IgE-antigen complexes enhance Fc epsilon R and Ia expression by murine B lymphocytes.

Murine monoclonal IgE interacts with B cells of BALB/c mouse spleen with greater efficiency in the presence of its specific antigen. Complexes of anti-DNP IgE and DNP-OVA not only resist elution from B lymphocytes by acid but have a substantially longer dissociation half-time when compared with monomeric IgE (440 vs. 8 min, respectively). Further, these IgE-antigen complexes induce Fc epsilon R expression in lymphoid cells more efficiently than IgE alone. Maximum levels of B cell Fc epsilon R were observed after a 24 h incubation with 1 microgram/ml IgE in the presence of 1 microgram/ml DNP-OVA, while 30 micrograms/ml monomeric IgE was needed to elicit a similar increase of Fc epsilon R expression. Most importantly, overnight incubation of B cell-enriched BALB/c spleen cells with IgE-antigen complexes resulted in an augmented membrane expression of class II MHC antigens. B cell surface expression of both I-A and I-E antigens responded to a comparable level after incubation with IgE-antigen complexes but did not occur in response to either IgE or antigen alone. The enhanced sIa expression occurred in parallel to IgE-antigen concentrations that gave rise to Fc epsilon R hyperexpression. Moreover, double staining for Fc epsilon R and surface Ia antigen shows that B cells exhibiting the highest density of Fc epsilon R also demonstrated the most surface I-A, suggesting that B lymphocytes are autonomous in their response to IgE-antigen complexes. Changes in class I MHC or sIgM were not observed after overnight incubation with IgE and antigen. These results demonstrate the importance of IgE-antigen complexes for intercellular signaling and further suggest that the IgE system plays a broader role in immune response than it has generally been credited.

Mechanism of macrophage reversal of Friend erythroleukemia: macrophage regulation of normal and leukemic erythropoiesis.

The acute erythroleukemia induced in mice by the anemia-inducing strain of the Friend virus complex is caused to regress by normal macrophages. We examined the possibility that reversal of leukemia is related to a macrophage regulatory function in erythropoiesis. We found that the ability of macrophages to induce leukemia regression correlates with nonimmunological, in vivo suppression of normal and susceptible leukemic erythroid progenitors. The macrophage effect on erythropoiesis appears to be due to changes in a humoral regulator, related to but independent of erythropoietin. The results suggest a novel regulatory system for erythropoiesis, operative in vivo, and involving macrophages as accessory or suppressor cells. This regulation appears to be disrupted in erythroleukemic mice, but can be restored, and the disease can be made to regress by treatment with normal macrophages.

The anti-herpes simplex virus activity of n-docosanol includes inhibition of the viral entry process.

n-Docosanol-treated cells resist infection by a variety of lipid-enveloped viruses including the herpesviruses. Previous studies of the mechanism of action demonstrated that n-docosanol inhibits an event prior to the expression of intermediate early gene products but subsequent to HSV attachment. The studies reported here indicate that n-docosanol inhibits fusion of the HSV envelope with the plasma membrane. Evidence suggests that antiviral activity requires a time-dependent metabolic conversion of the compound. Cellular resistance to infection declines after removal of the drug with a t1/2 of approximately 3 h. Reduced expression of viral genes in n-docosanol-treated cells was confirmed by a 70% reduction in expression of a reporter gene regulated by a constitutive promoter inserted into the viral genome. Inhibited release in treated cells of virion-associated regulatory proteins--an immediate post entry event--was indicated by a 75% reduction in the expression of beta-galactosidase in target cells carrying a stably transfected lacZ gene under control of an HSV immediate--early promoter. Finally, the fusion-dependent dequenching of a lipophilic fluorescent probe, octadecyl rhodamine B chloride, inserted into the HSV envelope was significantly inhibited in treated cells. Inhibition of fusion between the plasma membrane and the HSV envelope, and the subsequent lack of replicative events, may be the predominant mechanism for the anti-HSV activity of n-docosanol.

IL-10 inhibits lipopolysaccharide-induced murine B cell proliferation and cross-linking of surface antigen receptors or ligation of CD40 restores the response.

The effects of IL-10 on murine B cell proliferation in vitro were investigated. IL-10 inhibited LPS-induced B cell proliferation with an EC50 of approximately 500 pg/ml. IL-10-mediated inhibitory activity was not overtly associated with cytotoxicity or induction of apoptosis. The presence or the absence of T cells and mononuclear phagocytes did not affect the inhibitory activity of IL-10 on LPS-induced proliferation. LPS-stimulated, IL-10-exposed B cells progressed from G0 or from M to G1A of the cell cycle, but were inhibited from entry into subsequent phases. IL-10 had no discernible effect on B cell proliferation elicited with goat anti-mouse IgM plus IL-4. Moreover, cross-linking, but not mere ligation, of surface Ag receptors restored LPS-induced B cell proliferation in the presence of IL-10. The proliferative response to ligation of CD40 with anti-CD40 Abs was also not inhibited by IL-10, and as observed with goat anti-mouse IgM, the presence of such Abs in IL-10-containing B cell cultures allowed for the proliferative response to LPS. A variety of other Abs reactive with murine B cell surface markers were ineffective at modulating the response to IL-10. IL-1, IL-2, IL-4, IL-5, IL-6, IFN-gamma, and TGF-beta were also ineffective in this regard. These observations suggest that IL-10 has a role in the suppression of inappropriate B cell proliferation, i.e., proliferation by B cells that have not effectively interacted with relevant Ag or CD40 ligand.

FcR epsilon+ lymphocytes and regulation of the IgE antibody system. V. Preliminary physicochemical characterization of the T cell-selective IgE-induced regulant EIRT.

We previously showed that murine lymphoid cells exposed to elevated levels of IgE exhibit the de novo expression of Fc receptors for IgE (FcR epsilon), and the production of soluble mediators, which we have termed IgE-induced regulants (EIR). Described herein is the preliminary physicochemical characterization of one such regulant, that being the EIR responsible for the Lyt-2+ T cell-dependent expression of FcR epsilon and secretion of an IgE-binding factor (IgE-BF) which can potentiate IgE synthesis; the former activity has been denoted EIRT for its selectivity of action on T cells, and the latter activity has been termed enhancing effector molecule (EEM) for its presumed potentiating influence on IgE antibody synthesis. Characterized in parallel was the conventional lymphoid cell-derived cEIRT and a murine monoclonal T cell hybridoma-(MBI-2)-derived mcEIRT. EIRT from either source was shown to exhibit the characteristics of a protein with a molecular mass of 45 to 60 kd. Once enriched by gel filtration, neither cEIRT nor mcEIRT preparations displayed any other EIR-like activity except that of EIRT, as evidenced by the ability of these preparations to act selectively to induce the Lyt-2 T cell-dependent expression of FcR epsilon and the production of EEM, the lack of detectable SFA activity that could induce Lyt-1+ T cells to produce the IgE-BF denoted suppressive effector molecule (SEM), and the lack of detectable levels of the B cell-selective EIRB, as indicated by the incapacity of either preparation to induce B cell FcR epsilon expression. Neither cEIRT nor mcEIRT displayed IgE-binding affinity, in contrast to the EEM produced in response to stimulation with these regulants. The only EIR-like activity detected in the unfractionated supernatant fluid from cultures of the monoclonal T cell hybridoma MBI-2 was that of EIRT. Careful in vitro analysis established that such preparations did not contain enhancing factor of allergy (EFA), SFA, EIRB, or IgE-BF. Thus, the enhancement of IgE synthesis observed in animals given this mcEIRT preparation was most likely due to the activity of EIRT known to be present. During the course of these studies, clues as to the physicochemical nature of other EIR activities was obtained. Thus, upon molecular sieve analysis, two distinct molecular mass species of EIRB (one 15 to 20 kd and the other 30 to 35 kd) were demonstrated to be present in conventional lymphocyte-derived cEIR.(ABSTRACT TRUNCATED AT 400 WORDS)

FcR epsilon+ lymphocytes and regulation of the IgE antibody system. I. A new class of molecules, termed IgE-induced regulants (EIR), which modulate FcR epsilon expression by lymphocytes.

Exposure of unfractionated murine lymphoid cells to suitable amounts of IgE in vitro results in the de novo expression of Fc receptors for IgE (FcR epsilon) by both T and B lymphocytes. Monoclonal IgE, in the monomeric state, directly induces FcR epsilon expression by B cell-enriched but not T cell-enriched lymphoid cells. In contrast, the same monoclonal IgE in the aggregated state can directly induce FcR epsilon expression by either lymphocyte class independently. The fact that monomeric IgE induces almost equal proportions of FcR epsilon+ B and T cells unfractionated cell cultures, but can directly induce only FcR epsilon+ B cells when lymphocyte subpopulations are independently exposed to IgE, suggested the involvement of either soluble mediators or cognate interactions in this FcR epsilon expression process. Indeed, the studies presented demonstrate that IgE-stimulated lymphoid cells produce soluble mediators, termed IgE-induced regulants (EIR), which can induce FcR epsilon expression in cultures of fresh lymphoid cells. EIR-stimulated FcR epsilon expression is independent of IgE in either its native or processed state, and is largely by T cells of the Lyt-2+ subset. Thus, total depletion of T cells or more selective elimination of the Lyt-2+ subset prevented the development of FcR epsilon+ cells in cultures exposed to EIR but not in those exposed to monomeric IgE. Conversely, depletion of B cells had the opposite effect in that the remaining T cells retained the ability to express FcR epsilon in response to EIR but were unresponsive to monomeric IgE. Because of its selective activity in inducing FcR epsilon expression by T lymphocytes, EIR from unfractionated lymphoid cell cultures has been designated EIRT. This selectivity of EIRT inductive properties for T lymphocytes was additionally confirmed by analyses of the FcR epsilon+ lymphoid cells subsequent to induction with IgE or EIRT. Thus, unlike monomeric IgE, which induces FcR epsilon+ cells equally distributed among T and B lymphocytes, EIRT induces FcR epsilon+ cells in only the T cell class. These findings indicate that production of the EIRT and subsequent expression of FcR epsilon by Lyt-2+ T cells depends upon the initial interaction of IgE with B cells. Finally, an interesting paradox observed was that although IgE-induced FcR epsilon expression by B cells is unaffected by total T cell depletion, selective blocking of just Lyt-1+ T cells significantly diminishes this FcR epsilon induction process.(ABSTRACT TRUNCATED AT 400 WORDS)

FcR epsilon+ lymphocytes and regulation of the IgE antibody system. II. FcR epsilon+ B lymphocytes initiate a cascade of cellular and molecular interactions that control FcR epsilon expression and IgE production.

This study additionally explores the orderly sequence of events concerning the induction of Fc receptors for IgE (FcR epsilon) that are initiated by IgE and mediated by IgE-induced regulants (EIR). Thus, lymphoid cells exposed to monomeric IgE displayed an early phase of exclusive B cell FcR epsilon expression, followed by the progressive appearance of FcR epsilon+ T cells, ultimately resulting in equal proportions of FcR epsilon+ B cells and T cells. Parallel cultures of lymphoid cells stimulated with EIR derived from unfractionated lymphoid cells (EIRT) also manifested rapid induction of FcR epsilon+ cells, but these FcR epsilon+ cells were predominantly T cells from the outset. Data presented here demonstrate that IgE-induced FcR epsilon expression by B cells ultimately results in the production of EIRT, which then induces FcR epsilon expression by T cells. The existence of EIRT that selectively induce T cell FcR epsilon expression prompted us to search for an EIRB that is selectively active in inducing FcR epsilon+ B cells. Indeed, IgE-stimulated, T cell-depleted lymphoid cells produce an EIRB that selectively induces FcR epsilon expression by B cells. This EIRB, but not EIRT, can also be generated by IgE stimulation of Lyt-2+ cell-blocked lymphoid cells, indicating that Lyt-1+ cells are not inhibitory to EIRB production and that production of EIRT is dependent upon functionally competent Lyt-2+ cells. Similar to IgE, EIRB induces rapid FcR epsilon expression, first by B cells and then by T cells, so that by 16 hr post induction equal proportions of FcR epsilon+ B and T cells were observed. Although complete T cell depletion does not affect IgE-induced FcR epsilon expression, selective blocking of Lyt-1+ cells markedly diminishes such responses, suggesting that Lyt-2+ cells are antagonistic to the induction of FcR epsilon+ B cells. Studies involving sequential T cell subset depletion clearly demonstrated that in the absence of functionally competent Lyt-1+ cells, Lyt-2+ cells exert an inhibitory influence on IgE-induced FcR epsilon expression by B cells. Stimulation of Lyt-1+ cell-blocked cultures with EIRT, and to a lesser degree with IgE, resulted in the elaboration of an EIR (EIRI), which lacks direct FcR epsilon-inductive properties, but conversely, directly inhibits IgE-induced FcR epsilon expression in fresh B cell cultures.

FcR epsilon+ lymphocytes and regulation of the IgE antibody system. III. Suppressive factor of allergy (SFA) is produced during the in vitro FcR epsilon expression cascade and displays corollary physiologic activity in vivo.

Exposure of lymphoid cells to IgE induces the expression of Fc receptors for IgE (FcR epsilon) and the production of soluble mediators, termed IgE-induced regulants (EIR). Conventional suppressive factor of allergy (SFA) and enhancing factor of allergy (EFA), derived from mouse ascites fluids, both inhibit IgE-induced FcR epsilon expression in vitro in cultures of unfractionated and T cell-enriched, but not B cell-enriched, lymphoid cells. This indicates that the inhibitory activities of both entities are T cell dependent, and distinguishes them from the inhibitory EIRI, which inhibits FcR epsilon induction in the absence of T cells. Moreover, SFA and EFA can be distinguished from one another by differences in the T cell subsets required for the inhibitory activity of each respective mediator on in vitro IgE-induced FcR epsilon expression. Thus, SFA requires the presence of Lyt-1+ T cells, whereas EFA requires the presence of Lyt-2+ T cells. Supernatant fluids from IgE-stimulated unfractionated lymphoid cell cultures suppress in vivo IgE synthesis in mice, indicating that SFA is produced along with the other species of EIR. To define conditions required for SFA production in vitro, EIR-rich supernatant fluids were tested for the presence of SFA by using Lyt-2+ cell-blocked indicator cells in the in vitro FcR epsilon induction assay system (this eliminates the inhibitory activity of EFA). SFA production in vitro by IgE-stimulated lymphoid cells was shown to result from cooperative interactions between B cells and Lyt-1+ T cells. In addition, as observed with the induction of FcR epsilon in general, induction of SFA requires the initial interaction of B cells with IgE, and the release of the B cell-selective EIRB. Once produced, EIRB can directly stimulate Lyt-1+ cells, but not Lyt-2+ cells, to produce SFA. The physiologic significance of the in vitro induction of SFA by the action of EIRB on Lyt-1+ cells was confirmed by the demonstration that EIRB, devoid of detectable SFA, selectively suppressed in vivo IgE synthesis after administration to intact mice. This indicates that EIRB can stimulate resident T cells of irradiated SJL mice to produce SFA. Finally, as shown previously with conventional ascites-derived SFA, the SFA produced in vitro after stimulation of lymphoid cells with IgE is devoid of IgE-binding properties, because its inhibitory effects on in vivo IgE antibody synthesis are not removed by passage over IgE affinity columns.

FcR epsilon+ lymphocytes and regulation of the IgE antibody system. IV. Delineation of target cells and mechanisms of action of SFA and EFA in inhibiting in vitro induction of FcR epsilon expression.

SFA and EFA are derived from distinct mouse T cell hybridomas secreting one or the other (but not both) factor, and although both are capable of inhibiting FcR epsilon expression by unfractionated spleen cells induced by monomeric IgE, neither was inhibitory for EIRT-induced FcR epsilon expression by T cells in the same cell population. This suggests that the final target cell for the inhibitory effects of SFA and EFA is the FcR epsilon+ B lymphocyte. T cells are required for both SFA- and EFA-mediated FcR epsilon inhibition, and more precisely, as shown in this study, SFA stimulates Lyt-1+ cells in the presence or absence of IgE to produce a suppressive effector molecule (SEM), and EFA together with IgE stimulates Lyt-2+ cells to produce an enhancing effector molecule (EEM), both of which can directly inhibit FcR epsilon expression by B cells. SFA and SEM can inhibit both IgE- and EIRB-induced FcR epsilon expression by B cells, indicating that SFA may act by blocking the EIRB-mediated expansion of the FcR epsilon+ B cell population. EFA and EEM, in contrast, can inhibit IgE-induced but not EIRB-induced FcR epsilon expression, indicating that EFA may act at some point before the release of EIR, perhaps involving those FcR epsilon+ B cells that respond to IgE and produce EIRB. Finally, although neither SFA and EFA display IgE binding properties, both SEM and EEM, in contrast, are IgE binding factors (IgE-BF) and may be homologous to the suppressive IgE binding factor and potentiating IgE binding factor described by other investigators. The possible interrelationships between these various cells and factors are discussed.

Antigen concentration determines helper T cell subset participation in IgE antibody responses.

A recently developed in vitro system for antigen-stimulated primary and secondary murine IgE antibody responses has been used to define (a) the relative participation of the Th1 and Th2 cell-derived lymphokines IFN-gamma and IL-4, respectively, in such responses, and (b) the role of antigen concentration in determining functional helper T cell activity. These studies confirm that IL-4 and IFN-gamma exert regulatory effects on IgE synthesis, but the nature and extent of their respective effects on primary and secondary IgE responses differ. Thus, primary IgE responses are considerably more sensitive to and dependent on IL-4 than are secondary IgE responses since (1) anti-IL-4 monoclonal antibody totally inhibited primary IgE responses, but only partially affected secondary responses; and (2) exogenously added IL-4 could stimulate primary IgE responses to optimal antigen concentrations, but had no effect on secondary IgE production. Likewise, antigen-stimulated primary IgE responses are about eightfold more sensitive than are secondary responses to the inhibitory effects of IFN-gamma. Studying the effect of antigen dose on the quantity of IgE antibody produced revealed that although IFN-gamma could be detected by ELISA in cultures exhibiting high-dose antigen-dependent diminution of IgE production, anti-IFN-gamma monoclonal antibody could not reverse this phenomenon. Thus, IFN-gamma is not solely responsible for decreased IgE synthesis associated with high-dose antigen exposure. IL-4 activity was detected in the fluid from cultures stimulated with low, but not high, levels of antigen. Moreover, addition of exogenous IL-4 restored IgE production to normal levels in cultures exposed to high antigen concentrations. Therefore, it appears that high levels of antigen result in selective stimulation of Th1 cells which produce IFN-gamma, and diminished activation of IL-4-producing Th2 cells. These results help explain observations regarding the influence of antigen dose on the generation of experimental and clinical IgE antibody responses in vivo.

IL-10 stimulates murine antigen-driven antibody responses in vitro by regulating helper cell subset participation.

The effects of IL-10 on in vitro antigen-driven murine antibody responses and helper cell IL-4 and IFN-gamma secretory capacity were investigated. Low antigen concentrations stimulated high responses in all antibody isotypes examined; IgD was not assayed. Under these conditions, exogenous IL-10 minimally potentiated synthesis of antigen-specific IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA, but inhibited antigen-specific IgE secretion. High antigen levels stimulated antigen-specific IgM, IgG2a, and IgG2b responses, but inhibited synthesis of all other isotypes. In high antigen cultures, IL-10 augmented secretion of antigen-specific antibody in all isotypes except IgE. Essentially all of the antibody produced in the presence of high or low antigen concentrations was antigen-specific. Exogenous IL-10 substantially stimulated production of antigen-nonspecific antibody in all isotypes except IgG3. IL-10 allowed for greater Ig+ cell yield; comparable numbers of CD4+ and CD8+ cells were observed in the presence or absence of IL-10 in culture. The stimulatory effects of IL-10 for in vitro antibody responses were observed during a limited period of time after in vivo antigen priming of the responding cell populations. In contrast, IL-10 inhibited IgE synthesis at all time points tested. Low concentrations of antigen maintained the in vitro capacity of helper cells to secrete IL-4, while high antigen concentrations did not. Exogenous IL-10 potentiated IL-4 secretory capacity in high antigen cultures. The capacity for IFN-gamma secretion was comparable in high and low antigen cultures and exogenous IL-10 significantly inhibited such capacity under both sets of conditions. We conclude that IL-10 is generally stimulatory for murine antibody responses in vitro, with the possible exception of antigen-specific IgE. Such stimulatory effects appear to reflect increased activity of type 2 helper cells with concurrent decrease in type 1 helper cell activity.

Elicitation of antigen-induced primary and secondary murine IgE antibody responses in vitro.

Described herein are methods for eliciting and quantitating primary and secondary murine IgE antibody responses in vitro, and the important role of antigen concentration in determining the level of IgE produced during an immune response. The methods for quantitating IgE antibody levels in culture supernatant fluids and in serum by ELISA are presented in detail. The specificity of such methods was confirmed in that (1) no other isotype of antibody registered in the IgE-ELISA, and (2) parallel determinations of IgE antibody concentrations could be obtained by independent analysis using Fc epsilon RI-dependent basophil degranulation. We examined various parameters of cell donor immunization and lymphoid cell culture which allow for optimal in vitro primary and secondary IgE responses. High relative antigen doses result in diminished IgE antibody responses in experimental animals, a finding confirmed herein. High antigen concentrations in vitro also result in relative suppression of IgE antibody synthesis. This was also true for in vitro production of IgG1 and IgA antibodies. Conversely, IgM and IgG2a responses were elicited at both low and high antigen concentrations; IgG2b and IgG3 were not produced under the conditions of priming and culture used herein. Finally, production of IgE in vitro depended on the presence of carrier-primed CD4+ T cells and hapten-specific B cells. Generation of maximal IgE antibody secretion, and hence elicitation of an allergic reaction, is thus dependent on the amount of antigen acting as stimulant for the immune response.

The cellular lesion responsible for exaggerated IgE synthesis accompanying allergic breakthrough.

Appropriate levels of IgE are maintained by a cellular and molecular network composed of (1) a suppressive, Ly-1+, CD4+ T cell-dependent arm that is activated by inappropriate high levels of IgE and (2) an enhancing, CD8+ T cell-dependent arm that controls this suppression in a feedback regulatory manner. Ly-1+ T cells also function to counterbalance (inhibit) the activity of these latter CD8+ T cells. It has been previously shown that Ly-1+ T cells can reverse low-dose irradiation-induced enhancement of IgE antibody responses (i.e., allergic breakthrough). We have analyzed lymphocytes isolated from mice subjected to low-dose irradiation to determine which component of this network is defective in such animals. Stimulation of normal lymphocytes with IgE in vitro resulted in the release of lymphokines that suppress IgE antibody responses. In contrast, similar stimulation of lymphocytes from irradiated mice did not elicit secretion of such suppressive lymphokines, unless the cells were depleted of CD8+ T cells or reconstituted with normal Ly-1+ T cells. Because Ly-1+ T cells of irradiated mice could not reconstitute the response, we conclude that this functional subset of CD4+ T cells, which normally controls CD8+ T cell activity in this network, is defective in animals that exhibit irradiation-induced allergic breakthrough.

Infection of haematopoietic stem cells in mice with Friend virus induced erythroleukaemia.

Animals infected with conventional anaemia (FVA) or polycythemia-inducing (FVP) strains of the Friend virus develop lethal erythroleukaemia. A variant strain, RFV, induces an initially identical disease except that it spontaneously regresses in 50% of infected mice. To determine whether pluripotent stem cells as measured by spleen colony forming units (CFU-s) in leukaemic mice are productively infected with virus and whether their infection influences the outcome of the disease, we tested CFU-s from leukaemic mice for susceptibility to cytotoxicity by monospecific antiviral gp70 antiserum. Spleen CFU-s from progressively leukaemic (FVP, FVA and RFV) mice were productively infected with virus. CFU-s in RFV progressors became infected by 40 days post-virus inoculation. FVA and FVP progressors became infected between 15 and 21 days post virus. Infection of CFU-s was accompanied by an increase in the proportion of replicating (S phase) CFU-s in these populations. Spleen CFU-s from fully regressed RFV regressor mice were uninfected and remained so throughout the course of their disease. Bone marrow CFU-s in both regressors and progressors remained uninfected and were not induced to increased cell cycling.

B cell activator. Effects on B cell expression of CD23, proliferation, and antibody secretion.

The studies herein describe a B cell hybridoma-derived, low m.w. (less than 1000 Da), hydrophilic mediator denoted B cell activator (BCA). BCA stimulates B cell expression of IgE-specific FcR (Fc epsilon RII or CD23) in a manner similar to IL-4. However, BCA can be readily distinguished from IL-4 because it does not 1) enhance B cell Ia expression; 2) bind 11B11 anti-IL-4 mAb; or 3) elicit superinduction of Fc epsilon RII expression or IgE production in cultures of LPS-activated B cells. Moreover, BCA is considerably more mitogenic than IL-4 for LPS-activated B cells and, in contrast to IL-4, lacks mitogenicity for anti-mu-activated B cells. BCA can enhance IgG2b and IgG3 production by LPS-activated B cells, responses that are suppressed by IL-4. BCA alone did not stimulate IgE and IgG1 production by LPS-activated B cells, but exerted synergistic activity when combined with IL-4 in stimulating secretion of these antibody isotypes. Finally, secondary Ag-driven IgG1, IgE, and IgA antibody responses can be stimulated by BCA in vitro. Thus, BCA appears to be a novel mediator with broad B cell activation properties.

Collagen-induced arthritis in mice. Relationship of collagen-specific and total IgE synthesis to disease.

The relationship between production of IgE and collagen-induced arthritis in mice was examined. Collagen-specific IgE was produced as a consequence of immunization of DBA/1 mice with chicken type II collagen emulsified in CFA. We observed a rise in collagen-specific IgE antibody levels at the onset of CIA clinical and histologic signs in DBA/1 mice. This rise in IgE paralleled that of IgG2a anticollagen antibodies, an isotype implicated in the pathogenesis of CIA by other laboratories. The collagen-specific IgE contained in the plasma of mice with CIA could arm basophils for Ag- (collagen) dependent degranulation. Collagen-specific IgE may thus contribute to CIA by promoting mast cell degranulation in the synovia of susceptible mice immunized with chick type II collagen; but, further work is required to establish such a role for IgE in CIA. However, genetic differences in disease susceptibility could not be accounted for by quantitative differences in collagen-specific IgE production. Further, comparable levels of IgE anticollagen antibodies were observed in animals with active CIA and after spontaneous remission, thereby confirming that the presence of such antibodies is insufficient for disease. Total IgE levels peaked just before spontaneous remission indicating active production of IL-4. IL-4 was administered to animals with CIA to determine if this lymphokine could be involved in the remission process. IL-4 facilitated remission of CIA. Enhanced total IgE production may thus be a marker for activation of Th2 cells that produce lymphokines such as IL-4 and IL-10, factors that may be involved in the spontaneous remission process.

Purification of murine suppressive factor of allergy into distinct CD23-modulating and IgE-suppressive proteins.

The murine suppressive factor of allergy (SFA) has been purified from a T-cell hybridoma and found to consist of two functionally and biochemically distinct protein molecules. One protein (17 kDa) modulates the low-affinity Fc receptor for IgE on lymphocytes (i.e., CD23); it decreases the binding avidity of IgE to CD23-bearing B cells without affecting quantitative expression of CD23 and is thus designated epsilon-receptor-modulating protein. The second protein (30 kDa) suppresses IgE biosynthesis (i.e., SFA). This purified SFA suppresses interleukin 4-induced IgE and IgG1 synthesis by lipopolysaccharide-activated spleen cells but has no effect on other antibody isotypes; since the activity of SFA is not blocked by anti-interferon gamma monoclonal antibody, it is thus distinct from interferon gamma. The data presented indicate that epsilon-receptor-modulating protein and SFA are protein molecules that are involved in modulating the CD23 molecule and IgE antibody synthesis, respectively.