Pulmonary Alveolar Macrophages Coated Asbestos ...

Comparison of Phagocytosis of Uncoated Versus Coated Asbestos Fibers by Cultured Human Pulmonary Alveolar Macrophages Theodore L. McLemore, Paul M. Stevens, Victor Roggli, S. Donald Greenberg, Milton V. Marshall and E. Clinton Lawrence Chest 1981;80;39S-42S DOI 10.1378/chest.80.1_Supplement.39S The online version of this article, along with updated information and services can be found online on the World Wide Web at: http://chestjournal.chestpubs.org/content/80/1_Supplement/39S.citation

Chest is the official journal of the American College of Chest Physicians. It has been published monthly since 1935. Copyright1981by the American College of Chest Physicians, 3300 Dundee Road, Northbrook, IL 60062. All rights reserved. No part of this article or PDF may be reproduced or distributed without the prior written permission of the copyright holder. (http://chestjournal.chestpubs.org/site/misc/reprints.xhtml) ISSN:0012-3692

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achieved by prior culture for five days in DMEM + 0.5% FCS) and fibroblast cell counts determined after one, two, and four days. DMEM + 0.5% FCS asbestos incubated for four hours as controls. Supernatants from unexposed PAM had no influence on fibroblast replication rate. In contrast, supernatants from PAM exposed to asbestos significantly increased the rate of fibroblasts doubling; the doubling time decreasing five-fold from a control of 310±21 hours to 54±4 hours, PI.1

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CULTURE TIME (HOURS) Ficunx 1. Effect of uncoated asbestos (ASB) or ASB bodies on viability of cultured human pulmonary alveolar macrophages (PAMs). Cells were cultured 24-72 hours with 0-300 ,g/ml naked ainosite ASB or 0-250 g/ml ASB bodies. Viability was then determined by the trypan blue dye exclusion method. Uncoated ASB (n = 5): No cytotoxicity was observed following culture of cells with 100 Lg/ml ASB for 24 hours. (P ) 0.05 in all instances; paired, 2-tailed Student’s t-test). Significant cytotoxicity was observed for all other concentrations or longer culture intervals with naked ASB (P .( 0.006 in all instances). ASB bodies (N = 7): Decreased PAM viability was observed following 24 hours of culture with ASB bodies (P < 0.032). All other values were not significant (P > 0.05 in all instances). Bars = mean ± SE. MATERIALS

Study

METHODS

AND

and operating at 40 kV. Micrographs were a 2000 line CRT on polaroid P/N 55 film.

from

Subjects

PAMs were obtained from 15 normal healthy, non-smoking volunteers, 21-29 years of age, who had no previous history of cigarette smoking or occupational exposure to ASB or other particulates. ASB bodies were obtained from necropsy lung tissue from three ASB insulation workers, 62-78 years of age.

Preparation PAMS ml sterile choscope

of PAMs

RESULTS PAMs obtained were cultured for tions

of

by

by

nique,

ASB

were obtained employing 0.15 M NaC1 through and prepared as previously

saline lavage with 250 a flexible fiberoptic brondescribed.

of PAMs

with

ASB

toxicity However,

Scanning

Electron

Microscopy

(SEM)

Procedures

For SEM studies, PAMs were incubated for the appropriate time interval on glass cover slips and processed as previously described. Cells were then examined with a JOEL 100 CX electron microscope equipped with ASID

23RD ASPEN LUNG CONFERENCE

(P

When viability

method)

24 hours

of culture

signfficant

ASB

contrast

to

the

naked

250 trypan

ASB,

ASB

ASB in

blue the

dye initial

cytotoxicity

PAMs of 48 studies

of

decrease

further

when periods These

hours

,ig/nil

after No

At PAM

significant 24

a slight

by

however,

for the longer time 0.05 in all instances). in

with

0.032).

ig/ml), and

1),

48

t-test).

initial

demonstrated =

cytoculture. followfor

2-tailed (>100

be ASB

concentrations

paired,

(Fig

was

to

lower

hours of decreased

incubated

(P

At

24

determined

demonstrated,

bated (P>

1).

decreased

hours

tech-

shown

no

ASB

deter-

this

the first significanfly

of

(as

exclusion

that

(Fig

0.005,

were

24-72

as

was

was observed after the < 0.005 in all instances). PAMs

for

evaluated

Using

PAMs

proportionately

bodies

then

,Lg/ml),

low