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Development of Lupus in BXSB Mice Is
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J Immunol 2000; 164:38-42; ;
doi: 10.4049/jimmunol.164.1.38
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References
Dwight H. Kono, Dimitrios Balomenos, Miyo S. Park and
Argyrios N. Theofilopoulos
Development of Lupus in BXSB Mice Is Independent of IL-41
Dwight H. Kono,2* Dimitrios Balomenos,† Miyo S. Park,* and Argyrios N. Theofilopoulos*
Although systemic lupus erythematosus appears to be a humorally mediated disease, both Th1 and Th2 type responses have been
implicated in its pathogenesis. The Th1 response, as exemplified by IFN-g production, has been uniformly shown in mouse lupus
models to be critical for disease induction. The role of Th2 type responses, however, is more complicated, with some studies
showing detrimental and others beneficial effects of IL-4 in these models. To further address this issue, we generated and analyzed
IL-4 gene-deficient BXSB mice. Mice homozygous for this deletion had significantly lower serum levels of total IgG1 compared
with wild-type BXSB, consistent with the lack of IL-4. However, no significant differences were observed in mortality, spleen
weight, severity of glomerulonephritis, levels of anti-chromatin and anti-ssDNA Abs, or frequency of activated (CD44high) CD41
T cells. The anti-chromatin Ab isotype response was virtually all Th1 type in both the knockout and wild-type BXSB. These
findings directly demonstrate that IL-4 and, by inference, Th2 cells are not obligatory participants in the induction and maintenance of lupus in this strain. The Journal of Immunology, 2000, 164: 38 – 42.
C
*Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037;
and †Centro Nacional de Biotecnologia, Madrid, Spain
Received for publication June 21, 1999. Accepted for publication October 12, 1999.
The costs of publication of this article were defrayed in part by the payment of page
charges. This article must therefore be hereby marked advertisement in accordance
with 18 U.S.C. Section 1734 solely to indicate this fact.
1
This work was supported in part by National Institutes of Health Grants AR42242,
ES08666, and AR39555. This is Scripps Research Institute Publication 12535-IMM.
2
Address correspondence and reprint requests to Dr. Dwight H. Kono, Department of
Immunology-IMM3, The Scripps Research Institute, 10550 North Torrey Pines Road,
La Jolla, CA 92037. E-mail address: [email protected]
3
Abbreviations used in this paper: SLE, systemic lupus erythematosus; PerCP, peridinin
chlorophyll protein; EAMG, experimental autoimmune myasthenia gravis; GN, glomerulonephritis; Yaa, Y chromosome accelerated autoimmunity and lymphoproliferation.
Copyright © 2000 by The American Association of Immunologists
body-mediated diseases like SLE might involve Th2-type responses, whereas cell-mediated responses in organ-specific autoimmune diseases might be mediated by Th1-type responses. This
attractive but simplistic model, however, has not been supported
by studies of humoral autoimmune diseases, such as lupus (17–22)
and myasthenia gravis (23), nor by studies of certain organ-specific
diseases such as experimental autoimmune encephalomyelitis (24)
and experimental autoimmune uveitis (25, 26). In the case of lupus, considerable evidence points to the importance of the Th1
response for disease induction and acceleration. Cytokine profiles
of spleen cells in BXSB and MRL-Faslpr mice showed mainly
increases in the Th1 cytokine, IFN-g (27). Accelerated disease in
closely related lupus susceptible and nonsusceptible mouse strains
was associated with increases in both IFN-g and the Th1-mediated
IgG isotypes, IgG2a and IgG3 (28). IFN-g accelerated, while antiIFN-g Ab or soluble IFN-gR, prevented disease in spontaneous
murine lupus (17, 18). Moreover, gene knockout of IFN-g or
IFN-gR eliminated disease in spontaneous and xenobiotic-induced
models of lupus (19 –22, 29).
Th2-type responses have also been associated with the development of SLE. Increases in number of IL-4-producing cells have
been found in some lupus-susceptible strains (30, 31); treatment
with blocking anti-IL-4 Ab or soluble IL-4R reduced autoantibody
production and nephritis in (NZB 3 NZW)F1 and MRL-Faslpr
mice (32, 33), and IL-4 knockout (MRL-Faslpr 3 B6)F2 mice had
less lymphadenopathy and end-organ disease compared with IL-4
wild-type littermate controls. Overall these studies suggest both
Th1 and Th2 responses are important for the development of lupus.
Other findings, however, suggest that the Th2 response may be
less important than Th1 response in lupus or even protective. In
BXSB and MRL-Faslpr mice, the increases in IL-4 levels and IL-4
producing cells although present are much less than the increases
observed in IFN-g levels or IFN-g-producing cells (27, 30, 34).
Knockout of the IL-4 gene had no effect on disease induction in the
mercury-induced model of systemic autoimmunity, which was previously considered a prototypic Th2-mediated humoral disease
(29, 35). Finally, expression of an IL-4 transgene by B cells in
(NZW 3 C57BL/6.Yaa)F1 mice protected rather than enhanced
the development of lupus nephritis, and this protection appeared to
be due to deviation of the autoantibody response away from especially
pathogenic Th1-mediated IgG subclasses such as IgG3 (36).
0022-1767/00/$02.00
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D41 T cells following activation become polarized into
two main subsets, designated Th1 and Th2, that manifest
distinct cytokine profiles and effector functions (1, 2).
The Th1 subset, which secretes IL-2, IFN-g, and TNF-b, promotes
mainly cellular-mediated responses that provide protection from
intracellular pathogens, activation of phagocytes, and delayed-type
hypersensitivity (DTH), but also, through IFN-g, class switching
to the IgG2a isotype. In contrast, Th2 cells that secrete IL-4, IL-5,
IL-6, IL-10, and IL-13 protect against extracellular pathogens, induce IgE-mediated allergic responses, activate eosinophils, and
promote humoral responses, particularly of the IgG1 and IgE subclasses. The development of CD41 cells into Th1 or Th2 cells can
be influenced by a number of factors, including cytokine milieu,
type of APC, Ag affinity and concentration, costimulatory molecules, and duration of exposure. The molecular basis for this functional dichotomy has not been fully defined; however, a number of
key cytokine and chemokine receptors expressed on either Th1 or
Th2 cells have been identified that are important for differentiation
and effector functions (3– 6). Several protooncogenes, kinases, and
transcription factors have also been implicated in the development
of the Th subsets (7–12).
Systemic lupus erythematosus (SLE)3 is a humoral-mediated
disease that is nonetheless dependent on CD41 T cells for disease
induction (13), presumably because pathogenic autoantibodies require T-dependent affinity maturation (14 –16). The functional dichotomy of Th subsets suggests that T cell-dependent autoanti-
The Journal of Immunology
39
FIGURE 1. Survival, spleen weight, and GN in IL-4 wild-type and knockout BXSB mice. Left panel, Survival at 5 mo. Center panel, Average spleen
weights, p , 0.01 between IL-41/2 and IL-42/2 groups. Only spleen weights of mice surviving to 5 mo of age are included. Right panel, Average GN
score. 1/1, 1/2, and 2/2, IL-4 wild-type, heterozygous knockout, and homozygous knockout, respectively. p . 0.05 between groups unless otherwise
stated. Numbers of 1/1, 1/2, and 2/2 mice were 7, 10, and 9, respectively, for mortality and 2, 9, and 3, respectively, for GN and spleen weight.
Materials and Methods
Mice
Wild-type and IL-4 knockout (IL-42/2) BXSB mice were bred and maintained under specific pathogen-free conditions at The Scripps Research
Institute. IL-4 knockout mice of mixed 129 3 C57BL/6 background were
kindly provided by Dr. W. Müller (University of Cologne, Germany) (37).
BXSB IL-4-deficient mice were generated by seven backcrosses to the
BXSB strain, followed by intercrossing of the heterozygous IL-41/2 offspring. Wild-type or heterozygous IL-4 knockout littermates from this intercross at N7 were used as controls. The BXSB Y chromosome containing
the Yaa (Y chromosome-accelerated autoimmunity and lymphoproliferation) gene was bred into the backcross mice, and only male mice with the
Yaa gene were analyzed in this study. Genotyping of the IL-4 wild-type
and knockout mutation by PCR was performed as described (38).
Pathology
Autopsy and histologic examination of mice were performed as previously
described (38) at 5 mo or if mice developed severe disease. Tissues were
fixed in Bouin’s solution, and sections were stained with periodic-acid
Schiff reagent. GN was graded on a 0 – 41 scale (38).
Serology
Serum IgG1 and IgG2a subclass levels were determined by ELISA as
previously described (29), with standard curves generated using calibrated
mouse serum (Binding Site, Birmingham, England). Levels of Ig were
calculated by regression plots of standard values (Statview, SAS Institute,
Cary, NC). Abs against chromatin or ssDNA were assayed as previously
described (14). HRP-conjugated anti-mouse IgG1- (Caltag, Burlingame,
CA) or IgG2a-specific (PharMingen, La Jolla, CA) detecting Abs were
used for identifying IgG1 or IgG2a anti-chromatin Abs. Positive and negative control sera were included on each plate for normalization. The
amount of chromatin-bound IgG1 or IgG2a Ab was calculated from an
IgG1 or IgG2a subclass standard curve that was included on each plate.
This standard curve was generated as above for detecting serum IgG1 and
IgG2a subclass levels (29).
Flow cytometry
Spleen cell suspensions were stained with anti-CD8-FITC, antiCD4-peridinin chlorophyll protein (PerCP), anti-CD44-PE, and antiCD62L-biotin/streptavidin-APC (PharMingen). Cell data were acquired on
a FACSORT and analyzed with the Cell Quest program (Becton Dickinson, Mountain View, CA). Live cells were gated based on their forward
and side scatter characteristics.
Statistical Analysis
Unpaired comparisons between groups were analyzed by t test. Survival
was analyzed using Kaplan-Meier statistics by log rank (Mantel-Cox) test
with censored events. A value of p , 0.05 was considered significant.
Results
Survival of IL-42/2 and wild-type BXSB mice
BXSB background mice deficient in IL-4 expression and IL-41/1
and IL-41/2 littermate controls were produced by backcrossing
IL-41/2 mice to the BXSB strain and then intercrossing the N7
generation. When the resulting wild-type and IL-42/2 mice were
followed for disease, both developed early mortality with no significant difference in survival at 5 mo (43% and 44%, respectively,
Fig. 1). The IL-41/2 mice had greater survival (90%) compared
with both wild-type and homozygous knockout BXSB mice, but
this did not reach statistical significance ( p . 0.05). The overall
62% (16/26) combined survival at 5 mo of these mice was comparable to our colony of BXSB mice, which have a 50% mortality
around 5[1–2] mo of age. This suggests that the backcrossing has
been sufficient to fix most if not all of the BXSB lupus-predisposing alleles.
IL-4 knockout mice develop lymphoaccumulation and GN
Wild-type and both partial and complete IL-4 knockout mice all
developed lymphoaccumulation. The average spleen weight of the
IL-42/2 mice, however, was slightly lower than the IL-41/1 mice
( p . 0.05), and significantly smaller than the IL-41/2 mice ( p ,
0.01, Fig. 1). Histologic examination of the kidneys revealed that
IL-4 deficient BXSB mice, with either homozygous or heterozygous deletions, developed the typical features of GN observed in
wild-type BXSB mice. Glomeruli had periodic-acid Schiff-staining
deposits, cellular proliferation primarily mesangial, occasional
polymorphonuclear cells, and rare crescents (not shown). There
were no significant differences in the type and severity of GN
among the three groups (Fig. 1).
IgG1 levels are decreased in IL42/2 mice
IL-4-deficient BXSB had significantly lower serum levels of total
IgG1 compared with wild-type or heterozygous IL-41/2 mice
(1/1, 171 6 85 mg/ml; 1/2, 182 6 95 mg/ml; 2/2, 78 6 21
mg/ml; p , 0.05), but similar levels of IgG2a, a subclass that is
dependent on IFN-g (Th2 cytokine) and not IL-4 (Fig. 2). This is
consistent with the deficiency of IL-4.
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To further define the role of the Th2-type response in SLE we
backcrossed the IL-4 gene knockout mutation onto the BXSB lupus-prone background and assessed its effect on autoimmunity.
Strikingly, homozygous deletion of the IL-4 gene did not affect
autoantibody production, glomerulonephritis (GN) or mortality in
this strain.
40
IL-4 KNOCKOUT BXSB MICE
FIGURE 4. Serum levels of IgG1 and IgG2a anti-chromatin Ab in IL-4
wild-type and knockout BXSB mice. 1/1, 1/2, and 2/2, IL-4 wild-type,
heterozygous knockout, and homozygous knockout, respectively. IgG1
levels are in OD units because all values for 1:100 serum dilution were
below the lowest concentration of the IgG1 standard (,0.31 mg/ml). Numbers of mice in each group are 6, 8, and 5 for 1/1, 1/2, and 2/2,
respectively. p . 0.05 for all unpaired comparisons between groups.
Levels of autoantibodies in IL-4-deficient and wild-type BXSB
mice
of our standard curve at a serum dilution of 1:100, indicating a
concentration ,0.31 mg/ml (Fig. 4). In contrast, levels of IgG2a
were similar for all IL-4 groups, with levels in the 300 – 400 mg/ml
range. Thus, even in the IL-41/1 BXSB mice, the relative amounts
of anti-chromatin IgG2a to IgG1 is much greater than would be
expected from the ratio of total IgG2a to IgG1. This would
strongly suggest a greater role for the Th1-type responses in the
production of anti-chromatin Ab.
The effect of IL-4 deficiency on autoantibody production was also
examined. Levels of both IgM and IgG Abs to chromatin and
ssDNA were measured at 4 –5 mo of age (Fig. 3). Although there
was some variability in mean values among the IL-41/1, IL-41/2,
and IL-42/2 littermates, none of the differences reached statistical
significance ( p . 0.05).
Similar to what was observed with total IgG1 and IgG2a levels,
IL-4 deficiency should skew the IgG1 and IgG2a anti-chromatin
Ab response away from IgG1 production toward the IgG2a response. When this was examined, however, no difference in the
IgG1 anti-chromatin levels could be detected because the IgG1
anti-chromatin levels were exceedingly low, below the lower limit
IL-4-deficient BXSB exhibit similar levels of memory/effector
subset CD41 T cells
A characteristic feature of autoimmune BXSB mice is the accelerated accumulation of effector/memory CD41 T cells compared
with nonautoimmune strains of mice (30, 39). As shown in Table
I, splenocytes from 2-mo-old wild-type and IL-4-deficient BXSB
mice exhibited similar percentages of CD41 and CD81 T lymphocytes in the spleen ( p . 0.05). Importantly, there were no
significant differences ( p . 0.05) in the percentage of effector/
memory phenotype CD41 T cells in wild-type and IL-42/2 BXSB
mice, either by the presence of CD44hi (22.0 6 4.4% and 26.8 6
7.0%, respectively) or CD62Llow (13.8 6 3.2% and 16.0 6 5.7%)
cell surface expression. In contrast, non-autoimmune C57BL/6
mice had lower levels of CD41 and CD41CD44high T cells ( p ,
0.028) compared with both wild-type and IL-42/2 mutant
BXSB mice.
Discussion
FIGURE 3. Serum levels of IgM and IgG anti-chromatin and antissDNA Abs in IL-4 wild-type and knockout BXSB mice. 1/1, 1/2, and
2/2, IL-4 wild-type, heterozygous knockout, and homozygous knockout,
respectively. Sera are from 4- to 5-mo-old mice. Numbers of mice in each
group are 6, 8, and 5 for 1/1, 1/2, and 2/2, respectively. p . 0.05 for
all unpaired comparisons between groups.
In this study, no significant difference in a broad range of autoimmune manifestations, including early mortality, GN, hyperIgG2a,
anti-chromatin autoantibody, and increases in memory/effector
phenotype Th cells were observed between homozygous IL-4 gene
knockout BXSB mice and their IL-4 wild-type littermates. Interestingly, slight differences between these two groups and the IL-4
heterozygous group were noted for spleen weight and mortality,
although the latter was not statistically significant. Overall, however, any differences between the IL-41/2 mice and the other
groups were small as there was significant GN at 5 mo for most of
the IL-4 heterozygous mice and autoantibody levels between the
groups were similar. Because of the number of mice examined,
small effects of IL-4 deficiency on GN and spleen size cannot be
excluded. Nevertheless, the findings clearly demonstrate that IL-4
is not required for lupus-like disease in BXSB mice and contradict
the notion that the Th2 response is important for the development
of lupus.
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FIGURE 2. Total serum IgG1 and IgG2a subclass levels in IL-4 wildtype and knockout BXSB mice. 1/1, 1/2, and 2/2, IL-4 wild-type,
heterozygous knockout, and homozygous knockout, respectively. Levels of
IgG1 are significantly lower in IL-42/2 BXSB than either 1/1 or 1/2
mice (p , 0.05). No significant differences were observed in total IgG2a
levels between IL-4 wild-type, 1/2 or 2/2 BXSB mice (p . 0.05). Numbers of 1/1, 1/2, and 2/2 mice were 3, 10, and 5, respectively.
The Journal of Immunology
41
Table I. Splenic T cell populations in IL-42/2 BXSB micea
Strain
% T cells
% CD41
% CD41
CD44high
% CD41
CD62low
% CD81
CD44high
% CD81
CD62low
B6
BXSB
IL-42/2
40.1 6 2.4
40.3 6 1.8
39.7 6 1.5
63.8 6 0.7
67.9 6 0.6*
71.0 6 1.2**
10.2 6 2.1
22.0 6 2.6***
26.8 6 4.1***
9.4 6 1.2
13.8 6 1.9
16.0 6 3.3
14.0 6 1.8
11.3 6 0.4
11.6 6 1.9
1.9 6 0.5
2.1 6 0.1
3.5 6 0.4***
a
Mean and SE are shown. Percentages are of all live-gated cells (column 2), T cells (column 3), CD41 cells (columns 4 and 5), or CD81 cells (columns 6 and 7). T cells
included all CD4- or CD8-staining cells. Spleen cell suspensions from three 2 mo-old male mice were stained with anti-CD4-PerCP, anti-CD8-FITC, anti-CD44-PE, and
anti-CD62L-biotin/streptavidin-APC. BXSB mice in this experiment are from the main colony.
p, p 5 0.011; pp, p 5 0.007; ppp, p , 0.028 (compared with B6 mice).
6.Yaa)F1 lupus mice. Deletion of the IL-4 gene in BXSB mice,
however, did not result in greater disease severity, which would be
expected if normal levels of IL-4 were acting to down-regulate the
autoimmune response. This suggests that disease suppression by
transgenic expression of IL-4 may not represent the normal function
of IL-4, but a consequence of pharmacologic doses of this cytokine.
In this case, the reduction in GN may also be due to the antiinflammatory effects of IL-4 (53) in addition to the postulated deviation of
autoantibody IgG subclasses to less pathogenic forms (36).
It is evident that the separation of diseases into Th1 and Th2
types is overly simplistic and that careful analysis of individual
cytokines will be necessary to define their contributions and interactions. The present findings further indicate that future studies
must also take into account the effects of different susceptibility
backgrounds.
Acknowledgments
We thank Dana Kidd for technical assistance.
References
1. Mosmann, T. R., and S. Sad. 1996. The expanding universe of T-cell subsets:
Th1, Th2 and more. Immunol. Today 17:138.
2. Romagnani, S. 1997. The Th1/Th2 paradigm. Immunol. Today 18:263.
3. Xu, D., W. L. Chan, B. P. Leung, D. Hunter, K. Schulz, R. W. Carter,
I. B. McInnes, J. H. Robinson, and F. Y. Liew. 1998. Selective expression and
functions of interleukin 18 receptor on T helper (Th) type 1 but not Th2 cells.
J. Exp. Med. 188:1485.
4. Szabo, S. J., A. S. Dighe, U. Gubler, and K. M. Murphy. 1997. Regulation of the
interleukin (IL)-12R b 2 subunit expression in developing T helper 1 (Th1) and
Th2 cells. J. Exp. Med. 185:817.
5. Sallusto, F., A. Lanzavecchia, and C. R. Mackay. 1998. Chemokines and chemokine receptors in T-cell priming and Th1/Th2- mediated responses. Immunol.
Today 19:568.
6. Pernis, A., S. Gupta, K. J. Gollob, E. Garfein, R. L. Coffman, C. Schindler, and
P. Rothman. 1995. Lack of interferon g receptor b chain and the prevention of
interferon g signaling in TH1 cells. Science 269:245.
7. Rincon, M., and R. A. Flavell. 1997. T-cell subsets: transcriptional control in the
Th1/Th2 decision. Curr. Biol. 7:R729.
8. Rincon, M., H. Enslen, J. Raingeaud, M. Recht, T. Zapton, M. S. Su, L. A. Penix,
R. J. Davis, and R. A. Flavell. 1998. Interferon-g expression by Th1 effector T
cells mediated by the p38 MAP kinase signaling pathway. EMBO J. 17:2817.
9. Zheng, W., and R. A. Flavell. 1997. The transcription factor GATA-3 is necessary and sufficient for Th2 cytokine gene expression in CD4 T cells. Cell 89:587.
10. Ho, I. C., D. Lo, and L. H. Glimcher. 1998. c-maf promotes T helper cell type 2
(Th2) and attenuates Th1 differentiation by both interleukin 4-dependent and
-independent mechanisms. J. Exp. Med. 188:1859.
11. Chow, C. W., M. Rincon, and R. J. Davis. 1999. Requirement for transcription
factor NFAT in interleukin-2 expression. Mol. Cell. Biol. 19:2300.
12. Murphy, K. M., W. Ouyang, S. J. Szabo, N. G. Jacobson, M. L. Guler,
J. D. Gorham, U. Gubler, and T. L. Murphy. 1999. T helper differentiation proceeds through Stat1-dependent, Stat4- dependent and Stat4-independent phases.
Curr. Top. Microbiol. Immunol. 238:13.
13. Wofsy, D. 1986. Administration of monoclonal anti-T cell antibodies retards
murine lupus in BXSB mice. J. Immunol. 136:4554.
14. Burlingame, R. W., R. L. Rubin, R. S. Balderas, and A. N. Theofilopoulos. 1993.
The genesis and evolution of anti-chromatin autoantibodies in murine lupus implicates T-dependent immunization with self antigen. J. Clin. Invest. 91:1687.
15. Ray, S. K., C. Putterman, and B. Diamond. 1996. Pathogenic autoantibodies are
routinely generated during the response to foreign antigen: a paradigm for autoimmune disease. Proc. Natl. Acad. Sci. USA 93:2019.
16. Tillman, D. M., N. T. Jou, R. J. Hill, and T. N. Marion. 1992. Both IgM and IgG
anti-DNA antibodies are the products of clonally selective B cell stimulation in
(NZB 3 NZW)F1 mice. J. Exp. Med. 176:761.
Downloaded from http://www.jimmunol.org/ by guest on June 17, 2017
The findings herein differ from studies of IL-4-deficient (MRLFaslpr 3 B6)F2 mice, wherein IL-4-deficient mice generated similar levels of IgG2a and autoantibodies, but had significantly less
lymphadenopathy and GN compared with IL-4 wild-type controls
(21). This may reflect different immunopathologic mechanisms because of background gene differences between the (MRL-Faslpr 3
B6)F2 mice and the BXSB strain, or may possibly be due to a MRL
susceptibility gene in linkage dysequilibrium with the IL-4 gene on
chromosome 11. In fact, a suggestive MRL locus on chromosome
11 linked to autoantibody production and vasculitis has been recently reported (40). On the other hand, studies showing reduced
autoantibodies and nephritis in (NZB 3 NZW)F1 and MRL-Faslpr
mice after treatment with blocking anti-IL-4 Ab or soluble IL-4R
(32, 33) favors the argument that background genes are the determining factor. This implies the existence of IL-4-dependent and
-independent mechanisms for developing lupus.
The current finding that lupus-like disease in BXSB mice does
not require IL-4 is similar to what has been observed for several
other humoral-mediated autoimmune diseases, including mercuryinduced systemic autoimmunity (29, 35) and experimental autoimmune myasthenia gravis (EAMG) (41). In the case of EAMG,
IL-4 deficiency may in fact facilitate development of disease (42).
Thus, in most cases of Ab-mediated autoimmune diseases, knockout of the IL-4 gene has resulted in no reduction or even worsening
of disease.
The failure of the IL-4 deletion to alter disease severity in BXSB
mice could be due to compensatory mechanisms that are sometimes observed in gene knockout mice. This seems unlikely, however, since IL-4 has been shown to be required for the generation
of Th2 T cells in vitro (43, 44) and since the IL-4 knockout blocks
Th2 type cytokines and IgG isotype responses (37, 45). Furthermore, the virtual absence of IgG1 anti-chromatin autoantibody relative to IgG2a and the lack of differences in IgG1 anti-chromatin
autoantibody between the wild-type and IL-4 knockout BXSB
mice support the contention that Th2 type responses are not important for disease in this model. Recent evidence, however, indicates that IL-13 is also important for development of Th2 T cells
(46) and double knockout of IL-4 and IL-13 had more profound
effect on Th2 development than knockout of either gene alone
(47). Some of this overlap in function is due to shared IL-4 and
IL13 receptor components and signal transduction molecules (48,
49). Nevertheless, IL-4 and IL-13 have nonoverlapping functions
(48, 50), with IL-4 important for class switching to IgG1 (47) and
Th2 cell development and IL-13 important for expulsion of parasites from the gut and in experimental models of allergic asthma
(51, 52). Although it seems unlikely that IL-13 could completely
compensate for the lack of IL-4 in the BXSB model, the availability of a double IL-4/IL-13 knockout created by simultaneously
mutating these adjacent genes with a single construct (47) will
make it possible to directly address this issue.
Santiago, et al. (36) reported that overexpression of an IL-4
transgene in B cells reduced disease severity in (NZW 3 C57BL/
42
36. Santiago, M., L. Fossati, C. Jacquiet, W. Muller, S. Izui, and L. Reininger. 1997.
Interleukin-4 protects against a genetically linked lupus-like autoimmune syndrome. J. Exp. Med. 185:65.
37. Kuhn, R., K. Rajewsky, and W. Muller. 1991. Generation and analysis of interleukin-4 deficient mice. Science 254:707.
38. Kono, D. H., R. W. Burlingame, D. G. Owens, A. Kuramochi, R. S. Balderas, and
A. N. Theofilopoulos. 1994. Lupus susceptibility loci in New Zealand mice. Proc.
Natl. Acad. Sci. USA 91:10168.
39. Sabzevari, H., S. Propp, D. H. Kono, and A. N. Theofilopoulos. 1997. G1 arrest
and high expression of cyclin kinase and apoptosis inhibitors in accumulated
activated/memory phenotype CD41 cells of older lupus mice. Eur. J. Immunol.
27:1901.
40. Gu, L., A. Weinreb, X. P. Wang, D. J. Zack, J. H. Qiao, R. Weisbart, and
A. J. Lusis. 1998. Genetic determinants of autoimmune disease and coronary
vasculitis in the MRL-lpr/lpr mouse model of systemic lupus erythematosus.
J. Immunol. 161:6999.
41. Balasa, B., C. Deng, J. Lee, P. Christadoss, and N. Sarvetnick. 1998. The Th2
cytokine IL-4 is not required for the progression of antibody-dependent autoimmune myasthenia gravis. J. Immunol. 161:2856.
42. Karachunski, P. I., N. S. Ostlie, D. K. Okita, and B. M. Conti-Fine. 1999. Interleukin-4 deficiency facilitates development of experimental myasthenia gravis
and precludes its prevention by nasal administration of CD41 epitope sequences
of the acetylcholine receptor. J. Neuroimmunol. 95:73.
43. Le Gros, G., S. Z. Ben-Sasson, R. Seder, F. D. Finkelman, and W. E. Paul. 1990.
Generation of interleukin 4 (IL-4)-producing cells in vivo and in vitro: IL-2 and
IL-4 are required for in vitro generation of IL-4-producing cells. J. Exp. Med.
172:921.
44. Swain, S. L., A. D. Weinberg, M. English, and G. Huston. 1990. IL-4 directs the
development of Th2-like helper effectors. J. Immunol. 145:3796.
45. Kopf, M., G. Le Gros, M. Bachmann, M. C. Lamers, H. Bluethmann, and
G. Kohler. 1993. Disruption of the murine IL-4 gene blocks Th2 cytokine responses. Nature 362:245.
46. McKenzie, G. J., C. L. Emson, S. E. Bell, S. Anderson, P. Fallon, G. Zurawski,
R. Murray, R. Grencis, and A. N. McKenzie. 1998. Impaired development of Th2
cells in IL-13-deficient mice. Immunity 9:423.
47. McKenzie, G. J., P. G. Fallon, C. L. Emson, R. K. Grencis, and A. N. McKenzie.
1999. Simultaneous disruption of interleukin (IL)-4 and IL-13 defines individual
roles in T helper cell type 2-mediated responses. J. Exp. Med. 189:1565.
48. Chomarat, P., and J. Banchereau. 1998. Interleukin-4 and interleukin-13: their
similarities and discrepancies. Int. Rev. Immunol. 17:1.
49. Murata, T., N. I. Obiri, and R. K. Puri. 1998. Structure of and signal transduction
through interleukin-4 and interleukin-13 receptors. Int. J. Mol. Med. 1:551.
50. McKenzie, G. J., A. Bancroft, R. K. Grencis, and A. N. McKenzie. 1998. A
distinct role for interleukin-13 in Th2-cell-mediated immune responses. Curr.
Biol. 8:339.
51. Grunig, G., M. Warnock, A. E. Wakil, R. Venkayya, F. Brombacher,
D. M. Rennick, D. Sheppard, M. Mohrs, D. D. Donaldson, R. M. Locksley, and
D. B. Corry. 1998. Requirement for IL-13 independently of IL-4 in experimental
asthma. Science 282:2261.
52. Wills-Karp, M., J. Luyimbazi, X. Xu, B. Schofield, T. Y. Neben, C. L. Karp, and
D. D. Donaldson. 1998. Interleukin-13: central mediator of allergic asthma. Science 282:2258.
53. Saleem, S., Z. Dai, S. N. Coelho, B. T. Konieczny, K. J. Assmann,
F. K. Baddoura, and F. G. Lakkis. 1998. IL-4 is an endogenous inhibitor of
neutrophil influx and subsequent pathology in acute antibody-mediated inflammation. J. Immunol. 160:979.
Downloaded from http://www.jimmunol.org/ by guest on June 17, 2017
17. Jacob, C. O., P. H. van der Meide, and H. O. McDevitt. 1987. In vivo treatment
of (NZB 3 NZW)F1 lupus-like nephritis with monoclonal antibody to g interferon. J. Exp. Med. 166:798.
18. Ozmen, L., D. Roman, M. Fountoulakis, G. Schmid, B. Ryffel, and G. Garotta.
1995. Experimental therapy of systemic lupus erythematosus: the treatment of
NZB/W mice with mouse soluble interferon-g receptor inhibits the onset of glomerulonephritis. Eur. J. Immunol. 25:6.
19. Haas, C., B. Ryffel, and M. Le Hir. 1998. IFN-g receptor deletion prevents autoantibody production and glomerulonephritis in lupus-prone (NZB 3 NZW)F1
mice. J. Immunol. 160:3713.
20. Haas, C., B. Ryffel, and H. M. Le. 1997. IFN-g is essential for the development
of autoimmune glomerulonephritis in MRL/Ipr mice. J. Immunol. 158:5484.
21. Peng, S. L., J. Moslehi, and J. Craft. 1997. Roles of interferon-g and interleukin-4
in murine lupus. J. Clin. Invest. 99:1936.
22. Balomenos, D., R. Rumold, and A. N. Theofilopoulos. 1998. Interferon-g is required for lupus-like disease and lymphoaccumulation in MRL-lpr mice. J. Clin.
Invest. 101:364.
23. Balasa, B., C. Deng, J. Lee, L. M. Bradley, D. K. Dalton, P. Christadoss, and
N. Sarvetnick. 1997. Interferon g (IFN-g) is necessary for the genesis of acetylcholine receptor-induced clinical experimental autoimmune myasthenia gravis in
mice. J. Exp. Med. 186:385.
24. Krakowski, M., and T. Owens. 1996. Interferon-g confers resistance to experimental allergic encephalomyelitis. Eur. J. Immunol. 26:1641.
25. Jones, L. S., L. V. Rizzo, R. K. Agarwal, T. K. Tarrant, C. C. Chan, B. Wiggert,
and R. R. Caspi. 1997. IFN-g-deficient mice develop experimental autoimmune
uveitis in the context of a deviant effector response. J. Immunol. 158:5997.
26. Tarrant, T. K., P. B. Silver, J. L. Wahlsten, L. V. Rizzo, C. C. Chan, B. Wiggert,
and R. R. Caspi. 1999. Interleukin 12 protects from a T helper type 1-mediated
autoimmune disease, experimental autoimmune uveitis, through a mechanism
involving interferon g, nitric oxide, and apoptosis. J. Exp. Med. 189:219.
27. Prud’homme, G. J., D. H. Kono, and A. N. Theofilopoulos. 1995. Quantitative
polymerase chain reaction analysis reveals marked overexpression of interleukin1b, interleukin-1 and interferon-g mRNA in the lymph nodes of lupus-prone
mice. Mol. Immunol. 32:495.
28. Takahashi, S., L. Fossati, I. Iwamoto, J. Merino, R. Motta, T. Kobayakawa, and
S. Izui. 1996. Imbalance towards Th1 predominance is associated with acceleration of lupus-like autoimmune syndrome in MRL mice. J. Clin. Invest. 97:1597.
29. Kono, D. H., D. Balomenos, D. L. Pearson, M. S. Park, B. Hildebrandt,
P. Hultman, and K. M. Pollard. 1998. The prototypic Th2 autoimmunity induced
by mercury is dependent on IFN-g and not Th1/Th2 imbalance. J. Immunol.
161:234.
30. Chu, E. B., D. N. Ernst, M. V. Hobbs, and W. O. Weigle. 1994. Maturational
changes in CD41 cell subsets and lymphokine production in BXSB mice. J. Immunol. 152:4129.
31. Luzina, I. G., R. H. Knitzer, S. P. Atamas, W. C. Gause, J. C. Papadimitriou,
M. B. Sztein, C. E. Storrer, and B. S. Handwerger. 1999. Vasculitis in the Palmerston North mouse model of lupus: phenotype and cytokine production profile of
infiltrating cells. Arthritis Rheum. 42:561.
32. Nakajima, A., S. Hirose, H. Yagita, and K. Okumura. 1997. Role of IL-4 and
IL-12 in the development of lupus in NZB/W F1 mice. J. Immunol. 158:1466.
33. Schorlemmer, H. U., G. Dickneite, E. J. Kanzy, and K. H. Enssle. 1995. Modulation of the immunoglobulin dysregulation in GvH- and SLE-like diseases by
the murine IL-4 receptor (IL-4-R). Inflamm. Res. 44(Suppl 2):S194.
34. Shirai, A., J. Conover, and D. M. Klinman. 1995. Increased activation and altered
ratio of interferon-g: interleukin- 4 secreting cells in MRL-lpr/lpr mice. Autoimmunity 21:107.
35. Bagenstose, L. M., P. Salgame, and M. Monestier. 1998. Mercury-induced autoimmunity in the absence of IL-4. Clin. Exp. Immunol. 114:9.
IL-4 KNOCKOUT BXSB MICE