Download Report

therapy (ie, IV augmentation therapy with purified
arantiprotease) and by emerging data suggesting the
biochemical and clinical efficacy of augmentation therapy.
design and methods. Chest 1994; 106:1223-33
Kok-Jensen A, Dirksen A. Decline in FEV2
among patients with severe hereditary alpha 1-antitrypsin defi¬
cific
21 Seersholm N,
ciency type Pi Z. Am J Respir Crit Care Med 1995; 152:1922-25
EK, Pierce JA, Province MA, et al. Variability of
pulmonary function in alpha 1-antitrypsin deficiency: clinical
22 Silverman
References
correlates'. Ann Intern Med 1989; 111:982-91
1 Laurell
CB, Eriksson S. The electrophoretic a1-globulin
of
pattern serum in aj-antitrypsin deficiency. Scand J Clin
Lab Invest 1963; 15:132-40
2 Brantly M, Nukiwa T, Crystal RG. Molecular basis of alpha
1-antitrypsin deficiency. Am J Med 1988; 84(suppl 6A):13-31
3 Lomas DA, Evans DL, Finch JT, et al. The mechanism of Z
aj-antitrypsin accumulation in the liver. Nature 1992; 357:
23 Fallat
RJ. Reactive ainvay disease and 1AT deficiency. In:
Crystal RG, ed. Alpha 1-antitrypsin deficiency; biology:
pathogenesis, clinical manifestations, therapy. New York:
Marcel Dekker, 1996; 259-79
Gishen P, Saunders AJS, Tobin MJ, et al. Alpha 1-antitrypsin
deficiency: the radiological features of pulmonary emphy¬
sema in subjects of Pi-type Z and Pi-type SZ: a survey by the
British Thoracic Association. Clin Radiol 1982; 33:371-77
Shin MS, Ho K-J. Bronchiectasis in patients with alphar
24
y
605-07
Higgins MW, Thorn T. Incidence,
intra- and intercountry differences. In:
prevalence, and mortality:
Hensley MJ, Saunders
MA, eds. Clinical epidemiology of chronic obstructive pul¬
monary disease. New York: Marcel Dekker, 1989
5 Lieberman J, Winter B, Sastre A. Alpha 1-antitrypsin P-types
in 965 COPD patients. Chest 1986; 89:370-73
6 Sveger T. Liver disease in alpha 1-antitrypsin deficiency
detected by screening of 200,000 infants. N Engl J Med 1976;
294:1316-21
7 O'Brien ML, Buist NR, Murphey WH. Neonatal screening
for alpha 1-antitrypsin deficiency. J Pediatr 1978; 92:1006-10
8 Silverman EK, Miletich JP, Pierce JA, et al. Alpha 1-antitryp¬
sin deficiency: high prevalence in the St. Louis area deter¬
mined by direct population screening. Am Rev Respir Dis
4
'
9
1989; 140:961-66
Spence WC, Morris JE, Pass K, et al. Molecular confirmation
of alpha 1-antitrypsin genotypes in newborn dried blood
specimens. Biochem Med Metab Biol 1993; 50:233-40
MJ, Cook PJ, Hutchinson DC. Alpha 1-antitrypsin
deficiency: the clinical and physiological features of pulmo¬
nary emphysema in subjects homozygous for Pi type Z: a
survey by the British Thoracic Association. Br J Dis Chest
1983; 77:14-27
11 Stoller JK, Smith P, Yang P, et al. Physical and social impact
of alpha 1-antitrypsin deficiency: results of a mail survey of
the readership of a national newsletter. Cleve Clin J Med
1994; 61:461-67
12 Cox DW. Alpha 1-antitrypsin: a guardian of vascular tissue.
Mayo Clin Proc 1994; 69:1123-24
13 Hussain M, Mieli-Vergani G, Mowat AP. Alpha 1-antitrypsin
deficiency and liver disease: clinical presentation, diagnosis
and treatment. J Inher Metab Dis 1991; 14:497-511
14 Larsson C. Natural histoiy and life expectancy in severe alpha
1-antitrypsin deficiency, Pi Z. Acta Med Scand 1978; 204:345-51
15 Janus ED, Phillips NT, Carrell RW. Smoking lung function,
and alpha 1-antitrypsin deficiency. Lancet 1985; 1:152-54
16 Brantly ML, Paul LD, Miller BH, et al. Clinical features and
history of the destructive lung disease associated with alpha
1-antitrypsin deficiency of adults with pulmonary symptoms.
Am Rev Respir Dis 1988; 138:327-36
17 Gadek JE, Crystal RG. Alpha 1-antitrypsin deficiency. In:
Stanbury JB, Wyngaarden JB, Frederickson DS, et al, eds.
25
antitrypsin deficiency:
26 El-Kassimi
trypsin
McGraw-Hill, 1983; 1450-67
18
Brantly M. In: Report of a WHO meeting on alpha 1-anti¬
trypsin deficiency. Geneva: Human Genetics Programme
Division of Noncommunicable Diseases, World Health Or¬
ganization, March 18-20, 1996
19 Wu MC, Eriksson S. Lung function, smoking, and survival in
severe alpha 1-antitrypsin deficiency, Pi ZZ. J Clin Epidemiol
1988; 41:1157-65
20 Alpha1-Antitrypsin Deficiency Registry Study Group. A reg¬
istry of patients with severe deficiency of alpha 1-antitrypsin:
128S
28
29
30
levels in
widespread
et al. Alpha 1-anti¬
bronchiectasis. Respir
Colp
C, Pappas J, Moran D, et al. Variants of alphar
antitrypsin in Puerto Rican children with asthma. Chest 1993;
103:812-15
Wall M, Moe E, Eisenberg J, et al. Long-term follow-up of a
cohort of children with alpha 1-antitrypsin deficiency. J Pe¬
diatr 1990; 116:248-51
Sveger T, Plitulainen E, Arborelius M. Lung function in
adolescents with alpha 1-antitrypsin deficiency. Acta Paediatr
1994; 83:1170-73
Seersholm N, Kok-Jensen A, Dirksen A. Survival of patients
with severe alpha 1-antitrypsin deficiency with special refer¬
ence
to
non-index cases. Thorax 1994; 49:695-98
In Vivo Adenovirus-Mediated
Expression of Human
Pre-Elafin, a Potent Neutrophil
Elastase Inhibitor*
Jean-Michel Sallenave, PhD;f Zhou Xing, PhD;
Frank Graham, PhD; and Jack Gauldie, PhD
(CHEST 1997; 11L128S-129S)
TT uman neutrophil elastase is thought to be involved in
-¦-¦¦-lung diseases such as emphysema or cystic fibrosis.
We have recently characterized an elastase-specific inhib¬
itor
(elafin) as a potent human neutrophil elastase inhibi¬
tor in
the lung. The gene codes for a secreted active 10 kd
protein, pre-elafin (PE), which can be cleaved in secre¬
tions, giving rise to a 6 kd product, elafin. A 538-base pair
(bp) fragment containing the endogeneous leader and
polyA sequences of the PE gene were inserted into the El
deleted region (replication-deficient) of adenovirus (Ad)5
under the control of either the human cytomegalovirus
(HCMV) promoter (830 bp), the Ad2-major late promoter
(MLP), or the mouse cytomegalovirus (MCMV) promoter.
*From the
Department of Pathology, McMaster University,
at the University of Edinburgh Medical School,
Hamilton, Ontario, Canada.
Currently
Scotland.
Thomas L.
Downloaded From: http://journal.publications.chestnet.org/ on 10/28/2014
occurrence? Chest 1993; 104:
FA, Warsy AS, Uz-Zaman A,
serum
Med 1989; 83:119-21
27
10 Tobin
The metabolic basis of inherited disease. 5th ed. New York:
a rare
1384-86
Petty 39th Annual Aspen Lung Conference: Genes and Gene Therapy
We have previously shown in vitro that MCMV drives the
synthesis of PE more efficiently than HCMV or MLP,
both in human and rodent epithelial cells and fibroblasts.
The aim of this study was to assess in vivo the comparative
strength of these promoters.
Three replication-deficient recombinant Ad5 viruses
(HCMV-PE, MLP-PE, MCMV-PE) were isolated and ti¬
trated. Sprague-Dawley rats were injected intratracheally
with 0.3 mL of phosphate-buffered saline containing either
3X108 (n=2), 1.5X109 (n=2),
3X109 (n=2)
plaque
the lungs were lavaged
forming
days,
with phosphate-buffered saline. The BAL fluids were centri¬
fuged and supernatant content of PE was assessed by
enzyme-linked immunosorbent assay (ELISA). The left lung
was snap-frozen in liquid nitrogen for Northern blot analysis
of PE messenger RNA'(mRNA).
Northern blot analysis of PE mRNA from lungs infected
with HCMV-PE and MLP-PE (days 2 and 5) was negative.
In contrast, MCMV-PE-infected lungs showed significant
levels at day 5. These data were confirmed at the protein
level by ELISA whereby BAL recovered from MCMV-PEinfected lungs (increasing doses) showed very high levels of
PE at day 5 (71.1, 54.3, and 66.6 ng/mL, respectively).
Interestingly, levels at day 2 were much lower (3.1, 13.7, and
6.9 ng/mL, respectively). No detectable levels (<0.5 ng/mL)
of PE protein were present in BAL from lungs infected with
HCMV-PE and MLP-PE (day 2 or day 5).
Our data show that, compared to HCMV and MLP, MCMV
is a very potent promoter in rat lung tissue. We were able to
detect significant levels of bodi PE mRNA and protein. We
believe that our findings have important implications for the
efficiency of gene transfer of anti-inflammatory molecules such
as PE in rodent models of lung inflammation.
units. After either 2 or 5
Rat
or
Prostacyclin Synthase*
Cloning and Regulation of Gene
Expression in the Lung
Mark Geraci, MD; David
Shepherd, BS; Mark Moore, BA;
Jennifer Vernon, BS; Jenny Allard, BA; John Shannon, PhD;
and Norbert F.
Voelkel, MD
(CHEST 1997; 111:129S)
"D rostacyclin synthase (PGIS) is the final committed
-*- enzymatic
step in the pathway of prostacyclin (PGI2)
production, occurring at a branch point where substrate
(PGH2) can be directed either toward prostacyclin, which
affords protection, or thromboxane, which augments injury.
The imbalance of prostacyclin and thromboxane is involved
in the pathogenesis of several diseases, such as pulmonary
hypertension, acute lung injury, and atherosclerosis. These
studies were designed to pursue the molecular cloning and
characterization of rat prostacyclin synthase.
An 1153-base pair fragment of the rat PGIS gene was
cloned and sequenced in its entirety. This fragment repre*From the Division of Pulmonary Sciences, University of Colo¬
rado Health Sciences Center, Denver.
predicted full complementary DNA se¬
quence and contains a complete open reading frame. To
study regulation of PGIS gene expression, this fragment was
used as a probe for Northern blot analysis under a variety of
conditions. Studies were performed to determine rat tissuespecific expression and expression during rat lung develop¬
ment. The in vivo effects of steroid hormones on lung PGIS
expression were examined in animals treated with corticoste¬
roids, testosterone, and estrogen. The effects of antioxidant
and lipopolysaccharide (LPS) administration on lung PGIS
expression were evaluated. The isolated-perfused rat lung (a
model of shear-stress) was used to elucidate the outcome of
differing flow rates on lung PGIS expression.
The results of these studies indicate that the PGIS
sequence across species is highly conserved. The rat gene
is most strongly expressed in heart and lung, and recog¬
nizes a major messenger RNA species of 2.0 kilobase (kb)
with two minor bands of 2.9 and 5.6 kb. Skeletal muscle
shows a unique band of 4.4 kb. In lung development, PGI2
synthase is upregulated late in gestation at day 21. PGIS
gene expression is induced by corticosteroids, antioxidants, LPS, and shear stress. Testosterone increases gene
expression, while estrogen has the opposite effect. These
studies demonstrate the following: (1) rat PGI2 synthase is
closely homologous to the bovine and human genes; (2)
multiple forms of transcripts exist and may represent
alternative splicing or distinct isoforms; (3) the gene is
expressed in rat lung late in gestation; (4) PGIS expression
is induced by shear stress; (5) unlike eyclooxygenase, PGIS
is induced by corticosteroids and antioxidants; and (6)
testosterone increases gene expression while estrogen
decreases expression. Using the rat model will enable
examination of gene expression under a variety of condi¬
tions and offers the potential for gene therapy trials.
sents
77% of die
Surfactant Protein-B
Deficiency*
Lawrence M.
Nogee, MD
(CHEST 1997; 11L129S-135S)
TD ulmonary surfactant is the mixture of lipids and pro-
-¦"
teins needed to reduce alveolar surface tension at the
air-liquid interface and to prevent end-expiratory atelec¬
tasis. An inability to produce sufficient quantities of
surfactant due to immaturity is the primary cause of the
respirator)' distress syndrome (RDS) observed in infants
born prematurely, and diminished surfactant function
contributes to the pathophysiologic condition of ARDS.1-2
While the phospholipid components of pulmonary surfac¬
tant are important in its surface tension-lowering proper¬
ties, specific surfactant proteins have been identified that
*From the Department of Pediatrics, Johns Hopkins University
School of Medicine, Baltimore.
Supported
by a Basil O'Conner Research Award from the March
of Dimes, and National Institutes of Health grant HL-54703.
Reprint
requests: Lawrence M. Nogee, MD, Division ofNeonatology,
CMSC 210, The Johns Hopkins Hospital, 600 N Wolfe St, Baltimore
MD 21287-3200; entail:
lnogee@welchlink.welch.jhu.edu
CHEST/111/6/JUNE, 1997 SUPPLEMENT
Downloaded From: http://journal.publications.chestnet.org/ on 10/28/2014
129S