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Abstract |
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Abstract
Introduction
Materials and methods
Results
Discussion
References
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The maturation of eosinophils in bone marrow, their migration to pulmonary tissue, and their subsequent
degranulation and release of toxic granule proteins contributes to the pathophysiology observed in asthma.
Interleukin-5 (IL-5) is essential for these processes to occur. Therefore, much emphasis has been placed on attempts to inhibit the production or activity of IL-5 in order to attenuate the inflammatory aspect of
asthma. In this report, the immunological consequences of long-term exposure to an antibody recognizing
IL-5 (TRFK-5) were studied in a murine pulmonary inflammation model. A single dose of TRFK-5 (1 mg/
kg, intraperitoneally) reversibly inhibited antigen-dependent lung eosinophilia in mice for at least 12 wk
and inhibited the release of eosinophils from bone marrow for at least 8 wk. Normal responses to aerosol
challenge were attained after 24 wk. In mice treated acutely with antibody (2 h before challenge), 50% inhibition of pulmonary eosinophilia occurred when 0.06 mg/kg TRFK-5 was administered (intraperitoneally; ED50), resulting in 230 ng/ml (IC50) in serum. In mice treated with one dose of TRFK-5 (1 mg/kg)
and rested before challenge, the antibody exhibited a half-life of 2.4 wk. After 18 to 19 wk, antigen challenge-induced eosinophilia was inhibited by 50% and serum levels of TRFK-5 were 25 ng/ml. TRFK-5 remaining in mice 8 wk after a single injection of TRFK-5 was sufficient to inhibit at least 50% of the eosinophilia induced in blood 3 h after injection of recombinant murine IL-5 (10 µg/kg, intravenously). To
assess the biologic effect of long-term exposure of mice to antibody, several parameters of immune-cell
function were measured. Throughout the extended period of activity of TRFK-5 (
12 wk) there were no
gross effects on antigen-dependent increases in T-cell recruitment into bronchoalveolar fluid (BALF), in
IL-4 and IL-5 steady-state mRNA levels in lung tissue, or in immunoglobulin E (IgE) and IgG levels in serum. There was a small increase in IL-5 steady-state mRNA production in TRFK-5-treated mice after 2 h
or 2 wk, but this was not observed at other times examined. In untreated mice, IL-5 steady-state mRNA
production in response to antigen challenge decreased > 6-fold with age, although at all time points there was an increase in mRNA levels following challenge. Therefore, at later times, 25 ng/ml rather than 230 ng/ml of TRFK-5 inhibited BALF eosinophilia, probably because of reduced IL-5 levels. Twenty-four weeks after treatment with TRFK-5, when challenge-induced eosinophilia was restored, there was an excess of CD4+ T cells in BALF from challenged mice. However, these T cells had no measurable effects on
other responses to challenge, including cytokine production, B-cell accumulation, and immunoglobulin
production in serum. Thus, the biologic duration of TRFK-5 was several months, and its activity was due
to the presence of antibody above a therapeutic threshold rather than to any profound effect on the immune system.
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Introduction |
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Top
Abstract
Introduction
Materials and methods
Results
Discussion
References
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Asthma is thought to occur because of the complex interaction of several physiological and immune system defects. This disease is characterized by variable and reversible airway obstruction and by bronchial hyperreactivity in response to nonspecific stimuli (1, 2). In addition, asthma is often associated with prominent airway inflammation and airway epithelial cell damage (3, 4). Elucidation of the mechanisms by which each of these parameters contributes to the asthmatic response has required the use of several animal models, each of which has been useful in mimicking different components of the human disease (5).
The importance of pulmonary inflammation to the asthmatic response has been appreciated only recently. Several cell types contribute to the observed inflammation, including mast cells, macrophages, dendritic cells, eosinophils, and T cells (3). In particular, the often dramatic infiltration of eosinophils into pulmonary tissue and their subsequent degranulation and release of toxic granule proteins is thought to be one of the major causes of epithelial cell damage (4). Eosinophil infiltration in the lungs and concomitant edema formation also contributes to airway obstruction and may result in the development or exacerbation of airway hyperreactivity. Thus, inhibition of eosinophil maturation, migration into pulmonary tissue, or function is predicted to have a beneficial therapeutic effect in asthma.
The cytokine interleukin-5 (IL-5) is essential for the maturation, differentiation, migration, and effector function of eosinophils (14, 15). Unlike most other cytokines, IL-5 has limited activity on cells other than eosinophils. Therefore, the inhibition of IL-5 should attenuate most eosinophil activity but have little effect on normal immune function. If airway inflammation is an underlying cause or significant contributory factor to airway obstruction and hyperreactivity, then inhibiting eosinophil influx by neutralization of IL-5 is likely to attenuate the asthmatic process (16). Experimental evidence from studies with mice, guinea pigs, and monkeys supports this concept (5, 17, 18). In most cases, treatment of animals with antibodies to IL-5 attenuates pulmonary inflammation and airway reactivity.
The use of therapeutic antibodies has been tested in humans for several diseases, including arthritis, transplantation, septic shock, and cancer (19). In most cases, the antibodies have been "humanized" to prevent a host immune response to the molecule. With little anti-antibody formation, the duration of action is expected to be similar to that of endogenous immunoglobulin (Ig), exhibiting a serum half-life of 3 wk (20). Depending on their immunoreactivity, this also might be the case for xenobiotic antibodies. In fact, antigen-challenged monkeys treated once with a rat monoclonal antibody recognizing IL-5 (TRFK-5) display reduced airway eosinophilia and hyperreactivity lasting for at least 3 mo (6).
To understand more fully the biologic implications of using a relatively long-acting antibody to treat pulmonary disease, we have extended the observations made in monkeys to a more experimentally tractable mouse model. In this way, it was possible to assess the effect of antibody treatment on several parameters of immune function, including eosinophil influx and T- and B-cell activity. Briefly, we show that treatment of mice with an anti-IL-5 monoclonal antibody results in inhibition of antigen-induced pulmonary infiltration of eosinophils for long periods of time because of its long circulating lifetime with little effect on immune system function.
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Materials and Methods |
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Top
Abstract
Introduction
Materials and methods
Results
Discussion
References
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Sensitization and Antigen Challenge
The sensitization and challenge protocol was performed as
described (13). Briefly, young male B6D2F1 mice (6 to 10 wk of age; Jackson Labs, Bar Harbor, ME), were sensitized by intraperitoneal injection with 15 µg of ovalbumin (OVA; Sigma Chemical Co., St. Louis, MO) adsorbed
to 2 mg of alum gel (aluminum hydroxide; Rehies, Inc.,
Berkeley Heights, NJ) in saline. A booster dose of the
OVA/alum mixture was administered 5 d later. Nonsensitized control animals received only alum gel in saline.
Twelve days after sensitization, the mice were placed in a
Plexiglas chamber and exposed twice to aerosolized OVA
(0.5%) for 1 h both in the morning and afternoon of a single day. The aerosolized OVA was produced by an ultrasonic nebulizer (Ultra-Neb 99; DeVilbiss, Somerset, PA).
At the indicated times before challenge (Figure 1a) or before sensitization and challenge (Figures 1b to 1f), mice
were injected intraperitoneally with TRFK-5 (rat antimouse IL-5) or GL113 (isotype control; rat anti-
-galactosidase). In all experiments, time was measured from the
point of injection of antibody unless noted otherwise.
Monoclonal antibodies were produced at Schering-Plough
(TRFK-5; Union, NJ) or Verax, Inc. (GL113; Lebanon,
NH), and endotoxin levels were 
0.015 EU/µg protein.
Recombinant murine IL-5 (rmIL-5) was obtained from
R&D Systems (Minneapolis, MN).
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Bronchoalveolar Lavage and Histology
At 24 h after OVA challenge, mice were euthanized by CO2 asphyxiation and samples of bronchoalveolar lavage fluid (BALF) and lung tissue were collected as described (13). Briefly, lungs were perfused through the pulmonary artery and lavaged through the trachea. BALF cells were applied to slides by cytocentrifugation, then fixed and visualized with Leukostat stain (Fisher Scientific, San Francisco, CA). Differential cell counts were made on at least 200 cells using standard morphological criteria, and total cell counts were determined with a Neubauer hemocytometer.
For histologic evaluation of lung tissues, perfused and lavaged lungs were removed and fixed in 10% phosphate-buffered formalin. The left lobes of each lung were embedded in paraffin, sectioned to 5 µm, and stained with hematoxylin and eosin. Cellular infiltrates were assessed in five randomly selected fields with a Zeiss Axiophot microscope (Carl Zeiss Inc., Thornwood, NY) at ×500 magnification. Groups were analyzed in a blind fashion.
Femoral bone marrow cells were collected as described and the number of eosinophils in bone marrow aspirates was measured using standard morphologic criteria (13).
Phenotypic Analysis of T-Cell Subsets
BALF from groups of six mice were collected 24 h after
OVA challenge, pooled and stained as described (21).
Briefly, cells were incubated with unlabeled anti-Fc
II receptor monoclonal antibody (clone 2.4G2; PharMingen,
San Diego, CA) to block Fc-mediated and nonspecific
binding and then stained with the following monoclonal antibodies (PharMingen): anti-Thy1.2-fluorescein (clone 53-2.1), anti-B220-phycoerythrin (clone RA3-6B2), anti-CD4-phycoerythrin (clone RM4-5), anti-CD8a-fluorescein (clone
53-6.7), anti-CD44-biotin (pgp-1; clone IM7), and anti-CD45RB-biotin (clone 16A) followed by streptavidin-peridinin
chlorophyll protein. Stained cells fixed in 1% paraformaldehyde were analyzed on a FACSort flow cytometer (Becton-Dickinson Immunocytometry Systems, San Jose, CA). The percentage of cells expressing a given surface phenotype
and the total cell counts were used to calculate the absolute numbers of T cells per volume of BALF.
RNA Isolation and Semiquantitative Polymerase Chain Reaction
At 6 h after OVA challenge, lung tissue was prepared for the measurement of IL-4 and IL-5 steady-state mRNA by semiquantitative polymerase chain reaction (PCR) as described previously (22, 23). Perfused lungs were removed and immediately frozen in dry ice. Lung tissue from groups of four mice were pooled and solubilized in guanidinium isothiocyanate and RNA was purified by cesium chloride ultracentrifugation. RNA was transcribed into cDNA with avian myeloblastosis virus-reverse transcriptase and serial dilutions prepared for PCR amplification with murine IL-4- or murine IL-5-specific primers as described (23). Levels of steady-state mRNA for each cytokine were quantitated by comparison to standard curves generated from serial dilutions of cDNA from concanavalin A-stimulated BALB/c splenocytes, an abundant source of cytokine mRNA.
Measurement of Serum IgE, IgG, and TRFK-5
Total serum IgE and IgG from blood samples obtained at the indicated times were determined by enzyme-linked immunosorbent assay (ELISA) using procedures that have been described previously (24). Serum Ig levels were quantified by comparison with purified isotype standards. Standard IgE was purchased from PharMingen, and anti-mouse IgE and IgG were a gift from Dr. Robert Coffman (DNAX Research Institute, Palo Alto, CA). TRFK-5 in serum was determined by an ELISA using mouse antirat IgG (Biosource, Camarillo, CA) as the capture antibody and alkaline phosphatase-labeled mouse antirat IgG (Sigma Chemical) as the detecting antibody. These antibodies do not cross-react with murine IgG. Serum samples were diluted at least fourfold and quantitated by comparison with serial dilutions of a known amount of TRFK-5.
Statistical Analysis
Data are expressed as mean ± SEM and representative
experiments are shown. The significance of differences between experimental groups (P 0.05) was determined by
analysis of variance using Fisher's protected least significance test (StatView; Abacus Concepts Inc., Berkeley,
CA). The half-life of TRFK-5 was determined by regression analysis (Graphpad Prism; Graphpad Software Inc.,
San Diego, CA).
Animal Care and Use
This study was performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and the Animal Welfare Act in a program approved by the American Association for the Accreditation of Laboratory Animal Care.
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Results |
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Abstract
Introduction
Materials and methods
Results
Discussion
References
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Effect of One Treatment of TRFK-5 on Pulmonary Inflammation
To determine the duration of the therapeutic effect of anti-IL-5 monoclonal antibody treatment, young mice were treated once with TRFK-5 or control antibody GL113 (1 mg/kg, intraperitoneally) and rested for 0 to 22 wk (Figures 1a to 1f). At the indicated times, mice were sensitized and challenged as described in Materials and Methods. Saline-treated, sensitized/challenged mice exhibited significant infiltration of eosinophils into BALF (1.5 to 2.5 × 105 eosinophils/ml BALF), whereas BALF from nonsensitized/challenged mice contained virtually no eosinophils (Figure 2A; 13). Treatment of mice with TRFK-5 2 h prior to challenge resulted in significant (> 80%) inhibition of eosinophil recruitment into BALF (Figure 2A; 17). One dose of TRFK-5 given before sensitization and challenge was effective at inhibiting the infiltration of eosinophils into BALF for at least 12 wk. After 24 wk, the TRFK-5 antibody was no longer effective and the pulmonary eosinophilia in this group was not different from that of either saline- or GL113-treated mice (Figure 2A). The inhibition of pulmonary eosinophilia was dependent on the dose of TRFK-5. That is, when 10-fold less TRFK-5 antibody was administered as a single dose (0.1 mg/kg, i.p.), antigen challenge-dependent eosinophil infiltration into BALF was inhibited for 6 wk (data not shown). After 8 wk, TRFK-5 was no longer effective at inhibiting eosinophilia.
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Eosinophils in the bone marrow were significantly reduced by antigen challenge from 299 ± 36 × 103/femur in saline-treated mice to 166 ± 26 × 103/femur in sensitized/ challenged mice (Figure 2B; 17). Treatment of sensitized mice with TRFK-5 2 h before challenge almost completely inhibited the reduction of eosinophils in the bone marrow (Figure 2B; 17). Exposure of mice to one dose of TRFK-5 inhibited this challenge-induced reduction of eosinophils for at least 8 wk. By 12 and 24 wk after treatment, TRFK-5 no longer inhibited the reduction of eosinophils in the bone marrow of sensitized/challenged mice. Therefore, the inhibition of bone marrow eosinophils by TRFK-5 mirrored the inhibition of eosinophil influx into BALF.
Serum Concentrations of TRFK-5
TRFK-5 is a rat IgG1 monoclonal antibody (25) that can be measured in mouse serum samples by quantitating rat IgG1 with rat-specific antibodies. In this way, TRFK-5 concentrations in serum samples obtained 24 h after challenge at the indicated times were measured in parallel with the number of eosinophils in BALF. One day after a single dose of TRFK-5 (1 mg/kg, intraperitoneally; Figure 1a), the serum concentration of TRFK-5 was 6,560 ng/ml (Figure 3). At this time, there was a large (> 80%) inhibition of the BALF eosinophils after antigenic challenge. There was an exponential fall in serum concentrations of TRFK-5 over time, and the half-life was evaluated by nonlinear regression over a period of 2 to 24 wk to be 2.4 wk (95% confidence interval 4.03 to 1.68; R squared 0.9976). After 18 to 19 wk, eosinophilia was inhibited by 50% and the concentration of TRFK-5 in serum was approximately 25 ng/ml.
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To assess the acute dose of TRFK-5 that inhibited BALF eosinophils by 50% (ED50) and the inhibitory concentration of TRFK-5 in the serum producing this effect (IC50), young sensitized mice were administered decreasing TRFK-5 doses 2 h before antigen challenge (see Figure 1a protocol). The level of eosinophils in BALF and of TRFK-5 in serum were measured 24 h after challenge (Figure 4). These mice were 18 to 19 wk younger than the ones just described. The calculated ED50 for TRFK-5 was 0.06 mg/kg intraperitoneally, and the IC50 producing this effect was 230 ng/ml.
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Effect of TRFK-5 Treatment on IL-5-Induced Blood Eosinophilia
A model of IL-5-induced blood eosinophilia was used to assess the potency of TRFK-5 remaining in mice after a single pretreatment with antibody. Mice pretreated with one dose of TRFK-5 (1 mg/kg, intraperitoneally) were rested for 8 wk, after which the serum concentration of TRFK-5 had dropped 14-fold, but antigen-challenge-induced pulmonary eosinophilia was still inhibited (Figure 3). Intravenous administration of rmIL-5 (10 µg/kg) to saline-pretreated mice produced a 5.5- to 7.5-fold increase in blood eosinophils measured 3 h after injection (Figure 5). When mice were pretreated with one dose of TRFK-5 (1 mg/kg intraperitoneally) 8 wk before rmIL-5 administration, blood eosinophilia was significantly reduced (50% to 65%) compared with that observed in saline- or GL113-pretreated mice.
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Effect of Long-Term Exposure to TRFK-5 on Immune Function
To assess the effects of long-term exposure to TRFK-5, several indicators of immune function were analyzed. Sensitized mice respond to aerosolized antigen challenge within 24 h with an increase in the numbers of T cells found in BALF, but with little change in the numbers of B cells (21, 26). The T-cell response consists of activated memory cells of the Th2 phenotype producing the characteristic cytokines IL-4 and IL-5 (21, 23). In addition, aerosol challenge of allergic mice results in increased levels of IgE and IgG in serum when measured after 24 h (13). Therefore, the numbers of T cells and B cells in BALF, the accumulation of cytokines in lung tissue, and the levels of Ig in serum were measured in TRFK-5-pretreated mice.
The migration of T cells into pulmonary tissue assessable by BAL was unaffected by a single treatment of mice with TRFK-5 for at least 12 wk (Figure 6). However, 24 wk after a single dose of TRFK-5, the mice exhibited a dramatic increase in the number of T cells that were measured in BALF compared with untreated or GL113-treated mice 24 h after challenge. This observation was confirmed by histologic evaluation of lung tissue collected at the same time, which indicated a dramatic increase in mononuclear cells surrounding bronchi and bronchioles (data not shown). Further analysis of the T-cell subtype by flow cytometry indicated that the cells were predominantly CD4+ with an activated, memory phenotype (CD44+, CD45RBlo; data not shown). Treatment of mice did not result in any meaningful changes in the numbers of B220+ B cells measured in BALF (Figure 6).
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Although treatment of mice with TRFK-5 resulted in a dramatic increase in T cells after 24 wk, at that time there were no measurable affects on several indicators of T-cell function, including cytokine production (Figure 7) and T-cell-dependent Ig production (Figure 8). T cells are the predominant source of IL-5 steady-state mRNA in this model (23), and therefore antigen challenge-induced changes in IL-5 mRNA levels are an indication of T-cell function. IL-5 steady-state mRNA levels in pulmonary tissue from mice treated once with TRFK-5 for 24 wk were not different than those measured in untreated or GL113-treated challenged mice (Figure 7A). In fact, there was less IL-5 mRNA in pulmonary tissue from all groups after 12 and 24 wk, reflecting an age-dependent decrease in the ability of mice to produce IL-5 mRNA that occurred after 8 wk (Figure 7A, data not shown). However, at all time points there were challenge-induced increases in the amount of IL-5 steady-state mRNA levels consistent with that found in a previous study (21). There were increased IL-5 mRNA levels in the lungs of mice treated with TRFK-5 once for 2 h before challenge compared with those in control or GL113-treated mice (Figure 7A). Similar results were obtained when TRFK-5 was given 2 wk before challenge, but this effect was not observed when TRFK-5 was given 4, 8, 12, or 24 wk before challenge (Figure 7A and data not shown). At all times tested, one dose of TRFK-5 had no effect on the lung levels of IL-4 mRNA (Figure 7B).
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In this allergic model, increases in serum levels of IgE and IgG are observed after challenge (13). Pretreatment of mice with TRFK-5 before challenge had no meaningful effect on their ability to produce IgE or IgG (Figures 8A and 8B, respectively). Although the role of T cells in the Ig response is not definitively known for this model, it is clear from this experiment that the increase in T cells in BALF seen at 24 wk after TRFK-5 treatment did not influence the levels of Ig in serum, and therefore there was no effect on T-cell-dependent Ig production.
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Discussion |
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Abstract
Introduction
Materials and methods
Results
Discussion
References
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IL-5 plays an important role in the progression of the inflammatory response observed in asthma (15). Experiments using animal models of allergic disease have shown that the inhibition of IL-5 activity by administration of anti-IL-5 monoclonal antibodies results in ablation of pulmonary eosinophilic infiltration and, in many cases, reduction in airway hyperreactivity to non-specific stimuli, two of the hallmarks of asthma (5, 17, 18). In a majority of these studies, animals were treated just before antigen challenge. However, if IL-5 antibodies are to be developed for use as a therapy for asthma, the effects of antibody treatment over long periods of time need to be determined, especially because a prior study with cynomolgus monkeys demonstrated that such treatment was effective for at least 3 mo (6). Therefore, this study was undertaken to evaluate the long term effects of anti-IL-5 therapy in a well-characterized murine model in which only the acute effects of anti-IL-5 antibodies have been examined previously (17). Several characteristics of anti-IL-5 antibody treatment in this model are unknown, including correlations among the dose and serum concentrations of antibody required to inhibit eosinophilia, the duration of the inhibition, and the long-term effects of antibody treatment on related immune function. In brief, this study demonstrates that relatively low serum concentrations of anti-IL-5 result in long-term inhibition of lung eosinophilia with little measurable effect on B- or T-cell function.
After a single administration (1 mg/kg, intraperitoneally) of TRFK-5, the antibody exhibited a serum half-life of 2.4 wk and inhibition of pulmonary eosinophilia required serum levels of less than 230 ng/ml of antibody (Figure 4). Antibody treatment had an effect at very early stages of eosinophil recruitment, because migration out of the bone marrow was also affected for an extended period of time. Interestingly, in mice that were 4 to 5 mo older, less TRFK-5 was required for inhibition of pulmonary inflammation (approximately 25 ng/ml in serum; Figure 3). This was likely due to lower levels of IL-5 in the older mice, a reflection of the 6-fold lower IL-5 mRNA levels detected at the later times (Figure 7).
The 2-wk half-life of this antibody, coupled with the requirement for very low levels of antibody needed for biologic effect, accounted for the prolonged duration of action seen in this model. This conclusion was tested by demonstrating that antibody remaining in mice weeks after a single dose of TRFK-5 neutralized 50% to 65% of the blood eosinophilia induced by administration of rmIL-5.
Long-term exposure to TRFK-5 did not affect the sensitization process in these mice. They exhibited challenge-induced increases in serum levels of IgE and IgG at all
times tested. In addition, these results suggested that anti-IL-5 treatment was not working by inhibiting Ig production by B cells. Furthermore, we were unable to detect
anti-idiotype responses in TRFK-5 treated mice (data not
shown). Although not all isotypes were examined, the
ability to mount a relatively normal Ig response to antigen
stimulation suggests that IL-5 plays only a small role in in
vivo B-cell function, contrary to the initial identification of
this cytokine as "B cell growth factor II" (27, 28) and consistent with more recent studies using IL-5-deficient mice
(29). In addition, although not tested directly, the presence
of normal antigen-dependent Ig production suggests that
an antiparasite response should occur. Experiments with
IL-5-deficient mice are consistent with this hypothesis
(29). In IL-5
/ mice, the absence of IL-5 during the life of
the mice does not result in increased numbers of M. corti
in the peritoneal cavity or liver compared with those found
in IL-5+/+ mice. Although IL-5/ mice do not respond to
challenge with eosinophilia, baseline eosinophil levels remain the same as in IL-5+/+ mice. IL-5/ mice have normal T- and B-cell development and normal T-cell-dependent Ig responses.
Treatment with TRFK-5 for at least 12 wk resulted in no measurable effect on the number of T cells measured in BALF. Also, there were no dramatic effects on levels of the T-cell-derived cytokines IL-4 and IL-5. However, there was a small increase in antigen challenge-induced IL-5 steady-state mRNA production in TRFK-5-treated mice after 2 h or 2 wk, but this was not observed at other times examined. This might reflect a feedback mechanism that regulates IL-5 production.
Twenty-four weeks after treatment with TRFK-5, when challenge again resulted in eosinophilia, there was a dramatic increase in the number of T cells found in BALF that was not reflected in the number of B cells. However, there was no functional consequence to this observation. That is, there was no difference between control mice and TRFK-5-treated mice in the amount of IL-4 or IL-5 mRNA produced in the lungs. Also, the additional T cells did not influence the amount of IgE or IgG produced by B cells. The T cells were phenotypically the same as T cells seen in GL113-pretreated and -challenged mice, predominantly CD4+CD45RBloCD44+. However, because there were no observed increases in T-cell function, it is likely that these cells were either anergic or antigen-nonspecific.
In conclusion, these data indicate that low concentrations of anti-IL-5 antibody are sufficient for inhibiting pulmonary and blood eosinophilia and that long-term treatment does not result in significant effects on normal immune function. The long-term effects of anti-IL-5 antibodies are therefore not due to any profound effect on the immune system. They are the result of the circulating lifetime of the antibody and, much like other therapeutic entities with shorter half-lives, when their concentration falls below a threshold the biologic effect dissipates.
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Footnotes |
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Address correspondence to: Charles G. Garlisi, Ph.D., Allergy and Immunology, Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033-0539. E-mail: charles.garlisi{at}spcorp.com
(Received in original form February 9, 1998).
Abbreviations: bronchoalveolar lavage fluid, BALF; enzyme-linked immunosorbent assay, ELISA; immunoglobulin, Ig; interleukin, IL; ovalbumin, OVA; polymerase chain reaction, PCR; recombinant murine IL, rmIL.Acknowledgments: The authors thank Dr. Francis M. Cuss for helpful discussions and support, and Dr. Steven Swanson for preliminary determinations of TRFK-5 levels in serum.
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References |
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Introduction
Materials and methods
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Discussion
References
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