- Email: [email protected]
108. Pulmonary function in rabbits with BCG-induced lung disease—an animal model of hypersensitivity pneumonitis
.!. NLERGY
on of cell-mediated immunity eder’s lung (PBlJ using fresh opreserwed lymphocytes. J. Anderson, B.A8 London,
J. Toogood, M.D., N. Lefcoe, M.D., R. T., J. Mazza, M.D., B. Jennings, Ontario, Canada.
Proliferation of lymphocytes from PBL patients in response to antigen challenge in vitro has recently been suggested to be more diagnostically reliable than the precipitin test. Subjects were documented by clinical, x-ray, pulmonary function, and precipitin test criteria: 8 active PBL, 2 nondiseased seropositive breeders, 5 control subjects. Fresh and cryopreserved lymphocytes were tested for proliferation in response to a variety of antigens. A microculture technique and 3H-thymidine uptake quantitated the response. Antigens from pigeon serum and droppings, highly specific and sensitive m the detection of precipitins in these patients’ sera, failed to stimulate cells from 50% of the patients. When cryopreserved lymphocytes were used in this system, even fewer patients responded to antigens, suggesting a subpopulation of cells responding to these antigens was destroyed or inactivated by this procedure. The cells responded signilicantly to phytohemagglutinin confirming viability and responsiveness to mitogens. Controls and nondiseased breeders were all negative. The results indicate lymphocyte proliferation in response to pigeon antigens may be less sensitive than the precipitin test as an index of exposure to the antigen and no more discriminating for diagnosis of active PBL. §upported by Ontario Thoracic Society and London-Middlesex Lung Association.
od trimmer’s disease-an allergic to molds in Swedish saw mills. Lars Belin,
GGteborg,
Sweden.
Repeated febrile episodes with cough, dyspnoea, and malaise was observed in 6 of 34 workers in the trimming department of a Swedish saw mill two years ago. Species of Rhizopus and Paecilomyces were grown from the dust in the working area. Precipitating antibodies and immediate skin reactivity against antigens from these molds could be demonstrated in 5 of the symptomatic workers but also in 17 of those without fever symptoms. Data from the total number (379) of wood trimmers of 17 saw mills now reveal this mold problem at 10 of the places. Here 40% to 80% of the trimmers (166) have precipitating antibodies to the same antigens as compared with 7% of those at the other 7 saw mills. The incidence of fever reactions is 10% to 20%. Increased total IgG and A as well as C-reactive protein in serum and fibrinogen degradation products in the urine are noted after repeated reactions. Environmental studies in two trimming departments have
CLiN. IIWUIJNOL, MARCH i978
revealed mean dust levels of 5 to 1 mg/m3 and outgrowth of lO* colonies, mainly Rhizopus species per gram of dust, Continuous monitoring shows short intense spore peaks in the air with large variations over months as well as over day. A technologically advanced indoor hot air method whereby the wood is dehydrated within a week creates conditions for rapid outgrowth of the mold on the planks, with later release of spores during trimming and sorting.
107. Hypersensitivity pne wood dust contaminated subtiiis, I. L. Bernstein, Wl M.D., P. F. Bonventre, Ph.D., J” S. Gallagher, Ph.D., S. M. Brooks, M.D., Cincinnatir Ohio. We investigated a family, 5 members of which developed recurrent hypersensitivity pneumonitis (HP) symptoms severe enough to require hospitalization in 2 instances. Detailed environmental survey for the known causes of HP was unrevealing. However, in remodeling the house, a natural challenge to old wood dust caused simultaneous illness in 3 family members. Samples of this wood were cultured for bacteria, actinomycetes, fungi, and algae. The predominant isolate on 2 separate occasions was B. subtiZis. After this isolate was washed, sonicated, and mill&ore filtered, an aqueous extract containing 88 pg protein/ml was prepared for in vivo and in vitro tests. Positive immediate skin tests to this extract (0.008 pg protein) were obtained in 2 members. Controlled bronchoprovocation with a total dose of 17.5 pg protein was positive in all 5 family members. An acute obstructive response was observed in I case, while delayed patterns of obstruction, restriction, or diffusion were found in other members. Control skin and bronchial tests in nonatopic subjects were negative. Precipitating and radioallergosorbent test (RAST) antibodies to extracts of B. subtilis isolates or exoenzymes derived from B. subtilis were not present in these cases. Although the precise immunologic pathogenesis of this syndrome is as yet unproved, we believe that this miniepidemic of HP may extend the current panel of causes to include B. subtilis.
108. Pulmonary function in ra BCG-induced lung disease-an animal model of hypersensitivity p R. McGovern, M.D.‘ M. Maron, Ph.D., V. Moore, Ph.D., J. Gara J. Fink, M.D., Milwaukee, Wis. The intravenous injection of killed BCG in oil into rabbits results in chronic granulomatous pulmonary inflammation resembling that observed in several lung diseases in man, e.g., hypersensitivity pneumonitis and sarcoid. This model has been used in the past to study the mechanisms of cell-mediated immunity in the lung. However, tbe effect of
VOLUME 61 WMBER 3
this inflammation on pulmonary function has not been prepiously determined. We measured pulmonary function parameters in New Zealand rabbits 3 weeks after intravenous injection of 200 wg BCG (n = 11) and in normal rabbits (n = 13). Each rabbit was anesthetized with sodium pentobarbital and a tracheostomy and carotid artery catheterization was performed. The animals were then placed in a body plethysmograph for pulmonary function testing. Following the measurements, the lungs were removed for weighing and histologic evaluation. In the BCG-treated rabbits, me pulmonary resistance was 97% greater (p < O.Ol), dynamic compliance was 54% lower (p < 0.05), the A-apOz gradient was 13 torr greater (p < 0.001) and the lung wt/body wt ratios were 152% greater (p < 0.001) than those in the normal rabbits. Histologic evaluation revealed diffuse involvement with noncaseating granulomas. It appears that the pulmonary function changes are qualitatively similar to those observed in human lung diseases characterized by chronic granulomatous inflammation.
09. Thermophiiic actinomycetes in outdoor urban air. l-4. P. Burge, Ph.D., W. R. Solomon, M.D., and J. R. Boise,
B.S., Ann Arbor,
oakeoiar
iavage irk
r, M.D., H. Richard Richard Stankus, Ph.D., J. Kaimel J= E. Salvaggio, M.D., Cleveland, New Orleans, La. Silica inhaiation by animals alters pulmonary macrophage function. We studied the effects of silica inhalation in man on cells obtained by bronchoalveoiar lavage. Twelve limited bronchoalveolar lavages from patients with silicosis were compared with 11 lavages from age, sex, and cigarette smoking-matched control subjects. We determined the following characteristics of the cells: (I) number and type recovered; (2) percent viable; (3) percent which adbered to a plastic surface; (4) ability to phagocytose and kill (I and 2 hr later) Listeria monocytogenes; and (5) capacity of the nonadherent cellular population to cause macropbage migration inhibition (MMI) when mixed with guinea pig peritoneal macrophages and exposed to tuberculin and streptokinase-streptodomase (SK-SD). There were no significant group differences in any of the above parameters with the exception of MMI. Cells from only 1 of 5 patients with silicosis exposed to SK-SD induced MMI, compared with 7 of 11 control subjects (p < O.Q5). Thus, we found evidence of lymphocyte and not macrophage dysfunction in samples emanating from the lung in human silicosis.
Mich.
Although discrete sources of thermophilic actinomycetes (TA) are associated clearly with illness, broader occurrence patterns for implicated agents remain unexplored. Thus, an ongoing cultural survey of these organisms in free air on an urban rooftop was begun. All collections have used single plates of trypticase soy agar exposed beneath stage 6 of a wind-oriented Andersen sampler and incubation at 50’ C for 1 to 7 days. Subculture of all TA isolates on several media was attempted. Exposure periods, initially 30 min, were later reduced to 10 min to !imit bacterial overloading. Single samples were obtained between 1,000 and 1,200 hr on 36 days and in diurnal series at 0800? 1200, 1600, and 2000 hr on 18 additional days. Interval analysis of 112 resulting samples, evenly spaced from August, 1976, to September, 1977, has been completed. TA reproductive units were recovered in 45% of total samples and in 76% of 30 min collections. However, low ambient TA levels were indicated, with a peak of 17 units/m~. Of 84 isolates, 25 were Thermoactinomyces candidus, 14 T. vulgaris, 5 Micropolyspora faeni, and 1 Saccharomonospora viridis. In 28 cases involving at least 4 morphologic types, isolates were unlike all described taxa. Ten of 11 isolates that resisted subculture were clearly not of me 4 identified species. TA recoveries varied little seasonally and were most prominent from 1,000 to 1,200 hr. Disease-associated TAs are present perennially in free air. Since ambient levels are low, points of greater prevalence should suggest a nearby source of TAs and a search for health effects.
11 I. Comparison of oral ami~o~byili~e terbutaline, separately and in combina~~ in asthmatic patients. J. D. Wolfer .D.r and D. P. Tashkin, M.D., Baltimore, Los Angeles, Calif.
Md.? and
Using a double-blind crossover design, we compared in 17 adult asthmatic patients the acute cardiac (heart rate, blood pressure) and bronchial (spirometry, plethysmography) effects of the following drugs or drug combinations: terbutaline, 5 mg; terbulatine, 2.5 mg, plus aminopbylline, 200 mg; aminophylline, 400 mg; terbutaline, 5 mg, plus aminophylline, 400 mg; terbutaline, 2.5 mg, aminopbylline, 200 mg; terbutaline, 2.5 mg, plus a~nophylline, 200 mg; and placebo. Mean peak theophylline plasma levels during study days when patients received theophylline were 12 kg/ml and 6.4 pg/ml for the 4OQ mg and 200 mg doses of aminophylline, respectively. Terbutaline, 5 mg, and aminophylline, 400 mg, administered separately were found to produce comparable bronchodilator and chronotropic effects and a similar frequency of adverse side effects. Terbutaline, 2.5 mg, and aminophylline, 200 mg, alone were also found to have similar effects to one another. The combination of both drugs in higher doses produced a significantly greater degree of bronchodilatation and an increase in beart rate than either drug alone in the same dosage. The combination of botb drugs in the lower doses had cardiopulmonary effects which
on of cell-mediated immunity eder’s lung (PBlJ using fresh opreserwed lymphocytes. J. Anderson, B.A8 London,
J. Toogood, M.D., N. Lefcoe, M.D., R. T., J. Mazza, M.D., B. Jennings, Ontario, Canada.
Proliferation of lymphocytes from PBL patients in response to antigen challenge in vitro has recently been suggested to be more diagnostically reliable than the precipitin test. Subjects were documented by clinical, x-ray, pulmonary function, and precipitin test criteria: 8 active PBL, 2 nondiseased seropositive breeders, 5 control subjects. Fresh and cryopreserved lymphocytes were tested for proliferation in response to a variety of antigens. A microculture technique and 3H-thymidine uptake quantitated the response. Antigens from pigeon serum and droppings, highly specific and sensitive m the detection of precipitins in these patients’ sera, failed to stimulate cells from 50% of the patients. When cryopreserved lymphocytes were used in this system, even fewer patients responded to antigens, suggesting a subpopulation of cells responding to these antigens was destroyed or inactivated by this procedure. The cells responded signilicantly to phytohemagglutinin confirming viability and responsiveness to mitogens. Controls and nondiseased breeders were all negative. The results indicate lymphocyte proliferation in response to pigeon antigens may be less sensitive than the precipitin test as an index of exposure to the antigen and no more discriminating for diagnosis of active PBL. §upported by Ontario Thoracic Society and London-Middlesex Lung Association.
od trimmer’s disease-an allergic to molds in Swedish saw mills. Lars Belin,
GGteborg,
Sweden.
Repeated febrile episodes with cough, dyspnoea, and malaise was observed in 6 of 34 workers in the trimming department of a Swedish saw mill two years ago. Species of Rhizopus and Paecilomyces were grown from the dust in the working area. Precipitating antibodies and immediate skin reactivity against antigens from these molds could be demonstrated in 5 of the symptomatic workers but also in 17 of those without fever symptoms. Data from the total number (379) of wood trimmers of 17 saw mills now reveal this mold problem at 10 of the places. Here 40% to 80% of the trimmers (166) have precipitating antibodies to the same antigens as compared with 7% of those at the other 7 saw mills. The incidence of fever reactions is 10% to 20%. Increased total IgG and A as well as C-reactive protein in serum and fibrinogen degradation products in the urine are noted after repeated reactions. Environmental studies in two trimming departments have
CLiN. IIWUIJNOL, MARCH i978
revealed mean dust levels of 5 to 1 mg/m3 and outgrowth of lO* colonies, mainly Rhizopus species per gram of dust, Continuous monitoring shows short intense spore peaks in the air with large variations over months as well as over day. A technologically advanced indoor hot air method whereby the wood is dehydrated within a week creates conditions for rapid outgrowth of the mold on the planks, with later release of spores during trimming and sorting.
107. Hypersensitivity pne wood dust contaminated subtiiis, I. L. Bernstein, Wl M.D., P. F. Bonventre, Ph.D., J” S. Gallagher, Ph.D., S. M. Brooks, M.D., Cincinnatir Ohio. We investigated a family, 5 members of which developed recurrent hypersensitivity pneumonitis (HP) symptoms severe enough to require hospitalization in 2 instances. Detailed environmental survey for the known causes of HP was unrevealing. However, in remodeling the house, a natural challenge to old wood dust caused simultaneous illness in 3 family members. Samples of this wood were cultured for bacteria, actinomycetes, fungi, and algae. The predominant isolate on 2 separate occasions was B. subtiZis. After this isolate was washed, sonicated, and mill&ore filtered, an aqueous extract containing 88 pg protein/ml was prepared for in vivo and in vitro tests. Positive immediate skin tests to this extract (0.008 pg protein) were obtained in 2 members. Controlled bronchoprovocation with a total dose of 17.5 pg protein was positive in all 5 family members. An acute obstructive response was observed in I case, while delayed patterns of obstruction, restriction, or diffusion were found in other members. Control skin and bronchial tests in nonatopic subjects were negative. Precipitating and radioallergosorbent test (RAST) antibodies to extracts of B. subtilis isolates or exoenzymes derived from B. subtilis were not present in these cases. Although the precise immunologic pathogenesis of this syndrome is as yet unproved, we believe that this miniepidemic of HP may extend the current panel of causes to include B. subtilis.
108. Pulmonary function in ra BCG-induced lung disease-an animal model of hypersensitivity p R. McGovern, M.D.‘ M. Maron, Ph.D., V. Moore, Ph.D., J. Gara J. Fink, M.D., Milwaukee, Wis. The intravenous injection of killed BCG in oil into rabbits results in chronic granulomatous pulmonary inflammation resembling that observed in several lung diseases in man, e.g., hypersensitivity pneumonitis and sarcoid. This model has been used in the past to study the mechanisms of cell-mediated immunity in the lung. However, tbe effect of
VOLUME 61 WMBER 3
this inflammation on pulmonary function has not been prepiously determined. We measured pulmonary function parameters in New Zealand rabbits 3 weeks after intravenous injection of 200 wg BCG (n = 11) and in normal rabbits (n = 13). Each rabbit was anesthetized with sodium pentobarbital and a tracheostomy and carotid artery catheterization was performed. The animals were then placed in a body plethysmograph for pulmonary function testing. Following the measurements, the lungs were removed for weighing and histologic evaluation. In the BCG-treated rabbits, me pulmonary resistance was 97% greater (p < O.Ol), dynamic compliance was 54% lower (p < 0.05), the A-apOz gradient was 13 torr greater (p < 0.001) and the lung wt/body wt ratios were 152% greater (p < 0.001) than those in the normal rabbits. Histologic evaluation revealed diffuse involvement with noncaseating granulomas. It appears that the pulmonary function changes are qualitatively similar to those observed in human lung diseases characterized by chronic granulomatous inflammation.
09. Thermophiiic actinomycetes in outdoor urban air. l-4. P. Burge, Ph.D., W. R. Solomon, M.D., and J. R. Boise,
B.S., Ann Arbor,
oakeoiar
iavage irk
r, M.D., H. Richard Richard Stankus, Ph.D., J. Kaimel J= E. Salvaggio, M.D., Cleveland, New Orleans, La. Silica inhaiation by animals alters pulmonary macrophage function. We studied the effects of silica inhalation in man on cells obtained by bronchoalveoiar lavage. Twelve limited bronchoalveolar lavages from patients with silicosis were compared with 11 lavages from age, sex, and cigarette smoking-matched control subjects. We determined the following characteristics of the cells: (I) number and type recovered; (2) percent viable; (3) percent which adbered to a plastic surface; (4) ability to phagocytose and kill (I and 2 hr later) Listeria monocytogenes; and (5) capacity of the nonadherent cellular population to cause macropbage migration inhibition (MMI) when mixed with guinea pig peritoneal macrophages and exposed to tuberculin and streptokinase-streptodomase (SK-SD). There were no significant group differences in any of the above parameters with the exception of MMI. Cells from only 1 of 5 patients with silicosis exposed to SK-SD induced MMI, compared with 7 of 11 control subjects (p < O.Q5). Thus, we found evidence of lymphocyte and not macrophage dysfunction in samples emanating from the lung in human silicosis.
Mich.
Although discrete sources of thermophilic actinomycetes (TA) are associated clearly with illness, broader occurrence patterns for implicated agents remain unexplored. Thus, an ongoing cultural survey of these organisms in free air on an urban rooftop was begun. All collections have used single plates of trypticase soy agar exposed beneath stage 6 of a wind-oriented Andersen sampler and incubation at 50’ C for 1 to 7 days. Subculture of all TA isolates on several media was attempted. Exposure periods, initially 30 min, were later reduced to 10 min to !imit bacterial overloading. Single samples were obtained between 1,000 and 1,200 hr on 36 days and in diurnal series at 0800? 1200, 1600, and 2000 hr on 18 additional days. Interval analysis of 112 resulting samples, evenly spaced from August, 1976, to September, 1977, has been completed. TA reproductive units were recovered in 45% of total samples and in 76% of 30 min collections. However, low ambient TA levels were indicated, with a peak of 17 units/m~. Of 84 isolates, 25 were Thermoactinomyces candidus, 14 T. vulgaris, 5 Micropolyspora faeni, and 1 Saccharomonospora viridis. In 28 cases involving at least 4 morphologic types, isolates were unlike all described taxa. Ten of 11 isolates that resisted subculture were clearly not of me 4 identified species. TA recoveries varied little seasonally and were most prominent from 1,000 to 1,200 hr. Disease-associated TAs are present perennially in free air. Since ambient levels are low, points of greater prevalence should suggest a nearby source of TAs and a search for health effects.
11 I. Comparison of oral ami~o~byili~e terbutaline, separately and in combina~~ in asthmatic patients. J. D. Wolfer .D.r and D. P. Tashkin, M.D., Baltimore, Los Angeles, Calif.
Md.? and
Using a double-blind crossover design, we compared in 17 adult asthmatic patients the acute cardiac (heart rate, blood pressure) and bronchial (spirometry, plethysmography) effects of the following drugs or drug combinations: terbutaline, 5 mg; terbulatine, 2.5 mg, plus aminopbylline, 200 mg; aminophylline, 400 mg; terbutaline, 5 mg, plus aminophylline, 400 mg; terbutaline, 2.5 mg, aminopbylline, 200 mg; terbutaline, 2.5 mg, plus a~nophylline, 200 mg; and placebo. Mean peak theophylline plasma levels during study days when patients received theophylline were 12 kg/ml and 6.4 pg/ml for the 4OQ mg and 200 mg doses of aminophylline, respectively. Terbutaline, 5 mg, and aminophylline, 400 mg, administered separately were found to produce comparable bronchodilator and chronotropic effects and a similar frequency of adverse side effects. Terbutaline, 2.5 mg, and aminophylline, 200 mg, alone were also found to have similar effects to one another. The combination of both drugs in higher doses produced a significantly greater degree of bronchodilatation and an increase in beart rate than either drug alone in the same dosage. The combination of botb drugs in the lower doses had cardiopulmonary effects which