Thematic Review Series

Respiration 1999;66:211

Pulmonary Defence Mechanisms

Laurent P. Nicod
Pulmonary Division, University Hospital, Geneva, Switzerland

Abstract Lung defence involves a wide array of mechanisms needed to remove inhaled particles and organisms. The various innate immune processes which take place either in the central or in the more distant airways are reviewed. The recruitment and the development of an adaptive immunity following the innate response are described. This entails the production of secretory immunoglobulins in the airways and either the influx of polymorphonuclear neutrophils in the alveoli to phagocytose more or less opsonized organisms or the activation of the T lymphocyte response to improve the removal of intracellular pathogens.

major bronchi are protecting the lungs with the anatomic barriers they represent associated with the cough reflex, the mucociliary apparatus and the secretory immunoglobulin A (IgA). Below, the superficial layers of the mucosa, a tight network of dendritic cells will sense and catch any invading organisms and bring them to the lymph nodes around the airways or in the hilum. In the respiratory units beyond the respiratory bronchioles particles will be caught by alveolar macrophages in a milieu rich in elements such as IgG, complement, surfactant and fibronectin. In these units when needed various amounts of neutrophils and lymphocytes will be recruited (fig. 1).

Protection of the Upper Airways and Bronchi Introduction Anatomic Barriers

The upper and lower airways together represent the biggest epithelial surface exposed to the outside environment. The inspired air is the source of the necessary oxygen for the body, but also introduces numerous particles, toxic gases and microorganisms. In addition, the lungs are inoculated repeatedly with bacteria from the nasooropharynx. In order to allow for gas exchange, foreign substances and microorganisms need to be stopped and removed without undue inflammation. A complex defence system is present from the nostrils till the alveoli. Schematically, the components of the lung defence can be divided into those located in the upper airways and those in the alveoli (table 1). The upper airways and the

The nasopharyngeal anatomic barriers are important to efficiently prevent the penetration of particles or organisms bigger than 23 m into the lower airways. A part of the small particles and microbes in the inhaled air is stopped by impaction on the mucosal surface. Appropriate components of host defence are needed to deal with them. Cough A cough is classically described as a very deep breath, followed by a forced expiration against a closed glottis, which opens suddenly to produce the expulsive phase. This allows enough turbulence and shearing forces to be produced in the major bronchi and trachea to extrude

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Fig. 1. Circulation of dendritic cells (DC) or alveolar macrophages from the lung to lymphoid organs and recirculation of sensitized lymphocytes. LN = Lymph node; GALT = gastrointestinal-associated lymphoid tissue.

Table 1. Defences of the respiratory tree

Upper airways and bronchi Anatomic barriers Cough Mucociliary apparatus Airway epithelium Secretory IgA Dendritic cell network Lymphoid structure Host defences in alveolar spaces Alveolar macrophages Immunoglobulins and opsonins Lymphocyte-mediated immunity Neutrophils and eosinophils

material such as debris, infected mucus or products of epithelial damage. The cough is triggered by a very wide variety of stimuli: mechanical, chemical or in relation with inflammatory mediators [1]. Mucociliary Apparatus Approximately 90% of inhaled particles with a diameter bigger than 23 m are deposited on the mucus overlying the ciliated epithelium. The particles are transported

from the terminal bronchioles to the trachea by the ciliary beats with the mucus. This motion occurs at a speed varying between 100 and 300 m ! s 1. The airway mucus is composed of a sol phase, or periciliary liquid about 510 m deep allowing the cilia to beat and a gel phase on the surface of the cilia whose thickness varies between 2 and 20 m. The flow of the gel is referred to as the mucociliary transport. The physical properties of mucus are provided mainly by mucins, which are mucoglycoproteins and proteoglycans secreted from the surface of epithelial cells and from the glands. Phospholipids are also secreted by the epithelial cells and submucosal glands of the airways weakening the adhesion of the mucus, altering its physical properties. The mucus gel acts as a barrier for bacteria which adhere to it, but where they can multiply depending on the conditions [2]. Important elements contributing to lung defences are carried with the mucus. Among them are the secretory IgA, lysozyme, lactoferrin or peroxydases. Lysozyme is a muramidase that degrades a glycosidic linkage of bacterial membrane peptidoglycans. It is regularly found in the lung secretions and, when purified, it demonstrates bactericidal properties [3]. It may be derived from many sources, including epithelial cells, serous cells of submucosal glands, macrophages and neutrophils. The iron binding proteins such as lactoferrin may reduce the availabili-

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Table 2. Respiratory epithelial cell cyto-

kines [adapted from 75] Chemokines -Chemokine family IL-8 GRO GRO -Chemokine subfamily Monocyte chemoattractant protein 13 Colony-stimulating factors GM-CSF G-CSF M-CSF Colony stimulating factor 1 Pleiotropic cytokines IL-1 TNF IL-6 IL-11 IL-16 or lymphocyte chemoattractant factor

receptor protein which is responsible for mediating cyclic AMP-induced increases in chloride secretion [7]. The resulting alterations in the epithelial overlying fluid composition are believed to lead to poor bacterial clearance from the airways and, thus, to the recognized clinical syndrome. Airway Epithelium The epithelial cells from the lining of the luminal surface are attached to their neighbors by a number of cellcell junctions. Included among these are tight junctions, intermediate junctions, gap junctions and desmosomes [8]. These structures form a barrier segregating both the luminal space from the pulmonary parenchyma. In addition through the gap junctions cell-cell communication allows the epithelium to function as an electrically integrated unit and allows small molecules to travel between airway epithelial cells. Communication via gap junctions is believed to help maintain the uniform beating of the cilia [9, 10] and transport through gap junctions may be a means for the cells to provide their neighbors with defence molecules such as antioxidants [11]. Desmosomes mediate for the former mechanical adhesion of cells to their neighbors and tight junctions completely obliterate the intercellular space just below the luminal surface [12]. This organization has at least two functions in host defence. One is to create an effective mechanical barrier and the other is to allow for polarity in function and permit maintenance of an ionic gradient for directional secretion of many substances. Epithelial cells also actively participate in host defence against inflammation. These cells are capable of producing and responding to a variety of eicosanoids, cytokines and growth factors which form a complex network regulating inflammatory responses (table 2). They also express cell surface receptors that can interact directly with inflammatory cells. Epithelial cells recruit inflammatory cells. Thus they release arachidonic acid derivate 15-HETE, an active neutrophil chemoattractant that can also modulate immune cells [13]. There is evidence that epithelial cells can secrete IL-8 in a bidirectional manner with preferential secretion into the lumen [14]. A variety of stimuli have been demonstrated to induce the release of IL-8. These include cigarette smoke, endotoxin and other bacterial products [15], viral infections [16] and a variety of cytokines, including IL-1, TNF and IFN- [17]. Other chemokines include GRO, GRO and MCP-1. Epithelial cells also release a lymphocyte chemoattractant factor or IL16 [18]. IL-16 is also an activating factor for CD4 T cells.

ty of elemental iron which is a cofactor for bacterial replication. However, in addition, lactoferrin may also be bactericidal by binding to endotoxin [4]. The secretory peroxidases (lactoperoxidases) or those from leukocytes (myeloperoxidases) act on thiocyanate ions or produce oxygen radicals which are bacteriostatic or bactericidal. Serum proteins, such as albumin, fibrinogen and immunoglobulins, exude from the blood vessels in the mucosa with many active plasma components during airway inflammation [5]. The latter ones are part of the defence mechanisms such as complements and others protect the airways such as the protease inhibitors. Mucociliary clearance is altered in several airway diseases where structural damage to the epithelium or alteration of the mucus composition increases bacterial adhesiveness. Bacterial products may, furthermore, alter the mucociliary clearance as some of these products are ciliotoxic. This is true for the Pseudomonas aeruginosaderived elastase [6]. Mucociliary clearance is altered in asthma, chronic bronchitis or cystic fibrosis. An abnormal structure of the cilia has been observed in some airway diseases such as chronic bronchitis and in congenital diseases such as the primary ciliary dyskinesia syndrome with ciliary immobility and interciliary discoordination with virtually absent mucociliary clearance. In cystic fibrosis the most common mutation results in a defective transmembrane

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Epithelial cells release growth factors which can sustain recruited inflammatory cells. Among these GM-CSF appears to be particularly important to sustain not only eosinophil survival but also mononuclear cell activation such as macrophages or dendritic cells [19]. Epithelial cells in the lung are capable of upregulating the expression of the adhesion molecule such as intercellular adhesion molecule-1 (ICAM-1) in response to inflammatory stimuli that will favor the binding of neutrophils or monocytes to an inflamed area. In addition epithelial cells are capable of expressing the major histocompatibility complex (MHC) of class I and II upregulated by cytokines including IFN-. Epithelial cells may thus have a certain capacity for presenting antigens to lymphocytes and regulating an antigen-driven lymphocyte response [20]. In addition to recruiting lymphocytes, producing cytokines which can regulate lymphocyte activity and expressing MHC receptors, epithelial cells can transport IgA. Secretory IgA The IgA released by the epithelial cells is the first line of defence related to the adaptive immunity. IgA antibody synthesized in the lamina propria is secreted as an IgAdimeric molecule associated with a single J chain of 23,000 daltons. This polymeric form of IgA binds specifically to a molecule called the poly Ig receptor or the secretory component expressed on the basolateral surfaces of the overlying epithelial cells. The complex is internalized and carried through the cytoplasm of the epithelial cell in a transport vesicle to its apical surface and released in the airways. IgAs are particularly important as they neutralize toxins, viruses and block the entry of bacteria across the epithelium. IgAs are poor activators of the classical pathway of complement, but can activate the alternate pathway allowing a better opsonization of bacteria [21]. Dendritic Cell Network Dendritic cells are antigen-presenting cells that constitutively express a high level of MHC (class II) antigen. These cells reside in human and rat airways with branching processes that extend in the plane of the epithelial basement membrane, forming a network optimally situated to sample inhaled antigens [22, 23]. The intimate association of dendritic cells with epithelial basal cells is reminiscent of the interaction between Langerhans cells and keratinocytes of the skin [24]. There are 1,400 B 140 cells/mm2 in the rat trachea when counting those located above and below the lamina propria. Recent evidence

indicates that airway dendritic cells constitute a highly reactive population, whose numbers and MHC content change rapidly under experimental conditions. For example, dendritic cells become more numerous in response to inhaled antigens [25] and rapidly decrease in number after treatment with glucocorticoids [24]. Dendritic cells are highly efficient antigen-presenting cells especially after maturation under the influence of TNF- and GM-CSF. Then after catching antigens in peripheral locations, these cells migrate via lymphatics to the T-cell-dependant areas of regional lymph nodes, where they encounter naive lymphocytes and initiate a primary immune response [26]. As they express both MHC class I and II with a wide repertoire of costimulatory molecules to activate naive CD4 and CD8 T cells, airway dendritic cells are considered the most potent antigen-presenting cells [27]. Lymphoid Structures It is known that in clinically healthy subjects T lymphocytes are present within the surface epithelium and are predominantly of the CD8 subset [28] whereas the CD4 T lymphocytes normally predominate beneath the epithelium. Bronchial biopsies from subjects with mild stable asthma have an increased infiltrate of inflammatory cells in their subepithelium including activated CD4 T cells which outnumber the CD8 T lymphocyte subset by about 3:1 [29]. Smokers with chronic bronchitis show a trend to have an increased number of CD8 cells over CD4 cells even in the subepithelium [30]. T cells and NK cells may play a crucial role in lung immunity, being able to react to pathogens in the absence of preliminary priming. For the T cells the ensuing secretion of cytokines may induce a Th1 or Th2 response depending on the pathogen encountered [31]. However, there may be less than 1% of T cells [32]. Beyond the mucosa the respiratory tract contains a considerable amount of various lymphoid tissues such as centrally the lymph nodes positioned in the mediastinum and hilar areas of the lung and submucosal aggregates spaced along the airways at branching points. Considerable attention has been given to bronchus-associated lymphoid structures (BALT) similar to Peyers patches in the small intestine. However, similar lymphoid structures in the airways of dogs, primates and humans are not nearly so prominent [33] and possibly of less significance in airway host defence. Nevertheless, the BALT at branching points of the airways are likely important structures of the afferent pathway of local immunity. After antigen uptake and processing by stimulated dendritic cells and macro-

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Table 3. Macrophage-secretory products

Cytokines IL-1/IL-1ra TNF-/TNFsRs IL-6 IL-10 IL-12 Chemokines IL-8 MIP-1/ RANTES MCP-1-MCP-3 Defensins and lysozyme Reactive oxygen intermediates Reactive nitrogen intermediates Enzymes Metalloprotease/TIMPs Macrophage metalloelectase Urokinase Acid hydrolases Bioactive lipids Cyclooxygenase Products of arachidonate Complement proteins Most components of complement pathways and the inhibitor C1q IL-1ra = IL-1 receptor antagonist.

phages within BALT, an antibody response is likely to be initiated. BALT would then be part of a network where immunization could occur at a remote site including the gastrointestinal tract, and through the redistribution of lymphocytes or plasma cells to other mucosal surfaces, antibodies could be provided there [34]. In studies on the IgA system, reference is made to this phenomenon [35].

Host Defences in Alveolar Spaces

Particles and microorganisms in the inspired air can escape the defence mechanisms of the airways and reach the alveolar surface. Defence mechanisms must be able to neutralize the invading pathogens by recruiting if necessary mononuclear cells for an organized cell-mediated defence or neutrophils whenever necessary. Once the inflammatory process has been effective, cell debris and exudate must be removed in order to recover the alveolar architecture.

Alveolar Macrophages Alveolar macrophages represent 85% of the cells retrieved by bronchoalveolar lavages. They are bone marrow-derived cells that can be differentiated from blood monocytes after they have emigrated into the tissues. These cells populate the various compartments of the lung, differentiating into mature macrophages, enriching themselves in phagolysosomes once they engulf ingested material into the alveoli. Although it is known that macrophages have the capacity to divide, under normal conditions fewer than 0.5% of the pulmonary population would be dividing [32]. If most macrophages are removed by the mucociliary apparatus, it is known that alveolar macrophages or airway macrophages can travel back into the lung parenchyma and migrate through the tissues or through the lymphatic vessels to the bronchial lymph nodes with ingested particles or antigens [36]. Alveolar macrophages are normally the only phagocytic cells present within the lower respiratory tract. They avidly phagocytose inhaled particles, but ingest viable bacteria considerably less efficiently [37]. The ability of macrophages to interact with microbes is mediated by surface receptors capable of binding specific ligands including toxins, polysaccharides, lipopolysaccharides, complement proteins and immunoglobulins. The ability to recognize surface lectins or microbes is crucial to innate immune defences. The mannose receptor mediates phagocytosis of yeast, Pneumocystis carinii and many organisms with carbohydrate recognition-like domains [38]. The capacity of CR3 to bind directly to microbes in the absence of an opsonin may represent another mechanism whereby macrophages recognize potential pathogens before the onset of adaptive immunity [39]. Alveolar macrophages have multiple functions through the release of numerous mediators (table 3) with the following properties. (a) Initiation of inflammation by the release of IL-1 and IL-1 or TNF- will induce a cascade of events in the alveolar milieu such as the release of chemokines (IL-8, MIP-1/, RANTES, MCP-1/3), of growth factors such as GM-CSF, G-CSF or M-CSF, or the appearance of adhesion molecules on endothelial cells or epithelial cells. Inflammatory cytokines will favor the activation of neighboring cells and attract several inflammatory elements from the blood. In addition bioactive lipids mostly derived from cyclooxygenase products of arachidonate such as thromboxane A2, LTB4, 5-HETE, PGE2 and PGD2 will influence vasoactive mechanisms, the function of T and B lymphocytes as well as of the macrophages in an autocrine manner.

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(b) The control of inflammation by the release of an inhibitor of IL-1 or TNF- in the form of IL-1 receptor antagonist or TNF-soluble receptors (TNFsR55 or TNFsR75) [40]. Macrophages will also have the capacity to block IL-1 or TNF production by their own release of IL-10 [41]. (c) Bactericidal properties are possible by the production of lysozyme. Defensins are cationic proteins capable of killing a wide variety of gram-positive and gram negative bacteria and fungi. Reactive oxygen intermediates (superoxide anion, hydrogen peroxide, hydroxyl radicals) or reactive nitrogen intermediates (nitric oxide, nitrites or nitrates) are also involved in killing microorganisms and eventually tumor cells. (d) Macrophages are involved in lung remodeling and repair. Indeed they produce macrophage metalloelastases, collagenase, metalloproteases (MMP1, MMP9) and their inhibitors TIMPs under the tight regulation of cytokines [42]. Macrophages thus remodel matrix constituents by their own production of urokinase to remove fibrin deposition. They can also release fibroblast growth factors such as transforming growth factor or platelet-derived growth factor. (e) By their production of several components of complement as well as of the C1q inhibitor they are likely to facilitate the opsonization of bacteria while they are able to inhibit undue inflammation by controlling the effect of the activation of the complement cascades [43]. Alveolar macrophages can under yet poorly understood circumstances acquire some characteristics of dendritic cells and may thus be able to activate T cells [44]. Meanwhile they are known in normal subjects to prevent T cell activation [45, 46]. This capacity to enhance or inhibit T cell immunity remains a matter of intensive research. IL-12 production has lately been shown to play a major role to induce a Th1 response and macrophages are a major source when stimulated by bacterial lipopolysaccharide and IFN- or during the interaction of CD40 ligand on T cells and CD40 on macrophages [47]. Immunoglobulins and Opsonins Several components in normal bronchoalveolar lavage fluid have the capability of coating, in a nonspecific manner, certain bacteria that will enhance phagocytic uptake by alveolar macrophages, thus qualifying as nonimmune opsonins. Surfactant [48], fibronectin [49] and C-reactive protein may have opsonic activities. Immunoglobulin G which constitutes 5% of the total protein content of normal BAL fluid [50] seems to be the predominant immunoglobulin with the strongest opsonic activity in the alveoli

contrarily to IgA. Secretory-IgA in addition does not have obvious secretory mechanisms to be delivered in the alveoli. IgG1 and IgG2 are present in greatest concentrations (65 and 28%, respectively), whereas IgG3 and IgG4 together account for ! 10% [51]. In terms of host defence IgG1 and IgG3 are considered to be the most important as only these two antibodies fix complement. IgG2 is a typespecific antibody against pathogens such as Streptococcus pneumoniae or Haemophilus influenzae [52]. IgG4 acts as a reaginic antibody in allergic disease and increased IgG4 may lead to hypersensitivity pneumonitis, however, if absent there is a predisposition to sinopulmonary infections and bronchiectasis [53]. Most components of the complement can be produced by monocytes or macrophages in vitro, however, most of them come from the liver cells carried to the lung by the blood. Activation of the entire complement pathway in the presence of microbes can result in their lysis and killing. When bacteria activate the alternate pathway, C3b is released allowing a good opsonization of bacteria for neutrophils or macrophages. C5a has phlogistic properties and exerts a powerful chemoattractant effect for polymorphonuclear cells. The complement pathway is under the tight control of C1q released by alveolar macrophages or released from blood exudation. Complement and in particular the alternative complement pathway are likely to play an important role as the first line of defence against many extracellular microbes as part of the innate immune defences [53, 54]. Lymphocyte-Mediated Immunity Alveoli contain about 10% lymphocytes among which 50% are CD4, 30% CD8, 1015% killer or natural killer (NK) cells and 5% B lymphocytes. The CD4/CD8 lymphocyte ratio is 1.5 and thus similar to peripheral blood. In the alveolar milieu lymphocytes may have a slightly altered phenotype and function. Thus NK lymphocytes are Leu 7-positive in the alveoli and their cytotoxic function is slightly altered compared to interstitial Leu 11positive NK cells [55]. The term cell immunity was originally reserved to describe reactions to intracellular pathogens requiring the collaboration of lymphocytes and macrophages for destruction (table 4). Humoral immunity refers to the immune process mediated by immunogloblulins. However, both humoral and cellular immunity initially require the presentation of antigens to T and B lymphocytes which will be sensitized essentially in structures such as lymph nodes. Once primed T lymphocytes may be reactivated by dendritic cells around the vessels or around the airways or by less professional anti-

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Table 4. Microorganisms capable of surviv-

ing in macrophages in the absence of cellspecific immunity M. tuberculosis and other species Mycobacterium lepraemurium Toxoplasma gondii Leishmania donovani Salmonella typhi Listeria monocytogenes Legionella pneumophila P. carinii Cytomegalovirus

cells recognize these MHC class I-associated viral antigens on the surface of infected cells and lyse them. Neutrophils and Eosinophils Neutrophils are myelopoietic cells of the granulocyte lineage, which also includes eosinophils and basophils. Neutrophils are also called polymorphonuclear neutrophils (PMN) because of their characteristic multilobed nuclei. They are the most abundant type of leukocytes in the body. Their maturation process takes about 2 weeks in the marrow; then they are released into the blood where about half are believed to be marginated, that is pooled on the walls of the vessels. Their half-life is 8 h. In the bronchoalveolar lavage they normally represent less than 2% of the cells. However, if cells residing in the alveoli are unable to control infectious agents, a massive flux of neutrophils occurs. Thus in an experimental model, if only 4 ! 105 Staphylococcus aureus are inoculated, they will be neutralized by macrophages only. But if 107 S. aureus are applied, a massive flux of PMN occurs able to control the infection. With 108 S. aureus, mice do not survive, despite or because of an even more important influx of PMN [65]. PMN have been shown to be essential in the early appearance of many bacteria such as P. aeruginosa, Klebsiella pneumoniae and H. influenzae [66]. The migration of PMN into the lung involves first their weak binding to the vascular endothelium through interactions between carbohydrate ligands on the PMNs and selectins on the endothelium. Thus sialyl Lewis X moiety on the surface of PMNs recognizes the TNF- rapidly induced P-selectins and the E-selectins whose expression appears after a few hours to make possible the rolling of PMN, the loose connection of PMN on the endothelium allowing them to roll. The leukocyte migration needs the interaction of ICAM-1 on endothelial cells and the leukocyte function antigen-1 (LFA-1) on the surface of PMN. After the interactions PMNs are firmly attached on the endothelium. The extravasation of PMNs involves LFA-1 and platelet endothelial cell adhesion molecule, expressed on the leukocytes and at the junction of endothelial cells. PMN will then migrate following chemotactic gradients [67]. Chemotactic factors include C5 fragments generated by the activation of the alternative pathway by bacteria or cleaved by the proteases of alveolar macrophages. Alveolar macrophages generate products of arachidonic acid such as 5- or 11-monohydroxyeicosatetraenoic acid and LTB4 [68]. Chemokines are small polypeptides also critically involved in granulocyte recruitment. The C-X-C group of molecules, among others IL-8, GRO and GRO, promote migration of neutrophils and are found

gen-presenting cells such as all those on which MHC class II is inducible. The cellular and humoral response oriented towards a given pathogen is referred to as adaptive immunity. More and more evidence accumulates pointing out the initial role of innate immunity to condition the later adaptive immunity [56]. A lot of attention has been paid to the two types of lymphokine secretion by CD4 cells, which might condition the type of adaptive response. Th1 cells produce IFN- and IL-2 and Th2 cells produce IL-4, IL-5 and IL-10, whereas both subsets produce IL-3, GM-CSF and TNF-/. Th1 cells mediate delayed-type hypersensitivity reactions, activate macrophages for microbicidal function and induce IgG2a. Th2 cells provide help for antibody responses and induce IgG1, IgA and IgE. Meanwhile Th2 cytokines favor the growth and activation of eosinophils and basophils. The two subsets are mutually inhibitory [57]. A growing body of evidence indicates that Th1 and Th2 are derived from common precursors [58]. Distinct microbial antigens may favor dominance of Th1 or Th2 cells [31, 59]. IL-12 released by infected macrophages plays a pivotal role in favoring a Th1 response [60]. Both Th1 and Th2 cytokines have important roles in pulmonary defences. IFN- produced by CD4 T cells, CD8 T cells, cells and NK cells have a central role in microbicidal activity against most intracellular pathogens, in particular Mycobacterium tuberculosis [61]. IFN and TNF- are synergistic in activating the tuberculostatic capacities of murine phagocytes [62]. Th2 lymphocytes have important roles in some granulomatous diseases such as schistosomiasis [63]. Cell-mediated cytotoxic responses are important in the defence against pulmonary viral infections. Antigen-specific CD8-cytotoxic T lymphocytes appear in the lung parenchyma within 1 week after pulmonary viral infections [64]. Eradication of viral infections is accomplished when cytotoxic CD8

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in bronchoalveolar lavage of patients with various types of pneumonia [69]. Mononuclear phagocytes, endothelial cells and epithelial cells have the ability to generate chemokines in response to microbial products such as LPS. TNF- and IL-1 produced by the cells of the host are required for the production of chemokines by fibroblasts and smooth muscle cells. TNF- and IL-1 markedly increase gene expression of chemokines in endothelial cells, fibroblasts and epithelial cells. Chemokines bind to proteoglycan molecules both in the extracellular matrix and on parenchymal cells. Chemokines are thus displayed on a solid substrate along which leukocytes migrate. Activated neutrophils eliminate microorganisms by means of a range of mechanisms, which involve phagocytosis, the release of oxygen radicals and the production of cytotoxic peptides or proteins. The recognition and phagocytosis of bacteria by neutrophils are a necessary prelude to intracellular killing. Bacteria are either recognized by their own protein or carbohydrate component or after opsonization by immunoglobulin or complement. In addition to the cytotoxic effects of the respiratory burst, there is an array of components in the granules that have the ability to kill and degrade microorganisms. Carbohydrate components of bacteria are attacked by enzymes such as sialidase, -mannosidase, -glucurone Nacetyl- glucosaminidase and lysozyme. Cytotoxic proteins such as defensins and serine proteinases damage

bacterial membranes by still only partially understood mechanisms [70]. In the Chdiak-Higashi disease, a congenital immunologic defect known to be accompanied by severe pyogenic infections, the granules cannot package the protein elastase or the cathepsin G superfamily [71]. In chronic granulomatous diseases, affected individuals are susceptible to bacterial infections because their phagocytic cells are unable to generate the product of respiratory bursts [72]. Neutrophil migration itself is already impaired in leukocyte adhesion deficiencies (LAD), such in LAD-II, which is characterized by a failure to express sialyl Lewis X, the counterreceptor for E-selectin and Pselectin [73]. Eosinophils are nondividing granulated cells, characterized by their unique crystalloid granules which contain four basic proteins: the major basic protein, the eosinophil cationic protein, the eosinophil peroxidase and eosinophil-derived murotoxin. Three T cell-derived cytokines have been shown to promote eosinophil growth and differentiation: IL-3 and IL-5 as well as GM-CSF. Helminthic larvae coated with immunoglobulins or complement are particularly susceptible to eosinophil-mediated cytotoxicity [74]. Eosinophil products can provoke many of the pathological features of asthma and other rare diseases such as the acute or chronic eosinophilic pneumonia or the eosinophilic vasculitis of the Churg-Strauss syndrome.

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