Etiologic Agents and Pathogenic Mechanisms in the Acute Byssinotic

11 E t i o l o g i c A g e n t s and P a t h o g e n i c M e c h a n i s m s in

Downloaded by UNIV OF CALIFORNIA SAN DIEGO on January 24, 2017 | http://pubs.acs.org Publication Date: July 1, 1982 | doi: 10.1021/bk-1982-0189.ch011

the A c u t e B y s s i n o t i c R e a c t i o n STERLING K. AINSWORTH and PATRICIA A. PILIA Medical University of South Carolina, Department of Pathology (Immunopathology), Charleston, SC 29425 Inhalation of cotton dust has long been known to cause a pulmonary disease termed byssinosis. This chapter is a comprehensive review of etiologic agents and pathogenic mechanisms of the acute byssinotic reaction. The chemical properties of cotton mill dust are described, and the various active agents proposed in the literature to produce the acute byssinotic reaction reviewed, together with their pathogenic mechanisms. This includes studies of immunoglobulin levels in animals and man, complement activation by both immunologic and non-immunologic means and investigations of possible hypersensitivity to cotton dust. Also reviewed and discussed are the active agents found in extracts of cotton m i l l dust and cotton bract which might be responsible for histamine release, chemotaxis and bronchoconstriction. The p o p u l a t i o n of workers i n the c o t t o n i n d u s t r y a t r i s k of c o n t r a c t i n g b y s s i n o s i s i n c l u d e s workers i n areas f o r c a r d i n g , s p i n n i n g and other dust producing o p e r a t i o n s . The c a r d i n a l acute symptoms of chest t i g h t n e s s , shortness of b r e a t h , coughing and wheezing were described as e a r l y as the mid 1 8 0 0 s by Mareska and Heymann (1_). More r e c e n t l y the f i n d i n g s of a r e d u c t i o n i n FEV^ (2)» peripheral leukocytosis, slight elevations i n body temperature and leukocyte recruitment t o a i r passages (_3) have been added t o the l i s t of r e c o g n i z a b l e symptoms. These acute symptoms t y p i c a l l y develop a f t e r exposure t o the dust, and are seen p a r t i c u l a r l y on Mondays f o l l o w i n g a weekend away from the work environment; i n a d d i t i o n , they are r e v e r s i b l e and g r a d u a l l y subside over a p e r i o d of s e v e r a l days o f continued exposure o r upon l e a v i n g the work area. Acute symptoms d i f f e r p r i m a r i l y i n degree and d u r a t i o n from c h r o n i c symptoms, which appear a f t e r s e v e r a l years of o c c u p a t i o n a l exposure. The more severe chronic symptoms are i r r e v e r s i b l e and p e r s i s t even i n a d u s t - f r e e atmosphere. f

©

0097-6156/82/0189-0163$06.00/0 1982 American Chemical Society

Montalvo; Cotton Dust ACS Symposium Series; American Chemical Society: Washington, DC, 1982.


164

COTTON

DUST

In g e n e r a l , workers w i t h i r r e v e r s i b l e c h r o n i c b y s s i n o s i s or c h r o n i c o b s t r u c t i v e lung d i s e a s e smoke and have been employed i n the i n d u s t r y f o r 10 or more years. We can assume t h a t repeated, acute r e a c t i o n s i n the lungs have a cumulative damaging e f f e c t . At t h i s time, however, i n f o r m a t i o n l i n k i n g the acute r e a c t i o n and chronic b y s s i n o s i s i s l a c k i n g , as i s a thorough understanding of the lung pathology, c a u s a t i v e agent(s) and pathogenesis. Laboratory exposure t o dust clouds and dust e x t r a c t a e r o s o l s r e s u l t s i n chest t i g h t n e s s , dyspnea and decreases i n both e x p i r a t o r y flow and dynamic lung compliance ( 4 ) . Bouhuys et a l . showed t h a t a e r o s o l s of c o t t o n dust e x t r a c t i n h a l e d by man produced r e v e r s i b l e small-airway o b s t r u c t i o n w i t h i n 10 minutes (5^); the same e f f e c t s were noted i n cardroom workers exposed t o dust on Mondays, i . e . decreased maximum e x p i r a t o r y f l o w r a t e s and increased airway r e s i s t a n c e ( 6 ) . These changes are compatible w i t h narrowing of s m a l l airways as the p r i n c i p a l e f f e c t of acute dust exposure. The l a r g e m a j o r i t y of people a t r i s k are a f f e c t e d when t o x i c dust c o n c e n t r a t i o n s are h i g h . Healthy people exposed f o r the f i r s t time to dust or dust e x t r a c t respond as s e v e r e l y as long term workers ( 5 ) . Experiments have a l s o shown t h a t f o l l o w i n g a s i n g l e exposure to c o t t o n dust or a e r o s o l i n h a l a t i o n of an aqueous dust e x t r a c t , exposure repeated 24 hours l a t e r has no e f f e c t These f i n d i n g s i n d i c a t e t h a t the e t i o l o g i c agent(s) i s water s o l u b l e , and that prior immunologic sensitization i s not necessary. I n d i v i d u a l d i f f e r e n c e s are n o t a b l e ; f o r i n s t a n c e , i n dusty workrooms where r e a c t i v e workers experience chest t i g h t n e s s , marked t a c h y p h y l a x i s and acute lung changes, some workers show no symptoms. This suggests a genetic p r e d i s p o s i t i o n to e i t h e r t o x i c , pharmacologic or immunologic adverse mechanisms. W i t h i n 30 minutes of t h e i r a d m i n i s t r a t i o n , 8 -adrenergic drugs o f t e n r e v e r s e most of the f u n c t i o n a l d e f i c i t i n Monday morning b y s s i n o t i c s . As there i s no mucous s e c r e t i o n , airway smooth muscle c o n t r a c t i o n i s considered the primary response. Exposure of man t o histamine a e r o s o l s produces pulmonary f u n c t i o n changes s i m i l a r to those seen a f t e r exposure to dust e x t r a c t . However, exposure to histamine a e r o s o l i n v a r i a b l y initiates c o n s t r i c t i o n of smooth muscle more r a p i d l y than exposure t o c o t t o n dust ( < 15 minutes), and d i s s i p a t e s w i t h i n minutes, w h i l e the acute e f f e c t s of i n h a l a t i o n of c o t t o n dust and dust e x t r a c t s l a s t s f o r hours. The s l o w l y developing and prolonged e f f e c t s of dust and e x t r a c t s suggest that mediators other than histamine are i n v o l v e d . Small airway c o n s t r i c t i o n and r e c r u i t m e n t of leukocytes on pulmonary surfaces are prominent, documented responses t o the i n h a l a t i o n of c o t t o n dust. C u r r e n t l y , one or both of these e f f e c t s are g e n e r a l l y a s c r i b e d t o endotoxin (8-10), to antigen-antibody r e a c t i o n s ( 1 1 ) , to l a c i n i l e n e C-7 methyl ether (12^, 13), t o a low molecular weight (~1000 d a l t o n s ) , n e u t r a l , h i g h l y water s o l u b l e substance t h a t i s s t a b l e i n b o i l i n g water and found i n c o t t o n b r a c t s (14), to chemotaxins present i n c o t t o n m i l l dust e x t r a c t s (15, 16) or to histamine r e l e a s i n g substances (17).



11.

AINSWORTH AND

PILIA

Acute Byssinotic

Reaction

165

Work i n t h i s l a b o r a t o r y has been d i r e c t e d toward a s s e s s i n g the c a u s a t i v e agent(s) and pathogenic mechanisms i n the acute b y s s i n o t i c r e a c t i o n . Our i n v e s t i g a t i o n s of t o x i c , immunologic and pharmacologic mechanisms have l e d to the development of s e v e r a l bioassay techniques germane to the eventual understanding of t h i s disease process. These bioassays have been important in a s c e r t a i n i n g 1) histamine r e l e a s i n g substances, 2) smooth muscle b r o n c h o c o n s t r i c t o r s , 3) chemotaxins, 4) endotoxin and 5) complement a c t i v a t o r s . In p a r t i c u l a r , we have concentrated our e f f o r t s on three observed responses of the human lung f o l l o w i n g the i n h a l a t i o n of c o t t o n dust, namely lung smooth muscle c o n s t r i c t i o n , leukocyte recruitment i n t o airways (3) and the reputed histamine r e l e a s e (18). This chapter w i l l review our f i n d i n g s and the p e r t i n e n t f i n d i n g s of others regarding the e t i o l o g y and pathogenesis of b y s s i n o s i s , the use of e x t r a c t s of c o t t o n m i l l dust and c o t t o n b r a c t and the importance of bioassays f o r understanding the disease process. Chemical P r o p e r t i e s of Cotton Mill Dust Studies i n c o t t o n t e x t i l e m i l l s have shown that the i n c i d e n c e of b y s s i n o s i s can be c o r r e l a t e d w i t h the average c o n c e n t r a t i o n of f i n e c o t t o n dust (2, _1£, 20) and w i t h the number of years of o c c u p a t i o n a l exposure (19-2IT. The c a u s a t i v e agent i n b y s s i n o s i s has been shown to be a water e x t r a c t a b l e , f i l t e r a b l e , n o n v o l a t i l e agent (40°C) that can be r e t a i n e d on d i a l y s i s (22.). The a c t i v e byssinogenic agent appears to be present c h i e f l y i n the c o t t o n b r a c t , the l e a f - l i k e s t r u c t u r e surrounding the c o t t o n b o l l ( a l s o a prominent component of c o t t o n m i l l dust ( 2 0 ) ) , because 1) water s o l u b l e e x t r a c t s cause an FEV. drop (T8, 23), and 2) the byssinogenic agent can be p a r t i a l l y d e a c t i v a t e d by steam. Wakelyn et a l . (24) reviewed the complex chemical composition of c o t t o n m i l l dust (CMD) found i n v a r i o u s processing operations. S u b s t a n t i a l chemical v a r i a t i o n e x i s t s i n CMD c o l l e c t e d from d i f f e r e n t m i l l s and d i f f e r e n t processing rooms, and even from d i f f e r e n t areas w i t h i n the same processing room. Dust composition v a r i e s i n the p r o p o r t i o n of a l l p l a n t p a r t s , i . e . , l e a f , b r a c t , capsule and l i n t , as w e l l as of f i b e r , f u n g i , b a c t e r i a , i n o r g a n i c m a t e r i a l s and m a t e r i a l from other contaminating vegetation. Attempts to l i n k spores of b a c t e r i a or f u n g i to the pathogens of b y s s i n o s i s have not been s u c c e s s f u l (2j>, 26), although these m i c r o b i o l o g i c a l contaminants cannot be excluded as p o t e n t i a l sources of water s o l u b l e substances capable of c o n t r i b u t i n g to the acute b y s s i n o t i c response. Our s t u d i e s have shown that c o t t o n dust e x t r a c t s c o n t a i n d e o x y r i b o n u c l e i c a c i d (DNA) and r i b o n u c l e i c a c i d (RNA) (27). DNA i s present i n v a r y i n g concentrations i n cardroom c o t t o n dust (28) and emanates from b a c t e r i a , f u n g i , protozoa and p l a n t c e l l s . DNA might be important i n the pathogenesis of b y s s i n o s i s , as minute amounts can cause complement conversion.


COTTON

166

DUST


Immunological Studies Immunoglobulin Studies i n Man. Previous s t u d i e s of the e t i o l o g y of b y s s i n o s i s have proposed an immunological pathogenic mechanism. Massoud and Taylor (11) reported higher t i t e r s of serum a n t i b o d i e s i n b y s s i n o t i c c o t t o n m i l l workers than i n nonb y s s i n o t i c workers. They suggested that higher t i t e r s of antibody combine w i t h a c o t t o n dust a n t i g e n , and that symptoms are e i t h e r produced d i r e c t l y by antigen-antibody complexes or i n d i r e c t l y by r e l e a s e of pharmacological substances. Recent s t u d i e s by Noweir (29) showed e l e v a t e d IgG concentrat i o n s i n f l a x workers, whether f r e e of r e s p i r a t o r y symptoms, b y s s i n o t i c or emphysematous. IgM and IgD were a l s o s i g n i f i c a n t l y higher i n some groups of b y s s i n o t i c s . No changes were noted f o r IgE and IgA. S i m i l a r r e s u l t s were seen i n c o t t o n workers. No s i g n i f i c a n t d i f f e r e n c e i n c o n c e n t r a t i o n s were seen between sera samples drawn p r e - s h i f t or 2 hours post-dust exposure on Monday or Tuesday. IgG v a l u e s proved t o be h i g h e s t i n m i l l workers employed l e s s than 10 years, and showed no r e l a t i o n s h i p t o the l e v e l of dust exposure. Furthermore, IgG l e v e l s decreased i n workers exposed f o r g r e a t e r than 10 years; w i t h exposure g r e a t e r than 20 years, IgG l e v e l s returned to normal. Noweir suggests t h a t p r e c i p i t a t i n g a n t i b o d i e s i n grade I I b y s s i n o t i c s d e t e r i o r a t e due to the advanced stage of the d i s e a s e , but t h i s i s unconvincing as an e x p l a n a t i o n because a n t i b o d i e s have a short h a l f l i f e ( d a y s ) , are c o n s t a n t l y r e p l e n i s h e d by plasma c e l l s and show e s s e n t i a l l y no a l t e r e d s t r u c ture i n otherwise h e a l t h y i n d i v i d u a l s . Our s t u d i e s of serum l e v e l s of IgG, IgM, IgA, IgE and complement (30) drawn from b y s s i n o t i c and n o n b y s s i n o t i c m i l l workers a t d i f f e r e n t times during the work week (Monday a.m. preshift, 4 hours post-exposure, and F r i d a y p.m.) d i d not d e v i a t e from normal ranges (see Table 1 ) . This i s i n agreement w i t h r e s u l t s obtained by Edwards and Jones ( 3 1 ) , which do not suggest an immunological mechanism (Type I or I I I ) f o r the pathogenesis of b y s s i n o s i s . Our r e s u l t s a l s o agree w i t h Noweir*s f i n d i n g s of no d i f f e r e n c e s i n the immunoglobulin c o n c e n t r a t i o n s between samples obtained on the f i r s t p r e - s h i f t a f t e r the weekend, two hours post exposure or on the second day of the work week w i t h i n each worker group ( c o n t r o l , exposedsymptom free, byssinotic grades I and I I , and emphysematous). In a d d i t i o n , Noweir's r e s u l t s showed no c o r r e l a t i o n between immunoglobulins and dust exposure. Noweir suggested that high l e v e l s of IgG i n d i c a t e the presence of p r e c i p i t a t i n g a n t i b o d i e s i n the sera of workers exposed to c o t t o n dust (29). However, these suggestions and the i n f e r e n c e that a Type I I I r e a c t i o n i s r e s p o n s i b l e are h i g h l y u n l i k e l y , because both our data and Noweir s show no weekly change. Moreover, our r e s u l t s demonstrate no s i g n i f i c a n t serum complement change. This i s important f o r , i f a Type I I I antigen-antibody r e a c t i o n were the pathogenic mechanism, one would expect the following: a) antigens or haptens would e l i c i t h i g h t i t e r s of f


11.

AINSWORTH A N D PiLiA

Acute Byssinotic

Reaction

167

TABLE I Mean Immunoglobulin Concentrations i n Sera o f B y s s i n o t i c and Non-Byssinotic Cotton M i l l Workers At D i f f e r e n t Time I n t e r v a l s a

MAM Downloaded by UNIV OF CALIFORNIA SAN DIEGO on January 24, 2017 | http://pubs.acs.org Publication Date: July 1, 1982 | doi: 10.1021/bk-1982-0189.ch011

Subjects

Age(yrs) Number

MPM

FPM

IgG ( m g / d l )

b

Normal White Males

20-29 30-39 40-49

4 4 3

1223 1223 890

1119 1295 817

1020 1072 647

B y s s i n o t i c White Males

40-49 50

3 11

1242 1137

1130 1133

1323 1057

IgA ( m g / d l )

b

Normal White Males

20-29 30-39 40-49

4 4 3

276 270 265

229 229 218

228 218 186


40-49 50

3 11

307 289

227 288

292 272

IgM

(mg/dl)

b

Normal White Males

20-29 30-39 40-49

4 4 3

167 64 139

155 65 143

147 66 134


40-49 50

3 11

65 98

66 91

68 75

a

MAM, Monday A.M., MPM, Monday P.M., FPM, F r i d a y P.M.

Immunoglobulin l e v e l s i n a l l c a t e g o r i e s f a l l w i t h i n normal ranges (IgG, 616-1543; IgA, 71-417; IgM, 57-343). There are no s t a t i s t i c a l d i f f e r e n c e s (< 0.05) between b y s s i n o t i c and n o n - b y s s i n o t i c subjects o r between times samples were taken.


COTTON

168

DUST


s p e c i f i c a n t i b o d i e s ; b) f o l l o w i n g s p e c i f i c antibody production, a p r e c i p i t o u s drop i n serum complement should occur; c) sera from c o n t r o l s and n o n - b y s s i n o t i c s should not r e a c t ; and, d) c l i n i c a l r a d i o l o g i c and h i s t o l o g i c f i n d i n g s should c o r r e l a t e . None of these c o n d i t i o n s are met, however, and i t i s t h e r e f o r e h i g h l y unlikely that immune complexes (antigen-antibody-complement) m i t i g a t e the b y s s i n o t i c r e a c t i o n . In V i t r o and Animal Immunoglobulin Studies. When i n j e c t e d i n t r a m u s c u l a r l y , e x t r a c t s of c o t t o n are immunogenic i n r a b b i t s , and show both true and pseudoimmune p r e c i p i t a t i n g r e a c t i o n s (32, 33) . However, i n r a b b i t s exposed to c o t t o n dust by i n h a l a t i o n , as i n man, no p r e c i p i t a t i n g a n t i b o d i e s are detected (34). Taylor £t a l . (35) i s o l a t e d a condensed polyphenol which they considered to be a c o t t o n a n t i g e n . P r e c i p i t i n and passive agglut i n a t i o n t e s t s were used to show s i g n i f i c a n t d i f f e r e n c e s i n mean (serum) t i t e r s (not s p e c i f i c immunoglobulins) between cardroom workers and c o n t r o l s and between b y s s i n o t i c and n o n - b y s s i n o t i c cardroom workers. However, t h i s study d i d not assess the c o n t r i b u t i o n of each antibody c l a s s , and no attempt was made to e x p l a i n why high antibody t i t e r s were seen i n normal ( c o n t r o l ) subjects never exposed to c o t t o n . The authors d i d note the p o s s i b i l i t y that a l l i n d i v i d u a l sera would r e a c t w i t h the condensed tannin " a n t i g e n . " Edwards and Jones (31) i d e n t i f i e d the condensed tannin e x t r a c t e d from c o t t o n p l a n t b r a c t s as a t a n n i n - l i k e polymer of 5, 7, 3 , 4 t e t r a h y d r o x y f l a v e n 3-4 d i o l (THF). They demonstrated n o n s p e c i f i c p r e c i p i t a t i o n w i t h IgG, IgM, IgA, f i v e myeloma IgG's and p o s i t i v e g e l d i f f u s i o n r e a c t i o n s w i t h heavy and l i g h t c h a i n s , Fab and Fc pieces of IgG. Nevertheless, they r e f u t e d t h i s r e a c t i o n as a t r u e antigen-antibody r e a c t i o n , and subsequently suggested that b y s s i n o s i s was not an immune complex mediated pulmonary disease. Sera of c o t t o n m i l l workers have been found to y i e l d p o s i t i v e immune p r e c i p i t a t i o n r e a c t i o n s w i t h antigens of c o t t o n dust, p l a n t d e b r i s and m i c r o b i a l contamination i n dust, i n p a r t i c u l a r c o t t o n dust, c a r p e l and stem antigens (29). The h i g h e s t p o s i t i v e p r e c i p i t a t i o n occurred w i t h grade I I b y s s i n o t i c s , w h i l e a diminished response occurred w i t h grade I I I b y s s i n o t i c s e r a , suggesting t h a t a n t i b o d i e s might d e t e r i o r a t e during the course of disease. P r e c i p i t a t i n g a n t i b o d i e s to c o t t o n antigens were a l s o found i n nonb y s s i n o t i c s , w i t h the c a r p e l e x t r a c t e x h i b i t i n g the g r e a t e s t r e a c t i o n w i t h a l l sera t e s t e d . I t was suggested t h a t b r a c t and l e a f p l a y no r o l e i n antibody i n d u c t i o n , and p o s t u l a t e d t h a t " a n t i g e n s " present i n c a r p e l may be the c a u s a t i v e agent(s) i n producing the b y s s i n o t i c response. In 1971, Edwards and Jones showed t h a t n o n - s p e c i f i c nonimmunological r e a c t i o n s occurred between p l a n t e x t r a c t s and normal human sera (31). Noweir (29) repeated s t u d i e s i n b y s s i n o t i c s s i m i l a r to ones performed by Massoud and Taylor (11) and Taylor e_t a l . (35), and demonstrated s i m i l a r f i n d i n g s , i . e . normal, b y s s i f

f



11.

AINSWORTH A N D PILIA

Acute Byssinotic

Reaction

169

n o t i c and n o n - b y s s i n o t i c sera p r e c i p i t a t e d w i t h substances from c o t t o n dust, c a r p e l and stem antigens. Again, however, no attempts were made to a s s i g n the p r e c i p i t a t i n g sera to s p e c i f i c immunoglob u l i n s , or to determine complement l e v e l s and s p e c i f i c i t y of the r e a c t i o n s as recorded by Edwards and Jones (31) and Kutz e_t a l . (33). Moreover, n e i t h e r Kutz e_t a l . (33) nor Ainsworth £t a l . (30) could demonstrate t r u e p r e c i p i t a t i n g a n t i b o d i e s a g a i n s t c o t t o n p l a n t and dust antigens i n c o t t o n workers and b y s s i n o t i c s . Kutz f u r t h e r examined e x t r a c t s of stems and cardroom c o t t o n dust t o p r e c i p i t a t e c e r t a i n serum p r o t e i n s i n c o t t o n m i l l workers. A l l c o n t r o l worker and r a b b i t s e r a t e s t e d p r e c i p i t a t e d a g a i n s t the c o t t o n "antigens" and t e a l e a f " a n t i g e n . " Plant polyphenolic tannins were demonstrated t o be the l i p o p r o t e i n and gamma-globulin p r e c i p i t a t i n g substances (33). Thus our s t u d i e s and others show that f i n d i n g s are not c o n s i s t e n t w i t h an immunological pathogenesis of b y s s i n o s i s . Some f i n d i n g s remain s u s p i c i o u s , however, and need f u r t h e r study, e s p e c i a l l y i n the area of atopy. H y p e r s e n s i t i v i t y to Cotton Dust. An a l l e r g i c pathogenesis f o r b y s s i n o s i s was proposed by Bouhuys e_t a l . ( 6 ) . While the acute b y s s i n o t i c r e a c t i o n has few s i m i l a r i t i e s to asthma, a short p e r i o d of exposure f o l l o w e d by c o n s t r i c t i o n of lung smooth muscle, drop i n FEV^, chest t i g h t n e s s and symptoms that are p a r t i a l l y a b o l i s h e d by a n t i h i s t a m i n e s are experienced by both asthmatics and b y s s i n o t i c s . However, m i l l workers who have a proven a l l e r g y to c o t t o n dust by h i s t o r y and symptoms demonstrate an acute r e a c t i o n w i t h i n minutes of exposure, while b y s s i n o t i c c o t t o n m i l l workers u s u a l l y r e q u i r e four to s i x hours to demonstrate a much m i l d e r r e a c t i o n . This d i f f e r e n c e might be explained by the type of mediator r e l e a s e d , i . e . slower r e a c t i n g mediators i n the case of b y s s i n o s i s . I f an a l l e r g i c ( a t o p i c ) mechanism were i n v o l v e d i n the acute b y s s i n o t i c r e a c t i o n , serum IgE values would be e l e v a t e d . Our r e s u l t s w i t h sera from b y s s i n o t i c s and c o n t r o l s u b j e c t s ( n o n b y s s i n o t i c c o t t o n m i l l workers) showed no s t a t i s t i c a l d i f f e r e n c e between mean values of serum IgE l e v e l s f o r the two groups and three d i f f e r e n t time i n t e r v a l s (Monday a.m.-pre-shift, 4 hours post exposure and F r i d a y p.m.) during the work week (Table I I ) . S i m i l a r s t u d i e s by Noweir a l s o f a i l e d to demonstrate a s i g n i f i c a n t d i f f e r e n c e i n IgE concent r a t i o n (29). F u r t h e r assessment of atopy i n the pathogenesis of the acute byssinotic reaction necessitates purification of well c h a r a c t e r i z e d and standardized c o t t o n dust antigens, and the development of an a l l e r g y s p e c i f i c t e s t (RAST) to measure serum IgE a n t i b o d i e s to c o t t o n dust a l l e r g e n s . Such developments would a l l o w c o r r e l a t i o n of s k i n t e s t s , c l i n i c a l h i s t o r y , dust exposure, FEV^ and serum IgE l e v e l w i t h s p e c i f i c IgE t o c o t t o n dust allergens. P a s s i v e cutaneous anaphylaxis s t u d i e s of b y s s i n o t i c and nonb y s s i n o t i c sera i n our l a b o r a t o r y and s i m i l a r s t u d i e s i n r a b b i t s


170

COTTON


exposed t o i n h a l a t i o n of c o t t o n dust (34) u n i f o r m l y negative r e s u l t s . Thus, t o date, the e l i m i n a t e atopy from c o n s i d e r a t i o n ; however, the a l l e r g e n t e s t s p e c i f i c t o c o t t o n dust i s c r u c i a l f i n a l and d e f i n i t i v e answer.

DUST

have demonstrated r e s u l t s obtained development of an to o b t a i n i n g the

TABLE I I Mean IgE Concentration i n Sera of B y s s i n o t i c and Non-Byssinotic Cotton M i l l Workers a t D i f f e r e n t Time I n t e r v a l s Subjects Number IgE (U/ml) 3

Non-Byssinotics Byssinotics a

MAM, Monday A.M.;

29 28

MAM

MPM

FPM

82 81

82 76

78 70

MPM, Monday P.M.; FPM, F r i d a y , P.M.

Mechanisms o f Complement A c t i v a t i o n . Complement i s a major mediator of the inflammatory response. Complement r e c r u i t s and e n l i s t s the p a r t i c i p a t i o n of humoral and c e l l u l a r effector systems, induces histamine r e l e a s e from mast c e l l s and d i r e c t s m i g r a t i o n of leukocytes (chemotaxis), i n a d d i t i o n t o producing phagocytosis and the r e l e a s e of lysosomal c o n s t i t u e n t s from phagocytes. There a r e two p a r a l l e l , but e n t i r e l y independent pathways l e a d i n g t o a c t i v a t i o n of the t e r m i n a l , b i o l o g i c a l l y important port i o n o f the complement sequence. These mechanisms o f a c t i v a t i o n , termed the c l a s s i c a l and a l t e r n a t i v e pathways, a r e t r i g g e r e d by d i f f e r e n t substances. The two pathways converge a t C3 (midpoint) w h i l e the remainder of the r e a c t i o n sequence, i n v o l v i n g the r e a c t i o n s of C5 through C9, i s common to both pathways. There i s now an abundance of i n v i t r o data demonstrating t h a t the complement system can be a c t i v a t e d by s e v e r a l substances o f both low and h i g h molecular weight without the presence of s p e c i f i c antibody. Thus, i n a d d i t i o n to c l a s s i c IgE mediated mechanisms of histamine r e l e a s e from b a s o p h i l s and t i s s u e mast c e l l s , there are non-immunologic mechanisms capable of i n d u c i n g mediator r e l e a s e i n v i t r o , including: the B~adrenergic b l o c k i n g p r o p e r t i e s of simple chemicals, such as toluene d i i s o c y a n a t e (TDI), the histamine r e l e a s i n g p r o p e r t i e s of enzymes, p o l y p e p t i d e s , l i g a n d s , seed p r o t e i n s and p o l y c a t i o n i c amines; and the complement a c t i v a t i n g p r o p e r t i e s o f other low molecular weight substances, such as p l i c a t i c a c i d , tannins and r a d i o g r a p h i c c o n t r a s t media. The c l a s s i c a l pathway may be a c t i v a t e d immunologically by antigen-antibody complexes and aggregated immunoglobulins, and non-immunologically by a number of c h e m i c a l l y d i v e r s e substances, i n c l u d i n g DNA, C - r e a c t i v e p r o t e i n , S t a p h y l o c o c c a l p r o t e i n A, t r y p s i n - l i k e enzymes and c e r t a i n c e l l u l a r membranes (Table I I I ) .



11.


Acute Byssinotic

Reaction

111

I n t e r e s t i n g l y , many of the major a c t i v a t o r s of the c l a s s i c a l and a l t e r n a t i v e complement pathways are of b a c t e r i a l o r i g i n . Cotton dust i s known to c o n t a i n s i g n i f i c a n t b a c t e r i a l contamination, and a l s o contains p r o t e o l y t i c enzymes, DNA, p l a n t p o l y s a c c h a r i d e s , polyanions and p o l y c a t i o n s . Therefore, i t i s not i n c o n c e i v a b l e that a number of agents are r e s p o n s i b l e f o r complement a c t i v a t i o n by c o t t o n dust. A c t i v a t i o n occurs by d i r e c t b i n d i n g of CI to these substances o r , i n the case of such enzymes as the f i b r i n o l y t i c enzyme plasmin, by d i r e c t p r o t e o l y t i c a t t a c k on the C l molecules. Of s p e c i a l i n t e r e s t to those investigating e t i o l o g i c agents and mediators i n the acute b y s s i n o t i c r e a c t i o n i s the f a c t that a r e l a t i v e l y s m a l l stimulus to complement a c t i v a t i o n may lead to generation of these b i o l o g i c a l l y potent products. The a l t e r n a t i v e pathway (or properdin pathway) may be a c t i v a t e d immunologically by human IgA and some human IgG and IgE molecules, and non-immunologically by c e r t a i n complex p o l y s a c c h a r i d e s , l i p o p o l y s a c c h a r i d e s , t r y p s i n - l i k e enzymes and cobra venom.

Mediator

TABLE I I I A c t i v a t i o n of the Complement System Alternative Classical

Immunologic Non-immunologic

IgG, IgM and aggregated

f

Ig s

IgA, IgG, (some) IgE

Trypsin-like enzymes

Trypsin-like enzymes

DNA

L i popolysaccharides

Staphylococcal protein A

Plant polysaccharides

C-reactive protein

Cobra venom f a c t o r s

Viruses

Bacterial c e l l wall

Bacterial c e l l wall

Viruses

Polyanions and polycations Cleavage of C'3 i n t o C3a and C5 i n t o C5a r e s u l t s i n peptide fragments w i t h chemotactic and histamine r e l e a s i n g b i o l o g i c a l properties. These fragments have been demonstrated to be as e f f e c t i v e as IgE i n r e l e a s i n g histamine from human b a s o p h i l s and mast c e l l s . Further a c t i v a t i o n of C5, C6 and C7 (C567) i s a l s o chemotactic. The chemotaxins (C3a, C5a, C567) and anaphylatoxins (C3a, C5a) are e f f e c t i v e i n the recruitment of polymorphonuclear leukocytes through blood v e s s e l w a l l s , the r e l e a s e of histamine


172

COTTON

DUST

from blood borne b a s o p h i l s and t i s s u e mast c e l l s , and the p o s s i b l e c o n t r a c t i o n of lung smooth muscle. These f u n c t i o n s are summarized i n Table IV.


TABLE IV B i o l o g i c E f f e c t s of Complement A c t i v a t i o n

C4a C2

Kinins (increase vascular permeability c o n t r a c t smooth muscle)

and

C3a

Chemotactic a c t i v i t y Histamine r e l e a s e K i n i n - l i k e a c t i v i t y ( c o n t r a c t s smooth muscle)

C5a

Chemotactic a c t i v i t y Histamine r e l e a s e Lysosomal enzyme r e l e a s e

C567

Chemotactic a c t i v i t y

In v i t r o s t u d i e s i n d i c a t e that c o t t o n m i l l dust e x t r a c t s do a c t i v a t e complement by the a l t e r n a t i v e pathway. Kutz Kallikrein ^Kininogen ^Kinins Aggregated IgE and A (mucosal surface) l a c k s C probably Arachidonic a c i d can lead to s y n t h e s i s of PG's , thromboxanes, and l e u k o t r i e n e s i n the lung and aggregation of p l a t e l e t s and r e l e a s e of histamine from p l a t e l e t s

TABLE V I I P o t e n t i a l E t i o l o g i c a l Agents i n E x t r a c t s of Cotton M i l l Dust That E f f e c t Histamine Release A.

Histamine r e l e a s e v i a a c t i v a t i o n of c l a s s i c a l or a l t e r n a t e complement pathways Endotoxins (LPS) from gram negative organisms. P r o t e o l y t i c enzymes Polyanions Polycations DNA Staphylococcal A p r o t e i n

B.

Histamine r e l e a s e v i a d i r e c t a c t i v a t i o n Peptides (N-formylmethionyl) Polyphenols and Tannins Tannic a c i d Quercetin Catechin Ellagic acid Rutin Trimethylamine Scopoletin Metasilicates L a c i n i l i n e C-7 methyl ether

C.

Histamine r e l e a s e v i a IgE ( a t o p i c h y p e r s e n s i t i v i t y Type I ) Antigen (hapten or complete) v i a IgE mediated r e l e a s e of histamine from b a s o p h i l s and t i s s u e mast c e l l s (complement independent).



176

COTTON

DUST

D i r e c t histamine r e l e a s e r s present i n c o t t o n m i l l dust are l i s t e d i n Table VII-B. A major problem i n the study of byssinogenic agents i n CMD has been the q u e s t i o n of s u i t a b l e assay systems. These assays have ranged from i n h a l a t i o n by human v o l u n t e e r s , leukocyte recruitment i n the lungs of animals i n v i v o , chemotaxis i n Boyden chambers, to histamine r e l e a s e by chopped lung t i s s u e . The l a t t e r method has been used i n a number of productive s t u d i e s and has l e d to the f i n d i n g s that histamine i s one of the mediators i n the acute b y s s i n o t i c r e a c t i o n . U n f o r t u n a t e l y i n our l a b o r a t o r i e s (46) and i n other's (47), the use of chopped lung t i s s u e s has appeared e x c e s s i v e l y l a b o r i o u s , not always r e l i a b l e and extremely i n s e n s i t i v e (48). We subsequently devised a simple, s e n s i t i v e and r e l i a b l e bioassay procedure u s i n g p i g p l a t e l e t s f o r the assessment of histamine r e l e a s i n g f a c t o r s i n c o t t o n m i l l dust (46). The p l a t e l e t histamine r e l e a s e assay demonstrated that cotton m i l l dust e x t r a c t , c o t t o n b r a c t e x t r a c t , c o t t o n l e a f e x t r a c t , dialyzed CMD e x t r a c t , polyphenols, compound 48/80, r u t i n , trimethy1amine HC1, q u e r c e t i n , c a t e c h i n , t a n n i c a c i d , e l l a g i c a c i d and sodium m e t a s i l i c a t e a l l r e l e a s e histamine d i r e c t l y (48). Thus not only do tannin compounds induce histamine r e l e a s e , but they may a l s o form higher molecular weight polymers and c o n t a i n components that s u r v i v e a c i d h y d r o l y t i c c o n d i t i o n s (48). Tannins are w i d e l y d i s t r i b u t e d i n the p l a n t kingdom. S i l i c a t e i s another type of widely d i s t r i b u t e d substance that induces the r e l e a s e of histamine from p l a t e l e t s . I t i s a common c o n s t i t u e n t of p l a n t t i s s u e s i n the form of d i s s o l v e d hydrous s i l i c a or s i l i c i c a c i d (24), which may c o n t r i b u t e to the b y s s i n o t i c syndrome. Cotton b r a c t s c o n t a i n 0.4-0.8% s i l i c a (24). For many of the byssinogenic substances, nanogram concentrations are s u f f i c i e n t to r e l e a s e histamine (48). I t i s now known that N-formylmethionyl (NFMet) peptides are s t r o n g l y chemotactic f o r PMN and macrophages and cause the r e l e a s e of histamine i n human b a s o p h i l s . Thus, l i k e the complement derived C3a and C5a a n a p h l y l a t o x i c (histamine r e l e a s i n g ) fragments, N-FMet peptides a l s o possess d i r e c t histamine r e l e a s i n g p r o p e r t i e s . H i s tamine r e l e a s e proceeds through a mechanism u n r e l a t e d to IgE a c t i v a t i o n of b a s o p h i l s and t i s s u e mast c e l l s . The histamine r e l e a s i n g a b i l i t y of these peptides c o r r e l a t e s w e l l w i t h t h e i r chemotactic a c t i v i t y (49). Chemotactic Agents i n E x t r a c t s of Cotton M i l l Dust Table V I I I l i s t s a v a r i e t y of substances that are known chemotaxins. Chemotaxins a t t r a c t white blood c e l l s , p r i m a r i l y n e u t r o p h i l s , e o s i n o p h i l s and macrophages, to a s p e c i f i c s i t e by causing d i r e c t e d movement. Once these c e l l s a r r i v e they phagocytize and degrade invading microorganisms, and/or r e l e a s e h y d r o l y t i c enzymes which can cause subsequent t i s s u e i n j u r y . These c e l l s a l s o r e l e a s e b r o n c h o c o n s t r i c t o r s , which provides another mechanism worthy of c o n s i d e r a t i o n .



11.


Acute Byssinotic

Reaction

TABLE V I I I Chemotactic Factors Proteins Complement fragments C3a C5a C567 Coagulation

proteins

Plasminogen a c t i v a t o r Fibrinopeptide B Prekallikrein Other p r o t e i n s Random c o i l p r o t e i n s (e.g., IgG h y d r o l y s i s products Cell-bound antigen

casein)

L i p i d s and other molecules Products of l i p i d p e r o x i d a t i o n Prostaglandins Bacterial f i l t r a t e factors N-Formyl methionyl peptides C y c l i c purine n u c l e o t i d e s Transfer f a c t o r Lymphokines


111

COTTON


178

DUST

Gram negative b a c t e r i a are n a t u r a l contaminants o f f i b r o u s plant material. I n processes where c o t t o n i s a g i t a t e d , i . e . d u r i n g r a p i d machining, b a c t e r i a on the c o t t o n f i b e r s o f t e n become a i r b o r n e , and the workers i n v o l v e d i n dustVj areas may be^exposed t o c o n c e n t r a t i o n s of b a c t e r i a as h i g h as 10 bacteria/m (50. > 51). A number of f a c t o r s s t i m u l a t e m i g r a t i o n of leukocytes and r e s i d e i n CMD (Table I X ) . Various s t r a i n s o f gram negative b a c t e r i a , e s p e c i a l l y those of Enterobacter and K l e b s i e l l a , produce potent chemotaxins. These organisms, i s o l a t e d from c o t t o n dust and administered t o guinea p i g s , i n i t i a t e PMN m i g r a t i o n i n airways (52); however, i s o l a t e d gram p o s i t i v e b a c t e r i a or suspensions from these organisms and t o v a r i o u s molds produces no r e a c t i o n . Various gram negative b a c t e r i a possess l i p o p o l y s a c c h a r i d e s (LPS) which d i f f e r g r e a t l y i n t h e i r a b i l i t y t o r e c r u i t leukocytes. S e v e r a l b a c t e r i a l LPS i s o l a t e d from c o t t o n were t e s t e d by exposing guinea pigs t o a e r o s o l s and counting the number of f r e e lung c e l l s a t 24 hours. Chemotaxis was found to be dependent on dose and species of b a c t e r i a and LPS i n h a l e d (52).

TABLE IX Chemotaxins i n Cotton Dust E x t r a c t s Exert e f f e c t s i n d i r e c t l y Exert e f f e c t s d i r e c t l y : through complement a c t i v a t i o n : 1. B a c t e r i a l N-formylmethionyl peptides 2. L a c i n i l e n e C-7-methyl e t h e r

a

1.Endotoxins 2.LPS ( p u r i f i e d endotoxin) 3.DNA 4.Bacterial f i l t r a t e factors

Recently shown by Ainsworth and Neuman not t o be chemotactic (16)

Some f a c t o r s , such as NFMet peptides and other chemotaxins, exert t h e i r e f f e c t s d i r e c t l y on c e l l s , w h i l e chemotaxigens a t t r a c t c e l l s i n d i r e c t l y through a c t i v a t i o n of complement and production of C3a, C5a or C567. Studies i n our l a b o r a t o r y have shown that potent chemotaxins r e s i d e i n aqueous e x t r a c t s of CMD (15). Further s t u d i e s have shown these chemotaxins t o be a n i o n i c , m.w. 200-2000 d a l t o n s , s t a b l e i n b o i l i n g water and a c i d h y d r o l y z a b l e (16). These p r o p e r t i e s are c o n s i s t e n t w i t h c h a r a c t e r i s t i c s of p e p t i d e s , p a r t i c u l a r l y N-FMet peptides. These a r e potent chemoattractants of b a c t e r i a l o r i g i n and are p o t e n t i a l c o t t o n dust chemotactic e t i o l o g i c agents. We have suggested t h a t polypeptides might be the c a u s a t i v e agent i n b y s s i n o s i s (15). Cotton dust i s h e a v i l y laden w i t h b a c t e r i a (40, 4 1 ) , and t h e i r p r o j e c t s (peptides) are chemotactic a t c o n c e n t r a t i o n s as low as 10 M (on the order of 10 jug/ml) (53). B i o l o g i c a l e f f e c t s could e a s i l y be e f f e c t e d by



11.

AINSWORTH AND

PILIA

Acute Byssinotic

Reaction

179

i n s p i r a t i o n of such potent substances. A further significant property of the peptides i s t h e i r c a p a c i t y to induce s p e c i f i c s e c r e t i o n of lysosomal enzymes from leukocytes (53-55) w i t h the p o s s i b l e sequelae of p a t h o l o g i c l e s i o n s , i . e . b r o n c h i t i s and emphysema. In b y s s i n o s i s , however, emphysema i s probably a r e s u l t of the i n h a l a t i o n of c i g a r e t t e smoke, and not of c o t t o n dust (56). The s t r u c t u r e of NFMet peptides i s p e c u l i a r to b a c t e r i a l metabolism as i n t e r m e d i a r i e s i n p r o t e i n b i o s y n t h e s i s . The b i o l o g i c response to NFMet peptides may be p a r t of a defensive mechanism a g a i n s t b a c t e r i a l i n f e c t i o n . The chemotactic f a c t o r ( s ) i n c o t t o n dust appears to be a small molecular weight p e p t i d e , c o n s i s t e n t w i t h the view that peptides, i n g e n e r a l , are u s u a l l y low molecular weight b i o l o g i c a l e f f e c t o r s . D i r e c t evidence that t h i s n a t u r a l l y occuring b a c t e r i a l chemotaxin i s an N-formylated peptide awaits f u r t h e r work. The p u r i f i c a t i o n and i d e n t i f i c a t i o n of chemotaxin(s) i n c o t t o n dust are being pursued i n our l a b o r a t o r y , and these products are being s i m i l a r l y i n v e s t i g a t e d i n an animal model and compared w i t h NFMet peptides. Human plasma k a l l i k r e i n has a l s o been d e s c r i b e d as being d i r e c t l y chemotactic f o r human n e u t r o p h i l s (57). Kallikrein incubated w i t h the complement component C5 generates chemotactic a c t i v i t y , p r o v i d i n g a f u r t h e r mechanism f o r C5 a c t i v a t i o n . C5a i s the most a c t i v e and probably the most b i o l o g i c a l l y important of the complement derived chemotactic and anaphylatoxic peptides. Some agents are b i f u n c t i o n a l , causing the r e l e a s e of histamine and r e c r u i t i n g leukocytes. B i f u n c t i o n a l mediators i n c l u d e b a c t e r i a l p e p t i d e s , endotoxins, DNA, C3a, C5a and b r a d y k i n i n . Each of these substances can exert dual e f f e c t s . This may e i t h e r occur d i r e c t l y , as i n the case of b a c t e r i a l peptides and b r a d y k i n i n causing chemotaxis and b r o n c h i a l smooth muscle c o n t r a c t i o n , or indirectly, as endotoxin and DNA conversion of complement. C3a and C5a act i n d i r e c t l y as complement fragments to e f f e c t histamine r e l e a s e , which i n t u r n c o n t r a c t s b r o n c h i a l smooth muscle. However, both appear to act d i r e c t l y to e f f e c t chemotaxis w i t h C5a, the more potent fragment. B r a d y k i n i n , a chemotaxin and muscle c o n s t r i c t o r , i s a c t i v a t e d by enzymatic cleavage. At present, blood c l o t t i n g anomalies i n b y s s i n o s i s have not been d e s c r i b e d , i n d i c a t i n g that i t i s u n l i k e l y that t h i s substance plays a r o l e , although aggregation of RBC by CMD e x t r a c t s has been reported i n our l a b o r a t o r y (58). C l i n i c a l s t u d i e s of c o t t o n m i l l workers who had p r e v i o u s l y demonstrated a decreased e x p i r a t o r y flow measured by flow volume curves and FEV^ during c o t t o n dust exposure showed an increase i n WBC to 25.5% a f t e r 4 hours of exposure. Segmented n e u t r o p h i l s increased most (33%), while e o s i n o p h i l mean counts d i d not change. The r a t i o of segmented n e u t r o p h i l s to e p i t h e l i a l c e l l s from n a s a l mucosal swabs increased from 0.56 before to 1.84 a f t e r 4 hours of exposure. P e r i p h e r a l blood and PMN counts increased upon exposure to c o t t o n dust, and PMN were r e c r u i t e d to the n a s a l mucosa. Chest t i g h t n e s s and decreased flow were t e m p o r a r i l y c o r r e l a t e d w i t h leukocyte recruitment f o l l o w i n g c o t t o n dust exposure ( 2 ) .



180

COTTON

DUST

In s t u d i e s on hamsters and guinea pigs exposed t o a e r o s o l s of crude and r e f i n e d e x t r a c t s from c o t t o n m i l l t r a s h and CMD, K i l b u r n et a l . (59) has f u r t h e r demonstrated recruitment of PMN beneath the basement membrane and on l u m i n a l surfaces of intrapulmonary airways and tracheas. When administered as a e r o s o l s o r dust, p o l y p h e n o l i c e x t r a c t s from c o t t o n t r a s h and pure and o x i d i z a t i o n p o l y m e r i z a t i o n products of q u e r c e t i n a l s o r e c r u i t PMN from the trachea t o t e r m i n a l b r o n c h i o l e s i n hamsters (60). In a d d i t i o n , l a c i n i l e n e ( s y n t h e t i c and n a t u r a l ) , reputed by K i l b u r n t o be chemotactic and t o be the b y s s i n o t i c agent, has r e c e n t l y been evaluated i n our l a b o r a t o r y f o r chemokinetic and chemotactic a c t i v i t y ( J J 6 , 6 1 ) . In our s t u d i e s , l a c i n i l e n e does not possess chemotactic o r chemokinetic a c t i v i t i e s . Interestingly, the agent used t o s o l u b i l i z e l a c i n i l e n e i n K i l b u r n ' s s t u d i e s ( P l u r o n i c P o l y o l F 6 8 , a polypropylene g l y c o l ) does not possess chemotactic a c t i v i t y , but i s a new c l a s s of chemokinetic compounds (16). The chemokinetic a c t i v i t y of the p o l y o l probably e x p l a i n s why l a c i n i l e n e was r e p o r t e d as the "chemotactic-byssinogenic" agent by these i n v e s t i g a t o r s .

Bioassay of B r o n c h o c o n s t r i e t o r Substances M i l l Dust and Cotton B r a c t

i n E x t r a c t s o f Cotton

In 1 9 6 2 , Davenport and Paton ( 6 2 ) s t u d i e d the chemical and b i o l o g i c a l p r o p e r t i e s of e x t r a c t s of two d i f f e r e n t samples of c o t t o n dust. They d e s c r i b e d an a c t i v e component i n these e x t r a c t s which c o n s t r i c t e d smooth muscle of r a t g a s t r o i n t e s t i n a l t r a c t and guinea p i g trachea. The r a t stomach s t r i p s were the most s e n s i t i v e to c o t t o n dust e x t r a c t (CDE) and t o 5-hydroxytryptamine (5HT). Brom-lysergic a c i d p a r t i a l l y antagonized the e f f e c t of CDE over the stomach s t r i p s , but f a i l e d t o antagonize the CDE c o n s t r i c t o r e f f e c t over r a t duodenum and guinea p i g trachea. A t r o p i n e and Mepyramine maleate were i n e f f e c t i v e i n a l l t i s s u e s . The a c t i v e component d e s c r i b e d was s o l u b l e i n water, i n s o l u b l e i n methanol, chloroform, acetone, ethanol and petroleum ether and possessed high K levels. I t a l s o had both b a s i c and a c i d i c groups i n a r a t i o of 2 b a s i c : 1 a c i d i c , and was d i a l y s a b l e , r e s i s t a n t t o b o i l i n g and not destroyed by the a c t i o n of p r o t e o l y t i c enzymes. In blood bathed p r e p a r a t i o n s , animals i n j e c t e d w i t h CDE r e l e a s e d a substance i n t o the blood stream t h a t e l i c i t e d r e l a x a t i o n o f r a t duodenum muscle but had no e f f e c t on r a t stomach muscle. The onset was slow which, added t o other f i n d i n g s , l e d these i n v e s t i g a t o r s t o p o s t u l a t e a k i n i n - l i k e substance t o be the mediator r e l e a s e d . Mohammed ( 6 3 ) r e p o r t e d that acetone treatment of aqueous c o t t o n dust e x t r a c t caused the p r e c i p i t a t i o n of s e v e r a l sugars and an aminopolysaccharide. The sugars were present i n c o n c e n t r a t i o n s equal t o those which e x h i b i t e d c o n t r a c t i l e a c t i v i t y on guinea p i g



11.


Acute Byssinotic

Reaction

181

ileum. Our experiments to date have f a i l e d to demonstrate c o n t r a c t i l e a c t i v i t y i n the acetone p r e c i p i t a t e of e i t h e r the c o t t o n dust or c o t t o n b r a c t e x t r a c t s (64). N i c h o l l s and Skidmore (65) demonstrated t h a t dust c o l l e c t e d from m i l l s w i t h a higher prevalence of b y s s i n o s i s caused greater smooth muscle c o n t r a c t i l e a c t i v i t y than dust from m i l l s w i t h a lower prevalence. Recently, R u s s e l l et a_l. (66) used an i s o l a t e d t i s s u e bath to measure canine t r a c h e a l i s muscle c o n t r a c t i o n caused by c o t t o n b r a c t e x t r a c t s (CBE). Morey et a l . (67) showed t h a t c o t t o n bract represents 20-43% of the c o t t o n dust t o t a l ; thus, the f i n d i n g s of Paton and Davenport using c o t t o n dust agree i n p r i n c i p l e w i t h R u s s e l l e_t a l . , who used c o t t o n b r a c t . Davenport and Paton (62) found a percentage of the a c t i v i t y of CDE represented by a 5HT-like component, but they a l s o found at l e a s t one more a c t i v e substance i n the CDE than R u s s e l l e t a l . (66). We have gained considerable knowledge i n our l a b o r a t o r y of b r o n c h o c o n s t r i c t o r s i n b r a c t and dust. A comparison of r a t bladder, c o l o n , ileum and stomach s e n s i t i v i t y to 5-HT showed the stomach to be the most s e n s i t i v e t i s s u e t o e x t r a c t s as has been reported by others (62). Cotton dust and c o t t o n b r a c t e x t r a c t contracted smooth muscle w i t h forces e q u i v a l e n t to a 0.32+0.05 jug/ml c o n c e n t r a t i o n of 5HT and 0.7+0.1 jug/ml c o n c e n t r a t i o n of 5-HT, r e s p e c t i v e l y (68). However, c o t t o n b r a c t e x t r a c t s were blocked by methysergide, a 5-HT a n t a g o n i s t , w h i l e c o t t o n dust e x t r a c t s were not. Diphenhydramine, a histamine a n t a g o n i s t , f a i l e d to s i g n i f i c a n t l y a f f e c t e i t h e r of the e x t r a c t s (§.§). Acetone p r e c i p i t a t e d CDE and CBE caused minimal c o n t r a c t i o n of the r a t stomach smooth muscle. The supernatant r e t a i n e d most of the a c t i v e substance (64). Butanol, however, d i d e x t r a c t the a c t i v e components of both CBE and CDE (69). E t h y l a c e t a t e , which w i l l e x t r a c t 5HT from an aqueous s o l u t i o n (64), was u n s u c c e s s f u l i n removing the a c t i v e components. These f i n d i n g s are i n agreement w i t h R u s s e l l e_t a l . (66), who used t h i s procedure to demonstrate that the a c t i v e component of b r a c t was 5 H T - l i k e . " The a c t i v e agent(s) i n e x t r a c t s of the dust and b r a c t were demonstrated to have a molecular weight below 20,000. In comparing 5HT e q u i v a l e n t values f o r CDE and CBE, i t might be suggested t h a t part of the c o n t r a c t i o n r e s u l t i n g from CDE i s due to the 20-40% b r a c t i n the dust. We have shown that the c o n t r a c t i o n caused by c o t t o n dust e x t r a c t i s d i f f e r e n t from the c o n t r a c t i o n caused by c o t t o n b r a c t e x t r a c t . CBE was blocked by methysergide. A methysergide c o n c e n t r a t i o n t h a t would block 75+2% of 5 HT a c t i v i t y would block 59+6% of CBE a c t i v i t y and only 12.6+4% of CDE a c t i v i t y (68). This suggests t h a t the two e x t r a c t s operate at d i f f e r e n t receptor s i t e s on the smooth muscle, and t h a t the nature of their a c t i v e component(s) would be different. Histamine, which i s known to be present i n c o t t o n dust, i s not r e s p o n s i b l e f o r the c o n t r a c t i l e a c t i v i t y i n these preparations (68). n



182

COTTON

DUST

Though the c o n t r a c t i o n s caused by c o t t o n dust and c o t t o n b r a c t have important d i f f e r e n c e s , there a r e a l s o s i m i l a r i t i e s between t h e i r chemical p r o p e r t i e s . I t i s d o u b t f u l that e i t h e r of the a c t i v e agents are p r o t e i n s because pronase has no a f f e c t on the e x t r a c t s and they a r e l a b i l e even a t 4 C. In a d d i t i o n , both a c t i v e substances were e x t r a c t e d w i t h butanol (69). Recent smooth muscle c o n s t r i c t o r s t u d i e s performed i n our l a b o r a t o r y i n c o l l a b o r a t i o n w i t h Dr. Marion Buck a t Yale Univers i t y have y i e l d e d i n t e r e s t i n g r e s u l t s . Various f r a c t i o n s were f i r s t t e s t e d i n human v o l u n t e e r s f o r b r o n c h o c o n s t r i c t o r a c t i v i t y and then bioassayed by the r a t stomach s t r i p i s o l a t e d organ bath technique. The f r a c t i o n s demonstrating c o n s t r i c t o r a c t i v i t y i n man c o r r e l a t e d p r e c i s e l y w i t h the c o n s t r i c t o r a c t i v i t y i n the i s o l a t e d organ bath. This e f f e c t was antagonized by methysergide (69). As the major c l i n i c a l m a n i f e s t a t i o n of the acute b y s s i n o t i c r e a c t i o n i s a drop i n FEV^, i t i s i n t e r e s t i n g t o speculate that the b r o n c h o c o n s t r i c t i o n observed i n c o t t o n m i l l workers may be i n p a r t or i n f u l l the r e s u l t of c o n s t r i c t o r substances i n i n h a l e d c o t t o n dust. I f the r e c e p t o r s i n v o l v e d i n the acute b y s s i n o t i c r e a c t i o n can be determined, pharmacologic b l o c k i n g agents may be used t o t r e a t a f f e c t e d c o t t o n m i l l workers o r , conversely, preprocessing of c o t t o n t o e l i m i n a t e the c o n s t r i c t o r might prove f e a s i b l e .

Conclusions In recent y e a r s , r a p i d s t r i d e s have been made toward understanding the composition of c o t t o n m i l l dust, c a u s a t i v e agents and pathogenic mechanisms. Studies on composition and chemistry have demonstrated s i g n i f i c a n t d i f f e r e n c e s i n c o t t o n dust emanating from v a r i o u s types of c o t t o n p l a n t s , d i f f e r e n t harvest times, harvest techniques and geographical area o f c u l t i v a t i o n . The c a u s a t i v e agent i s seemingly u b i q u i t o u s i n f r i a b l e vegetable m a t e r i a l s which, when machined, r a p i d l y produce p a r t i c l e s of r e s p i r a b l e s i z e and thus b y s s i n o s i s i n f l a x , hemp, and c o t t o n workers. Bioassays have been i n s t r u m e n t a l i n f u r t h e r i n g our understanding of e t i o l o g i c agents and pathogenic mechanisms i n b y s s i n o s i s , i n p a r t i c u l a r , bioassays of histamine r e l e a s i n g substances, chemotaxins, and b r o n c h o c o n s t r i c t o r s i s o l a t e d from e x t r a c t s of dust. Bioassays have helped t o d e f i n e parameters of b i o l o g i c a l a c t i v i t y of d u s t s , as w e l l as the immunological, p h y s i o l o g i c a l , pharmacological and p a t h o l o g i c a l r e a c t i o n s of c e l l s and muscle t i s s u e . The acute b y s s i n o t i c r e a c t i o n w i l l be b e t t e r understood when such bioassays are more c l o s e l y c o r r e l a t e d w i t h human r e a c t o r s t u d i e s , and i n t u r n , both human and bioassay r e s u l t s c o r r e l a t e d w i t h an appropriate animal model.


11.


Acute Byssinotic

Reaction

183

Acknowledgments


This research was supported by funds from Cotton I n c o r porated, Cotton Foundation, and the South C a r o l i n a Lung A s s o c i a t i o n . Dr. P i l i a i s a r e c i p i e n t of a N a t i o n a l Kidney Foundation Postdoctoral Fellowship. The authors g r a t e f u l l y a p p r e c i a t e the e d i t o r i a l a s s i s t a n c e of Janet Vesterlund and the s e c r e t a r i a l a s s i s t a n c e of C h a r l o t t e Spain.

Literature Cited 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12.

13. 14. 15.

Mareska, J.; Heymann, J . Ann. Soc. Med. de Gand. 1945, 16, 5245. Merchant J . A . , Lumsden, J.C., Kilburn, K.H., O'Fallon, W.M., Ujda, J.R., Germino, V.H., Hamilton, J.D. Dose Response in Cotton Textile Workers. J . Occup. Med. 1973, 15, 222230. Merchant, J . A . , Halprin, G.M., Hudson, A.R., Kilburn, K . H . , McKenzie, W.N., Hurst, D . J . , Bermazohn, P. Responses to Cotton Dust. Arch. Environ. Health 1979, 30, 222-229. Bouhuys, A . ; Heaphy, L.J., J r . ; Schilling, R.S.F.; Welborn, J.W. N. Eng. J . Med. 1967, 277, 170-175. Bouhuys, A . ; Lindell, S.E.; Lundin G. Brit. Med. J . 1960, 1, 324-326. Bouhuys, A . ; Van Duyn, J.; Van Lennep, H.J. Arch. Environ. Health. 1961, 3, 499-509. McDermott, M.; Skidmore, J.W.; Edwards, J . "2nd International Conference on Respiratory Diseases in Textile Workers"; Alicante, Spain, 1968, p 13-36. Cavagna, G.; Foa, V .; Vigliani, E.C. Br. J . Ind. Med. 1969, 26, 314-321. Pernis, B.; Vigiliani, E.C.; Cavagna, G.; F i n u l l i , M. Br. J . Ind. Med. 1961, 18, 120-129. Bergstrom, R.; Haglind, P.; Rylander, R. "Proceedings, 1980 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1980, p 22-25. Massoud, A . ; Taylor, G. Lancet 1964, 2, 607-610. Kilburn, K.H.; Lynn, D.; McCormick, J.P.; Schaefer, T.R. "Proceedings, 1979 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1979, p 19-20. Lynn, W.S.; Munoz, S.; Campbell, J.A.; Jeffs, P.W. Ann. N.Y. Acad. Sci. 1974, 221, 163-173. Buck, M.G.; Bouhuys, A. "Proceedings, 1980 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1980, p 31-34. Ainsworth, S.K.; Neuman, R.E. "Proceedings, 1980 Beltwide Cotton Production Research Conferences"; National Cotton Council of America, Memphis, Tenn., 1980, p 14-18.



184

COTTON

DUST

16. Ainsworth, S.K.; Neuman, R.E. Am. Rev. Respir. Dis. 1981 124, 280-284. 17. Antweiler, H. Ann. Occup. Hyg. 1960, 2, 152-156. 18. Bouhuys, A. In Breathing, Grune and Stratton: New York, p 41619. Fox, A.J.; Tombleson, J.B.L.; Watt, A.; Wilie, A.G. Br. J . Ind. Med. 1973, 30, 48-53. 20. Criteria for Recommended Standard. . . Exposure to Cotton Dust. U.S. Dept. HEW, PHS. CDD, NIOSH, 1974. (HEW Publication No. (NIOSH) 75-118) Washington, D.C. 21. Imbus, H.R.; Suh, M.W. Arch. Environ. Health 1973, 26, 183191. 22. Hamilton, J.D.; Halprin, G.M.; Kilburn, K.H.; Merchant, J.A.; Ujda, J.R. Arch. Environ. Health 1973, 26, 120-124. 23. Antweiler, H. Ann. N.Y. Acad. Sci. 1969, 158, 136. 24. Wakelyn, P . J . ; Greenblatt, G.A.; Brown, D.F.; Fripp, V.W. Am. Indust. Hygiene Assoc. J . 1976, 37, 22-31. 25. Tuffnell, P. Br. J . Ind. Med. 1960, 16, 307-309. 26. Zakhdov, A . Z . ; Schraiber, L . B . ; Gol'eva, I.V.; Katsenovich, L.A.; Samsonov, A.P. Hyg. Sanit. 1965, 30, 344. 27. Ainsworth, S.K.; Neuman, R.E. "Proceedings, 1977 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1977, p 62-64. 28. Ainsworth, S.K.; Neuman, R.E. Unpublished results. 29. Noweir, M.H. Chest 1981, 79, 62S-67S. 30. Ainsworth, S.K.; Mundie, T.G.; Pilia, P.A. "Proceedings, 1981 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1981, pp 78. 31. Edwards, J.H.; Jones, B.M. J . Immunol. 1973, 110, 498-501. 32. Sekul, A.A.; Ory, R.L. "Proceedings, 1979 Beltwide Cotton Production Conference"; National Cotton Council of America, Memphis, Tenn., 1979, p. 30-31. 33. Kutz, S.A.; Mentnech, M.S.; Olenchock, S.A.; Major, P.C. Chest 1981, 79, 56S-58S. 34. Kutz, S.A.; Mentnech, M.S.; Mull, J.C.; Olenchock, S.A. Arch. Environ. Health 1980, 35, 205-10. 35. Taylor, G.; Massoud, A.A.E.; Lucas, F. Br. J . Ind. Med. 1971, 28, 143-151. 36. Kutz, S.A.; Olenchock, S.A.; E l l i o t t , J.A.; Pearson, D . J . ; Major, P.C. Envir. Res. 1979, 19, 405-414. 37. Wilson, M.R.; Sekul, A.; Ory, R.; Salvaggio, J . E . ; Lehrer, S.B. Clin. Allergy 1980, 10, 303-308. 38. Mundie, T.G.; Ainsworth, S.K.; Boackle, R.J. Society for Experimental Biology and Medicine Southeastern Section, Charleston, S.C., (abstr.) Abstracts 5, p 14, 1980. (Manuscript, In Press, 1981). 39. Morey, P.R.; Bethea, R.M.; Kirk, I.W.; Wakelyn, P.J. Cotton Dust, Proc. of a Topical Symp.; 1975, Cincinnati, p 237. 40. Fischer, J . J . "Proceedings, 1979 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1979, p 8-10.


11.

AINSWORTH AND PILIA

41.

Morey, P.R.; Fischer, J.J.; Sasser, P.E. "Proceedings, 1980 Beltwide Cotton Production Conference"; National Cotton Council of America, Memphis, Tennessee, 1980, p 68-9. Cogen, F.C.; Norman, M.E.; Dunsky, E . ; Hirschfield, J . ; Zweiman, B. J . Allergy Clin. Immunol. 1979, Suppl. 64:299303. Braun, D.C.; Scheel, L.D.; Tuma, J . ; Parker, L. J . Occupatonal Med. 1973, 15, 241-244. Cinkotai, F.F. Am. Industrial Hygiene Assoc. J . 1976, 37, 324-338. Ainsworth, S.K.; McCormick, J.P.; Neuman, R.E. "Proceedings, 1979 Beltwide Cotton Production Research Conference; National Cotton Council of America, Memphis, Tenn., 1979, p 21-29. Ainsworth, S.K.; Neuman, R.E.; Harley, R.A. Br. J . Ind. Med. 1979, 36, 35-42. Greenblatt, G.A. "Proceedings, 1977 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1977, p 71-72. Ainsworth, S.K.; Neuman, R.E. "Proceedings, 1977 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1977, p 76-79. Hook, W.A.; Schiffman, E . ; Aswanikumar, S.; Siraganian, R. P. J . Immunol. 1977, 117, 594-596. Fisher, J.J.; Battigelli, M.C.; Foarde, K.K. "Proceedings, 1979 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1979, p 13-14. Lundholm, M.; Rylander, R. "Proceedings, 1979 Beltwide Cotton Production Research Conference"; National Cotton Council of America, Memphis, Tenn., 1979, p 11-13. Helander, I . ; Salkinoja-Salonen, M.; Rylander, R. Infect. Immunity 1980, 29, 859-862. Showell, H . J . ; Freer, R . J . ; Zigmond, S.H.; Schiffmann, E . ; Aswanikumar, S.; Corcoran, B.; Becker, E.L. J . Exp. Med. 1976, 143, 1154-1169. Becker, E . L . ; Showell, H . J . ; Henson, P.M.; Long, S.H. J . Immunol. 1974, 112, 2047-2054 Becker, E . L . ; Showell, H.J. J . Immun. 1974, 112, 2055-2062. Pratt, P.C. Chest 1981, 79, 495-535. Wiggins, R.C.; Giclais, P.C. (abstr.) Fed. Proc. 1980, 39, 1049. Ainsworth, S.K.; Neuman, R.E. (abstr.) Am. Rev. Respir. Pis. 1978, 117, 217. Kilburn, K.H.; Lynn, W.S.; Tress, L.L.; McKenzie, W.N. Lab. Invest. 1973, 28, 55-49. Kilburn, K.H. Ann. N.Y. Acad. Sci. 1974, 221, 335-339. Neuman, R.E.; Ainsworth, S.K., J . Reticuloendothelial Soc. 1980, 28, 305-312. Davenport, A.; Paton, W.P.M. Br. J . Ind. Med. 1962, 29, 19-32.

42. 43. Downloaded by UNIV OF CALIFORNIA SAN DIEGO on January 24, 2017 | http://pubs.acs.org Publication Date: July 1, 1982 | doi: 10.1021/bk-1982-0189.ch011

44. 45. 46. 47. 48. 49. 50.

51. 52. 53. 54. 55. 56. 57. 58. 59. 60. 61. 62.

Acute Byssinotic

Reaction


185

186

63.

COTTON

DUST

Mohammed, Y . S . ; El-Guzzah, R.M.; Adamyova, K. Carbohydrate Res.

1971, 20, 431-435.

64. 65.

Ainsworth, S.K.: Unpublished results. Nicholls, P.S.; Skidmore, J.W. Br. J . Ind. Med. 1975, 32, 289-

66.

Russell, J.A.; Gilberstadt, M.L.; Rohrback, M.S. (abstr.) Am. Rev. Respir. Dis. 1981, 124, 252. Morey, P.R. "Proceedings, 1977 Beltwide Cotton Production Research Conference;" National Cotton Council of America, Memphis, Tenn. 1977, 10-11. Mundie, T.G.; Cordova-Salinas, M.A.; Bray, V.J.; Ainsworth, S.K. "Proceedings, 1982 Beltwide Cotton Production Research Conference," National Cotton Council of America, Las Vegas, Nevada, 1982. Mundie, T.G.; Buck, M.; Cordova-Salinas, M.A.; Neuman, R.E.; Ainsworth, S.K. "Proceedings, 1982 Beltwide Cotton Production Research Conference," National Cotton Council of America, Las Vegas, Nevada, 1982.

296.


67. 68.

69.

RECEIVED

February 24, 1982.


Etiologic Agents and Pathogenic Mechanisms in the Acute Byssinotic

Recommend Documents