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*
Veteran’s Administration Medical Center, St. Louis, MO 63106;
Division of Rheumatology, Department of Medicine, and
Department of Laboratory Medicine/Pathology, Washington University School of Medicine, St. Louis, MO 63110; and
§
Departments of Medicine and Immunology, University of Colorado Health Science Center, Denver, CO 80262
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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This effect of complement in the humoral immune response is mediated in part by the interaction of activated C3 with specific receptors on the surfaces of lymphocytes (1). Complement receptor 1 (CR1, CD35)3 is the C3b/C4b receptor (8, 9). This molecule is a 190-kDa membrane-bound glycoprotein present in human erythrocytes, macrophages, neutrophils, B cells, follicular dendritic cells (FDC), and a subset of T cells. On erythrocytes, CR1 is responsible for the immune adherence phenomenon by which C3b-coated circulating immune complexes (IC) are bound and transported to the reticuloendothelial system. On macrophages and neutrophils, this protein facilitates the phagocytosis of complement-opsonized particles. CR1 has also been implicated as a regulator of B cell proliferation and differentiation (10, 11). On FDC, CR1 provides a mechanism by which C3-coated IC are retained in the splenic and lymphoid follicles (12). In addition, this receptor has an important role in the regulation of complement activation by serving as a cofactor for Factor I-mediated cleavage and inactivation of C3 and C4 (8, 9). CR1 can also regulate C3 and C5 convertases by a process known as decay acceleration.
Complement receptor 2 (CR2, CD21) is the iC3b/C3d,g receptor. This molecule is a 150-kDa transmembrane glycoprotein present in human mature B cells, FDC, and some T cells. CR2 participates in the regulation of B cell activation and differentiation, the generation of immunologic memory, and in Ig class switching (13, 14). On B cells, CR2 can form a molecular complex with other proteins, such as CD19 or CR1, which mediate specific signal transduction events that facilitate B cell activation (15, 16). In addition, in vitro studies have shown that ligation of CR2 using polymeric C3d,g or anti-CR2 Abs enhances proliferation of human B cells when primed with suboptimal amounts of T cell-dependent factors, phorbol esters, or anti-IgM (3). In vivo, depletion of mouse CR1 and CR2 using mAb decreases the primary Ab response to T-dependent and T-independent Ags (17). Similar findings have been reported in studies using a soluble form of human CR2 to inhibit the binding of C3d to the membrane-bound mouse receptor (18).
The mouse B cell homologues of human CR1 and CR2 have been previously described (19). Mouse CR2 is 67% homologous to its human counterpart and binds with similar affinities to human and mouse C3d (20, 21). Mouse CR1 binds mouse C3b with high affinity, and shares with its human counterpart cofactor activity for the Factor I-mediated cleavage of C3b to iC3b and C3d (22, 23). Nevertheless, certain structural differences are evident between the two species. In humans, CR1 and CR2 are proteins encoded by distinct but related genes. Murine CR1 and CR2 are the alternatively spliced products of a common gene, designated Cr2 (24). In addition, mouse CR1 and CR2 are primarily expressed on B cells and FDC. Mouse CR1 is not present on the membranes of erythrocytes or platelets, and, thus, is not the mouse immune adherence receptor (25, 26).
We and others have generated Cr2 gene-targeted mice deficient in the expression of mouse CR1 and CR2 (27, 28). These mice demonstrate a dramatic impairment in the Ag-specific IgG response to T cell-dependent Ags. Two mechanisms could account for this decreased Ab response. CR1 and CR2 deficiency on B cells could directly result in abnormalities in costimulatory signals needed for their optimal activation (15). In contrast, CR1 and CR2 on FDC may be important for localization of Ag within the splenic and lymphoid follicles in the form of C3-containing IC (12). The absence of Ag trapping by FDC could impair B cell maturation into Ig-secreting or memory B cells (29).
To clarify the roles of CR1 and CR2 on FDC compared with B cells, we have performed bone marrow (BM) reconstitution experiments using the Cr2-/- mice. The Ab response to two different T cell-dependent Ags, SRBC and keyhole limpet hemocyanin (KLH), was analyzed. Mice expressing CR1 and CR2 on FDC but lacking B cell receptor expression generate a diminished but still detectable immune response compared to controls. On the other hand, mice expressing CR1 and CR2 on B cells but lacking expression on FDC have an initial rise in the Ag-specific IgG titer but are unable to maintain it, as shown by a substantial decrease in IgG titers by day 14. This marked decrease in the Ag-specific IgG response persists after secondary immunization. This finding correlates with impairment of IC trapping in the splenic follicles of mice lacking expression of the receptors in their FDC. These experiments indicate that CR1 and/or CR2 on FDC play a critical role in the generation of a normal immune response, especially during the later stages of the primary immune response and in the generation of a robust secondary Ab response.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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(129Sv x C57Bl/6) Cr2-/- mice were generated using standard gene-targeting techniques as previously described (27). They were maintained under specific pathogen-free conditions.
BM transfer
BM was prepared as previously described (30). Recipient mice were lethally irradiated with 1050 rad (10.5 Gy) and reconstituted with 2 x 106 donor BM cells by i.v. injection. Analysis of the BM chimeras was performed 8 wk after transplantation.
Abs and flow cytometry
7G6 is a rat anti-mouse CR1/CR2 mAb (25). FITC-conjugated 7G6, FITC-conjugated anti-mouse IgM, and phycoerythrin-conjugated anti-mouse B220 were obtained from PharMingen (San Diego, CA). Phycoerythrin-conjugated anti-mouse CD4 and FITC-conjugated anti-mouse CD8 were obtained from Becton Dickinson (San Jose, CA). For flow cytometric analysis, single cell suspensions were prepared from spleen. One microgram of Ab was added to 1 x 106 cells for 60 min in 100 µl of PBS/1% BSA at 4°C. Flow cytometry was performed on a FACScan (Becton Dickinson).
Immunohistochemistry
Spleens were removed and frozen quickly in OCT compound (Miles, Elkhart, IN). Sections (10 µm thick) were cut and fixed in acetone. Endogenous peroxidase was quenched with 0.2% H2O2 in methanol. For IC staining, 10 µl of fresh mouse serum was added to 5 µl of horseradish peroxidase/mouse anti-horseradish peroxidase (HRP/mouse anti-HRP) (Dako, Carpinteria, CA) IC diluted in 35 µl of PBS (31). The IC solution was then added to spleen sections and incubated at 37°C for 10 min. After washing in PBS, the color reaction was detected with diaminobenzidine. Sections were counterstained with 1% methyl green and covered with crystal-mount (Biomeda, Foster City, CA). For germinal center staining, a 1:100 dilution of peanut agglutinin (PNA) conjugated to biotin (Vector, Burlingame, CA) followed by alkaline phosphatase (AP) conjugated to streptavidin (Zymed, San Francisco, CA) was used. The spleen sections stained with PNA were then counterstained with a 1:100 dilution of rat anti-mouse IgD (Southern Biotechnology, Birmingham, AL) polyclonal antiserum followed by a 1:10 dilution of rabbit anti-rat IgG conjugated with HRP (Southern Biotechnology). Bound AP and HRP were detected with AP reaction (Vector) and diaminobenzidine.
The germinal center number was calculated by counting the PNA-positive germinal centers seen under x100 magnification. Three different regions of each slide were analyzed and an average of the amount of germinal centers was calculated. The germinal center area was calculated by taking photographs of spleen sections under a x2.5 magnification and scanning them in a UMAX scanner (UMAX Data System, Hsinchu, Taiwan) using ADOBE PHOTOSHOP software. The area was then calculated using National Institutes of Health IMAGE 1.59 software. The results were expressed as the ratio of the PNA+ area within the follicle to the total area of the follicle (28).
Immunization of mice
BM recipient mice were immunized i.v. with 100 µl of PBS containing 108 SRBC at day 0 and were then boosted at day 21. Alternatively, mice were immunized with 100 µg of KLH i.v. at day 0 and then boosted at day 21. Serum was obtained before and at the indicated intervals after the first immunization. For germinal center staining, mice were immunized with 108 SRBC i.v., and 10 days postimmunization the spleens were collected for further analysis.
ELISA
Anti-SRBC Abs were measured by the method of Heyman et al. (32). Serum anti-KLH levels were measured by coating Immulon 4 plates (Dynatech Laboratories, Chantilly, VA) with 5 µg/ml of KLH in PBS. The detecting Ab was 100 µl of a 0.2 µg/ml AP-conjugated goat anti-mouse IgM or AP-conjugated goat anti-mouse IgG Ab (Southern Biotechnology) added for 1 h, followed by AP substrate p-nitrophenyl phosphate (Sigma, St. Louis, MO) at 1 mg/ml. The mean OD at 405 nm from triplicate wells was compared with a standard curve of titrated serum to calculate relative units (RU) (33). Calculation of the percent inhibition in the Ab response was performed by substracting the average RU in a specific set of chimeric mice to the average RU of the control animals at day 14 and dividing by the average RU of the control animals. The control animals are the Cr2+/+ mice reconstituted with a Cr2+/+ BM.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In contrast to human CR1 and CR2, murine CR1 and CR2 expression is limited to B cells and FDC (19). This facilitates the analysis of the role of these CR in the humoral immune response by restricting the cell population under study. Furthermore, FDC are a long-lived radioresistant cell population that are not thought to be replaced by donor cells following BM transfer (34). Therefore, this technique provides an excellent tool to study the relative functions of CR1 and CR2 on B cells and FDC. Lethally irradiated mice reconstituted with a particular BM will have B cells with a phenotype corresponding to that of the donor, while the FDC phenotype corresponds to that of the recipient.
We transferred BM cells from Cr2+/+ donors into
Cr2-/- recipients and from
Cr2-/- donors into
Cr2+/+ recipients. As controls, we transferred
BM from Cr2+/+ and
Cr2-/- donors into
Cr2+/+ and Cr2-/-
recipients, respectively. Eight weeks after the transfer, PBL were
studied using two-color flow cytometric analysis with a mAb against
B220 and a mAb recognizing both mouse CR1 and CR2 (7G6) to determine
the level of CR1 and CR2 expression on the surface of B cells. As shown
in Figure 1
,
Cr2-/- mice reconstituted with a
Cr2+/+ BM (Fig. 1d) had
comparable expression of CR1 and CR2 in the peripheral blood B cells
compared with the Cr2+/+ mice reconstituted with
a Cr2+/+ BM (Fig. 1a). On the
other hand, the Cr2+/+ mice reconstituted with a
Cr2-/- BM (Fig. 1c) and the
Cr2-/- mice reconstituted with a
Cr2-/- BM (Fig. 1b) had no
expression of CR1 and CR2 on their B cells. The same results were found
in the splenic lymphocyte population (data not shown). In addition, the
splenic total cell number, the percentage of
IgM+B220+ B cells, and the percentage of CD4
and CD8 T cells was comparable between the different BM chimeric mice
(data not shown).
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One method to detect FDC is to stain spleen sections with
preformed HRP/anti-HRP mouse IC (31). These IC were first treated
with fresh mouse serum as a source of complement. A specific signal
corresponding to Ag-Ab IC is detected in a discrete area of the
follicle corresponding to the FDC cluster. Under these circumstances,
IC trapping is largely mediated by CR1 and CR2. As shown in Figure 2, binding of these IC was only detected
in Cr2+/+ recipient mice independent of the BM
used for transfer (Fig. 2, A and C). The
area in which the IC are located was a discrete portion of the follicle
corresponding to the FDC zone. There was no IC binding detected in
Cr2-/- mice reconstituted with either a
Cr2-/- or a Cr2+/+ BM
(Fig. 2, B and D). The lack of IC trapping
was not due to the absence of FDC in the splenic follicles, since
FDC-M1 reactivity was preserved (data not shown; see 28 . The
impairment in IC staining was also not secondary to abnormalities in
the splenic architecture (27). These results confirm that
Cr2-/- recipient mice lack receptor expression
in the surface of their FDC.
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To determine the relative requirement for CR1 and CR2 expression
on B cells vs FDC in the Ab response to Ags, the humoral response was
first examined in the different BM chimeras by immunizing the mice with
the particulate T cell-dependent Ag, SRBC (Fig. 3). IgM responses were indistinguishable
when comparing the different BM chimeric animals (data not shown). As
expected, mice lacking CR1 and CR2 expression in both B cells and FDC
had a substantial impairment in the Ag-specific IgG titer compared with
the normal controls (Fig. 3a). There was an 88%
reduction in the Ag-specific total IgG compared with the
Cr2+/+ controls (Table I). Cr2+/+ mice
reconstituted with Cr2-/- BM had a diminished
but still detectable immune response with a reduction of 56% in the Ab
response compared with the control animals. On the other hand, while
Cr2-/- mice reconstituted with
Cr2+/+ BM had a small initial Ab response by day
7, thereafter they were unable to maintain the Ag-specific IgG titers,
as shown by the dramatic 94% decrease in Ab titer at day 14 (Fig. 3a).
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b). After
secondary immunization, Cr2-/- mice
reconstituted with Cr2+/+ BM had a substantial
impairment in the Ab response. The IgG response was decreased by 92%
and was indistinguishable when compared with mice with dual B cell and
FDC CR1 and CR2 deficiency (Table I). Cr2+/+
mice receiving a Cr2-/- BM also showed a
substantially diminished anti-SRBC titer response. Nevertheless,
the 77% decrease in the Ag-specific total IgG titer was still less
than the decrease found in the CR2-/- mice
reconstituted with Cr2+/+ BM. Similar findings
were observed when the BM chimeras were immunized with a second T
cell-dependent Ag, KLH. As shown in Figure 4 and Table I,
Cr2-/- mice reconstituted with a
Cr2+/+ BM still exhibited a substantial decrease
in the KLH-specific IgG titers compared with controls. These
experiments clearly demonstrate that CR1 and/or CR2 on FDC play a
critical role in the generation of a normal immune response, especially
during the later stages of the primary immune response and in the
generation of a robust secondary Ab response.
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At day 28, we sacrificed the BM chimeric animals that were immunized with SRBC and stained their spleen with PNA to determine the presence or absence of germinal centers. Surprisingly, despite the substantial differences in IgG responses between the different BM chimeric groups, germinal center formation was retained (data not shown). These results differed from previously published observations reporting a marked decrease in the number of germinal centers in the Cr2-/- mice after immunization (28), but were in accordance with our previously published observation concerning the humoral immune response in Cr2-deficient mice (27).
Since the spleens in our initial analysis were collected during the
secondary immune response, we decided to perform a more comprehensive
analysis of germinal center formation in the
Cr2-/- mice by immunizing another set of BM
chimeric animals with SRBC and collecting their spleen 10 days into
their primary immune response. As control, we also immunized
Cr2+/+ and Cr2-/- mice.
In all circumstances, germinal center formation was still retained 10
days after immunization (Table II).
Furthermore, the average number of germinal centers was similar between
the different groups of animals. Nevertheless, the size of the germinal
centers in the CR2-/- mice was markedly
reduced compared with the CR2+/+ mice (Fig. 5, Table II). A similar trend was seen in
the CR2-/- mice reconstituted with
CR2-/- BM compared with the
CR2+/+ mice reconstituted with a
CR2+/+ BM. The decrease in the germinal center
size correlated with the markedly decreased IgG response found in this
set of chimeric animals. The size of the germinal centers in the
CR2+/+ animals receiving a
CR2-/- BM was comparable to that of the
controls. In contrast, the germinal centers tended to be smaller in the
Cr2-/- mice reconstituted with a
CR2+/+ BM. These results suggested that,
although germinal center formation is retained in the
CR2-/- mice, these germinal centers are
functionally impaired as demonstrated by their reduced size and by the
diminished IgG response following immunization. Moreover, this
phenotype appears to correlate best with the lack of CR1 and CR2
on FDC.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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). Second, we confirmed
the expression of CR1 and CR2 on FDC by incubating spleen sections with
IC pretreated with fresh mouse serum (Fig. 2). The staining of FDC with
these IC is dependent on the presence of CR1 and CR2 on these cells
(see below). Independent of the BM donor phenotype, spleen sections
from CR2+/+ mice, but not
CR2-/- mice, reacted with these IC. Finally,
in data not shown, we also stained spleen sections with the mAb 7G6, or
mAbs specific against B and T cell surface markers. As expected, in the
CR2-/- mice receiving a
CR2+/+ BM, 7G6 stained the region of the
follicle populated by IgM+B220+ B cells. In
contrast, Cr2+/+ mice receiving a
CR2-/- BM stained brightly with this mAb in a
limited region of the follicle corresponding to the area of the FDC
cluster. In addition, staining for B and T cells revealed that the
general architecture of the spleen as well as the number and relative
percentages of B cells and T cells were comparable between the
different BM transfer groups.
We immunized the BM chimeric animals with two different T
cell-dependent Ags (Figs. 3 and 4). Independent of the Ag used, the
results showed that expression of the receptors in both B cells and FDC
was needed to restore the normal phenotype. Moreover, there were
differences in the immune response of mice selectively expressing the
receptors on their B cells vs mice expressing the receptors on their
FDC. Mice with selective deficiency in CR1 and CR2 on their B cells had
a diminished, although still detectable, IgG response. The number and
size of their germinal centers were comparable to controls (Fig. 5, Table II). In contrast, mice with selective deficiency of CR1 and CR2
on their FDC had a marked abnormality in the production of Ag-specific
IgG titers. Interestingly, these mice had an initial rise in the Ab
response to the Ag, but this rise rapidly waned during the later stages
of the primary immune response. The IgG titers during the secondary
immune response were severely depressed and paralleled those seen in
the Cr2-/- mice. Furthermore, although
germinal centers were present in this group of BM chimeric animals,
these germinal centers tended to be smaller compared with normal
controls.
The results concerning the humoral immune response in the
Cr2+/+ mice reconstituted with a
Cr2-/- BM presented in this paper are in
general agreement with the findings of Croix et al. (35). These
investigators injected homozygous Cr2-deficient embryonal
stem cells into RAG-2-deficient blastocysts to generate chimeric mice
in which CR1 and CR2 expression was deleted only on B cells. CR1 and
CR2 expression was retained on FDC. These mice were unable to generate
a significant Ab response to (4-hydroxy-3-nitrophenyl)acetyl-KLH. In
our experiments, we found that Cr2+/+ mice
receiving a Cr2-/- BM have a diminished, but
still detectable, immune response. On the other hand, our findings
differ substantially from those of Ahearn et al. (28). They reported
that Cr2-/- mice reconstituted with
Cr2+/+ BM have a normal Ag-specific total Ig
response, suggesting a minimal role of CR1 and CR2 expressed on FDC in
the development of humoral immunity. In addition, Ahearn et al.
reported a more substantial defect in germinal center formation
manifested by the absence of these specialized follicular structures in
the Cr2-/--immunized mice. Although germinal
center size was reduced in the Cr2-/- mice
analyzed in this paper, germinal center formation was still retained.
Interestingly, lack of CR1 and CR2 expression on both B cells and FDC
was essential to detect the small germinal center phenotype in our
experiments (Table II).
One explanation for the differences between the results of other groups
and ours likely relates to the dosage and type of Ag used. The Ag used
in the experiments of Ahearn et al. was the bacteriophage 
X174.
Croix et al. used 10 µg of (4-hydroxy-3-nitrophenyl)acetyl-KLH
injected. The mice described in this paper were injected with either
108 SRBC or 100 µg of KLH, a relatively larger dose.
Furthermore, we used nonhaptenated KLH. It is likely that specific
immunogens have distinct requirements for complement activation that
influence the development of an immune response, germinal center
formation, and the trapping of Ag by FDC.
We interpret our results in the context of the following model. B cell Ag-specific responses are initiated when Ag binds membrane Ig. During this initial phase of the humoral immune response, additional signals are also generated through the recruitment of molecules on the surface of the Ag-binding B cells. These molecules promote B cell proliferation followed by differentiation into Ig-secreting cells (36). CR1 has been shown to regulate B cell proliferation and differentiation (10, 11). CR2 can also directly modify this initial cognitive phase of the activation pathway by interacting with C3 components bound to the Ag and delivering intracellular signals through the CD19/CD21 signal transduction complex (36). Therefore, the lack of CR1 and CR2 on B cells will cause a relative impairment in their activation, resulting in a diminished Ab response and the presence of abnormal germinal centers. However, as shown in this report, the lack of CR expression on B cells cannot fully account for the severe defect in the humoral response observed in Cr2-/- mice.
It is important to note that the Cr2-/- B cells are not completely immunocompromised. Cr2-/- B cells respond normally in vitro to polyclonal stimuli such as treatment with anti-IgM or CD40 ligand (28). In addition, the IgM responses are indistinguishable when comparing the different BM chimeric animals. This finding agrees with the original observations made by Pepys (2), and it correlates with our previously published analysis of the Cr2-/- mice (26). The severe impairment of IgG responses in the context of an adequate IgM response in the Cr2-/- mice argues that, besides the role of these receptors in initial B cell activation, CR1 and/or CR2 are also important in vivo for other events that are necessary for normal B cell maturation into IgG-secreting cells.
Two different components have been described during the unfolding of a T cell-dependent B cell response. One involves extrafollicular events that lead to the initial production of Abs (37). The second component consists of intrafollicular events that result in the development of high affinity B cells (38). The extrafollicular component occurs in the periarteriolar lymphoid sheath and is characterized by the generation of Ab-forming cell (AFC) foci. These foci are first seen in the spleen 2 to 3 days after immunization. The AFC within these foci initially secrete IgM, but later some of these cells class switch and secrete Abs of the IgG type. AFC foci peak in size at day 7 postimmunization, but by day 14 they are barely detectable.
Intrafollicular events begin when, during the primary immune response, some Ag-primed B cells migrate to the germinal center (38, 39). In the germinal center, B cells proliferate and undergo somatic hypermutation and Ig class switching. These events are followed by a selection process in which B lymphocytes with an increased affinity for Ag are selected. Afterward, these selected B cells receive further maturation signals leading to the generation of IgG-producing cells or memory cells.
FDC are in part responsible for the germinal center events detailed above (40). FDC can provide signals that aid in the proliferation and differentiation of B lymphocytes. Furthermore, FDC have the ability to trap Ag on their surface as IC. It is likely that, after undergoing somatic hypermutation, only B cells that can recognize the trapped Ag on the FDC surface will be selected to continue this maturation pathway. In addition, the FDC can also serve as a reservoir of Ag for the long-term development of memory B cells.
The rise in Ag-specific IgG titers at day 7 seen in the Cr2-/- reconstituted with Cr2+/+ BM is likely to be secondary to the production of Abs by the extrafollicular, FDC-independent, AFC foci. The rapid fall in the IgG response at day 14 can be ascribed to the disappearance of these foci associated with an impairment in the ability of germinal center B cells to differentiate into IgG-secreting cells. This could explain the tendency for smaller germinal centers in this group of BM chimeric animals and in the CR2-/- mice. Furthermore, this impairment in the generation of IgG-secreting cells extends to the secondary immune response, implying a defect in the formation of Ag-specific memory B cells within the germinal center. Our data suggest that these abnormalities could be directly associated with the absence of CR in the surface of the FDC.
There are several ways in which the expression of CR1 and CR2 on FDC contributes to the development of a strong humoral immune response. As described above, FDC can retain Ag in the form of IC via CR (31). The retained Ag can interact with the surface Ig and deliver stimulatory signals that will aid in the further maturation of germinal center B cells (40). Alternatively, CR1 and CR2 can bind other molecules that are present on the surface of activated B cells (14). C3 bound to Ag might cross-link CR1 and/or CR2 with surface Ig and alter the signal that would be transduced through surface Ig by Ag binding alone. One result of the altered signal could be the rescue of B lymphocytes from apoptosis that would otherwise be induced by the binding of certain types of Ags (41). In addition, engagement of these IC can promote changes within the FDC that improve their ability to interact with B lymphocytes (31, 42). These associations can facilitate the survival, growth, and differentiation of B lymphocytes.
In summary, we have shown that CR1 and CR2 on both B cells and FDC are important in the generation of a humoral immune response to a particular set of Ags. Moreover, these experiments are the first to demonstrate an absolute requirement for CR1 and CR2 expression on FDC in humoral immunity. Depending on the antigenic stimulus, different aspects of humoral immune responses can be affected by a lack of CR1 and CR2 expression on B cells and/or FDC. These include B cell activation, B cell development into Ag-specific IgG-secreting cells, and germinal center formation. The CR2-/- mice described herein provide an excellent experimental model in which to study in detail the function of these receptors during each phase of the humoral immune response.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Hector Molina, Washington University School of Medicine, Division of Rheumatology Box 8045, 660 South Euclid Avenue, St. Louis, MO 63110. E-mail address:
3 Abbreviations used in this paper: CR, complement receptor; AFC, Ab-forming cell; AP, alkaline phosphatase; BM, bone marrow; FDC, follicular dendritic cell; HRP, horseradish peroxidase; IC, immune complex; KLH, keyhole limpet hemocyanin; PNA, peanut agglutinin; RU, relative units.
Received for publication January 12, 1998. Accepted for publication January 30, 1998.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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(LT)-deficient mice is due to abnormal organ development, not defective lymphocyte migration. J. Inflamm. 45:72.[Medline]
II-mediated immune-complex trapping. Immunology 80:34.[Medline]
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Y. Chen, D. Perry, S. A. Boackle, E. S. Sobel, H. Molina, B. P. Croker, and L. Morel Several Genes Contribute to the Production of Autoreactive B and T Cells in the Murine Lupus Susceptibility Locus Sle1c J. Immunol., July 15, 2005; 175(2): 1080 - 1089. [Abstract] [Full Text] [PDF]
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L. Birrell, L. Kulik, B. P. Morgan, V. M. Holers, and K. J. Marchbank B Cells from Mice Prematurely Expressing Human Complement Receptor Type 2 Are Unresponsive to T-Dependent Antigens J. Immunol., June 1, 2005; 174(11): 6974 - 6982. [Abstract] [Full Text] [PDF]
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Y. Aydar, S. Sukumar, A. K. Szakal, and J. G. Tew The Influence of Immune Complex-Bearing Follicular Dendritic Cells on the IgM Response, Ig Class Switching, and Production of High Affinity IgG J. Immunol., May 1, 2005; 174(9): 5358 - 5366. [Abstract] [Full Text] [PDF]
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 D. Gatto, T. Pfister, A. Jegerlehner, S. W. Martin, M. Kopf, and M. F. Bachmann Complement receptors regulate differentiation of bone marrow plasma cell precursors expressing transcription factors Blimp-1 and XBP-1 J. Exp. Med., March 21, 2005; 201(6): 993 - 1005. [Abstract] [Full Text] [PDF]
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 S. T. Test, J. K. Mitsuyoshi, and Y. Hu Depletion of Complement Has Distinct Effects on the Primary and Secondary Antibody Responses to a Conjugate of Pneumococcal Serotype 14 Capsular Polysaccharide and a T-Cell-Dependent Protein Carrier Infect. Immun., January 1, 2005; 73(1): 277 - 286. [Abstract] [Full Text] [PDF]
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 A. R. Ferguson, M. E. Youd, and R. B. Corley Marginal zone B cells transport and deposit IgM-containing immune complexes onto follicular dendritic cells Int. Immunol., October 1, 2004; 16(10): 1411 - 1422. [Abstract] [Full Text] [PDF]
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E. C. Whipple, R. S. Shanahan, A. H. Ditto, R. P. Taylor, and M. A. Lindorfer Analyses of the In Vivo Trafficking of Stoichiometric Doses of an Anti-Complement Receptor 1/2 Monoclonal Antibody Infused Intravenously in Mice J. Immunol., August 15, 2004; 173(4): 2297 - 2306. [Abstract] [Full Text] [PDF]
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T. D. de Stahl, J. Dahlstrom, M. C. Carroll, and B. Heyman A Role for Complement in Feedback Enhancement of Antibody Responses by IgG3 J. Exp. Med., May 5, 2003; 197(9): 1183 - 1190. [Abstract] [Full Text] [PDF]
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R. R. Reid, S. Woodcock, A. Shimabukuro-Vornhagen, W. G. Austen Jr., L. Kobzik, M. Zhang, H. B. Hechtman, F. D. Moore Jr., and M. C. Carroll Functional Activity of Natural Antibody is Altered in Cr2-Deficient Mice J. Immunol., November 15, 2002; 169(10): 5433 - 5440. [Abstract] [Full Text] [PDF]
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M. Gadjeva, A. Verschoor, M. A. Brockman, H. Jezak, L. M. Shen, D. M. Knipe, and M. C. Carroll Macrophage-Derived Complement Component C4 Can Restore Humoral Immunity in C4-Deficient Mice J. Immunol., November 15, 2002; 169(10): 5489 - 5495. [Abstract] [Full Text] [PDF]
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R. A. Barrington, O. Pozdnyakova, M. R. Zafari, C. D. Benjamin, and M. C. Carroll B Lymphocyte Memory: Role of Stromal Cell Complement and Fc{gamma}RIIB Receptors J. Exp. Med., November 4, 2002; 196(9): 1189 - 1200. [Abstract] [Full Text] [PDF]
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K. J. Marchbank, L. Kulik, M. G. Gipson, B. P. Morgan, and V. M. Holers Expression of Human Complement Receptor Type 2 (CD21) in Mice During Early B Cell Development Results in a Reduction in Mature B Cells and Hypogammaglobulinemia J. Immunol., October 1, 2002; 169(7): 3526 - 3535. [Abstract] [Full Text] [PDF]
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 C. H. Nielsen and R. G. Q. Leslie Complement's participation in acquired immunity J. Leukoc. Biol., August 1, 2002; 72(2): 249 - 261. [Abstract] [Full Text] [PDF]
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X. Wu, N. Jiang, C. Deppong, J. Singh, G. Dolecki, D. Mao, L. Morel, and H. D. Molina A Role for the Cr2 Gene in Modifying Autoantibody Production in Systemic Lupus Erythematosus J. Immunol., August 1, 2002; 169(3): 1587 - 1592. [Abstract] [Full Text] [PDF]
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 A. Kobayashi, T. Darragh, B. Herndier, K. Anastos, H. Minkoff, M. Cohen, M. Young, A. Levine, L. Ahdieh Grant, W. Hyun, et al. Lymphoid Follicles Are Generated in High-Grade Cervical Dysplasia and Have Differing Characteristics Depending on HIV Status Am. J. Pathol., January 1, 2002; 160(1): 151 - 164. [Abstract] [Full Text] [PDF]
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M. Hasegawa, M. Fujimoto, J. C. Poe, D. A. Steeber, and T. F. Tedder CD19 Can Regulate B Lymphocyte Signal Transduction Independent of Complement Activation J. Immunol., September 15, 2001; 167(6): 3190 - 3200. [Abstract] [Full Text] [PDF]
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A. Verschoor, M. A. Brockman, D. M. Knipe, and M. C. Carroll Cutting Edge: Myeloid Complement C3 Enhances the Humoral Response To Peripheral Viral Infection J. Immunol., September 1, 2001; 167(5): 2446 - 2451. [Abstract] [Full Text] [PDF]
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S. T. Test, J. Mitsuyoshi, C. C. Connolly, and A. H. Lucas Increased Immunogenicity and Induction of Class Switching by Conjugation of Complement C3d to Pneumococcal Serotype 14 Capsular Polysaccharide Infect. Immun., May 1, 2001; 69(5): 3031 - 3040. [Abstract] [Full Text] [PDF]
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L. G. Hannum, A. M. Haberman, S. M. Anderson, and M. J. Shlomchik Germinal Center Initiation, Variable Gene Region Hypermutation, and Mutant B Cell Selection without Detectable Immune Complexes on Follicular Dendritic Cells J. Exp. Med., September 25, 2000; 192(7): 931 - 942. [Abstract] [Full Text] [PDF]
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X. Wu, N. Jiang, Y.-F. Fang, C. Xu, D. Mao, J. Singh, Y.-X. Fu, and H. Molina Impaired Affinity Maturation in Cr2-/- Mice Is Rescued by Adjuvants Without Improvement in Germinal Center Development J. Immunol., September 15, 2000; 165(6): 3119 - 3127. [Abstract] [Full Text] [PDF]
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L. Kacani, W. M. Prodinger, G. M. Sprinzl, M. G. Schwendinger, M. Spruth, H. Stoiber, S. Döpper, S. Steinhuber, F. Steindl, and M. P. Dierich Detachment of Human Immunodeficiency Virus Type 1 from Germinal Centers by Blocking Complement Receptor Type 2 J. Virol., September 1, 2000; 74(17): 7997 - 8002. [Abstract] [Full Text]
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K. J. Marchbank, C. C. Watson, D. F. Ritsema, and V. M. Holers Expression of Human Complement Receptor 2 (CR2, CD21) in Cr2-/- Mice Restores Humoral Immune Function J. Immunol., September 1, 2000; 165(5): 2354 - 2361. [Abstract] [Full Text] [PDF]
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S. E. Applequist, J. Dahlstrom, N. Jiang, H. Molina, and B. Heyman Antibody Production in Mice Deficient for Complement Receptors 1 and 2 Can Be Induced by IgG/Ag and IgE/Ag, But Not IgM/Ag Complexes J. Immunol., September 1, 2000; 165(5): 2398 - 2403. [Abstract] [Full Text] [PDF]
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D. Qin, J. Wu, K. A. Vora, J. V. Ravetch, A. K. Szakal, T. Manser, and J. G. Tew Fc{gamma} Receptor IIB on Follicular Dendritic Cells Regulates the B Cell Recall Response J. Immunol., June 15, 2000; 164(12): 6268 - 6275. [Abstract] [Full Text] [PDF]
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Z. Chen, S. B. Koralov, M. Gendelman, M. C. Carroll, and G. Kelsoe Humoral Immune Responses in Cr2-/- Mice: Enhanced Affinity Maturation but Impaired Antibody Persistence J. Immunol., May 1, 2000; 164(9): 4522 - 4532. [Abstract] [Full Text] [PDF]
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 X. J. Da Costa, M. A. Brockman, E. Alicot, M. Ma, M. B. Fischer, X. Zhou, D. M. Knipe, and M. C. Carroll Humoral response to herpes simplex virus is complement-dependent PNAS, October 26, 1999; 96(22): 12708 - 12712. [Abstract] [Full Text] [PDF]
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A. M. Oliver, F. Martin, and J. F. Kearney IgMhighCD21high Lymphocytes Enriched in the Splenic Marginal Zone Generate Effector Cells More Rapidly Than the Bulk of Follicular B Cells J. Immunol., June 15, 1999; 162(12): 7198 - 7207. [Abstract] [Full Text] [PDF]
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 D.T. Fearon Innate immunity and the biological relevance of the acquired immune response QJM, May 1, 1999; 92(5): 235 - 237. [Full Text] [PDF]
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N. Baumgarth, O. C. Herman, G. C. Jager, L. Brown, L. A. Herzenberg, and L. A. Herzenberg Innate and acquired humoral immunities to influenza virus are mediated by distinct arms of the immune system PNAS, March 2, 1999; 96(5): 2250 - 2255. [Abstract] [Full Text] [PDF]
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M. Kopf, S. Herren, M. V. Wiles, M. B. Pepys, and M. H. Kosco-Vilbois Interleukin 6 Influences Germinal Center Development and Antibody Production via a Contribution of C3 Complement Component J. Exp. Med., November 16, 1998; 188(10): 1895 - 1906. [Abstract] [Full Text] [PDF]
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