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. 

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ImmuQhrest Laboratories, Inc. 

'> ~W '`'~ ~~~r .- . 

1600 East Gude Drive FEB 619E 

FYr -©7~.~ - a~ ~ y- o ~ 9 a 12 

Rockville, Maryland 20850 0111ce of Vie !+a.rtan. .4' -

301- 762- 7193 1 :or 11esuciac;s ;Ana c ux : :: zj,. .. .. --... . . . . -- 

January 26, 1984 

Dr . John Moore 

Assistant Administrator 

Office of Pesticide and Toxic Substances TS788 

EPA 

401 M Street, S .W . 

Wasington, D,C . 20460 

Dear jack, 

p. 1 

":~~ I 

.020 

First, congratulations on your new position . I know NIEHS feels 

your loss but your contributions there, especially with NTP, will 

undoubtedly be appreciated for years . 

Enclosed is a draft manuscript describing the immunotoxicology 

studies on acetonitrile . As you recall, when I left the program 

I briefly discussed the possibility of some manuscript generation 

with you on the compound . 

I have forwarded a copy of the draft to Gene McConnAll for 

internal review . 

Jack, good luck to you at EPA and I look forward to seeing you at 

some point in the future . 

Sincerely, 

JiW' McCoy 

Director of Research 

.- 

M R1I13o1% 

000002 
LIMITED TOXICITY OF INHALED ACETONITRILE 

W ON THE IMMUNE SYSTEM OF MICE 1 

BowlJpsQ C .A .2, McCartney, D .M .2, Coate, W .B .3, and McCoy, 

J .L . 

1 . Supported in part by Contract No . NO1-CP-95655-02 with 

the National Toxicology Program . 

2 . ImmuQuest Laboratories, Inc ., 1600 East Gude Drive, 

Rockville, Maryland 20850 . 

3 . Hazleton Laboratories, America, Vienna, Virginia 22180 . 

4 . Formerly with the National Toxicology Program, National 

Institute of Environmental Health Sciences, Research 

Triangle Park, North Carolina . 
SUMMARY 

Hematological, cellular and humoral immune responses as well as 

clinical chemistry ;arameters of mice exposed to acetonitrile by 

inhalation were examined following observations that 800 ppm (10 

treatment days) of this agent induted thyttic atrophy in mice and rats . 

B6C3F1 mice received ten 6-hour exposures to 0, 100, 200, or 400 ppm 

of acetonitrile during ;, 'ourteen day period and were evaluated by a 

battery of immunological assays . Hematologic parar~,-~ters including 

hematocrit, hemoglobin, RBC and WBC were significantly depressed 

(P<0 .05) at increasing concentrations of the chemical . Lymphocyte 

proliferative responses to T-cell mi :ogens were depressed along with a 

somewhat depressee host susceptibility co rra^zpLaitable tumor cell, 

cha ::lenge, although neither of these responses were dose dependent . 

Studies of B-lymphocyte function indicated that levels of serum IgG 

were significantly depressed (P<0 .05) with increasing concentrations 

of acetonitrile while concentrations of IgA and IgM were unchanged . 

Other assays of B-cell fun,:tion, including Cunningham plaqiie assay 

responses to sheep RBC and B-cell proliferative responses to mitogens, 

were not significantly altered by acetonitrile ., Delayed hypersens : 

tj "" i`y :,Rsponses to keyhole limpet hemocyanin responses were not 

affected b ; acetonitrile treatment . In a concurrent 90-day prechronic 

study, liver, kidney and thyroid functions as evidenced by serum 

clinical chemistry evaluation appeared to remain intact in mice 

e4vn,eri:ving 65 6-hour exposures at up to 400 ppm inhaled acetonitrile . 

'F-u~~~*-e%r, tto significant lesions were observed by histopathological 

1!#,Wtio.n of Oese tissues or other body organs . Some treatment 

t;e`ILg*+ed- Astol,og-ical changes were observed in female animals receiving 

000004 
400 ppm acetonitrile consisting~of slight cytoplasmic vacuolization of 

ttepatocytes . The data demonstrate that with these doses, inhalation 

of acetonitrile in mice appears to induce limited toxic effects on 

lymphoid cell function and significant alterations in WBC and IgG 

levels in a dose response relationship generally not seen in other 

organs . 

INTRODUCTION 

Acetonitrile is a low molecular weight volatile agent that is 

used as a catalyst in chemical reactions and as a solvent in 

extractive distillation and crystalization o .' pharmaceutical and 

agricultural products including vitamins, bacter :cides, hormones, 

insecticides, plant growth regulators and fungicit:es (1) . The 

production volume of acetonitrile is estimated at 80 million pounds , 

per year in the United States and it is estimated that 23,000 

occupational workers may be exposed annually by dermal and inhalation 

routes (2) . 

A series of studies have demonstrated a variety of toxicological 

dysfunctions following injection or inhalation of high ,doses of 

acetonitrile . Marian et al . (3) observed that daily intramuscular 

injection of acetonitrile induced exopht.thalmos in rabbits, the 

severity of which was proportional to the degree of thyroid 

hyperplasia . Daily subcutaneous injections of acetonitrile into 

Mtf3tttile zacs have been reported to cause -, -rest in growth and 

T *d:la~C°g,!V;me,tit of thyroids, histological changes of the hypophysis, fatty 

e~c~,artioz of the liver and kidney and atrophy of the thymus (4) . 

Jc,OoO,ie:d to acetonitrile vapor by inhalation for ninety days 

,no'. ' Fac~~c effects : at 166 or 330 p:pm but did demonstrate 
p. 5 

ptlbondry inflammatory changes and minor abnormalities in the kidney 

htaid liver at a concentration of '665 ppm . Rats inhaling 2800 ppm of 

Adetonitrile vapors two hours per day for five days displayed 

difficulty in breathing, impaired renal function, paralysis of 

eittremities, severe loss of body weight and hemorrhages in the lung 

and brain (S) . Finally, Drew et al ., (7) observed increases in serum 

glutaroine oxalacetic transaminase (SGOT), glutamic pyruvic 

transaminase (SGPT) and ornithine carbamyl transferase activity in 

tats receiving a single four hour inhalation exposure to 3000 ppm of 

,$cetonitrile, indicating acute hepatic injury . 

. '.Metabolism and pharmacok~inetic investigations of acetonirritz~ are 

tomplex and have been somewhat unreliable . It was initially presumed 

that in vivo , acetonitrile degrades to formic acid and thiocyanate 

but the finding of serum hydrogen cyanide (HCN) by a number of 

Investigators suggests other pathways may be involved . Major toxic 

r±~~f:f:ec:t~ may result from the production of HCN by its action in 

. .,-

Th ~hibxting cytochrome oxidase and impairing cellular respiration . c 

r,va-t.iotz of cyanide after acetonitrile exposure 

ze:.ports of human poisonilig victims which 

is supported by 

showed high levels of 

kh ,ra7rious organs, blood and urine ~(9) and by studies in rats, 

:~!~tid, .~d .~gs showing urinary thiocyanates in all three speciez! 

~1:#,?'t-ion of acetonitrile and free cya. " ide in the blood of 

d=ags . :~'S) . After intraperi :oneal injection of or 

o-~~-tf~~e t~Q d,4eto-ritrile, free and conjugated hydrogen 

~OFt-,.a.tiittile, were fvu.nd in urine and various 

-ghost tancentrartlob-A of accumulated 

riive.r ;t: :5t?fit* Wnd lung iind f ree and 

3.n 

1 
Recent interest in specific toxic effects of chemical agents on 

the immune system (11-14) has arisen since it may be possible by using 

short-term assays to provide predictive toxicity information of 

chemicals using r-Apidly proliferating populations of lymphoid cells . 

Further, chemical damage to the immune system may result in hosts with 

depressed abilities to combat infectious disease or carcinogenic 

processes . The early work of Dessau (4) who demonstrated atrophy of 

thymus tissue coupled with the finding that acetonitrile accumulates 

in the spleen (6, 10) suggested to us that this chemical may express 

toxic action on normal lymphoid cell functions . The importance of 

possible acetonirrile~toxicity to lymphoid tissue is obvious 

considering its diverse use and the possible exposure of workers 

during its synthesis and use or possible exposure to others outside 

these operations . 

MATERIALS AND METHODS 

Mice 

Female B6C3C1 mice (6 weeks of age) wer_ received from the 

Frederick Cancer Research Facility, Frederick, Maryland, quarantined 

for 14 days prior to initiation of exposures . and held for an 

additional 6-8 weeks . Between chemical exposures, the animals were 

housed in stainless-steel cages suspended in 6-M3 inhalation chambers 

with food pellets (Ziegler NIH-07 diet) and tap water available ad 

libi tum . 

Chemical 

Acetonitrile was obtained from Vistron Corporation (Lot No . 

70413g1) . Gas chromatography analysis verified the purity of the 

compound was 99 .6% . The chemical was stored at 4oC until use . 

000007 
Chemical Exposure and Generation 

In preliminary studies we demonstrated that over a two week 

period, 10 repeated 6-hour inhalation exposures to acetonitrile were 

lethal to B6C3F1 mice at 3000 and 1500 ppm concentrations . A 

concentras.ion of 750 ppm resulted in death of 4/10 animals and caused 

significant body weight decreases in survi~ring mice . These 

concentrations, as well as 375 and 188 ppm exposures, induced thymic 

atrophy and scme hydropic degeneration of the liver .. In view of these 

toxicological data we further tested inhaled acetonitrile in 

immunotoxicity studies at 100, 200 and 400 ppm where no or only subtle 

toxicity might be expected in other organs . 

Test mice received 10 exposures of acetonitrile (100, 200 or 400 

ppm) six hours/day during a 14-day period in the immunology study . 

Other mice, evaluated by clinical chemistry and histopathology, were 

exposed to these same concentrations of compound six hours/day, 5 days 

per week, during a concurrent 90-day prechronic study . Acetonitrile 

vapor was generated into 6-M3 glass and stainless-st(.el inhalation 

chambers by sweeping the head space of glass flasks containing the 

liquid compound . The chambers were ventilated with filtered air at a 

flow of 1200L/cin . All flasks were set in water baths and held at 

approximately 26oC with immersion heaters . Control animals werc 

e.xposed to filtered air only . Acetonitrile vapor concentrations were 

ti7clifored every 30 minutes during 6-hour exposures using a Miran 801 

~T,wfft=rvd Vapor Analyzer calibration checked daily . An analytical 

.:'0;i~rgth of 11 .1 microns was used for all analyses . Animals wer- 

-v~bd twice daily for evidence of clinical signs of chemical 

000008 
Tumor .Susceptibility Assay 

Twenty control mice and 10 mice per test group were inoculated by 

the subcutaneous route with a TD10-20 dose of transplantable PYB6 

tumor cells two days following the last exposure to acetonitrile . 

Animals were individually housed and palpated for tumor formation 

twice weekly . Tumor incidence and latency period to tumor detection 

were determined up to sixty days post-ti .nor chall~nge . Tumors were 

confirmed after sacrifice on day 67 . 

Clinical Chemistry Evaluations 

Serum from each mouse was evaluated for liver, kidney and thyroid 

function including SGOT, SGPT, SDH, BUN and creatinine as measurPd by 

Centrifi Chem(R) autoanalyzer and determination of T3 and T5 hormone 

levels using radioimmunoassays with specific anti T3 and T5 sera . 

RESULTS 

Our initial prechronic studies of acPtonitrile demonstrated that 

mice exposed to a concentration of 750 ppm or higher (inhaled six 

hours daily for ten days) resulted in significant weight losses, 

atrophy of thymus, some hydropic degeneration of liver ti~,sue or 

death . Further, lower concentrations of compound (375 and 188 ppm) 

also induced thymus atrophy . In studying possible toxicological 

actions of inhaled acetonitrile on the immune system we chose 

concentrations of 400, 200 and 100 ppm administered ten times (six 

hours/day) during a fourteen day period . Since :.he immunology 

experiments were performed concurrently with a larger 90-day 

P.teehronic inhalaticn study of acetonitrile, clinical chemistry 

evalu.ations of liver, kidney, and thyroid function as well as necropsy 

wmd hi.stopathological evaluation findings from the 90 day study are 

6lso included in this report . . 0000 
A . Clinical Signs of Chemic P t Toxicity All chemically treated mice 

remained normal in phvsical appearance during the 14 day, as well 

as the 90 day, exposure periods of the compound . No significant 

weight losses were seen in any treatment group up to 90 days of 

chemical exposure . 

B . Necropsy and Histopathology No gross lesions were observed in 

animals in any treatment group during the 14 or 90 day exposure 

periods . Histologically, thymic atrophy was observed in the 400 

and 200 ppm treated mice during these time periods . Further, 

some sliqht vacuolization of hepatocytes and hydropic 

degeneration of lirer tissue was observed in animals exposed to 

400 ppm acetonitrile over 90 days . 

C . Clinical PatholoRY 

1 . Liver, Kidney and Thyroid Function Evaluation All animals 

exposed to acetonitrile, including the 400 ppm levels up to 90 

days, had SGOT, SGPT, SDH, BUN, creatinine and T3 and TS hormone 

levels within normal ranges and demonstrated no significa 

differences from control animal values in this study . 

2 . Hematology The results of hematology profiles of animals 

treated 10 times during a 14-day period are presented in Table 1 . 

Suppressions of hematocrit, hemoglobin, RBC and WBi, counts, as 

well as proportional numbers of neutrophils and monocytes were 

significant as analyzed by Dunnett's two-tailed t-test with the 

400 and 200 ppm concentrations . The suppiessions appeared 

compound-related as indicated by further analysis by linear 

regression . Hematocrits and WBC counts at the 100 ppm 

concentration were not significantly different from the untreated 

control group . 

000012 
D . Organ/Brain Weights Table 2 presents organ weight data (mean 

, 

values of 6 animals per treatment qroup) . The data are expressed 

as organ/brain weight ratios (the brain being one of the most 

constant organs in terms of weight loss-gain in chemical treated 

animals) . Although no statistically (as evaluated by the twotailed Dunnett's t-test) significant depression in spleen, liver, 

kidney, or thymus/brain weight ratios were observed, there was a 

trend for the thymus weights to be depressed at the 200 and 400 

pQm dose levels . Spleen/brain weight ratios tended to increase, 

~~~~cca~~a2~~2~.cc ~ra~~c4~-3Q.n ~-`~~LAbY7D2S DTi9 Df S~G~ ~g4aS. as~d ~gA 

1~SC Cc~w- tCCQ ¢z D3,~ 2.,S--

presented in Table 3 . Mead IgG levels in the sera of animals 

exposed to 100, 200, and 400 ppm of acetonitrile were decreased 

by 25 .8%, 33 .1%, and 48 .1% respectively, compared to untreated 

animals . The IgG levels were demonstrated (by Dunnett's twotailed t-test) to be significantly depressed as compared to , 

controls and a dose-response effect of the compound was indicated 

by linear regression analysis . 

Mean IgA concentrations in the sera of acetonitrile-treated 

animals were not significantly different from untreated animals . 

Dose effects in linear regression analysis weiE not observed 

between treated and untreated animals . 

Mean IgM concentrations were depressed by 11 .5% and 6 .0% at 

the 100 and 200 ppm levels and although these reductions were 

significant, linear regression analysis showed no dose related 

effects of acetonicrile . 

0000 13 

16 
., . 

F . Cunning ham Plaque Assay The antibody plaque forming response to 

SRBC is presented in Table 4 . Geometric and arithmetic means of 

PFC and PFC per 106 spleen cells were analyzed with Dunnett's ttpst . There was no significant difference between any of the 

chemical exposed and control mice . It is of interest, however, 

that the number of plaques formed per spleen was reduced by 

12 .9%, 22 .2% and 18 .5% for treatment groups at 100, 200, and 400 

ppm, respectively . These decreases in PFC per spleen is 

contrasted to the increa&ed mean spleen weights for animals in 

these respective groups (Table 2) and is somewhat suggestive of a 

depression in this immune function . 

G . Lymphocyte B_las togenesis Response Results of lymphocyte blastogenesis studies are presented in Table 5 . Dunnett's two-tailed 

t-test was performed on the mean (total) cpm for each mitogenic 

response assessed . There was no significant difference in 

lymphocyte responses to PHA, Con-A, or LPS between acetonitrileexposed and control animals . 

Analysis of the percentage change in net cpm revealed a 

depression in all groups examined at each exposure level . At 100 

ppm of acetonitrile, lymphoproliferative responses to PHA, Con-A, 

and LPS were depressed by 23 .3%, 26 .4% and 11 .4%, respectively, 

as compared to untreated animals . Lymphoproliferative responses 

to the three mitogens by spleen cells of mice exposed to 400 ppm 

were also depressed although to a lesser extent than observed in 

the 100 ppm group . Although there was a trend for some 

dap~,.Ossion in responsiveness to the three mitogens suggesting 

tame possible functional loss of T' and B lymphocyte activity, 

., : . 11 000014 
linear regression analysis showed no dose-related trends in these 

responses . 

H . Delaved Hy"rsensitivitv Responses Results of the delayed 

hypersensitivity response assay are presented in Table 6 . Mean 

response indicies calculated from counts per minute of KLH 

challenged ear versus saline challenged ear of six mice per group 

ranged from 1 .52 to 2 .19 .' Dunnett's two-tailed t-test analysis 

of these indices failed to reveal a significant difference 

between treated and control gro?!ps of mice . Further, doseresponse effects were not demonstrated using the TerpatraJoncfcheere test for trend . 

for saline inoculated ears (from six mice per 

from 1193 to 2107 while KLH inoculated ear mean cpm 

Mean cpm 

group) ranged 

I . 

for the acetonitrile treated and control 

to 4178 . 

Tumor Suscepti bilitv Susceptibility of 

significantly different in 

chemically treated animals 

independent t-test . There 

one-tailed exact test for proportional comparison revealed 

significant difference between treated and control groups . 

latency time to tumor detection and mean tumor volume were 

groups ranged from 2964 

acetonitrile-treated and 

untreated animals to PYB6 tumor challenge is presented in Table 

7 . Tumor incidence in untreated mice challenged with an LD10-20 

of tumor was 11% . Tumor incidence in animals exposed to 100 or 

400 ppm of acetonitrile was 44% and 33%, respectively . Fishers 

comparisons made 

as determined by 

no 

Mean 

n .) t 

between control and 

a two-sample 

was a trend, however, for the mean 

latency period to tumor detection to be somewhat deceased in the 

acetonitrile-treated mice . 000015 

11 
DISCUSSION 

Previous studies employing large doses of acetonitrile have 

demonstrated thyroid (3), liver (4, 5, 7), kidney (4, 5, 6), and lung 

(6) abnormalities, as well as thymus atrophy (4) in experimental 

animals receiving compound intramuscularly (3), subcutaneously (4) or 

by inhalation (5, 6, 7) . Pozzani et al ., (5) further reported that 

rats exposed seven hours per day for ninety days to 665 ppm 

acetonitrile by inhalation exhibited pulmonary inflammatory changes 

and minor changes in the kidney and liver . No toxic lesions were 

obsPrved in their animals exposed to 330 or 166 ppm acetonitrile . Our 

initial prechronic inhalation studies of exposures to acetoni.trile 

over a fourteen day period are consistent with the findings of Pozzani 

et al . (5) . We observed some hydropic degeneration in the liver in 

animals receiving 750 or greater ppm inhaled acetonitrile . 

Furthermore, we also observed thymic atrophy in mice as earliec 

reported for rabbits (3) at levels as low as 188 ppm of compoun~., 

suggesting a high degree of sensitivity of this organ to the chemical . 

As in Pozzani's earlier studies (5) we observed no other toxicity at 

the 375 and 188 ppm concentrations . 

In investigating possible systemic immunotoxicological actions of 

inhaled acetonitrile, we selected levels (400 ppm as the highest 

level) that presumably would not induce general toxicity to other 

major target organs in the animal, but levels that might result in 

toxicity to the immune system . 

We chose a battery c.f in vivo and in vitro assays of humoral and 

cellsrLar immunity to thoroughly evaluate possible acetonitrile 

toxicity on various immune compartments as previously described (13, 

I) 000016 
14) . These assays have been successfully used in invest+3ating 

suppressive immuno-modulation by such agents as polybrominated 

byphenyls (15), benzene (lb), diethylstilbestrol (14, 17) and heavy 

metals (18) . ' 

With the concentrations of acetonitrile used here, we were able 

to quickly demonstrate relatively specific toxicity on the immune 

aystem, with no other major target organ toxicity, even after the 90-

day exposure period, with the exception of some slight to moderate 

hydropic degeneratio,j of the liver . Short-term toxicity to lymphoid 

cells was manifested by statistically significant decreased total WBC 

,and neutrophils and-IgG levels . WBC counts and neutrophils were not 

significantly depressed in exposei groups at the end of the 90-day 

subchronic study, but were depressed after 10 exposures . In 

evaluations of thymus/brain weight ratios, Cunningham SRBC plaque, 

lymphocyto blastogenesis to T and B cell mitogens, and susceptibility 

to tumor challenges, although statisticaly significant changes were 

not observed, there were trends with each assay that suggested some 

immune suppression . It would appear that at the highest concentration . 

used (400 ppm) over a ten-day treatment period, toxic effects on some 

parameters measured (i .e ., WBC, neutrophils, and IgG Levels) vere 

evident, but that toxicity as measured by the other humoral and 

cellular immunity assays was marginal . Increased levels of 

acetonitrile might have magnified some of these effects . Such a 

treatment regimen, however, might have also caused a more pronounced 

general toxicity to the host, a situation we attempted to avoid . 

Even though there was a depre :sion in neutrophil (considered the 

first line of defense against microbial infections (19)) numbers anV 

000017 

.~.= 
proportions following the 10-day. series of acetonitrile exposures 

there was no evident increased infecLious diseases in the animals . 

This disease-free status included the 90 day treatment period of the 

concurrent subchrc.nic study . The statistically significant short-term 

depression of neutrophil numbers was obviously not sufficiently severe 

to cause a recognized increased susceptibility to intercurrent 

infections . 

It is of interest that only IgG levels were depressed ; a 

phenomenon not observed with IgM and IgA measures . One possible 

explanation is that plasma cells secreting IgG are more sensitive to 

toxic actions of aceLonitrile than are those cells that secrete IgM 

and IgA . Acetonitrile may either be directl-y-41-ytotoxic to the IgG 

secreting cells, disrupt their differentiation processes or may 

interfere with secretion of IgG . 

Delayed hypersensitivity reactions as measured by the ear 

swelling technique were,generally normal in acetonitrile treated mice . 

This finding coupled with tti- continual IPA of clinical signs of 

chemical toxicity and lack of any observable infections suggests that 

in vivo immune competence remained resonably intact in compoundtreated mice . 

Finally, 

found, due to 

suggests that 

although apparent life-threatening effects were not 

the limited immune toxicity of acetonitrile, this study 

by using short-term immunological measures of toxicity 

that a more generalized toxicity of chemicals 

would not be surprising since lymphoid tissue 

, 

may be predicted . This 

revresents a population 

of rapidly dividing cells that may be one of the f :.rst target 

populations to be damag "!d by many chemical or en :iro,:mental agents . 
Whether such a battery of immunological tests as employed in this 

study will allow predictior of more generalized chemical toxicity in 

other cases is a question calling for further application of thsis 

approach . 

REFERE NCES 

1 . Hawley, G .G ., The Condensed Chemical Dictionary . 8th Ed . Van 

Nostrand Reinhold Company-, New York, New York, 1971 . 

2 . NIOSH, Criteria for Recommended Standard Occupational Exposure to 

Acrylonitrile . DHEW Rpbl . 78-116 . U .S . Government Printing 

Office, Wash .ngton, D .C . 

3 . Marian, D ., Rosen, S .H . and Cipra, A ., Further Studies on the 

Exophthalmos in Rahbits Produced by Methyl Cyanide . Proc . Soc . 

E :.V . Biol . Med ., 30 : 649-651, 1933 . 

4 . Dessau, F ., Chronic Acetonitrile Poisoning in Rats . Acta Brevia 

Neerland . Physiol . Pharmacol . Microbiol ., 7 : 57-60, 1937 . 

5 . Pozzani, U .C ., Carpenter, C .P ., Palm, P .E ., Weil, C .S . and Nai, 

J .H ., An Investigation of che Mammalian Toxicity of Acetonitrile, 

J . Occup . Med ., 1 : 634-642, 1959 . 

6 . Haguenoer, J .M ., Dequidt, J . and Jacquemont, M .C ., Experimental 

Toxicology of Acetonitrile : Acute Toxicity by Inhalation, Europ . 

J . Tox . 8 : 102, 1975 . 

7 . Drew, R .T ., Patel, J .M . and Lin, F .N ., Changes in Serum Enzymes 

in Rats After Inhalation of Organic Solvents Singly and In 

Combination . Toxicol . Applied Pharmacol . 45 : 809-819, 1978 . 

8 . Williams, R .T ., Detoxication Mechanisms, 2nd Ed . John Wiley and 

Sons, Inc ., New York, 1959 . 

9 . Amdur, M .L ., Accidental Group Exposure to Acetonitrile . J . 

Occup . Med . 1 : 627, 1959 . 

10 . Haguenoer, J .M ., Dequidt, J . and Jacquemont, M .C ., Experimental 

Toxicology of Acetonitrile : Acute Intoxication by the 

Intraperitoneal Route, Eur . J . of Tox . 8 : 94, 1975 . 

11 . Moore, J .A ., The Immunotoxicology Phenomenon, Drug and Chem . 

Toxicol . 2 : 1, 1979 . 

12 . Lucier, G .W . and Hook, G .E .R ., Target Organ Toxicity : Immune 

System, Environ . Health Perspec:t . 43 : 1-135, 1982 . 

13 . Yos, J .G ., Immune Suppression as Related to Toxicology CRC Crit 

Rev . Toxicol . 5 : 67-101, 1977 . 

16 000019 
14 . Dean, J .H ., Luster, M .I ., Boorman, G .A . and Lauer, L .D ., 

Procedures Available to Examine the Immunotoxicity of Chemicals 

and Drugs, Pharm . Reviews, 34 : 137-148, 1982 . 

15 . Luster, M .I ., Faith, R .E . and Moore, J .A ., Effects of 

Polybrominated Biphenyls (PBB) on Immune Response in Rodents . 

Environ . Health Prospect . 23 : 227-232, 1978 . 

16 . Irons, R .D ., Heck, H ., Moore, B .J . and Murihead, K .A ., Effects of 

Short-Term Benzene Administration on Bone Marrow Cell Cycle 

Kinetics in the Rat, Toxicol . Appl . Pharmacol . 51 : 399-409, 1979 . 

17 . Boorman, G .A ., Luster, M .I ., Dean, J .H . and Wilson, R .E ., The 

Effect of Adult Exposue to Diethylstilbestrol in the Mouse on 

Macrophage Function . J . Reticuloendothel . Soc . 28 : 547-559, 

1980 . 

18 . Koller, L .D ., Immunotoxicology of Heavy Metals, Int . J . 

Immunopharmacol . 2 : 269-279, 1980 . 

19 . Gadebusch, H .H ., Phagocytes and Cellular Immunity, CRC Press, 

Bora, Florida, 3, 1979 . 
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. . 

4A. 0 

FJ 

N 

i 

; 

W 

01 

w 

+ 

.r 

V 

w 

~p. 

w 

v 

"sV 

w 

OD 

Js 

w 

w 

w ro o 

w w 

a o~ 

. . 

"J o 

W 

w 

4" 

rI 

1 

W 

w 

W 

w 

v 

w .t 

O 

1 

v 

w 

O 

N 

V 

w 

+ 

VI 

+J CD ..1 

00 w N 

. s " 

N ~ N 

. 

w w 

" 

Vl'' .Y 

f'. i V 

ao 

tn ~w b z w 

ro 

N 

" 1 V" 

a 

O 

ti 

ro 

n 

H 

H 

H QHl 

N 

H 

N 

H 

v 

z rt v 

rt 

0 to 

1~I 
Table 3 

EFFECT OF ACETONITRILE INHALATION ON THE CONCENTRATION 

OF CIRCULATING IgG, IqA, AND IcM IN B6C3F1 MICE1 

Acetonitrile Mean Immuno lobulin Concentration + SEM2 (8 Change) 

Treatment m 100m ($ Chance) 

(PPM) IgG - Id `4-- - ,- - - -- 

gM 

----- 

0 168 .2 + 10 .1 (-) 44 + 0 (-) 24 .3 + 0 .4 (-) 

100 124 .8 + 12 .9 45 .2 + 0 .8 21 .5 + 0 .3 (25 .8%-+) (2 .7%-+) (11 . 5T + ) 

200 112 .5 + 7 .5 47 .2 + 1 .0 22 .8 + 0 .4 

(13 .1$ i) (6 .78 { ) (6 .0$ i ) 

400 87 .2 + 7 .0 42 .2 + 0 .8 24 .8 + 0 .3 

(48 .1$ F ) (4 .1%-+) (2 .0%-+) 

1 . Numbers represent Mean + SEM Concentration of Immunoglobulin 

Levels from pools of 6 .mice . 

2 . Immunoglobulin Concentraticn expressed as ma/dl .