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Assessment of immunotropic effects of long-termexposure to radiofrequency and microwave
radiation.
S. Szmigielski, E. Sobiczewska, M.P. Dąbrowski, W. Stankiewicz,
Department of Microwave Safety, Military Institute of Hygiene and Epidemiology,
Warsaw, Poland.
Main functions and responsibilities of the immune system for homeostasis of the organism.
Type of the immuneresponse
Function of the immune system Immune cells andtransmitters engaged
IMMUNOTROPIC factors
Anti-infective, anti-neoplasticRESISTANCEIMMUNE
COMPETENCE
PROCREATIVE potency
Supervision ofMETABOLISM
Proper course ofREGENERATIVE PROCESSES
IMMUNO-GENICITY
Supervision of genetically-determined
QUALITY OF TISSUES
phagocytic cells
antigen-presen-ting cells (APC)
TCD4
effector cells
monokines
lymphokines
macrophageactivators;
activators oflymphopoiesis andlymphocytefunctions;
thymic extracts
Immunologic competence
Immunogenicity of tissues
Monocytes, Macrophages, Dendritic cells
PHAGOCYTOSIS
Presentation of antigens
MONOKINES
HLA I i II Class + conventional antigens
BONE MARROW
TCD8
TCD4
NK
B
Mf
THYMUS
INDUCTION of response
Immunoregulation Lymphokines
EFFECTOR Response RECOGNITION
Ig receptors, TCR
BONE MARROW
MICROWAVE RADIATION and the IMMUNE SYSTEM.
Authors Effects Exposure conditions Clinicalrelevance
Chou, Guy et al.,1985 Slightly lowered mitogenic response oflymphocytes to mitogens (PHA) invitro
Life-time exposure of rats,2450 MHz, pulsed,
SAR 0.15 – 0.4 W/kg??
Smialowicz, 1984 Increased mitogenic response oflymphocytes to mitogens (PHA) invitro
rats, 2450 MHz, continous andpulsed SAR 1 – 5 W/kg ??
Veyret et al., 1991 Enhancement or lowering of humoralresponse (antobody levels), dependingon modulation of carrier wave
mice, 9400 MHz, SAR 0.015W/kg ??
Stankiewicz et al.,2000
Different response of humanlymphocytes to exposures in vitro ,depending on pulse modulation of thewave
human lymphocytes, 1320MHz, continous and pulsed (5sec), 10 W/m2)SAR 1 W/kg ??
Several publications(1975 – 2005) fromthe Soviet Union andEast Europeancountries
Reports on fragmentaric and/orpoorly documented abnormalities insingle parameters of the immunesystem: shifts in blood picture; lowered serum Ig levels; weaker response of lymphocytes to
mitogens in microcultures.
Laboratory examinations ofrelatively small groups ofworkers exposed to RF/MWradiation, frequently withoutcomparison with valid controlgroups
No evidencefor existence of shifts inimmune statusof exposedworkers
Assessment of functional state of immunocompetent cells (PBMC) after exposure to low-level pulse-modulated microwave radiation.
Schedule of the study.
Isolation of PBMC
(1) Microculture 24 hrs 48 hrs 72 hrs
(2) Transfer of medium Harvestingbetween microcultures, of cultures, according to scheme counting of
radioactivity 1300 MHz, 5 sec pulse,
10 W/m2, SAR = 0.18 W/kg 1 hour 3H- TdR
Methods for assessment of functional state of Peripheral Blood Mononuclear Cells (PBMC).
1). Isolation of PBMC;2). Microcultures of PBMC;3). 24 hrs – transfer of medium,
according to scheme;4). 48 hrs – addition of 3H-TdR;5). 72 hrs – harvesting of cultures
(1)Spontaneous
(2)Spontaneous
100 l medium
100 l medium
Suppressive Activity of T cells(SAT)
SAT = 100 – {(6) : [½ (8) + (4)] }%
(3)PHA
(4)PHA
Lymphocyte – Monocytecooperation index (LM)
LM = (2) : (1)
(5)Con - A
(6)PHA
Normal values (Healthy men)
SAT > 30%LM 3 –6
IL-2 90 – 100%
100 l medium + cells
Saturation of IL-2 receptorsIL-2 = (5) : (8) x 100 %
(7)Con – A
(8)Con - A
100 l medium
Assessment of functional state of PBMC exposed to pulse-modulated 1300 MHz radar radiation(10 W/m2, 1 hr, SAR 0.018 W/g)
Sponataneous 3H-TdR
(103 DPM/105 cells)
PHA-induced3H-TdR
(103 DPM/105 cells)
Con-A – induced3H-TdR
(103 DPM/105 cells)
PHA/Con-A (P/C)ratio
Exposed Non-exp Exposed Non-exp Exposed Non-exp Exposed Non-expMean 1.16 1.86 83.24 69.22 37.95 35.19 2.15 2.06SD 0.41 0.75 11.60 9.21 2.47 9.19 0.36 0.65Significance of difference p < 0.05 NS NS NS
SAT index(Con-A-induced
suppresive activity of T lymphocytes)
Saturation of IL-2(Interleukin-2receptors on T
lymphocytes %)
LM index(lymphocyte- monocyte
cooperation index)
IFN - concentration in culture medium
(pg/ml)
Exposed Non-exp Exposed Non-exp Exposed Non-exp Exposed Non-expMean 22.73 29.71 91.13 96.67 17.15 4.83 510.5 630.4SD 9.25 3.95 9.16 5.74 5.96 1.18 118.0 92.4Significance of difference NS NS p < 0.01 p < 0.05
TNF concentration
(pg/ml)
IL-1 concentration
(pg/ml)
IL- 1 raconcentration
(pg/ml)
IL-10concentration
(pg/ml)Exposed Non-exp Exposed Non-exp Exposed Non-exp Exposed Non-exp
Mean 2421.1 1986.6 740.9 287.8 670.4 1312.5 631.5 311.3SD 478.9 985.8 129.2 131.6 256.9 691.5 193.9 122.7Significance of difference NS p < 0.01 p < 0.05 p < 0.01
RF/MW exposure levels of military personnel in Poland.
Type of equipmentPercent
of exposedpopulation
Frequen cyof EM
radiation
Highest fieldlevels during
shift
Permissible levels forworkers
(8 hr shift)
Dailyradiation
dose(W/m2 x h)
(1) (2) PL EC
RTC- stationary 2 % 1 - 20MHz
30 V/m 70V/m
77V/m
1.5
RTC- mobile 50 % 1 - 100MHz
100 V/m 70 V/m 77 V/m 5
Defence Radars 30 % 1 - 20GHz
50 W/m2 10W/m2
100W/m2
2
Fire Control Radars 15% 1 - 20GHz
50 W/m2 10W/m2
100W/m2
5
EM RepairWorkshops
3 % 1 MHz 20GHz
50-100W/m2
10W/m2
100W/m2
> 8
Assessment of immune status in personnel with multiyear occupationalexposure to pulse-modulated microwave (radar) radiation.
WorkersExposed Control
Number of subjects20 20
Age (mean SD) 39,3 8.6 37.9 9.6
Medical examination passed passedYears ofexposure(mean SD) 8.4 4.3 0Range of powerdensity duringshift (W/m2) 4 - 22 0
Mean dailyexposure (W/m2)
6.4 3.6 0
ExposureAssessment
Range of dailyexposure dosis(W x h/m2) 16 - 46 0
Basic evaluation of immune status
(general medical examination and routine laboratory tests)
Questionnairing (or interview) which includes frequency of infections and use of antibiotics,
occurance of allergic symptoms, anaemias, tissue distrophias, autoaggressive diseases, neoplastic
diseases and other deficiences of the immune system;
Blood picture with differential count of leukocytes;
Serum levels of immunoglobulins (IgA, IgG, IgM, IgE);
Delayed hypersensitivity to various antigens in skin tests;
Quantitative analysis of subpopulations of peripheral blood mononuclear cells (PBMC), including
T lymphocytes (TCD4 and TCD8), monocytes (CD14), B lymphocytes (CD19) and NK cells
(CD56).
Specialistic immunodiagnostic methodsallow to diagnose type of immune abnormalities and localize the abnormalities
on the scheme of organization and structure of the immune system,
Microculture of peripheral blood mononuclear cells (PBMC) with determination of:
Spontaneous transformation of lymphocytes in 72-hr culture;
Mitogenic response of lymphocytes to phytohaemagglutinin (PHA), concanavalin A (Con-A) and pokeweed
mitogen (PWM) or lipopolysaccharide (LPS);
Con-A-induced suppressor activity of T lymphocytes (SAT);
Lymphocyte-monocyte cooperative index (LM);
Saturation of interleukin-2 (IL-2) receptors on T lymphocytes;
Activity of monokines (IL-1, IL-1ra) and selected lymphokines (e.g. IL-2, IFN, IL-6, IL-8, IL-10) in culture
supernatants;
Flow cytometry of peripheral blood mononuclear cells (PBMC) with detailed fenotypic characterization
of immucompetent cells;
Analysis of cell cycle of stimulated lymphocytes in culture (optional);
Leukocytosis (109/l) Granulocytes (109/l) Lymphocytes (109/l) Ratio CD4/CD8Exposed Non-exp Exposed Non-exp Exposed Non-exp Exposed Non-exp
Number of subjects (N) 20 20 20 20 20 20 20 20Mean 8.32 7.96 5.23 5.01 3.08 2.96 2.58 2.46SD 1.47 1.56 1.12 0.96 0.82 1.22 0.32 0.24Significance of difference NS NS NS NSAnalysis ofindividualdifferences
Increased*UnchangedDecreased*
1172
1163
0182
0191
1163
2153
2144
3134
Significance of difference NS NS NS NS
Sponataneous 3H-TdR
(103 DPM/105 cells)
PHA-induced3H-TdR
(103 DPM/105 cells)
Con-A – induced3H-TdR
(103 DPM/105 cells)
PHA/Con-A (P/C)ratio
Exposed Non-exp Exposed Non-exp Exposed Non-exp Exposed Non-expNumber of subjects (N) 20 20 20 20 20 20 20 20Mean 1.15 1.86 81.27 69.22 38.21 35.19 2.02 2.06SD 0.31 0.75 11.60 9.21 7.21 9.19 0.56 0.65Significance of difference p < 0.05 NS NS NSAnalysis ofindividualdifferences
Increased*UnchangedDecreased*
1136
2162
3161
1145
5114
4106
4124
3116
Significance of difference p < 0.05 NS NS NS
* difference > 2SD from mean value in non-exposed
SAT index(Con-A-induced
suppresive activity of T lymphocytes)
Saturation of IL-2(Interleukin-2receptors on T
lymphocytes %)
LM index(lymphocyte- monocyte
cooperation index)
IFN - concentration
in culture medium(pg/ml)
Exposed Non-exp Exposed Non-exp Exposed Non-exp Exposed Non-expNumber of subjects (N) 20 20 20 20 20 20 20 20Mean 24.26 29.71 90.28 96.67 16.47 4.83 462.5 630.4SD 5.37 3.95 8.32 5.74 6.21 1.18 106.4 92.4Significance of difference NS NS p < 0.01 p < 0.05Analysis ofindividualdifferences
Increased*UnchangedDecreased*
0164
1172
0164
0191
1451
1181
1145
1190
Significance of difference NS NS p < 0.01 p < 0.05
TNF concentration
(pg/ml)
IL-1 concentration
(pg/ml)
IL- 1 raconcentration
(pg/ml)
IL-10concentration
(pg/ml)
Exposed Non-exp Exposed Non-exp Exposed Non-exp Exposed Non-expNumber of subjects (N) 20 20 20 20 20 20 20 20Mean 22.85.1 1986.6 831.6 287.8 632.4 1312.5 748.3 311.3SD 637.3 985.8 165.2 131.6 273.1 691.5 148.6 122.7Significance of difference NS p < 0.01 p < 0.05 p < 0.01Analysis ofindividualdifferences
Increased*UnchangedDecreased*
6122
2144
1631
1163
1910
2171
1451
2171
Significance of difference NS p < 0.01 p < 0.05 p < 0.01
* difference > 2 SD from mean value in non-exposed
Conclusions.
1. Measurable shifts in functional state of human immunocompetent cells wasnoted after in vitro exposure to pulse-modulated 1300 MHz MWs at 10 W/m2
(SAR = 0. 18 W/g);
2. Slightly lower spontaneous mitogenic response of lymphocytes (3T-dRincorporation), accompanied by stimulation of macrophage activity werenoted in microcultures;
3. Assessment of immune status in workers exposed occupationally to pulsemodulated MW (radar) radiation at power densities of 4- 22 W/m2 (daily dosis3 – 10 W x h/m2) during 5 – 15 years revealed symptoms of stimulation ofmacrophage functions (increeased LM cooperation index, increased IL-1 andlowered IL-1ra serum levels, high IL-10 level);
4. Immunotropic effects of MWs were noted at exposure levelsconsiderably below the limits recommended by ICNIRP.