HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Morimoto, S. Articles by Kobata, T. Search for Related Content PubMed PubMed Citation Articles by Morimoto, S. Articles by Kobata, T.

CD134L Engagement Enhances Human B Cell Ig Production: CD154/CD40, CD70/CD27, and CD134/CD134L Interactions Coordinately Regulate T Cell-Dependent B Cell Responses¹

Shinji Morimoto^*,,, Yumiko Kanno^*, Yuetsu Tanaka^§, Yoshiaki Tokano, Hiroshi Hashimoto, Serge Jacquot^||, Chikao Morimoto^||, Stuart F. Schlossman^||, Hideo Yagita^,¶, Ko Okumura^,¶ and Tetsuji Kobata^2,*,

^* Division of Immunology, Institute for Medical Science, Dokkyo University School of Medicine, Tochigi, Japan; Departments of Immunology and Rheumatology, Juntendo University School of Medicine, Tokyo, Japan; ^§ Department of Infectious Disease and Immunology, Okinawa-Asia Research Center of Medical Science, Faculty of Medicine, University of the Ryukyus, Okinawa, Japan; ^¶ Core Research for Evolutional Science and Technology of Japan Science and Technology Corporation, Tokyo, Japan; and ^|| Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, MA 02115

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
CD134 is a member of the TNFR family expressed on activated T cells, whose ligand, CD134L, is found preferentially on activated B cells. We have previously reported that the CD70/CD27 interaction may be more important in the induction of plasma cell differentiation after the expansion phase induced by the CD154/CD40 interaction has occurred. When CD134-transfected cells were added to PBMCs stimulated with pokeweed mitogen, IgG production was enhanced in a dose-dependent fashion. Addition of CD134-transfected cells to B cells stimulated with Staphylococcus aureus Cowan I strain/IL-2 resulted in little if any enhancement of B cell IgG production and proliferation. We found that while CD134-transfected cells induced no IgG production by themselves, it greatly enhanced IgG production in the presence of CD40 stimulation or T cell cytokines such as IL-4 and IL-10. The addition of CD134-transfected cells showed only a slight increase in the number of plasma cells compared with that in the culture without them, indicating that an increased Ig production rate per cell is responsible for the observed enhancing effect of CD134L engagement rather than increase in plasma cell generation. These results strongly suggest different and sequential roles of the TNF/TNFR family molecules in human T cell-dependent B cell responses through cell-cell contacts and the cytokine network.

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion References

 
Tcell-dependent B cell responses require both cytokines and physical cell-cell contacts driving the different steps of B cell maturation: activation, clonal expansion, eventually Ig class switching and V gene hypermutation, and finally differentiation to memory B cells or to Ab-producing cells (1). A number of stimuli have been identified that are important for initiation of the differentiation events occurring during a T-dependent, Ag-driven B cell differentiation (e.g., costimulation of T cells through CD28 (2, 3, 4), CD154 (CD40 ligand) on activated T cells (5), indicating their possible role in early B cell activation).

The B cell molecule CD40 and its ligand CD154 on the T cell are believed to play a central role in T cell-dependent B cell responses (6). CD40 is a member of the TNFR family, and CD154 belongs to the reciprocal family of TNF-related ligands and is expressed on activated CD4⁺ T cells. CD40-mediated signaling appears to be critical for B cell proliferation, isotype switching, germinal center formation, and memory B cell commitment. However, the concept of the central role of CD40 in T cell-dependent B cell responses may somewhat require modification because of previous contradictory finding that disruption of the interaction between CD40 and CD154 does not impair differentiation to Ab-producing cells. Patients with hyper-IgM immunodeficiency, in whom CD154 expression is deficient, usually have normal or elevated levels of IgM (7, 8, 9). These observations suggest that there might be CD154-independent and -specific signals that drive a B cell toward terminal differentiation to Ab-producing cells.

We previously demonstrated that the CD70/CD27 interaction, a ligand-receptor pair of the TNF/TNFR family, enhanced IgG production by human B cells (10, 11). CD70 is expressed on activated T cells, and its receptor CD27 is expressed by a subset of B cells. In contrast to the CD154/CD40 interaction, the CD70/CD27 interaction played a minor role in B cell proliferation and its major function was to contribute to the formation of Ig-producing cells (12), suggesting that the CD70/CD27 interaction may be more important in the induction of plasma cell differentiation at a time when the expansion phase induced by the CD154/CD40 interaction has already occurred.

Previous studies in the mouse have reported that the cross-linking of CD134 (OX40) ligand (CD134L),³ a relatively novel member of the TNF family, on B cells resulted in a dramatic B cell proliferation and Ig secretion (13). In addition, the CD134/CD134L interaction was found to be critical for the T cell-dependent IgG production and its interaction did not affect the germinal center formation nor memory B cell development (14). Although the cDNAs encoding human CD134 and CD134L have been cloned (15, 16, 17, 18), little is known about their roles in human T cell-dependent B cell responses. Recently, it was reported that ligation of CD134 on T cells increases IL-4 production by naive T cells and promotes their development into effector cells producing higher levels of the IL-4 (19).

It is well known that many cytokines derived from T cells affect B cell proliferation and differentiation (20). In particular, IL-4 appears to be involved in B cell growth and switch toward IgG4 and IgE. IL-2 is involved in the growth and differentiation of activated B cells but appears to have no effect on isotype switch. IL-10 acts as a cofactor for B cell proliferation and is particularly important in Ig secretion by inducing differentiation of B cells in the plasma cell pathway. IL-10 is also involved in isotype switch toward IgG1, IgG3, and IgA1.

In this study, to clarify the role of the CD134/CD134L interaction and regulatory mechanisms by TNF/TNFR family molecules in human T cell-dependent B cell responses, we have compared the effects of CD134L, CD27, and CD40 ligation on B cell Ig production and B cell proliferation in T cell-dependent and -independent systems. We demonstrate here that CD134/CD134L interactions induced an increase in B cell Ig production per cell but not B cell proliferation nor plasma cell generation in the presence of CD40, IL-4, or IL-10 stimulation. These results strongly suggest different and sequential roles of the CD134/CD134L, CD70/CD27, and CD154/CD40 interactions in human T cell-dependent B cell responses through cell-cell contacts and the cytokine network.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
Monoclonal Ab and cytokines

The following mAbs were used in this study and have been described elsewhere: anti-CD3 (OKT-3, IgG2a), anti-CD14 (63D3, IgG1), anti-CD57 (HNK-1, IgM), anti-CD70 (2F11, IgG1), anti-CD134L (5A8; IgG1), and anti-CD154 (5c8, IgG2a) (11, 17, 21, 22). Purified anti-CD134, FITC-conjugated anti-CD3, anti-CD4, anti-CD8, anti-CD14, anti-CD38, PE-conjugated anti-CD19, anti-CD56, anti-CD154, anti-CD134, and isotype-matched control mAbs (mouse IgG1 and mouse IgG2a) were obtained from PharMingen (San Diego, CA). Recombinant human IL-4, IL-10, neutralizing anti-human IL-4 Ab, and anti-human IL-10 Ab were obtained from PharMingen.

Cell preparation

Human PBMCs were isolated from healthy donors by Ficoll-Hypaque (Pharmacia, Piscataway, NJ) density-gradient centrifugation. PBMCs were separated into the E rosette-positive and the E rosette-negative populations with 5% sheep erythrocytes. The E rosette-negative cells were depleted of monocytes by adherence to the plastic surface of culture dishes and further purified into B cells by complement (Cedarlane, Ontario, Canada) lysis with OKT-3, HNK-1, and 63D3 plus rat anti-mouse IgG1 mAb (PharMingen). The resultant B cell population was <2% CD14⁺, <1% CD3⁺, <2% CD57⁺, and >95% CD19⁺. All cultures were conducted in RPMI 1640 medium supplemented with 10% FCS, 2 mM L-glutamine, penicillin G (200U/ml), and gentamicine (10 µg/ml).

Transfectant cells

The murine pre-B cell line, 300-19 (23), was transfected by electroporation with the CD134 cDNA cloned into pMKITneo vector (pMKITneohgp34) (kindly gift from Dr. T. Uchiyama, Kyoto University, Kyoto, Japan). Transfected cells were selected by growth in medium with the neomycin analogue G418 (Life Technologies, Gaitherburg, MD), and cells with high-density CD134 expression were cloned by FACS. The CD70 cDNA-transfected cells, the CD154 cDNA-transfected cells, and the vector alone-transfected cells (mock) have been previously described (12).

Detection and quantification of IgG by ELISA

PBMCs (1 x 10⁵/well) were cultured with pokeweed mitogen (PWM) (Life Technologies) at a final dilution of 1:100, and B cells (1 x 10⁵/well) were cultured with Staphylococcus aureus Cowan I strain (SAC) (Sigma, St. Louis, MO) at a final concentration of 1:10,000 (v/v) and IL-2 (50 U/ml) in 96-well round-bottom plates in 0.2 ml of culture medium for 10 days at 37°C in a humidified atmosphere with 5% CO₂. Various amounts of mAb (0.5, 1, and 2 µg/ml) or irradiated (10,000 rad) transfected cells (5 x 10³, 1 x 10⁴, and 2 x 10⁴ per well) with or without IL-4 (100 U/ml) or IL-10 (100 ng/ml) were added at the beginning of the experiment. In some experiments, CD134-transfected cells were pretreated with anti-CD134 mAb at 5 µg/ml for 30 min on ice and then added to the cultures after extensive washing. The culture supernatants were harvested and added to goat anti-human Ig (Southern Biotechnology Associates, Birmingham, AL)-coated 96-well flat-bottom ELISA plates overnight. After discarding supernatants and washing with Tween 20 PBS, the bound human IgG was detected with HRP-labeled goat anti-human IgG at a dilution of 1:2000 followed by addition of p-nitophenyl phosphate substrate (Sigma), and the amount of IgG present was assessed by spectrophotometric analysis at 490 nm.

B cell proliferation assay

B cells (10⁵/well) were cultured with SAC (0.01%) and IL-2 (50U/ml) in 96-well round-bottom plates in 0.2 ml of culture medium for 4 days at 37°C in a humidified atmosphere with 5% CO₂. Various amounts of irradiated (10,000 rad) transfectant cells with or without IL-4 (100 U/ml) or IL-10 (100 ng/ml) were added at the beginning of the experiment. After 3 days, the cultures were pulsed with 1 µCi/well of [³H]thymidine. Eighteen hours later, the cells were harvested, and [³H]thymidine incorporation was measured in a liquid scintillation beta counter (Wallac, Turuku, Finland).

Flow cytometric analysis

Flow cytometric analysis was performed using FACScalibur (Becton Dickinson, San Jose, CA), and data were processed using the CellQuest program (Becton Dickinson). In some experiments, FITC-conjugated goat anti-mouse Ig (Southern Biotechnology Associates) was used as a second Ab in this study. Isotype-matched mouse IgG control was used throughout the studies and always reacted with <5% of the cells.

Statistical analysis

The paired t test was used to determine statistical significance of the data. Values of p < 0.05 were considered significant.

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
Effect of anti-CD134L mAb on T cell-dependent B cell IgG production

Recent studies in the mouse have demonstrated that CD134/CD134L interactions play an essential role in T cell-dependent Ab production (13, 14). In vitro, cross-linking of CD134L induced B cell proliferation and differentiation. In vivo, blockage of CD134/CD134L interaction by means of a polyclonal anti-CD134 Ab strongly inhibited primary and secondary IgG, but not the IgM Ab response to T cell-dependent Ag, suggesting that murine CD134/CD134L interaction is crucial importance for the terminal differentiation of activated B cells into plasma cells producing high levels of Ig. First, to investigate whether CD134/CD134L interaction is also involved in human T cell-dependent B cell responses, we examined the effect of anti-CD134L mAb on a T cell-dependent, PWM-driven B cell Ig synthesis system. The addition of anti-CD134L mAb caused a significant inhibition of IgG production in a dose-dependent manner (Fig. 1A). Consistent with our and other reports (6, 11), anti-CD70 mAb and anti-CD154 mAb also inhibited IgG production and mixture of these Abs blocked IgG production by 90% (Fig. 1B). These results strongly suggest that CD134/CD134L interactions are involved in human T cell-dependent B cell Ig production.

View larger version (15K): [in this window] [in a new window]   FIGURE 1. Inhibition of T cell-dependent B cell IgG production by mAb to CD70, CD154, and CD134L. PBMCs (1 x 105/well) were cultured with PWM (1/100) in the presence or absence of varying amounts of anti-CD134L mAb (0.5, 1, and 2 µg/ml) (A) or indicated mAb (1 µg/ml) (B). The IgG concentration in the supernatant was measured by ELISA after 10 days of culture. The data shown are representative of three different experiments. *, p < 0.05; **, p < 0.01.

In the previous mouse study, it was shown that activated B cells, such as peritoneal B cells expressing high levels of MHC class II Ag or B cells in the periarteriolar lymphoid sheath following immunization with specific Ag, express CD134L (24). Therefore, we examined the expression of CD134 and CD134L in human PBMCs. We found that PWM stimulation preferentially induced CD134 on T cells and CD134L on B cells (Fig. 2).

View larger version (23K): [in this window] [in a new window]   FIGURE 2. Expression of CD134 and CD134L on activated T and B cells. T cells (CD3+, CD4+, or CD8+) and B cells (CD19+) were analyzed by two-color flow cytometry after 3 days of culture with PWM (1/100). The data shown are representative of three different experiments.

To further clarify the role of CD134/CD134L interactions in B cell activation, we undertook a series of experiments examining the effect of CD134- and mock-transfected cells on Ig production in the T cell-dependent, PWM-driven system. In our previous studies (10, 11, 12), CD70-transfected cells but not CD154-transfected cells enhanced B cell Ig production in the T cell-dependent system. When CD134-transfected cells were added to PBMCs stimulated with PWM, IgG production was enhanced in a dose-dependent fashion as well as upon addition of CD70-transfected cells, whereas CD154- and mock-transfected cells had no effect (Fig. 3A). Moreover, the enhancing effect of CD134-transfected cells on IgG production could be blocked by pretreatment of these cells with anti-CD134 mAb (Fig. 3B). These results clearly indicate that enhancement of IgG production by CD134-transfected cells was specifically mediated by CD134/CD134L interactions.

View larger version (16K): [in this window] [in a new window]   FIGURE 3. Effect of CD70-, CD154-, or CD134-transfected (t) cells on T cell-dependent B cell IgG production. PBMCs (1 x 105/well) were cultured with PWM (1/100) in the presence of varying amounts of irradiated (10,000 rad) transfected cells (5 x 103, 1 x 104, and 2 x 104 per well) (A) or CD134-transfected cells (2 x 104 per well) with or without pretreatment with anti-CD134 mAb or control mAb (B). The IgG concentration in the supernatant at 10 days of culture was measured by ELISA. The data shown are representative of three different experiments. *, p < 0.05.

To determine whether CD134-transfected cells interacted primarily with CD134L⁺ B cells and resulted in induction of IgG production, we examined the effects of these transfected cells in a T cell-independent, SAC/IL-2-driven B cell activation system. As shown in Fig. 4, addition of CD134-transfected cells in the absence of T cells resulted in little if any enhancement of B cell IgG production and B cell proliferation, whereas CD154-transfected cells enhanced B cell proliferation but not IgG production, and CD70-transfected cells moderately enhanced both, consistent with our previous studies (10, 11, 12).

View larger version (15K): [in this window] [in a new window]   FIGURE 4. Effect of CD70-, CD154-, or CD134-transfectant (t) cells on T cell-independent B cell proliferation and IgG production. B cells (1 x 105/well) were cultured with SAC (0.01%) and IL-2 (50 U/ml) in the presence of varying amounts of irradiated (10,000 rad) transfected cells (5 x 103, 1 x 104, and 2 x 104 per well) for 4 days (A) or with irradiated transfected cells (2 x 104 per well) for 10 days (B). [3H]Thymidine incorporation of B cells was measured during the last 18 h of a 4-day culture (A). The IgG concentration in the supernatant at 10 days of culture was measured by ELISA (B). The data shown are representative of three different experiments. *, p < 0.05.

View larger version (17K): [in this window] [in a new window]   FIGURE 5. Effect of CD134L engagement with CD40 ligation or T cell cytokines on B cell IgG production and proliferation. B cells (1 x 105/well) were cultured with SAC (0.01%) and IL-2 (50 U/ml) in the presence or absence of irradiated (10,000 rad) transfected (t) cells (1 x 104 per well each), IL-4 (100 U/ml), or IL-10 (100 ng/ml) for 10 days (A) or 4 days (B). The IgG concentration in the supernatant at 10 days of culture was measured by ELISA (B). The data shown are representative of three different experiments. *, p < 0.05; **, p < 0.01.

These results strongly suggest that CD134L engagement can directly mediate a signal for B cell IgG production in the presence of CD154-transfected cells, IL-4, or IL-10, which is not accounted for by the increase in B cell proliferation nor up-regulation of CD134L on B cells.

Increase in IgG production per cell by CD134L engagement

Finally, we decided to examine whether the enhancement of IgG production by CD134L engagement could have been due to increased Ig production per cell or to production by a larger portion of the plasma cells. We stained with anti-CD19 and anti-CD38 mAbs B cells cocultured with CD134-transfected cells together with or without CD154-transfected cells, IL-4, or IL-10 in the presence of SAC/IL-2. As shown in Fig. 6, the presence of CD134-transfected cells with CD154-transfected cells, IL-4, or IL-10 showed an almost same percentage of cells with a plasma cell phenotype (CD19^-CD38⁺) as that in culture without CD134-transfected cells. To confirm the increased per cell Ig production by CD134L engagement, we measured the number of Ig-producing cells in the culture with CD134L engagement. As shown in Table I, the addition of CD134-transfected cells showed an almost equal number or a slight increase in the number of viable B cells, thus indicating only a slight increase of CD19^-CD38⁺ plasma cells. Therefore, when calculated, per cell IgG production was significantly higher in the presence of CD154-transfected cells, IL-4, or IL-10 after CD134L engagement. These results clearly indicate that an increased Ig production rate per cell is responsible for the observed enhancing effect of CD134L engagement on IgG production. Supporting this result, CD134-transfected cells together with CD70-transfected cells enhanced B cell IgG production induced by CD134-transfected cells alone or CD70-transfected cells alone in the presence of SAC/IL-2 plus IL-4 or IL-10 (data not shown).

View larger version (38K): [in this window] [in a new window]   FIGURE 6. Effect of CD134L engagement on plasma cell generation. B cells (1 x 105/well) were cultured with SAC (0.01%) and IL-2 (50 U/ml) in the presence of irradiated (10,000 rad) CD154-transfectant cells (1 x 104 per well), IL-4 (100 U/ml), or IL-10 (100 ng/ml) with or without irradiated (10,000 rad) CD134-transfectant cells (1 x 104 per well) for 10 days. The cells were stained with FITC-labeled anti-CD38 mAb and PE-labeled anti-CD19 mAb. Dead cells were removed by propidium iodide staining. Cells were and then enumerated by flow cytometric analysis. The data shown are representative of three different experiments.

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

In our previous and present studies, CD40 engagement induced B cell proliferation but not Ig production, and CD27 engagement induced moderate B cell proliferation and Ig production (6, 10, 11, 12). CD134L engagement induced Ig production but not B cell proliferation in the presence of additional stimuli such as CD40 ligation, IL-4, or IL-10. We have previously reported that the CD70/CD27 interaction may be more important in the induction of plasma cell differentiation at a time when the expansion phase induced by the CD154/CD40 interaction has already occurred (12). B cells express CD134L only upon activation, while CD40 and CD27 are expressed by resting and activated B cells. In contrast, CD134, CD154, and CD70 are only expressed on activated T cells. Thus, it is more likely that CD154/CD40, CD70/CD27, and CD134/CD134L interactions exhibit their functions in the relatively early, intermediate, and late phases of T cell-dependent B cell responses, respectively. In fact, the additive and sequential expression of CD154 and CD70/CD134 on T cells after activation appears to correlate with B cell expansion and plasma cell generation/Ig production, respectively, in the course of T cell-dependent B cell responses. It should be noted that CD134/CD134L interactions may play a critical role in the regulation of sufficient and effective Ig production because both CD134 and CD134L are expressed on T and B cells only after activation, and CD134L engagement appeared to exhibit its enhancing effect on B cell IgG production only when additional stimuli are provided.

Early stimulatory signals derived from CD40 ligation, IL-4, or IL-10 seem to be required for human B cell Ig production induced by CD134L engagement. This notion was supported by the finding that neutralizing anti-IL-4 or IL-10 Ab reduced Ig production enhanced by CD134L engagement. Previous studies have shown that the expression of CD154 and the production of several cytokines in vivo reach their maximum in the same time period in the early phase of B cell responses (25). IL-4 is reported to inhibit the differentiation but not the proliferation of IL-2-dependent primary Ag-specific B cell responses (26, 27). IL-10 synergizes with IL-2 for B cell Ig production (28). CD134L-mediated signal alone appeared to be insufficient to induce high level of B cell Ig production in the absence of additional signal(s) provided by CD40 or T cell cytokines even though B cell receptor is cross-linked with SAC and IL-2 is provided. CD40 signal or T cell cytokine stimulation is required to couple the CD134L signal to the signal pathway for production of higher levels of Ig in addition to basal Ig production by SAC plus IL-2. It is possible that CD40 ligation induces B cells to secrete IL-10 (29). It was demonstrated that resting B cells differ from activated B cells in their stimulation requirements for terminal differentiation (30). Resting B cells need activated T cell contact with Th2-type cytokines such as IL-4 and IL-10 to initiate Ig production. Activated B cells differentiate by activated T cell contact with Th1-like cytokine such as IL-2. B cells responding to CD134L stimulation might be a distinct subpopulation because 10–30% of B cells can express CD134L after activation.

At present, the precise mechanisms of synergistic effect of CD134/CD134L interaction and CD40 ligation, IL-4, or IL-10 on B cell Ig production is still unknown. CD134/CD134L interactions may require Th2-type cytokines such as IL-4 and IL-10 for exhibiting its activity of B cell Ig production. Recent study in the mouse has shown that CD134 engagement on T cells induced IL-4 expression and inhibited IFN- expression, resulting in Th2 immune responses (31). In humans, ligation of CD134 also increased IL-4 production by naive T cells and promoted their development into effector cells producing higher levels of the Th2 cytokines (19). Taken together, CD134/CD134L interactions appear to provide a costimulatory signal for the Th2 immune responses in a two-signaling system, with CD134L on activated B cells having the potential to costimulate T cells, and CD134 on activated T cells providing a signal for B cell Ig production. It is possible that CD134L engagement increases responsiveness to IL-4 or IL-10. The precise biological nature of CD134L-mediated B cell activation and its signaling events remain to be clarified in future studies.

The mechanism by which CD134L engagement induced Ig production is apparently different from that of CD27 engagement: Ig production by CD27 engagement was mainly due to an increased number of Ig-producing cells, while that by CD134L engagement was due to an increased per cell production rate. This notion was supported by the fact that addition of both CD70- and CD134-transfected cells showed the additive effect on B cell IgG production. Thus, CD70/CD27 and CD134/CD134L interactions might be a T cell-dependent mechanism stimulating B cells to produce large amount of Ab in the immune responses at the relatively early and late phase, respectively. Because it has been recently reported that CD27 is also a marker for human memory B cells (32, 33), the CD134/CD134L interaction may be more important in the sufficient and effective Ab production after plasma cells were generated from memory B cells by CD70/CD27 interactions. We have previously reported that the extent of T cell-dependent B cell responses may be regulated by the ratio of CD27⁺ T cells and CD70⁺ T cells and that CD27⁺ T cells may serve as regulatory T cells (11). Because only CD27 is expressed on T cells among these molecules (CD40, CD27, and CD134L) expressed on B cells, these findings elucidate that the CD70/CD27 interaction may play a pivotal role in regulating T cell-dependent B cell Ig production.

The ability of human CD134/CD134L interaction to mediate a signal for B cell Ig production but not B cell proliferation is of considerable interest because murine CD134/CD134L interaction is reported to play an important role both in B cell proliferation and Ig production (13, 14). In addition, Ig production by murine CD134/CD134L interactions appears to be independent on T cell-derived cytokines (13), while human CD134/CD134L interactions require them. CD27 expression was not observed on resting and activated B cells in the mouse (34), and no overt impediment in T cell-dependent B cell responses could be observed in CD27-deficient mice (J. Borst, unpublished observations). Thus, there might be a difference in regulatory mechanisms of T cell-dependent B cell responses by TNF/TNFR family molecules between humans and mice.

In summary, these observations fit our current models of the kinetic dynamics of human B cell terminal differentiation during the response to T cell-dependent Ags, which is summarized in Fig. 7. CD154/CD40 interaction is first required to stimulate the clonal expansion of Ag-specific B cells and induce Ig isotype switching. In the second step, CD70/CD27 interaction induces further B cell proliferation to lesser extent and induces plasma cell generation. In the third step, CD134/CD134L interaction enhances Ab production by plasma cells in the presence of IL-4 or IL-10. These TNF/TNFR family molecules appear to coordinate T cell-dependent B cell responses through cell-cell contacts and the cytokine network. At the present, it is not clear whether CD27 and CD134L engagement induce Ig class switching. It is possible that the enhanced IgG production is due to an increase IgG production by already isotype-switched cells. Nagumo et al. have reported that CD27 engagement augmented IgE production in cooperation with IL-4 and CD40 signal (35), in which CD70/CD27 interactions do not appear to induce IgE class switching. In addition, Morio et al. have demonstrated that CD27 did not interact with Ku, which is required for Ig class switching (36). Our present data are not only important to clarify the exact steps in T cell-dependent B cell responses, but also are of interest in considering the mechanisms of autoantibody production and hypergammaglobulinemia observed in autoimmune diseases such as systemic lupus erythematosus.

View larger version (18K): [in this window] [in a new window]   FIGURE 7. Model for the role of TNF/TNFR family in human T cell-dependent B cell maturation. See Discussion for details.

	Acknowledgments

 
We thank Dr. Toshiyuki Hori and Dr. Takashi Uchiyama for providing pMKITneohgp34; Dr. Hiroyasu Nakano for his help in the preparation of the transfectant cells; and Ms. Yukiko Ikeda and Mrs. Junko Sugimoto for their secretarial assistance.

	Footnotes

 
¹ This work was supported in part by grants from the Ichiro Kanehara Foundation; the Japan Rheumatism Foundation; the Kanagawa Academy of Science and Technology; the Ministry of Education, Science, Sports, and Culture; and the Ministry of Health, Japan.

² Address correspondence and reprint requests to Dr. Tetsuji Kobata, Division of Immunology, Institute for Medical Science, Dokkyo University School of Medicine, 880 Kitakobayashi, Mibu, Tochigi 321-0293, Japan.

³ Abbreviations used in this paper: CD134L, CD134 ligand; PWM, pokeweed mitogen; SAC, staphylococcus aureus Cowan I strain.

Received for publication September 20, 1999. Accepted for publication February 9, 2000.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 

1. Clark, E. A., J. A. Ledbetter. 1994. How B and T cells talk to each other. Nature 367:425.[Medline]
2. Lane, P., C. Burdet, S. Hubele, D. Scheidegger, U. Muller, F. McConnell, M. Kosco-Vilbois. 1994. B cell function in mice transgenic for mCTLA4-H1: lack of germinal centers correlated with poor affinity maturation and class switching despite normal priming of CD4⁺ T cells. J. Exp. Med. 179:819.[Abstract/Free Full Text]
3. Linsley, P. S., P. M. Wallace, J. Johnson, M. G. Gibson, J. L. Greene, J. A. Ledbetter, C. Singh, M. A. Tepper. 1992. Immunosuppression in vivo by a soluble form of the CTLA-4 T cell activation molecule. Science 257:792.[Abstract/Free Full Text]
4. Ronchese, F., B. Hausmann, S. Hubele, P. Lane. 1994. Mice transgenic for a soluble form of murine CTLA-4 show enhanced expansion of antigen-specific CD4⁺ T cells and defective antibody production in vivo. J. Exp. Med. 179:809.[Abstract/Free Full Text]
5. Foy, T. M., J. D. Laman, J. A. Ledbetter, A. Aruffo, E. Claassen, R. J. Noelle. 1994. gp39-CD40 interactions are essential for germinal center formation and the development of B cell memory. J. Exp. Med. 180:157.[Abstract/Free Full Text]
6. Van Kooten, C., J. Banchereau. 1996. CD40-CD40 ligand: a multifunctional receptor-ligand pair. Adv. Immunol. 61:1.[Medline]
7. Allen, R. C., R. J. Armitage, M. E. Conley, H. Rosenblatt, N. A. Jenkins, N. G. Copeland, M. A. Bedell, S. Edelhoff, C. M. Disteche, D. K. Simoneaux, et al 1993. CD40 ligand gene defects responsible for X-linked hyper-IgM syndrome. Science 259:990.[Abstract]
8. Aruffo, A., M. Farrington, D. Hollenbaugh, X. Li, A. Milatovich, S. Nonoyama, J. Bajorath, L. S. Grosmaire, R. Stenkamp, M. Neubauer, et al 1993. The CD40 ligand, gp39, is defective in activated T cells from patients with X-linked hyper-IgM syndrome. Cell 72:291.[Medline]
9. DiSanto, J. P., J. Y. Bonnefoy, J. F. Gauchat, A. Fischer, G. de Saint Basile. 1993. CD40 ligand mutations in X-linked immunodeficiency with hyper-IgM. Nature 361:541.[Medline]
10. Agematsu, K., T. Kobata, F. C. Yang, T. Nakazawa, K. Fukushima, M. Kitahara, T. Mori, K. Sugita, C. Morimoto, A. Komiyama. 1995. CD27/CD70 interaction directly drives B cell IgG and IgM synthesis. Eur. J. Immunol. 25:2825.[Medline]
11. Kobata, T., S. Jacquot, S. Kozlowski, K. Agematsu, S. F. Schlossman, C. Morimoto. 1995. CD27-CD70 interactions regulate B-cell activation by T cells. Proc. Natl. Acad. Sci. USA 92:11249.[Abstract/Free Full Text]
12. Jacquot, S., T. Kobata, S. Iwata, C. Morimoto, S. F. Schlossman. 1997. CD154/CD40 and CD70/CD27 interactions have different and sequential functions in T cell-dependent B cell responses: enhancement of plasma cell differentiation by CD27 signaling. J. Immunol. 159:2652.[Abstract]
13. Stuber, E., M. Neurath, D. Calderhead, H. P. Fell, W. Strober. 1995. Cross-linking of OX40 ligand, a member of the TNF/NGF cytokine family, induces proliferation and differentiation in murine splenic B cells. Immunity 2:507.[Medline]
14. Stuber, E., W. Strober. 1996. The T cell-B cell interaction via OX40-OX40L is necessary for the T cell-dependent humoral immune response. J. Exp. Med. 183:979.[Abstract/Free Full Text]
15. Baum, P. R., 3rd R. B. Gayle, F. Ramsdell, S. Srinivasan, R. A. Sorensen, M. L. Watson, M. F. Seldin, E. Baker, G. R. Sutherland, K. N. Clifford, et al 1994. Molecular characterization of murine and human OX40/OX40 ligand systems: identification of a human OX40 ligand as the HTLV-1-regulated protein gp34. EMBO J. 13:3992.[Medline]
16. Godfrey, W. R., F. F. Fagnoni, M. A. Harara, D. Buck, E. G. Engleman. 1994. Identification of a human OX-40 ligand, a costimulator of CD4⁺ T cells with homology to tumor necrosis factor. J. Exp. Med. 180:757.[Abstract/Free Full Text]
17. Imura, A., T. Hori, K. Imada, T. Ishikawa, Y. Tanaka, M. Maeda, S. Imamura, T. Uchiyama. 1996. The human OX40/gp34 system directly mediates adhesion of activated T cells to vascular endothelial cells. J. Exp. Med. 183:2185.[Abstract/Free Full Text]
18. Latza, U., H. Durkop, S. Schnittger, J. Ringeling, F. Eitelbach, M. Hummel, C. Fonatsch, H. Stein. 1994. The human OX40 homolog: cDNA structure, expression and chromosomal assignment of the ACT35 antigen. Eur. J. Immunol. 24:677.[Medline]
19. Ohshima, Y., L. P. Yang, T. Uchiyama, Y. Tanaka, P. Baum, M. Sergerie, P. Hermann, G. Delespesse. 1998. OX40 costimulation enhances interleukin-4 (IL-4) expression at priming and promotes the differentiation of naive human CD4⁺ T cells into high IL-4-producing effectors. Blood 92:3338.[Abstract/Free Full Text]
20. Banchereau, J., F. Rousset. 1992. Human B lymphocytes: phenotype, proliferation, and differentiation. Adv. Immunol. 52:125.[Medline]
21. Morimoto, C., N. L. Letvin, A. W. Boyd, M. Hagan, H. M. Brown, M. M. Kornacki, S. F. Schlossman. 1985. The isolation and characterization of the human helper inducer T cell subset. J. Immunol. 134:3762.[Abstract]
22. Lederman, S., M. J. Yellin, A. Krichevsky, J. Belko, J. J. Lee, L. Chess. 1992. Identification of a novel surface protein on activated CD4⁺ T cells that induces contact-dependent B cell differentiation (help). J. Exp. Med. 175:1091.[Abstract/Free Full Text]
23. Streuli, M., C. Morimoto, M. Schrieber, S. F. Schlossman, H. Saito. 1988. Characterization of CD45 and CD45R monoclonal antibodies using transfected mouse cell lines that express individual human leukocyte common antigens. J. Immunol. 141:3910.[Abstract]
24. Calderhead, D. M., J. E. Buhlmann, A. J. van den Eertwegh, E. Claassen, R. J. Noelle, H. P. Fell. 1993. Cloning of mouse OX40: a T cell activation marker that may mediate T-B cell interactions. J. Immunol. 151:5261.[Abstract]
25. Van den Eertwegh, A. J., R. J. Noelle, M. Roy, D. M. Shepherd, A. Aruffo, J. A. Ledbetter, W. J. Boersma, E. Claassen. 1993. In vivo CD40-gp39 interactions are essential for thymus-dependent humoral immunity. I. In vivo expression of CD40 ligand, cytokines, and antibody production delineates sites of cognate T-B cell interactions. J. Exp. Med. 178:1555.[Abstract/Free Full Text]
26. Llorente, L., M. C. Crevon, S. Karray, T. Defrance, J. Banchereau, P. Galanaud. 1989. Interleukin (IL) 4 counteracts the helper effect of IL2 on antigen- activated human B cells. Eur. J. Immunol. 19:765.[Medline]
27. Llorente, L., F. Mitjavila, M. C. Crevon, P. Galanaud. 1990. Dual effects of interleukin 4 on antigen-activated human B cells: induction of proliferation and inhibition of interleukin 2-dependent differentiation. Eur. J. Immunol. 20:1887.[Medline]
28. Fluckiger, A. C., P. Garrone, I. Durand, J. P. Galizzi, J. Banchereau. 1993. Interleukin 10 (IL-10) upregulates functional high affinity IL-2 receptors on normal and leukemic B lymphocytes. J. Exp. Med. 178:1473.[Abstract/Free Full Text]
29. Burdin, N., C. Peronne, J. Banchereau, F. Rousset. 1993. Epstein-Barr virus transformation induces B lymphocytes to produce human interleukin 10. J. Exp. Med. 177:295.[Abstract/Free Full Text]
30. Coffman, R. L., B. W. Seymour, D. A. Lebman, D. D. Hiraki, J. A. Christiansen, B. Shrader, H. M. Cherwinski, H. F. Savelkoul, F. D. Finkelman, M. W. Bond, et al 1988. The role of helper T cell products in mouse B cell differentiation and isotype regulation. Immunol. Rev. 102:5.[Medline]
31. Flynn, S., K. M. Toellner, C. Raykundalia, M. Goodall, P. Lane. 1998. CD4 T cell cytokine differentiation: the B cell activation molecule, OX40 ligand, instructs CD4 T cells to express interleukin 4 and upregulates expression of the chemokine receptor, Blr-1. J. Exp. Med. 188:297.[Abstract/Free Full Text]
32. Klein, U., K. Rajewsky, R. Kuppers. 1998. Human immunoglobulin (Ig)M⁺IgD⁺ peripheral blood B cells expressing the CD27 cell surface antigen carry somatically mutated variable region genes: CD27 as a general marker for somatically mutated (memory) B cells. J. Exp. Med. 188:1679.[Abstract/Free Full Text]
33. Tangye, S. G., Y. J. Liu, G. Aversa, J. H. Phillips, J. E. de Vries. 1998. Identification of functional human splenic memory B cells by expression of CD148 and CD27. J. Exp. Med. 188:1691.[Abstract/Free Full Text]
34. Oshima, H., H. Nakano, C. Nohara, T. Kobata, A. Nakajima, N. A. Jenkins, D. J. Gilbert, N. G. Copeland, T. Muto, H. Yagita, K. Okumura. 1998. Characterization of murine CD70 by molecular cloning and mAb. Int. Immunol. 10:517.[Abstract/Free Full Text]
35. Nagumo, H., K. Agematsu, K. Shinozaki, S. Hokibara, S. Ito, M. Takamoto, T. Nikaido, K. Yasui, Y. Uehara, A. Yachie, A. Komiyama. 1998. CD27/CD70 interaction augments IgE secretion by promoting the differentiation of memory B cells into plasma cells. J. Immunol. 161:6496.[Abstract/Free Full Text]
36. Morio, T., S. H. Hainssian, L. B. Bacharier, H. Teraoka, S. Nonoyama, M. Seki, J. Kondo, H. Nakano, S.-K. Lee, R. S. Geha, J. Yata. 1999. Ku in the cytoplasm associates with CD40 in human B cells and translocates into the nucleus following incubation with IL-4 and anti-CD40 mAb. Immunity 11:339.[Medline]

This article has been cited by other articles:

				H. Xu, Y. Huang, P. M. Chilton, L.-R. Hussain, M. K. Tanner, J. Yan, and S. T. Ildstad Strategic Nonmyeloablative Conditioning: CD154:CD40 Costimulatory Blockade at Primary Bone Marrow Transplantation Promotes Engraftment for Secondary Bone Marrow Transplantation after Engraftment Failure J. Immunol., November 1, 2008; 181(9): 6616 - 6624. [Abstract] [Full Text] [PDF]

				J. M. van Laar, M. Melchers, Y. K. O. Teng, B. van der Zouwen, R. Mohammadi, R. Fischer, L. Margolis, W. Fitzgerald, J.-C. Grivel, F. C. Breedveld, et al. Sustained Secretion of Immunoglobulin by Long-Lived Human Tonsil Plasma Cells Am. J. Pathol., September 1, 2007; 171(3): 917 - 927. [Abstract] [Full Text] [PDF]

				A. Zingoni, T. Sornasse, B. G. Cocks, Y. Tanaka, A. Santoni, and L. L. Lanier Cross-Talk between Activated Human NK Cells and CD4+ T Cells via OX40-OX40 Ligand Interactions J. Immunol., September 15, 2004; 173(6): 3716 - 3724. [Abstract] [Full Text] [PDF]

				H. Hase, Y. Kanno, M. Kojima, K. Hasegawa, D. Sakurai, H. Kojima, N. Tsuchiya, K. Tokunaga, N. Masawa, M. Azuma, et al. BAFF/BLyS can potentiate B-cell selection with the B-cell coreceptor complex Blood, March 15, 2004; 103(6): 2257 - 2265. [Abstract] [Full Text] [PDF]

				I. R. Humphreys, L. Edwards, G. Walzl, A. J. Rae, G. Dougan, S. Hill, and T. Hussell OX40 Ligation on Activated T Cells Enhances the Control of Cryptococcus neoformans and Reduces Pulmonary Eosinophilia J. Immunol., June 15, 2003; 170(12): 6125 - 6132. [Abstract] [Full Text] [PDF]

				L E Schiffer, N Hussain, X Wang, W Huang, J Sinha, M Ramanujam, and A Davidson Lowering anti-dsDNA antibodies--what's new? Lupus, December 1, 2002; 11(12): 885 - 894. [Abstract] [PDF]

				H. Hase, Y. Kanno, H. Kojima, C. Morimoto, K. Okumura, and T. Kobata CD27 and CD40 Inhibit p53-independent Mitochondrial Pathways in Apoptosis of B Cells Induced by B Cell Receptor Ligation J. Biol. Chem., November 27, 2002; 277(49): 46950 - 46958. [Abstract] [Full Text] [PDF]

				K. Warnatz, A. Denz, R. Drager, M. Braun, C. Groth, G. Wolff-Vorbeck, H. Eibel, M. Schlesier, and H. H. Peter Severe deficiency of switched memory B cells (CD27+IgM-IgD-) in subgroups of patients with common variable immunodeficiency: a new approach to classify a heterogeneous disease Blood, March 1, 2002; 99(5): 1544 - 1551. [Abstract] [Full Text] [PDF]

				X. He, C. M. Weyand, J. J. Goronzy, W. Zhong, and J. M. Stuart Bi-directional modulation of T cell-dependent antibody production by prostaglandin E2 Int. Immunol., January 1, 2002; 14(1): 69 - 77. [Abstract] [Full Text] [PDF]

				L. C. Ndhlovu, N. Ishii, K. Murata, T. Sato, and K. Sugamura Critical Involvement of OX40 Ligand Signals in the T Cell Priming Events During Experimental Autoimmune Encephalomyelitis J. Immunol., September 1, 2001; 167(5): 2991 - 2999. [Abstract] [Full Text] [PDF]

				Y. Takahashi, Y. Tanaka, A. Yamashita, Y. Koyanagi, M. Nakamura, and N. Yamamoto OX40 Stimulation by gp34/OX40 Ligand Enhances Productive Human Immunodeficiency Virus Type 1 Infection J. Virol., August 1, 2001; 75(15): 6748 - 6757. [Abstract] [Full Text]

				S. Jacquot, L. Macon-Lemaitre, E. Paris, T. Kobata, Y. Tanaka, C. Morimoto, S. F. Schlossman, and F. Tron B cell co-receptors regulating T cell-dependent antibody production in common variable immunodeficiency: CD27 pathway defects identify subsets of severely immuno-compromised patients Int. Immunol., July 1, 2001; 13(7): 871 - 876. [Abstract] [Full Text] [PDF]

				S. Nakae, M. Asano, R. Horai, N. Sakaguchi, and Y. Iwakura IL-1 Enhances T Cell-Dependent Antibody Production Through Induction of CD40 Ligand and OX40 on T Cells J. Immunol., July 1, 2001; 167(1): 90 - 97. [Abstract] [Full Text] [PDF]

				V. Malmstrom, D. Shipton, B. Singh, A. Al-Shamkhani, M. J. Puklavec, A. N. Barclay, and F. Powrie CD134L Expression on Dendritic Cells in the Mesenteric Lymph Nodes Drives Colitis in T Cell-Restored SCID Mice J. Immunol., June 1, 2001; 166(11): 6972 - 6981. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Morimoto, S. Articles by Kobata, T. Search for Related Content PubMed PubMed Citation Articles by Morimoto, S. Articles by Kobata, T.