environmental exposures and the immune system: gender specific differences and consequences allen...
TRANSCRIPT
Environmental Exposures and the
Immune System: Gender Specific Differences and
consequences
Allen Silverstone, SUNY Upstate Medical Allen Silverstone, SUNY Upstate Medical University Dept. of Microbiology and University Dept. of Microbiology and
Immunology and University of Rochester School Immunology and University of Rochester School of Medicine, Dept. of Environmental Medicineof Medicine, Dept. of Environmental Medicine
ON THE EVE OF THE REISSUANCE OF THE GREAT
1970’S CLASSIC THE JOY OF SEX
IT GIVES ME GREAT PLEASURE TO TALK TO YOU, ABOUT
THE JOY OF IMMUNOLOGY
Which can last longer than sex.
THE FUNCTION OF THE THE FUNCTION OF THE IMMUNE SYSTEM IS FIRST IMMUNE SYSTEM IS FIRST
AND FOREMOST TO AND FOREMOST TO PROTECT US FROM THE PROTECT US FROM THE
MICROBIAL WORLD (THE MICROBIAL WORLD (THE PATHOGENIC PART PATHOGENIC PART
ANYWAY)ANYWAY)
IT DOES THIS BY UTILIZING A IT DOES THIS BY UTILIZING A LARGE NUMBER OF DIFFERENT LARGE NUMBER OF DIFFERENT CELL TYPES WHICH IN VARIOUS CELL TYPES WHICH IN VARIOUS
INTERACTIONS DELIVER A INTERACTIONS DELIVER A RESPONSE THAT IS MORE OR RESPONSE THAT IS MORE OR
LESS SPECIFIC, AND, WITH LESS SPECIFIC, AND, WITH LUCK CONTROLS AND LUCK CONTROLS AND
ELIMINATES THE MICROBIAL ELIMINATES THE MICROBIAL AGENT BEFORE THE ORGANISM AGENT BEFORE THE ORGANISM
IS KILLED.IS KILLED.
THESE DIFFERENT CELL THESE DIFFERENT CELL TYPES DEVELOP WITHIN TYPES DEVELOP WITHIN
THE ORGANISM IN SPECIFIC THE ORGANISM IN SPECIFIC TISSUES AND IN SPECIFIC TISSUES AND IN SPECIFIC
STAGES.STAGES.
IMMUNOTOXICOLOGY
A SUBDISCIPLINE OF IMMUNOPHARMACOLOGY
SORT OF A NEGLECTED CHILD OF IMMUNOLOGY
PHARMACOLOGICAL AGENTS CAUSE IMMUNOSUPPRESSION
1- Steroids: Affect cell traffic (lymphocytopenia; monocytopenia; neutrophilia); Cell function ( Inhibit T and B cell maturation; inhibit IL-1 and TNF production, IL-2,4,6, and 10 production reduced).
2-Alkylating agents for Chemotherapy (kill stem cells; useful in establishing tolerance) e.g. Cyclophosphamide; nitrogen mustards.
3- Nucleotide Synthesis Inhibitors: Azathioprine, allopurinal, methotrexate.
4-Cyclosporin, FK506, Rapamycin
IMPROPER NUTRITION CAN LEAD TO REDUCED IMMUNE RESPONSES
1-Malnutrition reduces lymphatic tissue, lymphoid atrophy, reduced CD4/CD8 ratio; reduced Th-2 activity, reduced IgA production to vaccines (problems in developing countries); reduced opsonization (reduced complement production )
2-Zinc deficiency: (reduced CD4/8; T-cell anergy)
3-Vitamin A deficiency (reduced mitogen responses)
4-Iron deficiency (reduced neutrophil activity; ferritin-transferrin binding vs. bacterial iron binding proteins).
ENVIRONMENTAL AGENTS CAN AFFECT IMMUNITY
1-Dioxins/PCBs: Reduced CMI, humoral responses; increased sensitivity to viral, bacterial, parasitic infections.
2- Heavy Metals: Cadmium, mercury , lead(Dietert and Associates have shown that immunosuppression by lead is gender affected
3-Diesel Fuel Particulates: (opsonizing functions; neutrophil activity, CMI effects
4- Many pesticides and herbicides:
TCDD (the most toxic dioxin): Arises as a byproduct inmanufacture and in combustion. Its toxicity is such thatit is the reference standard for toxicity of other PHAH’S
These Compounds are relatively resistant to environmentaland metabolic accumulation, and are thus highly persistent.They are very lipophilic and are persistent in human tissues.
• Fatal Cachexia
•Chloracne
•Hepatotoxicity
•Repro Toxicity
•Cancer
Dioxins & Dioxin Like PCBsWhat do they do?
•Decreased T-cell responses to tumors•Decreased primary and memory responses • Increased susceptibility to bacterial, parasitic and viral infections
Toxicities: Immunotoxicity:
THYMIC ATROPHYTHYMIC ATROPHY
UNLIKE DEXAMETHASONE, DIOXIN CAUSESTHYMIC ATROPHY SLOWLY, BUT ALSO IN
BCL-2 TRANSGENIC MICE
HO
CH3
OH
C = CHO OH
C2
H5
C2
H5
-Estradiol Diethylstilbestrol (DES)
Important for normal physical development, maintain reproductive cycles and regulation for liver metabolism and immune function, and ensure balance of normal body systems
Physiological levels
Hormone replacement therapy and treatment for prostate cancer
Pharmaceutical therapy
*
HO
CH3
OH
C = CHO OH
C2
H5
C2
H5
-Estradiol Diethylstilbestrol (DES)
* Related to autoimmune diseases* Related to breast and reproductive organ cancer* Influence on the development and functions of
reproductive system* Influence on neural-endocrine system and behavior* Effects on immune system
High levels
The effects of estrogen on immune system (1)
Induction of thymic atrophy and decrease of all CD4/8 subsets of thymocytes
Inhibition of spleen T-cell proliferation response to stimulants in vitro
Increase of INF- , IL-4 and shift immune response to Th2 from Th1
Decrease of numbers of peripheral T cells
Impairment of T-cells
Inhibition of differentiation, proliferation and survival of BM B cell precursor
Increase of number of plasma cells, immuno-globulins and autoantibodies.
Reduction of NK cell number and activity Reduction of delayed type hypersensitivity
Increase of susceptibility and death to infection
The effects of estrogen on immune system (2)
Inhibition of BM B-cell precursor
Hyperactivity of B-cells
0 2 4 6 8 10 12
0
20
40
60
80
100
120P
erce
nt
of c
ontr
ol (
%)
Days after E2 injection
number of total cells*****
ESTROGEN ALSO CAUSES ATROPHYESTROGEN ALSO CAUSES ATROPHY
BESIDES SUPPRESSIVE BESIDES SUPPRESSIVE EFFECTS ON THE IMMUNE EFFECTS ON THE IMMUNE SYSTEM, MANY ESTROGEN SYSTEM, MANY ESTROGEN AFFECTING XENOBIOTICS AFFECTING XENOBIOTICS
AND EVEN PHYSIOLOGICAL AND EVEN PHYSIOLOGICAL LEVELS OF ESTROGEN ARE LEVELS OF ESTROGEN ARE
ASSOCIATED WITH ASSOCIATED WITH HYPERIMMUNITY HYPERIMMUNITY
(ASTHMA,ALLERGY, ETC.)(ASTHMA,ALLERGY, ETC.)
OR AUTOIMMUNE DISEASEOR AUTOIMMUNE DISEASE
AUTO-IMMUNE MODELS SHOW THE DEPENDENCE OF BOTH
GENETICS AND SEX
Immune System
Environmental Triggers/
Potentiators
Sex Hormones Genetics
Immune System
Hyper
Immunity
Auto- Immune Disease
Immune Suppression
Sex Hormones
Immune System
Hyper
Immunity
Auto- Immune Disease
Immune Suppression
Environmental Triggers/
Potentiators
Immune System
Hyper
Immunity
Auto- Immune Disease
Immune Suppression
Genetics
GETTING BY WITH A LITTLE HELP GETTING BY WITH A LITTLE HELP FROM MY FRIENDSFROM MY FRIENDS
TOM GASIEWICZTOM GASIEWICZ OF THE OF THE UNIVERSITY OF ROCHESTER WHO UNIVERSITY OF ROCHESTER WHO INTRODUCED ME TO DIOXIN AND INTRODUCED ME TO DIOXIN AND ITS RECEPTOR AND WITH WHOM I ITS RECEPTOR AND WITH WHOM I HAVE CREATED A PARADIGM FOR HAVE CREATED A PARADIGM FOR
STUDYING HOW RECEPTOR STUDYING HOW RECEPTOR MEDIATED ACTIONS AFFECT THE MEDIATED ACTIONS AFFECT THE
IMMUNE SYSTEMIMMUNE SYSTEM
& John MacLachlan who insisted we study E2& John MacLachlan who insisted we study E2
AhR-KO Chimeras Eliminate Epithelial Targets
DP
CD44+CD25-
CD44+CD25+HSA+
CD44-CD25hi
HSA+
CD44-CD25int
HSA+
CD4+CD8+HSA+/-
TCRCD3+CD4+
TCRCD3+CD8+
RIP
Stem cellfrom bonemarrow orfetal liver
CD8- CD4- DN
SP
CD44-CD25lo
HSA+
=DN 1
=DN 2
=DN 3
=DN 4A
=DN 4B
Cl Cl
O
TCDD
O
Cl Cl
RAG Block
HSA+ LY 5.1+or LY 5.2+
CD44+CD25+HSA+
CD44-CD25hi
HSA+
CD44-CD25-HSA+
BrdU incorporation in the CD44-CD25hi HSA+ Cell Population after TCDD Treatment
CD44+CD25-
CD44-CD25int
HSA+
Hours after TCDD Treatment
Per
cen
t of
Cel
ls I
nco
rpor
atin
g B
rdU
p<0.01
Oil 30 g/kg TCDD
***
***
*** **
*
0 6 12 18 24 40 60 80 100 120 1400
5
10
15
20
25
30
100
300
500
700
900
1100
0 h 3 h 6 h 24 h 1 w 2 w 7 w
exposure time
adseverin
HPRT
Svensson C. and Lundberg K., In manuscript
*
**
*
*
*
*
AADSEVERIN INDUCTION OCCURS WITHIN
THREE HOURS OF TCDD EXPOSURE IN VIVO
0
20
40
60
80
V T V T V T
% TUMOR CELL CYTOTOXICITY
-/- Ÿ +/+ +/+Ÿ +/+ B6 W.T.
SPLEEN CELLS FROM MICE INNOCULATED WITH P815TUMOR TESTED AT 100/1 FOR TUMOR CYTOTOXICITY
Hemopoietic Expression of ER is required for maximum response to E2
WT >WT KO >WT WT >KO KO >KO
Th
ym
ic c
ellu
lari
ty (
x 1
0
7 )
0
5
10
15
20
25
30
controlestradiol
ER- ER- is is required for a full sized thymus!! required for a full sized thymus!!
Wild Type ER KO ER KO ER KO
Th
ymu
s W
eigh
t (m
gs)
0
10
20
30
40
50
60
70
Oil E2
ERKO ARE PARTIALLY SENSITIVE TO E2
. ERKO ARE FULLY SENSITIVE TO E2
Ovx (cst) results in full size thymuses!Ovx (cst) results in full size thymuses!
GETTING BY WITH A LITTLE GETTING BY WITH A LITTLE HELP FROM MY FRIENDSHELP FROM MY FRIENDS
KEN KORACH, DENNIS LUBAHN, KEN KORACH, DENNIS LUBAHN, AND SYLVIA HEWITT FOR HELP AND SYLVIA HEWITT FOR HELP AND SUPPLYING MICE LACKING AND SUPPLYING MICE LACKING
ESTROGEN RECEPTOR ALPHA ESTROGEN RECEPTOR ALPHA (AERKO’S) AND BETA (BERKO’S)(AERKO’S) AND BETA (BERKO’S)
And for only being able to give us males in And for only being able to give us males in sufficient number … that lead us to realize how sufficient number … that lead us to realize how important the ER is for Male Immune system.important the ER is for Male Immune system.
GETTING BY WITH A LITTLE GETTING BY WITH A LITTLE HELP FROM MY FRIENDSHELP FROM MY FRIENDS
Emilie RissmanEmilie Rissman who gave us Tfm AERKO’s who gave us Tfm AERKO’s to rule out Androgen receptor involvement to rule out Androgen receptor involvement in certain estrogen effectsin certain estrogen effects
Ken Olden and NIEHSKen Olden and NIEHS for their support for for their support for the last 13 years for our work in this area.the last 13 years for our work in this area.
******
*** ***a
***
***
0 20 40 60 0 1 2 3 4 0 4 8 12
Thymus weight number of cells BrdU positive cells
mg/thymus 108/thymus % of thymocytes
Oil
1 E2
3 E2
Estradiol (E2) inhibits BrdU incorporation in DNAand induces thymic atrophy
Effects of E2 on proliferation activity of subpopulations
*CD3-CD8+
*****
******
***
***
**
CD3-DP
CD3-DN*
** ***
*********
*****
***
***
CD44+CD25+
CD44-CD25+
CD44-CD25-
A B
0 2 4 6 8 10 0 2 4 6 8 10
100
80
60
40
20
0
Days after E2 injection
Per
cen
t of
con
trol
(%
)
Control Estradiol Estradiol DES 24 Hours 24 Hours 60 Hours 24 Hours
0% Serum5% Charcoal
Stripped Serum 5% Full Serum
18-81 Proliferation and DNA Profile
0% Serum
DNA content
0% Serum
1.E+04
1.E+05
1.E+06
1.E+07
0 12 24 36 48 60 72
Hours
Nu
mb
er o
f V
iab
le C
ells
Dioxane (Control)Estradiol (20 uM)DES (20 uM)
107
106
105
104
5% Charcoal-Stripped Serum
1.E+04
1.E+05
1.E+06
1.E+07
0 12 24 36 48 60 72Hours
Nu
mb
er
of
Via
ble
Ce
lls
Dioxane
Estradiol (20 uM)
DES (20 uM)
107
106
105
104
5% Full Serum
1.E+04
1.E+05
1.E+06
1.E+07
0 12 24 36 48 60 72Hours
Nu
mb
er o
f V
iab
le C
ells
Dioxane
Estradiol (20 uM)
DES (20 uM)
107
106
105
104
G2/M build up
Wild Type ER KO
Per
cen
t of
Oil
Tre
ated
Con
trol
0
20
40
60
80
100
120
Oil E2 Prog Prog + E2 DHT
PROGESTERONE POTENTIATES THE ER KO RESPONSE TO E2
GETTING BY WITH A LITTLE HELP GETTING BY WITH A LITTLE HELP FROM MY FRIENDS (CORNELL)FROM MY FRIENDS (CORNELL)
INFORMATION AND DISCUSSIONS INFORMATION AND DISCUSSIONS WITH WITH ROD DIETERTROD DIETERT (GENDER (GENDER
AFFECTS ON LEAD ALTERATIONS OF AFFECTS ON LEAD ALTERATIONS OF THE IMMUNE SYSTEMTHE IMMUNE SYSTEM))
THE SAME DISCUSSIONS WITH THE SAME DISCUSSIONS WITH FRED QUIMBYFRED QUIMBY ON GENDER ON GENDER
EFFECTS ON DEVELOPMENT OF EFFECTS ON DEVELOPMENT OF DOGGIE LUPUS in high PCB areasDOGGIE LUPUS in high PCB areas..
GETTING BY WITH A LITTLE GETTING BY WITH A LITTLE HELP FROM MY FRIENDSHELP FROM MY FRIENDS
JERRIE GAVALCHIN: NOW AT JERRIE GAVALCHIN: NOW AT CORNELL UNIVERSITY FOR HER CORNELL UNIVERSITY FOR HER FEMALE SPECIFIC LUPUS-LIKE FEMALE SPECIFIC LUPUS-LIKE
AUTOIMMUNE MODEL … WHICH AUTOIMMUNE MODEL … WHICH ALLOWED US TO SHOW ALLOWED US TO SHOW
EXOGENOUS ESTROGENS AND EXOGENOUS ESTROGENS AND PCB’S COULD MAKE MALES HAVE PCB’S COULD MAKE MALES HAVE
DISEASEDISEASE
NoNormal (A) and Successively worse nephrotic damage
HISTOLOGICAL EVALUATIONOF KIDNEY DAMAGE
11
TCDD Increases Memory TCDD Increases Memory Phenotype in Autoimmune MicePhenotype in Autoimmune Mice
RECENT RESULTS WITH SNF-1 MODELRECENT RESULTS WITH SNF-1 MODEL
1- ESTROGEN TREATMENT OF NON 1- ESTROGEN TREATMENT OF NON AUTOIMMUNE INTACT FEMALE DB-1 AUTOIMMUNE INTACT FEMALE DB-1 MICE PRODUCES AUTOIMMUNE MICE PRODUCES AUTOIMMUNE DISEASEDISEASE
2- CHIMERIC FEMALE MICE (DB 1 2- CHIMERIC FEMALE MICE (DB 1 GIVEN HSCs FROM SNF-1) GIVEN HSCs FROM SNF-1)
DEVELOP AUTOIMMUNE DISEASEDEVELOP AUTOIMMUNE DISEASE
3- ER ALPHA KO’S IN ABOVE ARE 3- ER ALPHA KO’S IN ABOVE ARE RESISTANT TO E-2 TREATMENTRESISTANT TO E-2 TREATMENT
THE BOTTOM LINE SEEMS TO THE BOTTOM LINE SEEMS TO ME THE FOLLOWINGME THE FOLLOWING
1- WE MUST DO MORE GENDER 1- WE MUST DO MORE GENDER SPECIFIC STUDIES ON SPECIFIC STUDIES ON
IMMUNOTOXIC EFFECTSIMMUNOTOXIC EFFECTS
2- WE MUST IDENTIFY THE 2- WE MUST IDENTIFY THE SPECIFIC CELLULAR SPECIFIC CELLULAR
TARGETS FOR TOXIC AGENTS TARGETS FOR TOXIC AGENTS THAT CAUSE IMMUNE THAT CAUSE IMMUNE
EFFECTSEFFECTS
3- WE MUST DETERMINE HOW 3- WE MUST DETERMINE HOW ESTROGEN ITSELF PLAYS A NORMAL ESTROGEN ITSELF PLAYS A NORMAL
AND NECESSARY AS WELL AS AND NECESSARY AS WELL AS ABNORMAL AND DAMAGING ROLE IN ABNORMAL AND DAMAGING ROLE IN
THE IMMUNE SYSTEMTHE IMMUNE SYSTEM
4- THIS MEANS IDENTIFYING THE 4- THIS MEANS IDENTIFYING THE CELLULAR TARGETS FOR EFFECTS, CELLULAR TARGETS FOR EFFECTS, AND THEN WHAT GENE AND THEN WHAT GENE ALTERATIONS OCCUR TO CAUSE ALTERATIONS OCCUR TO CAUSE THOSE EFFECTSTHOSE EFFECTS