Eur. J. Immunol. 1991. 21: 1445-1451

IL4-induced motility in B cells

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Birgitta Clinchy, Carina Elenstrom, Eva Severinson and Goran Moller

T and B cell collaboration: induction of motility in small, resting B cells by interleukin 4*

Department of Immunology, Arrhenius Laboratories for Natural Sciences, Stockholm University, Stockholm

In this report we investigate if I L 4 can work as a chemoattractant factor by inducing locomotion in B cells. We found that murine recombinant IL 4 (rIL 4) induced motile morphology and migration through polycarbonate micropore filters of murine, splenic B cells at an optimal concentration of 3 ng/ml. Kinetic studies revealed optimal migration at 8-16 h, although a significant response could be detected already after 1 h. Flow cytometric studies confirmed that the migrated cells were indeed B cells.We also compared the activity of small, dense B cells and large, low-density B cells, based on Percoll gradient separation. We found no difference in IL 4-induced motility among the two groups. Furthermore, we looked at B cells activated in vitro by preculture in lipopolysaccharide (LPS) or IL 4. Our data indicate that both LPS and IL 4 can increase the general capacity for motility in B cells after preculture for 24 h. Tand B cell collaboration requires close cell-cell contacts in order for Tcell help to be administered to the B cell. One way of enhancing such cell contacts could be through directional cell migration induced by helper factors (chemotaxis). We suggest that IL 4 can play a role as a chemoattractant factor that enhances cell contacts between T helper cells and B cells.

1 Introduction The exact mechanisms whereby T cells help B cells are presently unknown. One aspect of Tand B cell collaboration, that has been demonstrated by many investigators, is the need for close cell-to-cell contacts between these two cell types during the antibody response [l-41. This is probably a reflection of reciprocal activation, where the T cells that deliver helper factors to B cells are themselves stimulated by antigen bound to the B cell. A major problem is how the “right” B and T cells find each other. The frequency of antigen-specific lymphocytes is initially very low, likewise the probability of two rare cells getting sufficiently close to each other. One mechanism that could enhance the chances for contact formation betweenTand B cells is directional migration induced by chemoattractant factors. It is becoming increasingly clear that chemotaxis plays an important role in directing lymphocyte migration in the body. Several of the defined interleukins have been found to possess chemotactic properties. For example, IL 1, IL 2 and IL 8 (neutrophil-activating protein) have all been implicated in the recruitment of lymphocytes to sites of inflammation [5-71, although the effects of IL 1 remain somewhat controversial. SN from mitogen- or antigenactivated leukocytes contain additional factors that can stimulate lymphocyte motility. Several laboratories have

[I 86551

* This work was supported by grants from the Swedish Medical Research Council and the Swedish Natural Science Research Council.

Correspondence: Birgitta Clinchy, Dept. of Immunology, Arrhenius Laboratories for Natural Sciences, Stockholm University, S-10691 Stockholm, Sweden 0 VCH Verlagsgesellschaft mbH. D-6940 Weinhcim, 1991

characterized an attractant for CD4+ T lymphocytes, referred to as Lymphocyte Chemoattractant Factor (LCF) from such SN [8-111. Factors that selectively attract Tcell precursors have also been isolated from mouse [ 121 and rat [13] thymic stromal cells, extending the role of chemoattractants to include influences on cell differentiation. Both T and B cells have the capacity for locomotion. However, since most studies o n lymphocyte migration have focused onTcells, little information is available regarding B cell migration. Early studies by Unanue et al. showed that anti-Ig antibodies could induce motility in rat and mouse B cells [ 14-17]. Staphylococcus aureus, Cowan strain [ 181and IFN-y [19] are two other compounds reported to activate locomotor capacity in B cells. In this report we examine IL 4, a helper factor derived from activated CD4+ Tcells, as a possible chemoattractant for B lymphocytes. IL 4 has multiple effects on B and Tcells, as well as other cells of the immune system [20, 211. It is an important differentiation and isotype switch factor for both mouse and human B cells [22-251. It also influences cell surface expression of several molecules on B cells, for example, MHC class I1 antigens [26]. Recent studies in this laboratory have revealed a new property of IL 4 in its effect on B cells. It was found that IL 4 induced adhesion when added alone and that it increased cell aggregation among LPS-stimulated murine B cells. We have also observed IL 4-induced appearence of cells with elongated protrusions, which is characteristic of cells with locomotor capacity, after culture of B cells in IL 4 for 48 h [27]. Similar findings have recently been reported by Wilkinson and Islam [19] who observed a change from spherical, non-motile shapes to polarized, motile morphology among B cell-enriched human PBL after 24 h exposure to human rIL 4. 0014-2980/91/0606-1445$3.50+ .2510

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Eur. J. Immunol. 1991. 21: 1445-1451

B. Clinchy, C. Elenstrom, E. Severinson and G. Moller

This has led us to investigate if I L 4 , released as a chemoattractant factor fromTcells, can induce motility in B cells.

2 Materials and methods 2.1 Animals and media CBA, B1O.BR and (C57BL/6 x CBA)FI mice, of both sexes, were obtained from our own breeding facility at Stockholm University.The mice were used at 6-14 weeks of age. RPMI 1640 supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, penicillin 50 IU/ml, streptomycin 50 pg/ml (Gibco BRL, Grand Island, NY) and 5% of heat-inactivated FCS (Gibco, Ltd, Paisley, GB) was used in all experiments, unless otherwise indicated. 2.2 Cytokines and reagents Highly purified (> 98% according to SDS-PAGE) murine rIL 4 obtained from E. coli transfected with cDNA clone pKCR.IL4.Neo coding for mouse IL 4 [20] was a generous gift from Dr. Hama (On0 Pharmaceutical Co., Osaka, Japan). This preparation was used in all experiments. The biological activity of 1 U was defined as the concentration inducing half maximal response in the IgGl induction assay. One unit corresponded to 0.6ng of rIL4. LPS was extracted from E. coli 055:B5 (Dept. of Bacteriology, Karolinska Institute, Stockholm, Sweden), it wasused at 50 mg/ml, which was the concentration giving optimal proliferation of splenocytes after 2 days of culture. 2.3 Cell preparations To purify B cells, single-cell suspensions of splenocytes were first incubated with Gey's solution to lyse red blood cells and then washed twice in Earle's balanced saline solution (BSS, Gibco, BRL, Grand Island, N.Y.). T cells were depleted from the cell suspension by incubation with monoclonal anti-Thy-1.2 (AT83A), anti-CD4 (RL172.4) and anti-CD8 (3IM) antibodies together with low-tox rabbit complement (Buxtedt Rabbit Company Ltd., GB) for 1 h at 37 "C in BSS. The mAb were used as hybridoma culture SN. The hybridomas were all gifts from Dr. H. R. MacDonald, Lausanne, Switzerland. Small B cells were then isolated by Percoll (Pharmacia, Uppsala, Sweden) density centrifugation as previously described [28]. The cells in the band between 70% and 60% Percoll were collected. They consisted of 89%-96% Ig+ und 0%-5% Thy-1.2+ cells as determined by immunofluorescence staining. In experiments where high- and low-density B cells were compared, cells from the layer between 60% and 50% Percoll were also collected. Cell viability, based on trypan blue exclusion, was always > 95%.

presence of 50 pg/ml LPS or 5 U/ml rIL 4. Since it has been reported, in several different systems, that prolonged culture of cells in serum can result in increased motile behavior [29,30],we reduced the concentration of FCS in the medium from 5% to 0.5% during the preculture. We confirmed the activity of LPS in the precultures by separately measuring [3H]dThd uptake (data not shown). Viable cells, recovered after centrifugation on LympholyteM (Cedarlane Laboratories Ltd, Hornby, Canada), were washed and resuspended in regular media at 2.5 x 106/ml. Migration was then tested in the cell migration assay, as described below, with medium containing 5% FCS. 2.5 Motile shapes Purified B (0.5 x 106-2 x 106) cells were cultured for the indicated time in 24-well tissue culture plates (Costar) in medium with or without 50 pg/ml LPS or 5 U/ml rIL 4 at 37°C and 5% COf. Using a Nikon 114 inverted phase contrast microscope with a 30 x magnification, cells were scored as either spherical or polarized, the latter having a tapered body and uropod. Ten individual, randomly chosen fields were counted and the percentage polarized cells was calculated. At least 500 cells were counted for each experimental condition. 2.6 Cell migration assay Migration of B lymphocytes was assessed employing a modification of the Boyden chamber technique [31]. A total of 5 x lo4 cells, suspended in 20 pl medium, were placed in the upper compartment of a 48-well microchemotaxis chamber [32] (Neuro Probe, Inc., Cabin John, MD) separated by two polyvinylpyrrolidone-freepolycarbonate filters, 5 pm pore size, (Nuclepore, Co., Pleasanton, CA) from 25 pl of medium alone (control) or different concentrations of rIL4 in medium. The cells were allowed to migrate through the pores in the filters for the time indicated in each experiment at 37 "C in 5% CO2. Examination of the filters revealed very few adherent cells; therefore, lymphocyte locomotion was quantified by harvesting and counting the cells that had completely traversed the filters and collected on the bottom of the chamber.This approach has previously been used both with human neutrophils [33] and murine splenocytes [34]. For kinetics experiments, remaining cells were removed from the top wells at indicated time points and all cells in the bottom wells were counted at the end of the experiment. All conditions were set up in quadruplicates in each experiment and the mean was calculated. In some figures, migration is represented as % of control.The total number of cells that had migrated in medium alone was considered the background and was set to 100%. The effect of each variable is then expressed relative to the background response.

2.4 Preculture conditions 2.7 Phenotyping of migrated lymphocytes In some experiments 5 x lo6 B cells were precultured in tissue culture flasks (Costar, Cambridge, MA) in 10 ml medium with 0.5% FCS and 1 x lo6 irradiated (1200 rds), syngeneic spleen cells as accessory cells, in the absence or

In experiments where cells were stained for immunofluorescence after migration, 4.7 mm diameter blind well chambers (Neuro Probe) were used instead of a microchemotaxis

Eur. J. Immunol. 1991.21: 1445-1451

IL4-induced motility in B cells

chamber. Of a cell suspension (1 x 106) 400 pl was loaded in the top compartment and 200 pl of rIL 4, at a concentration of 2.5 or 5 U/ml, was added to the lower compartment. Otherwise the experimental conditions were the same as in the cell migration assay. Cells that had reached the bottom of the chamber were collected after the indicated time, counted and stained.

2.8 lmmunofluorescence staining FITC-conjugated rabbit F(ab')z anti-mouse Ig (Dakopatts, Copenhagen, Denmark), culture SN from the rat hybridoma M1/70. 15.11.5 (Mac-1, alpha subunit, American Type Culture Collection, Rockville, MD) and FITC-conjugated mouse anti-rat Ig x light chain (MARK1, Immunotech S.A., Marseille, France) were used. Cells were stained by the original technique [35] and analyzed by FCM using a Becton Dickinson (Mountain View, CA) FACScan.

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3.2 Induction of lymphocyte migration A more direct measurement of cell motility is to study cells that migrate through a matrix or filter. Fig. 2 shows the dose-response curve of migrating B lymphocytes to rIL 4 after 19 h, utilizing a microchemotaxis chamber with two 5 pm pore size polycarbonate filters. The response was optimal at 5 U/ml and declined at higher concentrations of rIL 4. Similar results have been obtained using culture SN from H28 and X63 cells transfected with the gene for murine IL 4 (data not shown). The background migration, in medium alone, varied from 0.6%-5.6% of input cells. The time kinetics up to 24 h of the response is presented in Fig. 3. A notable effect was seen already after 2 h. It increased until 8-16 h after which only a slight increase, or even decrease, was seen. Ap value of < 0.05 was obtained when significance values were calculated from 8 independent experiments with concentrations of rIL 4 ranging from 2 to 20 U/ml and migration times ranging from 6 to 24 h.

3 Results IL 4 alone does not induce proliferation in B cells [38, 391 and we found no uptake of [3H]dThd or increase in total cell numbers among B cell cultured in rIL 4 for 24 h (data not

3.1 Induction of motile shapes by IL 4

Motile cells display a characteristic morphology with an anterior pseudopod or lamellipodia and a trailing uropod (polarized cells). Chemotaxis is achieved by orienting the direction of locomotion along a chemoattractant gradient, such that the pseudopod extends towards the higher chemoattractant concentration [36]. Scoring of polarized cell morphology with phase contrast microscopy has been widely used as one method to measure cellular capacity for locomotion [37]. Recently,Wilkinson et al. [19] showed that such polarization could be seen in human B cells exposed to IL 4. We cultured purified murine B cells with rIL 4 or in medium alone and inspected the cultures in an inverted microscope at different time points. As can be seen in Fig. 1, a significant increase in the number of polarized cells was evident already after 30 min. By 20 h 37% of the B cells showed polarized morphology. This data confirms the results by Wilkinson et al., although the kinetics shown here are faster than those previously published.

40

1

0.6

25 5 10 20 Unitdm1 rIL 4

12

Figure 2. Dose-response curve of IL 4-induced B cell migration. Purified B cells (5 x 104) were placed in the top compartment of a &-well micro chemotaxis chamber. The cells were separated from the bottom chamber, containing varying concentrations of rIL 4, by two polycarbonate filters (pore size 5 pm). The chambers were incubated for 19 h at 37 "C, 5% CO2 and the cells that had migrated through the filters and reached the bottom chamber were harvested and counted. Data is expressed as percent of migration in medium alone. One representative experiment out of four is shown. Experiment 1

a

051

2

3

4

40

Experiment 2

400,

400,

20

Time (hours) Figure 1. Time-course of polarization of B cells. Purified B lymphocytes were cultured in medium alone (0),50 p g h l LPS (A) or 5 Ulml r I L 4 ( 0 ) .The percentage of polarized cells were evaluated with phase-contrast microscopy at the indicated time points. Mean and SE from three separate experiments.

8

16

hours

24

2

4

6

0

24

hours

Figure 3. Time kinetics of IL 4-induced B cell migration. The experiment was performed as described in Fig. 1. The cells were allowed to migrated for the indicated time before harvesting. ( 0 ) 10 U/ml (0)2 U/ml rIL4.

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B. Clinchy. C. Elenstrom, E. Severinson and G. Moller

shown). Hence, the cell harvested from the rIL 4-containing lower half of the chamber were all the result of migration and not due to proliferation of a few migrated cells.

(data not shown). Consequently, we conclude that B cells are mostly responsible for the migration observed here. 3.4 Comparison of migration in small and large B cells

3.3 Phenotype of migrated cells Only a fraction (maximum 15%) of the total B cell population, loaded in the top compartment of the chamber, actually migrated through the filters into the rIL 4containing lower half, even under optimal conditions. Therefore, it was important to establish that the migrated cells truly were B cells and not contaminating cells of other types. Immunofluorescence staining of our purified B cells, before the migration assay, routinely gave 89%-96% Igf cells and 2%-14% contaminating cells positive for the marker Mac-1. This marker is present on myeloid cells but absent on lymphoid cells [40]. It was therefore conceivable that some of the migrating cells were of M@ lineage, rather than B cells.Very few (0%-5%) Tcells were present among our purified B cells, hence we found it unlikely that they should contribute significantly to the migratory response. We collected migrated cells from the lower wells, containing various concentrations of rIL4, and compared the staining with that of the original B cell population. As can be seen inTable 1, no enrichment of Mac-l+ cells was found among the migrated population.The majority (71%-90%) was still Ig+. Culture of B cells in rIL 4 for 24 h did not alter their phenotype with regard to the markers tested here

5 UniWml

10 Unitdm1

rIL 4

Figure 4. Migration of small and large B cells in response to IL 4. B cells were separated into a high- and a low-density population on Percoll gradients. Small, high-density cells (open bars) and large. low-density cells (filled bars) were placed in a micro chemotaxis chamber with 5 or 10 U/ml of rlL 4. as described in the legend to Fig. 1 and allowed to migrate for 17 h. Mean and SD from two separate experiments.

It has been suggested that only activated lymphocytes can respond to chemoattractant factors. This has been demonstrated for Tcells [6, 41, 421. Whether B cells also require preactivation is less clear. To examine this question we separated T-depleted splenocytes into a large and a small cell fraction on Percoll gradients. De Franco et al. [28] have reported that the small, dense population represents resting, unactivated cells, whereas the larger, low-density population contains in vivo preactivated cells. In our assay (Fig. 4) we did not find an improved migration response to rIL 4 among the large B cells. On the contrary, the small, dense population appeared to be slightly more responsive than the large cells. 3.5 Migration of B cells precultured in LPS or IL 4 We also wished to examine locomotor responses to I L 4 among B cells preactivated in vitro.For these experiments

P r c c u l N ~ i a Medium

LPS

PrcculNR time:

25h

K-4

Medium

Lps

n-4

Medium

24h

LB

K-4

48h

Figure 5. Effects of preculture with LPS or IL 4 on subsequent migration of B cells. Purified B cells ( 5 x loh)were precultured in medium with 0.5% FCS and 1 X loh irradiated syngeneic spleen cells either alone or in the presence of 50 pglml LPS or 5 U/ml rlL 4 for the indicated time. Viable cells were recovered by density gradient centrifugation and washed extensively. The cells were resuspended in medium and 5 x 10" cells added to micro chemotaxis chambers containing either medium alone (open bars) or 5 U/ml rIL 4 (hatched bars). Migrated cells were recovered after 16-18 h and counted.

Table 1. Phenotype of migrating cells

Exp.

Migration time (h)

rlL J"J (Ulml)

Original cell population Mac- 1h, Ig

(Yo1 3 Ih I9

2.5 5 5

3 11

ND

1 84 80 03

a) Amount o f rIL 4 placed in lower compartment of blind-well chamber. b) Results are given as perccntages of positive cells out of 5000 or loo00 cells analyzed by FCM.

Migrated cclls Mac- I 'g (Yo) (74,) I 3 ND

ND 90 71

Eur. J. Immunol. 1991. 21: 1445-1451

we used LPS, a polyclonal B cell activator known to induce proliferation and differentiation in murine B cells. IL 4R, although present on resting B cells, are up-regulated by LPS and by IL 4 itself [43]. In the same experiments we also looked at B cells preactivated by rIL 4. In Fig. 5 purified, small, high-density B cells were cultured for 2.5, 24 or 48 h with LPS, r I L 4 or medium, and then tested in the migration assay. B cells precultured in medium alone retained normal background migration and responded with increased migration to rIL 4, at the same level as would be expected of fresh B cells. B cells stimulated with LPS or rIL 4, on the other hand, showed a dramatic increase in migration even when only medium was present in the chemotaxis chambers. Migration in I L 4 was also elevated, but not much above migration in medium alone. This induction was not immediate and required preculture for more than 2.5 h. The cells migrating after LPS stimulation were verified to be B cells by immunofluorescence staining (data not shown).

3.6 Induction of motile shapes by LPS We also tested the ability of LPS to directly induce motile morphology in B cells. As can be seen in Fig. 1, LPS induced motile morphology in 34% of the examined B cells after 20 h of exposure. The level of polarization was the same as that induced by rIL 4 after 20 h, but the kinetics of the response was much slower.

4 Discussion We have previously reported that IL 4 can induce cellular aggregation and appearence of cells with the morphological characteristics of motile cells among B cells [27]. This, taken together with other findings that several interleukins and T cell-derived factors can elicit chemotactic responses in lymphocytes, prompted us to investigate if IL 4 could function as a chemoattractant for B cells. To our knowledge, this has been suggsted in only one previous study where human, B cell-enriched PBL displayed motile morphology after exposure to IL 4 [ 191. Rather than using changes in morphology as the only indicator of locomotor behaviour,we have, in addition, studied migration of B cells through polycarbonate filters in a microchemotaxis chamber. A model for how lymphocytes aquire motility has been proposed [44]. In this model the locomotor process is divided into two stages. A resting lymphocyte is first induced by growth activators to enter the cell cycle. The growth activator can be a mitogen or a cytokine. The cell has now aquired locomotor capacity and can, in the second stage, respond to exogenous chemoattractants, such as IL 8, or to locomotor activators made by the cell itself. A cell with locomotor capacity can respond to an attractant by increasing the speed of locomotion (chemokinesis) or by directional migration (chemotaxis). The growth-dependent induction of locomotor activity, in stage one, requires protein synthesis, but not necessarily DNA synthesis. It is a time-consuming process that takes several hours, whereas the attractant-induced response, in stage two, is immediate and noticeable within < 1 h.

IL4-induced motility in B cells

1449

We shall here try to discuss our findings in relation to this proposed model. We found that murine rIL 4 could induce dose-dependent migration of purified, murine, splenic B cells (Fig. 2). The dose-response curves showed inhibition at high concentrations of rIL4. Many chemotactic substances have this kind of dose-response curve and it is consistent with the requirement that cells undergoing chemotaxis are more sensitive to lower concentrations of a chemoattractant, far from the source, whereas at the active site, where concentrations are higher, they may become immobilized [5, 71. Kinetics experiments revealed a rather fast response with detectable migration after 1 h (Fig. 3). Maximal numbers of migrating cells was reached by 8-16 h. In addition, cells with motile morphology could be detected as early as after 30 min exposure to IL 4 (Fig. 1).These time-kinetics are not in accordance with those in the previous study 1191, where no significant effect of IL 4 on human B cell motility was observed until after 6 h and maximal response was not reached until after 24 h of culture. Based partly on the slow kinetics for polarization, IL 4 has previously been considered to fit in the category of cytokines capable of growthinduced locomotor activation. Our data, on the other hand, indicate that IL 4 could also function as an attractant. This discrepancy could, of course, be attributed to a species difference between mice and man or to the different origin, blood vs. spleen, of the B cells used in these two studies. In general, T lymphoblasts seem to be more effective in their migratory responses than resting T cells [6, 41, 421. Whether the same conditions apply to B cells is not evident. We wanted to establish if preactivation of B cells could further enhance the migratory response to I L 4 and addressed this question using two approaches. First, we compared B cells separated on Percoll density gradients, and second, we compared the responses of LPS- or IL4-stimulated B cells to those of B cells cultured in medium alone. Our results show that in vivo derived B cell blasts, separated by Percoll, responded at the same level as small B cells, both in migration in medium alone and in IL 4 (Fig. 4). Hence, separation based on cell density cannot distinguish B cells that have aquired locomotor capacity from those who have not. B cells stimulated in vitro with LPS for 24 or 48 h, on the other hand, enhanced their background migration in medium, but showed very little additional increase in migration when IL 4 was added (Fig. 5). Exposure to LPS for 2.5 h was not sufficient for this response. To our knowledge, LPS has not before been described as a direct inducer of locomotion in B cells. However, earlier work has indicated that LPS can affect lymphocyte migration since injection of LPS recruits lymphocytes in vivo [45]. It would be expected that LPS should behave as a growth activator and our data, both from the studies on LPS-induced polarization and the preculture experiments, supports this assumption. Surprisingly, in the same preculture experiments, IL 4 could also enhance overall migratory ability of B cells after 24 or 48 h of culture (Fig. 5). If IL 4 was only working as attractant, as some of our data has already indicated, one would expect the activity of IL 4 to dissap-

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B. Clinchy, C. Elenstrom, E. Severinson and G. Moller

pear after washing of the precultured cells. However, these findings place IL 4 together with LPS as a growth activator. In summary, in this report we show evidence that IL 4 can behave both as a growth activator and as an attractant; that is, it activates both stage one and stage two in the model discussed above. A previous study has labeled I L 4 as a growth activator only. With the methods employed in this study we cannot determine if the attractant induced motility is chemotactic or chemokinetic. It has been suggested that cellular adhesion molecules play an important role in regulating cell motility in tissue, for example during homing to LN. It is presently not known if chemoattractants influence cell motility through up-downregulation of such adhesion molecules, but several reports indicate that this may be the case [46,47]. Masinovskyet al. [48] recently showed that IL 4 promoted lymphocyte adhesion to endothelial cells by induction of vascular cell adhesion molecule 1.We have, so far, been unable to detect inhibition of IL 4-induced B cell aggregation or migration with antibodies specific for the adhesion molecule lymphocyte function-associated antigen 1. ([27], and unpublished data.) The physiological role of cell contacts during the Tdependent antibody response is still unclear. It could have a multifunctional role, where close proximity of cells is necessary both for triggering of signals via cell surface bound receptorslligands and for local secretion of soluble helper factors in sufficiently high concentrations. Lassila et al. [49] have proposed that Tcells must be preactivated by APC, other than B cells, in order to direct antibody responses. Our proposal, that IL 4, released from activated Th cells, attract B cells, fits well within this hypothesis. We suggest that chemotaxis regulated by Th factors could be one mechanism whereby contact formation between Th cells and B cells is facilitated. It is also tempting to speculate that IL 4 from Th cells is instrumental in maintaining the architecture of germinal centers through enhanced influx and cellular adhesion of B cells. The authors wish to thank Dr. Julius Klomiriek for use of the blind-well chambers.

Received June 26. 1990; in revised form January 18, 1991.

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IL4-induced motility in B cells

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T and B cell collaboration: induction of motility in small, resting B cells by interleukin 4.

In this report we investigate if IL 4 can work as a chemoattractant factor by inducing locomotion in B cells. We found that murine recombinant IL 4 (r...
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