence of the stress response. Experimental
mechanistic approaches from this perspective are based on the awareness
that the immune system has levels of complexity that rival those of the
nervous system. This chapter will discuss our recent findings using Stressors
to alter antigen-driven events that lead to cellular and humoral immune
responses; illustrate some of the interactions between the variables listed in
Table 1; and highlight some of the possible immunological mechanisms
involved in stress-induced changes in immune function.

II. THE EFFECTS OF HANDLING ON IMMUNE FUNCTION AND ON TUMOR METASTASES

Early stress studies often used a simple, yet effective, method of psycho-
social stimulation, that is, picking up and holding an animal for a short
period. Many of these studies examined the effects of handling neonatal rats
or mice (Solomon, Levine, &c Kraft, 1968; Friedman, Glasgow, &c Ader,
1970; LaBarba, 1970; Michaut, Dechambre, Doumerc, Lesourd, Deville-
chabrolle, & Moulias, 1981; Lown &c Dutka, 1987). However, many stud­
ies that have investigated handling of neonatal or adult rodents have not ex­
amined immune function per se, but, for example have looked at changes in
survival time following tumor or viral challenge (Newton, Bly, & McCrary,
1962; Ader, 1965; Friedman et al., 1970). Our approach has been to study the
effects of repeated daily handling of adult mice on a number of specific and
nonspecific immune effector mechanisms or an antibody response, as well as
a tumor challenge (Moynihan, Brenner, Koota, Breneman, Cohen, &c Ader,
1990; Brenner, Cohen, Ader, &c Moynihan, 1990).
4 Jan A. Moynihan et al.

^.D -
Handled Day 5
2--«·--
F
e
° - —o— Unhandled
in
§ 1.5-
Φ
Ü
S LO­
IS

h—
O
5 0.5-

0.0- | 1 | 1 | '

150 225

750 150 225

Reciprocal Serum Dilution
FIGURE 1 The IgG anti-KLH antibody response in handled versus control BALB/c female
mice. Mice in both groups (n = 12/group) were immunized following two weeks of daily
handling (2 min/day) with 100 μg of KLH. Mice were bled to measure serum antibody titers on
Days 5, 10, and 15 following immunization (Moynihan et al., 1990). Data are expressed as
mean ± SEM.

Group-housed adult BALB/c.ByJ female mice were held gently without

restraint for 2 min/day for 2 weeks prior to intraperitoneal (ip) injection of
the T cell-dependent protein antigen keyhole limpet hemocyanin (KLH).
Mice were bled at various times following immunization to measure specific
anti-KLH antibody responses. Mice that were handled for two weeks prior
to immunization had a significant and reproducible decrease in lgG anti-
KLH antibody responses compared to unhandled control mice. The results
of one such experiment are shown in Figure 1. Handling was also associated
with a significant decrease in antigen-induced in vitro spleen cell production
of the cytokine interleukin (IL)-2, produced by a subset of mature T helper
cells designated T helper 1 (TH1) cells (Street & Mosmann, 1991) (Figure
2). Additionally, the proliferative response of spleen cells from nonim-
munized mice to the T cell mitogen concanavalin A (ConA) was diminished
in cell cultures from handled mice. The proliferative response to the B cell
mitogen lipopolysaccharide (LPS) was unaffected. Thus, handling mice for 2
weeks resulted in both depressed T cell function and diminished lgG anti­
body production to a T cell-dependent antigen.
 1 ■ Stress-Induced Modulation of Immune Function in Mice 5

Days Post Immunization

FIGURE 2 In vitro KLH-induced IL-2 production in spleen cell cultures from handled versus
control BALB/c female mice. Mice {n = 6/group/time point) were handled as described earlier,
immunized following handling with 100 μg of KLH, and sacrificed 6, 9, 12, and 15 days
following immunization. Spleen cells were cultured in 96-well plates at 2 X 10 5 cells/well with
80 μg/ml of KLH. Supernatants were removed 24 h later and assayed for IL-2 using the IL-2-
dependent cell line CTLL (Mosmann, Cherwinski, Bond, Giedlin, & Coffman, 1986). Data are
expressed as mean ± SEM.

We have also examined the effects of 2 weeks of daily handling on the

response to intravenous injection of a tumor cell line, resulting in lung
métastases. The line 1 alveolar carcinoma (Yuhas, Toya, &c Wagner, 1975;
Yuhas, 1977) was injected into syngeneic BALB/c female mice following 2
weeks of handling. Mice were sacrificed 8 to 21 days later (the variability
was between, not within, experiments) and the numbers of lung tumors
were enumerated. Figure 3 illustrates that numbers of lung métastases were
reproducibly increased in handled mice, and that this increase was statis­
tically significant regardless of the overall numbers of métastases (i.e., an
average of 149 métastases in Experiment 3 versus 5.4 in Experiment 2).
The line 1 tumor is reported to be sensitive to lysis by NK cells, and is
not particularly immunogenic, presumably due to poor expression of class I
major histocompatibility antigens (Cerosaletti, Bleiden, Harwell, Welsh,
Frelinger, & Lord, 1990). In a number of studies in which we examined
both in vivo and in vitro NK cell cytotoxicity against either the YAC-1
lymphoma line or line 1 cells, we did not observe any decrease in NK cell
function in handled mice. Thus, although immune function (as evidenced
by decreased T cell function and antigen-specific lgG production) is altered
in handled mice, these immunological changes may not necessarily be in­
volved in the increased métastases following tumor challenge.
6 Jan A. Moynihan et al.

b.

140- ■■
^^1
105- ^^^^l
^Λ \
70-
H
35- H
0- _JHL

FIGURE 3 Pulmonary métastases in handled or unhandled control BALB/c female mice. Mice
(„ = 7-12/group/experiment) were injected intravenously following 2 weeks of handling with
10 5 line 1 cells. Mice were sacrificed 8 to 21 days later (the variability was between, not within,
experiments) and pulmonary métastases were enumerated (Brenner et al., 1990). Four replica­
tions are shown (a-d). Data are expressed as mean±SEM.

HANDLING RESULTS IN HABITATION OF THE STRESS RESPONSE

We have speculated that handling results in a classic stress response in

mice, resulting in activation of the HPA axis and the sympathetic nervous
system (SNS). We began these experiments with the bias that the handled
mice were the "stressed" group, and that the unhandled controls were
 1 ■ Stress-Induced Modulation of Immune Function in Mice 7

150-

0)

Ä100-
a>
si
a.
0)
c
'5. 50-J
ω
CO
E Unhandled
en
i2
û. Handled

Time post injection (min)

FIGURE 4 Plasma epinephrine and corticosterone responses in handled versus control
BALB/c female mice. Mice (n = 6/group/time point) were injected with saline intraperitoneally
and sacrificed at the indicated times to measure plasma epinephrine and corticosterone. The
mice sacrificed at time zero did not receive an injection. Data are expressed as the mean± SEM.

"nonstressed." However, we have observed that following the administra­

tion of an ip injection (a Stressor in itself), previously handled mice have
attenuated plasma corticosterone and catecholamine responses compared to
previously unhandled mice (Figures 4a and 4b). Indeed, it has been reported
in the literature that habituation to handling is associated with higher gam-
mabutyric acid (GABA) receptor binding in certain brain areas associated
with an increase in number of GABA receptors (Biggio, Corda, Concas,
Demontis, Rossetti, & Gessa, 1981), lower levels of cerebellar cGMP and
8 Jan A. Moynihan et al.

Chronic
-16,-9,-2,+5
Injection period (day)
FIGURE 5 Pulmonary métastases in chemically sympathectomized versus control BALB/c
female mice. Mice (« = 18-20/group) were injected intraperitoneally with either 125 mg/kg
of the neurotoxin 6-hydroxydopamine (6-OHDA) or vehicle at the times indicated. Mice
were injected intravenously with 10 5 line 1 tumor cells on Day 0, sacrificed 14 days later,
and pulmonary métastases were enumerated (Brenner et al., 1992). Data are expressed as
mean ± SEM.

cAMP (Corda, Biggio, & Gessa, 1980), reduced opioid-induced analgesia

(Fanselow & Sigmundi, 1986), and reduced glucocorticoid responses com­
pared to unhandled animals (Ader, Friedman, Grota, & Schaefer, 1968;
Ader & Grota, 1969; File, 1982; Moynihan et al., 1990). Indeed, ablation
of the peripheral SNS using the drug 6-hydroxydopamine (6-OHDA) prior
to, but not following, injection of line 1 tumor cells resulted in increased
numbers of pulmonary métastases (Brenner, Feiten, Feiten, & Moynihan,
1992; see Figure 5). Thus, the increase in pulmonary métastases observed in
either handled or sympathectomized mice may be a function of the attenua­
tion, rather than the elicitation, of a stress or catecholamine response. Al­
though increased métastases are associated with neuroendocrine changes,
there is no evidence that increased métastases in either handled or sym­
pathectomized mice are associated with altered immune effector mecha­
nisms (Brenner et al., 1990, 1992). Thus, it may be that not all stress-
induced changes in tumor or disease progression are associated with altered
immunity.
 1 ■ Stress-Induced Modulation of Immune Function in Mice 9

IV. THE EFFECTS OF STRESS ON VIRAL INFECTION AND IMMUNITY

Although the increased number of lung métastases associated with han­

dling does not appear to be immunologically mediated, there are clear in­
stances in which stress-induced changes in disease pathogenesis have been
correlated directly with immune function. For example, recent studies have
demonstrated that the viral-specific immune responses of C57B1/6 female
mice following primary infection in the footpad with herpes simplex virus-
type 1 (HSV-1) can be altered by either restraint stress (Bonneau, Sheridan,
Feng, & Glaser, 1991a,b) or mild electric footshock (Kusnecov, Grota,
Schmidt, Bonneau, Sheridan, Glaser, & Moynihan, 1992). Using a foot-
shock model, we (Kusnecov et al., 1992) have observed that Stressor admin­
istration results in depression of spleen and popliteal lymph node cell num­
ber in infected mice; in vitro cytotoxic T cell responses to virally-infected
cells; IL-2 production following stimulation with virally-infected cells in
vitro; and humoral immune responses. In addition, infectious virus recov­
ered from the footpads of the shocked mice differs dramatically from both
the apparatus control and the home cage mice (44.5 ± 1.9, 9.1 ± 3.5, and
1.0 ± 0 plaque-forming units X 10 5 , respectively, for the shock, apparatus
control, and home cage mice). Thus, impaired antiviral responses are associ­
ated with impaired clearance of virus from the site of infection.

V. THE CENTRAL NERVOUS SYSTEM RESPONSE OF A RECIPIENT MOUSE TO THE ODORS

EMITTED BY A STRESSED CONSPECIHC ALTERS CELL-MEDIATED AND HUMORAL IMMUNE
FUNCTION DIFFERENTLY

Thus far, this chapter has discussed the effects of physical Stressors on
immune function. In addition to these studies, we have recently focused on a
paradigm that does not involve physical pain or trauma, and is of clear
ethologic relevance to the rodent. The paradigm involves pheromonal stim­
ulation of conspecifics. The olfactory system of a rodent is highly developed,
and is critical for communication among these animals (Harrington & Ro­
sario, 1992). Other investigators have determined that odors (such as bo­
tanical odors) can be immunoregulatory (Shibata, Fujiwara, Iwamoto, Mat-
suoka, & Yokoyama, 1990). Further, there is some evidence that the odors
of predators or of stressed conspecific animals can alter the number of
antibody forming cell to sheep red blood cells (SRBC, Zalcman, Kerr, &
Anisman, 1991). We have tested the hypothesis that odors or pheromones
produced by footshock-stressed mice result in altered immune function in
conspecific odor recipients. We hypothesize that this ethologically relevant
model of stress mimics components of a natural predator-prey situation.
Initially, we examined the responses of BALB/c male mice to odors
10 Jan A. Moynihan et al.

0.4
a.

0.3 H
E
e
<o
o
0.2 H
m
Φ
o
e
(0
o 0.1 H
Stress odor-exposed
n
< Control odor-exposed

0.0

Con A (ug/ml)

</>
Φ
o
Ü
<
>-
«*-
o
<Λ
0)

o
<D
a.
e/)

200

E:T Ratio
FIGURE 6 (a) IL-2 production by spleen cells from stress versus control odor-exposed
BALB/c male mice. Mean (±SEM) absorbance for IL-2 produced by Con A-stimulated (0.3-5
μg/ml) spleen cells obtained from control odor-exposed (n = 12) and footshock stress odor-
exposed (n = 12) mice following a 24 h odor exposure (Coclee et al., 1993). IL-2 was assayed
using the CTLL bioassay. (b) NK cell cytotoxicity of spleen cells from odor-exposed mice.
Varying numbers of spleen cells from either stress odor-exposed (n = 12) or control odor-
exposed (n = 12) mice were incubated with YAC-1 lymphoma cells in a standard 6-h 51Cr-
release assay. Data are expressed as mean ± SEM.