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Review Upper Respiratory Tract Immunity: Adrian W. Zuercher
Review Upper Respiratory Tract Immunity: Adrian W. Zuercher
Review
Upper Respiratory Tract Immunity
ADRIAN W. ZUERCHER
INTRODUCTION
occur via mucosal surfaces like the respiratory, gastrointestinal, or genital epithelium. The mucosal immune system is an important component of the bodys defense against such
infections and consequently induction of mucosal, in addition to systemic immunity, might improve vaccine efficacy. Several orally administered vaccines, for example, against poliovirus and gastrointestinal bacterial infections, have been developed and are widely used. In contrast, to date most vaccines against respiratory pathogens are applied parenterally and thus do not induce significant mucosal immunity. For the
development of effective mucosal vaccines a more profound understanding of the immune mechanisms operative at mucosal surfaces and of the interplay between different mucosal compartments is needed. Moreover, factors like the dose, form of application, and type of mucosal adjuvants are critical to the induction
of effective mucosal immunity. This brief review will focus mainly on the nasal route and will summarize
some recent findings concerning the function of the mucosal immune system of the upper respiratory tract.
Furthermore, routes of cross-immunization between distinct mucosal compartments and how they might be
relevant to vaccine development will be addressed. Finally, I will outline critical factors for the rational design of nasal vaccines and in this context highlight some recent preclinical and clinical developments in the
field.
OST VIRAL INFECTIONS
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FIG. 1.
Model of mucosal immune response in the upper respiratory tract of the mouse.
T-cell areas and consists mainly of naive B-cells (5075%, depending of the strain) (88) and naive (CD45RBhigh)
CD41 T-cells (1530%) (85). But whereas PP in naive, conventionally reared mice are in a state of constant (low level) activation, which is reflected by chronic germinal center reactions, NALT in nave mice
appears to be more quiescent and germinal centers develop only after direct stimulation with antigen, such
as viral infection. In this respect NALT resembles more PP of germ-free mice (83). Thus, even though both
tissues are primary sites of antigen contact, the lower antigenic burden encountered by NALT compared to
PP might explain these differences.
Similar to PP, IgA is the major isotype of antibody produced by NALT (41,86). Specific IgA producing
B-cells are induced in NALT after intra-nasal infection with viral pathogens like influenza virus (2,77,85)
or reovirus (88,89). Production of IgA antibodies is preceded by the induction of germinal centers and the
expansion of IgA1 B-cells in NALT (88). Moreover, it has been shown functionally (88) as well as on the
molecular level (72) that significant proportions of IgG1 B-cells are generated in NALT. Additionally,
NALT is a site of induction of specific T-cells. We have shown that upon intra-nasal infection with reovirus, virus-specific CD81 CTL are generated in NALT, characterized by a higher precursor frequency
compared to draining cervical LN (88). Others have shown induction and persistence of memory CD41
T-cells in NALT after influenza virus infection (85).
After the initial induction in NALT, immune responses are amplified in the draining LN (Fig. 1). Even
though clear anatomical evidence is lacking, functional observations indicate that the submandibular or posterior cervical LN serve this purpose in the upper respiratory tract, analogous to the mesenteric LN in the
gut-associated lymphoid tissue (GALT). The more pronounced expansion of lymphocytes in these LN, compared to NALT, after viral infection (11-fold vs. twofold), the more protracted response and the delayed
onset of responses support this hypothesis (88). Even more strongly than NALT, the draining LN show high
production of specific IgG antibodies. This represents an important mechanism by which mucosal infection
via the nasal route leads to systemic (serum IgG) in addition to mucosal (IgA) immunity. This is a feature
particularly important for the development of nasal vaccines, since an important prerequisite for them is to
induce systemic immunity comparable to a parenterally applied vaccine.
From the draining LN, specific effector cells, by passing through the systemic circulation, home to mucosal effector sites in the upper respiratory tract, such as the lacrimal glands (16), the palatine salivary
glands (88,89) and non-lymphoid tissue of the nasal passage, or as sometimes called, the diffuse-NALT
(Fig. 1). These non-lymphoid sites, particularly the nasal passage tissue, appear to be the major sites of persistence of memory B-cells (50) and T-cells (84) in the upper respiratory tract. This is in agreement with
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no significant antibody responses in the upper respiratory tract were observed. But after local nasal challenge
of intestinally primed mice B-cell responses with the characteristics of memory responses were detected in
NALT, submandibular LN and palatine salivary glands (89). These included rapid onset and peak of specific
IgA and IgG antibody production as well as increased magnitude of responses compared to primary responses
upon intranasal infection without previous intestinal priming. We concluded that trafficking of memory B-cells
from the GALT to the upper respiratory tract was responsible for protection, thus indicating a gut-to-upper respiratory tract homing pathway. Interestingly, the ability to home to the upper respiratory tract was restricted to
memory B-cells, whereas no trafficking of (effector) memory CTL was observed.
Coffin et al. have shown that intra-nasal but not oral immunization of mice with inactivated rotavirus led
to specific B-cell responses in NALT and bronchial LN as well as in intestinal lymphoid tissue. Importantly, intra-nasal priming afforded protection from subsequent intestinal challenge with live virus (21).
Likewise, it was shown in a mouse model that intranasal immunization of pregnant mothers with virus-like
particles conferred protection from intestinal rotavirus infection and diarrhea in the offspring (23). These
data thus suggest an upper respiratory tract to gut axis of immune trafficking and it was proposed that expression of aE integrin might play a role in homing of nasally induced effector B-cells to the intestinal lamina propria (26). Another particularly interesting route of cross-immunization within the integrated mucosal
immune system is induction of vaginal immunity by intranasal immunization (68,69,87). It was shown in
mice that herpes simplex specific immunity in the genital tract could be established by nasal immunization
with a recombinant adenoviral vector (3335). Similarly, nasal vaccination with human papilloma virus 16
virus-like particle resulted in specific immunity in the genital tract (3). Interestingly, lymphoid compartments in the lower respiratory tract (trachea, lung, tracheobronchial LN) appeared to be the sites of induction of specific responses whereas NALT played a minor role only (4).
In summary, the above examples document various routes of cross-protection between distant mucosal
compartments. However, the underlying mechanisms remain largely elusive. Particularly little is known
about the cellular components responsible for cross-protection and whether their relocation to distant mucosal compartments is guided specifically by homing receptors or alternatively occurs randomly. Some attempts have been undertaken to elucidate such mechanisms. As outlined above, our studies indicated that
after intestinal priming protection of the lower respiratory tract is achieved by effector memory CTL, whereas
specific memory B-cells relocate to the upper respiratory tract and are responsible for rapid secondary responses and protection (89). This represents an example showing distinct cellular components induced in
the GALT relocating to different distant mucosal compartments. Another approach has been the study of
expression of integrins in the upper respiratory tract and their potential role in homing. Csencsits et al. have
demonstrated that the pattern of integrins expressed by NALT high endothelial venules differs significantly
from PP (29). Notably, it is clear that the interaction of a4b7 integrins with MadCAM is a major mechanism of nave lymphocyte homing to PP as well as homing of effector cells to the intestinal lamina propria
(8). In contrast, a4b7MadCAM interactions played a minor role for homing to NALT, but binding to
NALT HEV mainly involved L-selectin and peripheral node adressin (PNAd). These findings were confirmed by studies in L-selectin2/2 mice, which displayed a marked reduction of lymphocytes homing to
the upper respiratory tract resulting in decreased local immune responses (27,28). Also involvement of the
CCR9-TECK/CCL25 interaction for homing to the intestinal lamina propria is well established (13,46).
Whether similar interactions are operative in guiding mucosally induced lymphocytes to non-intestinal mucosal compartments remains to be investigated. Recently a chemokine ligand termed MEC/CCL28 that is
specifically expressed on mucosal epithelial tissue (60) and shares homology with TECK/CCL25 as well
as with CTECK/CCL27 was identified. All of these appear to be involved in homing to mucosal sites (47).
This may indicate a network of integrin and chemokine receptors and ligands controlling homing to nonintestinal mucosal sitesa hypothesis that awaits deeper investigation.
ZUERCHER
munity induced. Indeed, it is known that local cytokine patterns in NALT greatly vary depending of the
type of antigen used. The quiescent NALT is characterized by a non-committed Th0 cytokine environment
(42) that can be biased towards either a Th1- or Th2-like cytokine pattern dependent of the antigen. Intranasal infection of BALB/c mice with live influenza virus induced a Th1like cytokine pattern with production of IFNg, IL-2, and IL-6, whereas intranasal immunization with inactivated influenza in combination with CT induced strong IL-4 and IL-6 responses. These distinct cytokine patterns were reflected in the
IgG isotype subclasses that predominated, that is, a major IgG2a response in the former and a strong IgG1
response in the latter case (52).
Despite the large quantity of preclinical data and obvious advantages of intranasal vaccines, currently
none are licensed for the use in humans. Nevertheless, some promising developments have been clinically
tested, some of which have been summarized in an excellent review by Davis (30). Several nasally administered influenza virus vaccines have been evaluated in humans. Among these, Tomoda et al. used cold
adapted, live attenuated strains of influenza for immunization via the nasal route (79). They found that secretory nasal IgA as well as IFNg production by specific CD41 T-cells was critical for protection against
challenge during a subsequent epidemic outbreak. Similarly, in a comparative study using a trivalent intranasal influenza virus vaccine the effectiveness of cold-adapted, live attenuated influenza virus vaccine at
preventing the shedding of virus was proven (6). The safety and efficacy of this nasal influenza virus vaccine was subsequently confirmed in children (7) as well as HIV-patients (45). However, a possible disadvantage of this vaccine is the low level of specific serum IgG1 achieved. In an alternative approach, a virosome based influenza vaccine, containing HLT as a mucosal adjuvant, proved to be immunogenic in
humans and induced strong specific mucosal and systemic antibody responses (36,37). In another recent
study, Greenbaum et al. reported the effects of intranasal vaccination with an inactivated trivalent influenza
virus vaccine in humans (39). In their study, a single immunization resulted in seroconversion in 4070%
of vaccinees (strain dependent) and an overall shift from non-immune to immune in about 50% of the volunteers. Taken together, these studies demonstrate that intranasal influenza vaccineseither in live attenuated or virosomal formulationsare effective for the use in humans.
In contrast to these advanced developments in the influenza virus field, no human trials for nasally applied RSV vaccines have been performed so far. Nevertheless, a parenterally administered RSV subunit
vaccine consisting of a fragment of RSV G-protein coupled to a carrier protein (BBG2Na) (61) has been
evaluated in a phase III clinical trial in humans. The same vaccine was tested via the nasal route in mice
and induced robust systemic and mucosal immunity that conferred protection from respiratory challenge,
importantly without signs of lung pathology (38). No nasally administered parainfluenza vaccines are currently evaluated clinically, however, in a green monkey model it was demonstrated that parenteral immunization with Sendai virus prevented subsequent nasal infection with parainfluenza virus (43). No nasal vaccines for genital infections such as herpes or papilloma virus have progressed to human trials to date.
ACKNOWLEDGMENTS
I would like to thank John J. Cebra for his continuing support and for his helpful suggestions in writing this
manuscript. Furthermore, I thank Guido Dietrich, Christian Moser, and Jean-Francois Viret for critically reading the manuscript. This work was supported by grant AI-23970 from the National Institutes of Health.
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