Increased Expression of Leukocyte Function Associated Antigen-1 (LFA-1) and Intercellular Adhesion Molecule-1 (ICAM-1) by Alveolar Macrophages of Patients with Pulmonary Sarcoidosis

doi:10.1378/chest.100.4.910

Chest

Volume 100, Issue 4, October 1991, Pages 910-916

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Leukocyte function associated antigen-1 (LFA-1) and its ligand intercellular adhesion molecule-1 (ICAM-1) are cell adhesion molecules that play an important role in the capacity of mononuclear phagocytes (MPs) to present antigens to T lymphocytes. Since in pulmonary sarcoidosis (PS) this capacity is increased at sites of disease activity, we studied the expression of LFA-1 and ICAM-1 on peripheral blood monocytes (BMs) and alveolar macrophages (AM) obtained by bronchoalveolar lavage (BAL) from normal subjects (n = 7) and patients with PS (n = 14). To accomplish this, immunocytochemical stainings were made on cytocentrifuge preparations using anti-LFA-1 (anti-CD 11a) and anti-ICAM-1 (anti-CD 54) monoclonal antibodies (MoAbs). Normal and sarcoid BMs displayed a high percentage of positivity with both MoAbs with no difference between study groups (LFA-1: control BM 87.8 ± 8.8 percent; sarcoid BM 84.7 ±9.5 percent; ICAM-1: control BM 80.8 ±10 percent; sarcoid BM 88.0 ±4.2 percent; p = NS for all comparisons). In both groups the percentage of cells expressing LFA-1 and ICAM-1 molecules among AMs was lower than among autologous BMs (LFA-1: control AM 46.5 ±13.2 percent, p<0.001 vs control BM; sarcoid AM 64.2 ± 15.9; p<0.001 vs sarcoid BM) (ICAM-1: control AM 42.7 ±8.5 percent, p<0.001 vs control BM; sarcoid AM 72.1 ±10.6, p<0.001 vs sarcoid BM). AMs from patients with PS showed a higher degree of positivity for LFA-1 and ICAM-1 than normal AMs (p<0.02 and p<0.001, respectively). The positivity for LFA-1 and ICAM-1 molecules on sarcoid AMs was not correlated with the positivity for two different BM-associated markers (te, the CD 11b and the CD 14 molecules) and was not correlated with the percentage of T lymphocytes in BAL, selected as a marker of the intensity of the alveolitis. These results suggest that the increased ability of sarcoid AMs to induce the proliferation of T lymphocytes may be related, at least in part, to the increased expression of LFA-1 and ICAM-1 molecules on their surfaces.

Section snippets

Study Population

The diagnosis of PS was established in 14 consecutive nonsmoking patients by using previously described criteria, including transbronchial or open lung biopsy specimens.²³ As controls, seven normal nonsmoking volunteers were studied. They had no history of pulmonary disease and had normal chest roentgenograms and pulmonary function test results. Details on the two groups of subjects are given in Table 1.

Isolation of Lung and Blood MP

Bronchoalveolar lavage (BAL) cells were obtained by using previously described procedures,

Total and Differential Cell Counts

Table 3 shows the data on BAL fluid recovery, total and differential cell counts. As expected, patients with PS had increased numbers of total cells and increased percentage of T lymphocytes in BAL (p<0.01, for both comparisons). Total cell counts and percentage of T cells in BAL fluid did not correlate with any clinical parameter.

Immunocytologic Stainings of BM and AM

BMs and AMs were identified by morphologic criteria and positive cells were recognized by red staining of the cell membrane. The data are expressed as percentage of

DISCUSSION

The leukocyte adhesion molecules LFA-1 and ICAM-1 play an important role in the intercellular communications during immune responses. The main finding of this study was that in PS, the percentage of AMs expressing LFA-1 and ICAM-1 molecules is increased compared with AMs of control subjects. The increased expression of these adhesion molecules by sarcoid AMs was not correlated with the expression of two different monocyte lineage markers and was not correlated with the percentage of T

ACKNOWLEDGMENTS:

The authors thank Dr. T. A. Springer (Department of Pathology, Harvard Medical School, Boston, MA) for providing the anti RR 1/1 MoAb, Dr. M. R. Bonsignore for critically reviewing the manuscript, and Drs. F. Tinè and A. Melodia for help in the statistical evaluation of the results.

REFERENCES (35)

SpataforaM et al.
Lung compartmentalization of increased TNF releasing ability by mononuclear phagocytes in pulmonary sarcoidosis.
Chest
(1989)
MartzE.
LFA-1 and other accessory molecules functioning in adhesion of T and B lymphocytes.
Hum Immunol
(1987)
MaliziaG et al.
Expression of leukocyte adhesion molecules by mucosal mononuclear phagocytes in inflammatory bowel disease: immunohistological evidence for enhanced antigen presenting capacity.
Gastroenterology
(1991)
CrystalRG et al.
Interstitial lung disease of unknown cause: disorders characterized by chronic inflammation of the lower respiratory tract.
N Engl J Med
(1984)
SemenzatoG.
The immunology of sarcoidosis.
Semin Respir Med
(1986)
HunninghakeGW et al.
Pulmonary sarcoidosis: a disorder mediated by excess helper T lymphocyte activity at sites of disease activity.
N Engl J Med
(1981)
DanieleRP et al.
Pathogenesis of sarcoidosis: state of the art.
Chest
(1986)
BittermanPB et al.
Mechanisms of pulmonary fibrosis: spontaneous release of alveolar macrophage derived growth factor in interstitial lung disorders.
J Clin Invest
(1981)
HunninghakeGW.
Release of interleukin 1 by alveolar macrophages of patients with pulmonary sarcoidosis.
Am Rev Respir Dis
(1984)
RobinsonBWS et al.
Gamma interferon is spontaneously released by alveolar macrophages and lung T lymphocytes in patients with pulmonary sarcoidosis.
J Clin Invest
(1985)

LemVM et al.

Bronchoalveolar cells from sarcoid patients demonstrate enhanced antigen presentation.

J Immunol

(1985)

VenetA et al.

Enhanced alveolar macrophage mediated antigen induced T lymphocyte proliferation in sarcoidosis.

J Clin Invest

(1985)

SchwartzRH.

T lymphocyte recognition of antigen in association with gene products of the major histocompatibility complex.

Annu Rev Immunol

(1985)

UnanueER et al.

The basis for the immunoregulatory role of macrophages and other accessory cells.

Science

(1987)

Sanchez-MadridF et al.

A human leukocyte differentiation antigen family with distinct alpha subunits and a common beta subunit: the lymphocyte function associated antigen 1 (LFA-1), the C3bi complement receptor (OKM 1-Mac 1), and the p 150,95 molecule.

J Exp Med

(1983)

KnappW et al.

CD antigens 1989.

Int J Cancer

(1989)

RothleinR et al.

A human intercellular adhesion molecule (ICAM-1) distinct from LFA.

J Immunol

(1986)

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The natural course of sarcoidosis is variable, but no single parameter has been generally accepted as a good marker for disease activity. Adhesion molecules are required for the migration of inflammatory cells; thus, they may be markers of activity in sarcoidosis.
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Our data suggest that the serum sICAM-1 level and the ICAM-1 expression on AM may be good markers of disease activity and also a predictor of outcome in sarcoidosis.
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Alveolar macrophages (AM) originate from blood monocytes and, during the maturation process, undergo functional and morphological changes which are also reflected in their phenotypic pattern. Among the macrophage membrane antigens, adhesion molecules of the integrin family are particularly important for effector functions and cell-cell interactions. The aim of this study was to analyse the membrane expression of selected integrins by AM recovered from bronchoalveolar lavage (BAL) as compared to their precursors, peripheral blood monocytes (PBM). The cells were stained using a sensitive immunoperoxidase assay with 10 different monoclonal antibodies. The data showed a higher expression by AM than PBM of all but one of the studied adhesion molecules. The only exception was CD11b (Mac-1, CR3) which showed a higher expression in PBM than in AM. Several molecules, for example, CD49d (VLA-4), CD51 (vitronectin receptor), and CD54 (intercellular adhesion molecule-1, ICAM-1) were found to be upregulated by AM in patients with a lymphocytic pattern of BAL. In contrast, the phenotype of PBM does not show any changes in these patients. In conclusion, we have demonstrated differences in the expression of integrins between AM and PBM which can be partially responsible for some of their functional differences.
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ICAM-1 antigen expression, induced on AMs after 24 h of culture, was significantly inhibited by fusafungine in a concentration-dependent manner, as measured by flow cytometry analysis using an anti-CD54 monoclonal antibody. TNF-α production, but not RANTES release (measured by ELISA), was significantly inhibited. mRNA studies, by means of polymerase chain reaction amplification of complementary deoxyribonucleic acids (RT-PCR), showed no significant modification of mRNA levels, suggesting that fusafungine acts mainly at a post-transcriptional level. In the same conditions, dexamethasone significantly inhibited the release both of TNF-α and RANTES by AMs, mainly acting at the mRNA level, but had no effect on ICAM-1 expression.
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To investigate the effect of chronic smoke exposure on pulmonary macrophages (PM), the expression of seven different surface and intracellular molecules of PM was studied in induced sputum (IS) samples from healthy volunteers — nine smokers and seven non-smokers. Sputum was induced by inhalation of nebulized saline (3·5% NaCl). Cell viability and total cell counts (TCC) were performed immediately. Cell differentials were determined on May-Grünwald Giemsa-stained cytospin preparations. The PM were immunologically characterized by use of the following monoclonal antibodies: RFD1, RFD7, CD11b, CD54, CD68, CD71 and HLA-DR. The stainings were performed with a three-step, indirect immuno-alkaline phosphate method. Viability and TCC did not differ between the groups. Smokers had a higher percentage of macrophages (P<0·05) and a lower proportion of neutrophils (P<0·05). The percentage of macrophages expressing RFD1, HLA-DR, CD71 (P<0·01 for all) and CD54 (P<0·05) was significantly lower in smokers, whereas the remaining markers were expressed equally in the two groups. The results indicate that smoking induces a decrease in the expression by PM of surface molecules known to be associated with the antigen-presenting function.
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1998, Allergology International
Interstitial lung disease (ILD) can be separated into two groups: (i) that in which the causes are known; and (ii) that in which the etiologies have not been determined. Meanwhile, complement is essential for the inflammatory response and complement systems have been recognized as factors involved in organizing ILD. In the present study we investigated whether there were any differences in complement-related membrane proteins on lymphocytes and alveolar macrophages in bronchoalveolar lavage fluids (BALF) between different ILD. Bronchoalveolar lavage fluid samples were obtained from 12 patients and eight healthy individuals. Bronchoalveolar lavage fluid cells were incubated with monoclonal antibodies (mAbs) against C3b/C4b receptor (CR1), C3bi receptor (CR3), membrane cofactor protein (MCP) and decay accelerating factor (DAF) and were then labeled with fluorescein isothio-cyanate-conjugated anti-mouse IgG. Cells were analyzed using a flow cytometer.The results demonstrate for the first time that CR3 and DAF are elevated on alveolar macrophages, and that MCP and DAF are increased on BALF lymphocytes in sarcoidosis. The possible roles of complement-related membrane proteins in the pathogenesis of immune processes that are ongoing in sarcoidosis are still obscure, but our results may provide some useful information on the mechanisms underlying sarcoidosis.

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Increased Expression of Leukocyte Function Associated Antigen-1 (LFA-1) and Intercellular Adhesion Molecule-1 (ICAM-1) by Alveolar Macrophages of Patients with Pulmonary Sarcoidosis

Section snippets

Study Population

Isolation of Lung and Blood MP

Total and Differential Cell Counts

Immunocytologic Stainings of BM and AM

DISCUSSION

ACKNOWLEDGMENTS:

Chest

Hum Immunol

Gastroenterology

Interstitial lung disease of unknown cause: disorders characterized by chronic inflammation of the lower respiratory tract.

N Engl J Med

The immunology of sarcoidosis.

Semin Respir Med

Pulmonary sarcoidosis: a disorder mediated by excess helper T lymphocyte activity at sites of disease activity.

N Engl J Med

Pathogenesis of sarcoidosis: state of the art.

Chest

Mechanisms of pulmonary fibrosis: spontaneous release of alveolar macrophage derived growth factor in interstitial lung disorders.

J Clin Invest

Release of interleukin 1 by alveolar macrophages of patients with pulmonary sarcoidosis.

Am Rev Respir Dis

Gamma interferon is spontaneously released by alveolar macrophages and lung T lymphocytes in patients with pulmonary sarcoidosis.

J Clin Invest

Bronchoalveolar cells from sarcoid patients demonstrate enhanced antigen presentation.

J Immunol

Enhanced alveolar macrophage mediated antigen induced T lymphocyte proliferation in sarcoidosis.

J Clin Invest

T lymphocyte recognition of antigen in association with gene products of the major histocompatibility complex.

Annu Rev Immunol

The basis for the immunoregulatory role of macrophages and other accessory cells.

Science

A human leukocyte differentiation antigen family with distinct alpha subunits and a common beta subunit: the lymphocyte function associated antigen 1 (LFA-1), the C3bi complement receptor (OKM 1-Mac 1), and the p 150,95 molecule.

J Exp Med

CD antigens 1989.

Int J Cancer

A human intercellular adhesion molecule (ICAM-1) distinct from LFA.

J Immunol