Selective enhancement of human IgE production in vitro by synergy of pokeweed mitogen and mercuric chloride

IgE production in vitro was investigated in cultures of human peripheral blood mononuclear cells (MNC) from non-atopic donors with pokeweed mitogen (PWM), mercuric chloride (HgCl2), or both. PWM alone induced a few IgE immunoglobulin secreting cells (ISC) detected by reverse plaque forming cells (PFC) and many IgG, IgM, and IgA PFC. HgCl2 alone failed to produce significant numbers of ISC of any class. PWM plus HgCl2 caused a selective increase of IgE PFC without affecting IgG, IgM, and IgA PFC. Co-cultures of B cells plus mitomycin C (MMC) treated T cells stimulated by PWM alone produced more IgG, IgM, IgA and IgE PFC than those of B cells plus T cells. However, PWM plus HgCl2 produced significantly more IgE PFC selectively in those cultures. This effect was observed in the cells of most of the donors, but a few donors showed different responses.     

Distribution of Ig Classes and IgG Subclasses among Human B Cells Activated by Nocardia opaca-Delipidated Cell Mitogen or by Pokeweed Mitogen

The relative proportions of cells synthesizing the three major Ig classes or one of the four IgG subclasses in cultures stimulated with pokeweed mitogen (PWM) or Nocardia-delipidated cell mitogen (NDCM) were investigated. In cultures of human peripheral blood mononuclear cells (PB MNC) stimulated with PWM, the number of IgG-containing cells (CC) was higher than the number of IgM-CC, and a substantial number of IgA-CC was found. Conversely, in NDCM-stimulated PB MNC cultures IgM-CC outnumbered IgG-CC and only few IgA-CC were detected. In those cultures, the removal of T cells resulted in an increase in the number of IgM-CC concomitant with a decrease in the number of IgG-CC. A substantial number of cells containing simultaneously IgG or IgA in addition to IgM could be found in PWM-stimulated cultures. These cells were virtually absent in NDCM-stimulated cultures. The relative proportions of IgG subclass-CC were IgG1-CC greater than IgG2-CC greater than IgG3-CC greater than or equal to IgG4-CC in PWM-stimulated and IgG2-CC greater than IgG1-CC greater than IgG3-CC greater than or equal to IgG4-CC in NDCM-stimulated cultures. The removal of T cells from NDCM-stimulated cultures did not result in major alteration of this distribution. The role of T cells and of the genomic order of the Igh-C genes in their phenotypic expression triggered in vitro by PBA is discussed.     

Lipopolysaccharide- and cholera toxin-specific subclass distribution of B-cell responses in cholera.

The immunoglobulin subclass responses to homologous lipopolysaccharide (LPS) and to cholera toxin (CT) in adult patients infected with Vibrio cholerae O1 and V. cholerae O139 were studied. LPS-specific antibody-secreting cells (ASC) of both the immunoglobulin A1 (IgA1) and IgA2 subclasses were seen, with the IgA1 ASC response predominating in both V. cholerae O1- and O139-infected patients. For antibodies in plasma, by day 11 after onset of disease, all V. cholerae O1- infected patients responded to homologous LPS with the IgA1 subclass (P = 0.001), whereas fewer (68%) responded with the IgA2 subclass (P = 0.007). About 89% of V. cholerae O139-infected patients responded with the IgA1 subclass (P = 0.003), and only 21% responded with the IgA2 subclass (not significant [NS]). Both groups of cholera patients showed significant increases in LPS-specific IgG1, IgG2, and IgG3 antibodies in plasma. In feces, the response to homologous LPS occurred in both groups of patients with the IgA1 and IgA2 subclasses, with 55 to 67% of patients showing a positive response. V. cholerae O1- and O139-infected patients showed CT-specific ASC responses of the different IgG and IgA subclasses in the circulation, and the pattern followed the order IgG1 > IgA1 > IgG2 > IgA2, with low levels of IgG3 and IgG4 ASC. Plasma anti-CT antibody responses in all subclasses were seen by day 11 after onset of disease. Although there were no increases in CT-specific ASC of the IgG3 (NS) and IgG4 (NS) subtypes, there were significant increases of these two subclasses in plasma (P 相似文献   

Abnormal behavior of gamma-committed B lymphocytes probed by a lymphocyte blastogenesis inhibitory factor in autoimmune MRL mice

Recently, we have characterized a lymphocyte blastogenesis inhibitory factor (LBIF) which was purified from the culture supernatant of a human histiocytic lymphoma U-937 (Sugimura, K. et al., Eur. J. Immunol. 1989. 19: 1357). In this study, we investigated the effect of LBIF on the antibody production of autoimmune MRL mice in vitro. We demonstrated here that (a) LBIF inhibited the IgM, IgG and IgA antibody responses of lipopolysaccharide (LPS)-stimulated spleen cells of normal BALB/c mice, (b) in the case of old autoimmune MRL/Mp-lpr/lpr (MRL/l) and MRL/Mp-(+)/+ mice, however, LBIF inhibited IgM and IgA but not IgG responses of LPS-stimulated spleen cells, (c) the antibody production of all IgG subclasses, IgG3, IgG1, IgG2b and IgG2a, was not sensitive to LBIF inhibitory activity in these autoimmune mice, (d) in young MRL mice (3-5-week-old MRL/l), which were phenotypically normal, LPS-induced antibody production of all isotypes (IgM, IgG and IgA) was strongly inhibited by LBIF as shown in normal BALB/c mice and (e) in the case of 7-week-old MRL/l the insensitivity to LBIF was concomitant with the appearance of gamma + B lymphocytes. Thus, by employing LBIF as a probe, this study showed a correlation between the pathogenesis of MRL autoimmune disease and the lack of LBIF sensitivity of hyperactive B lymphocytes and suggested that the intrinsic abnormality of autoimmune MRL B lymphocytes might be confined to gamma- but not mu- or alpha-committed B cells.     

Specificity of immediate skin reaction with streptokinase-streptodornase (SK-SD) in humans.

Five patients with no detectable serum IgA (less than 20 mug/ml) and one patient with low serum IgA were compared to normal subjects. The number of circulating E-RFC was normal as was the lymphocyte DNA synthesis induced by PHA, Con A, and streptokinase-streptodornase. The patients had normal numbers of IgA-bearing lymphocytes and normal or increased numbers of B cells. Purified anti-immunoglobulin antibodies specific for IgG, IgA and IgM induced a normal lymphocyte DNA synthesis as did PWM. The patients' lymphocytes were able in vitro to transform into actively secreting IgA plasmocytes. This transformation was determined by counting the IgA and immunoglobulin-containing cells and then measuring the IgA and IgG secretion in the cultures. In some patients PWM was selectively suppressive in IgA B-cell transformation into IgA secreting cells; in the other patients PWM had no effect on the IgA B-cell differentiation. PWM enhanced the IgG secretion in the patients' cultures as well as IgA and IgG secretion in the normal controls.     

Ontogenic development of proliferative and polyclonal antibody and autoantibody responses to staphylococcal peptidoglycan, protein A and cell walls in mice

In the spleens of newborn mice, polyclonal IgM, IgG and IgA responses were low to protein A, cell walls (CW) and PWM, intermediate to peptidoglycan (PG), and high to LPS. Small, not significant increases in Ig responses to LPS (which occurred during the first 2 wks) were observed, whereas, the responses to PG, protein A, CW and PWM continued to increase for 8 wks, and the increases were high and significant. In most cases, there was no change in the dose response and kinetics patterns (characteristic for each stimulant) during ontogeny. Ontogenic development of autoantibody-secreting cells was different from the development of cells secreting all Ig and anti-SRBC antibodies. The increases in the numbers of cells secreting IgM anti-DNA and IgM anti-bromelin-treated mouse RBC antibodies in response to LPS and PG during postnatal development were larger than the increases of all IgM-secreting cells. In contrast, the increases of IgM anti-DNA secreting cells in response to protein A and PWM were smaller than the increases of all IgM-secreting cells. The frequencies of cells secreting anti-DNA antibodies in LPS- and PG-stimulated cultures were low in newborns and continued to increase until 8 wks of age, but they were high and did not change throughout ontogeny in protein A-stimulated cultures. Changes in the frequencies of anti-RBC antibody-secreting cells were less distinct and mostly insignificant. Postnatal changes in mitogenic responses were smaller and not correlated with the development of polyclonal Ig responses. Our data indicate different modes of ontogenic development of the ability of cells to produce polyclonal Ig, autoantibodies and heteroantibodies, and to synthesize DNA, in response to different stimulants.     

Isolation of rat IgM to IgG hybridoma isotype switch variants and analysis of the efficiency of rat Ig in complement activation

Sequential sublining was used in combination with enzyme-linked immunosorbent assays to isolate mu----gamma isotype switch variants of the rat IgM secreting mouse-rat B cell hybridoma line BA1.8. Switch variants to all four subclasses of IgG were obtained. The variant antibodies retained the antigen specificity of the parental IgM for the O18 (lipopolysaccharide) antigen of Escherichia coli. In sodium dodecyl sulfate-polyacrylamide gels the apparent molecular mass of the gamma heavy chains decreased in the order gamma 2b greater than gamma 1 greater than gamma 2c greater than gamma 2a. IgM, IgG1, IgG2a, IgG2b and IgG2c of the BA1.8 variant family and IgG2b, IgE and IgA of the previously described BA1.2 family were used for a comparative analysis of the capacity of rat Ig to activate complement. Efficient lysis of sheep erythrocytes coated with the O18 antigen was observed with IgM and all IgG subclasses, but no lysis was triggered by IgE or IgA. One hundred to 1000 IgG molecules were required to mediate the same hemolytic activity as one IgM molecule. The four IgG subclasses were equally efficient at mediating lysis by rat or human complement, while IgG2a was less efficient with guinea pig complement than the other three IgG subclasses. Antibody-triggered binding of C3 to pathogenic O18:K1 E. coli bacteria was measured using serum containing 125I-labeled C3. K1-encapsulated strains did not fix C3 efficiently in the absence of specific antibodies while acapsular mutants fixed C3 via the alternative pathway. IgM and all IgG subclasses triggered C3 binding to the K1 encapsulated bacteria. The capacity of IgM to mediate C3 fixation was not greater than that observed with IgG.     

In vitro immunoglobulin production by mononuclear cells in rheumatoid arthritis

Since patients with rheumatoid arthritis (RA) exhibit serum hypergammaglobulinemia and autoantibody (rheumatoid factor) production, we compared elaboration and control of in vitro RA mononuclear cell (MNC), Ig assayed by enzyme-linked immunoassays or by hemolytic plaque formation, in 37 RA patients and 17 normal subjects. We found (1) RA spontaneous plaque-forming cells were significantly reduced (RA 344 vs normal 627 PFC/10(6) MNC, P less than 0.002); (2) RA spontaneous IgG and IgM (but not IgA) elaboration was significantly diminished (IgG RA 339, normal 776; IgM RA 255, normal 869 ng/ml, P less than 0.001; IgA RA 87, normal 124); (3) RA stimulated IgG and IgM production (but not IgA) was also decreased (IgG RA 2434, normal 3862, P less than 0.06; IgM RA, 1676, normal 3323, P less than 0.005; IgA RA 1859, normal 2315); (4) reduced RA Ig elaboration was not clearly due to altered numbers of T or non-T cells, age, medications, clinical features of disease, or response kinetics; (5) relative improvement of RA in vitro IgG, but not usually IgM, secretion followed removal of adherent cells, addition of indomethacin or addition of mitomycin C-treated T cells; (6) MNC from synovial fluids, but not bone marrows, exhibited spontaneous Ig production in excess of stimulated synovial fluid cellular or peripheral blood Ig elaboration. These observations indicate selective impairment of peripheral blood MNC IgG and, particularly, IgM secretion in RA. This defect appears to reflect accessory cell influences which differ from normal as well as the sequestration of primed or activated cells in the synovial fluid.     

Repair of Immunoglobulin Response in B Cell Line (JK32.1) Originating from Immunodeficient Patient via Implantation of Functional Plasma Membranes

Human-human B cell hybridoma JK32.1, constructed from B lymphocytes of a common variable immunodeficient patient and nonsecreting cell line, retains the defects of B cell immunodeficiency. Efforts to clarify whether the defect is located within the plasma membranes of this cell line were carried out by implanting them with plasma membrane fraction derived from normal functional cells via intact noninfectious Sendai virus. The implanted cells were activated with various mitogens and their Ig responses and isotype switching were examined. Restoration of IgM secretion was achieved in the implanted JK32.1 cells following stimulation with SAC, PWM, or retinoic acid. Augmented IgM response was also obtained in the implanted cells treated with retinoic acid and lipopolysaccharide (LPS) despite their unresponsiveness to LPS alone. No IgG or IgA response could be detected in the implanted JK32.1 cells. These data suggest that this immunodeficient cell line possesses at least two different malfunctions, one located within the plasma membrane moiety of the cells and the other located within the cytoplasmic and/or nucleic components. The plasma membrane moiety defect can be repaired temporarily by delivering proper signals via the implanted plasma membranes. However, this manipulation of the cells could not overcome the intrinsic defect of the cells which blocks isotype switching and secretion of IgG, IgE, and IgA antibodies.     

Bacteroides-specific IgG and IgA subclass antibody-secreting cells isolated from chronically inflamed gingival tissues

The emergence of cells that produce IgG and IgA subclass antibodies to Bacteroides gingivalis (Porphyromonas gingivalis) fimbriae and lipopolysaccharide (LPS) antigens was examined in mononuclear cells isolated from inflamed gingiva of different stages (slight, moderate or advanced) of adult periodontitis (AP). Antigen-specific IgM, IgG (including IgG1, IgG2, IgG3 and IgG4) and IgA (including IgA1 and IgA2) producing cells were enumerated by the ELISPOT assay and were compared with total Ig-producing cells of each isotype or subclass. In advanced AP, the B. gingivalis fimbriae-specific IgG- and IgA-secreting cells represented 5% of total IgG- or IgA-secreting cells, while those from the moderate stage comprised approximately 1% of these two isotypes. Cells producing antibody specific for B. gingivalis LPS were observed at frequencies of 0.1% and 0.4% for IgG and IgA cells, respectively in the advanced stage. When IgG subclasses were analysed in moderate AP, the anti-fimbriae subclass responses were largely IgG1 (60%), followed by IgG2 (20%), IgG3 (10%) and IgG4 (10%). Fimbriae-specific IgG subclass responses were elevated in the advanced stage of AP, and IgG4 (40%) and IgG1 (30%) were dominant, followed by IgG3 (20%) and IgG2 (10%). IgA1 cells predominated in both the moderate and advanced stages, however a relative increase in IgA2 cells occurred in advanced AP. Mononuclear cells isolated from gingiva of AP patients did not contain cells producing antibody to antigens such as Escherichia coli K235 LPS, cholera toxin or the hapten dinitrophenyl coupled to bovine serum albumin. These results show that local IgG and IgA subclass responses occur to a protein antigen of a major periodontal disease (PD)-associated pathogen, B. gingivalis, and the increase in IgG4 and IgA2 responses may be associated with host protection.     

IgM rheumatoid factor elaboration by blood, bone marrow, and synovial mononuclear cells in patients with rheumatoid arthritis

IgM rheumatoid factor (RF) elaboration by rheumatoid arthritis (RA) synovial, bone marrow, and blood mononuclear cells (MNC) is reported. IgM RF was prepared from RF-positive sera by sequential euglobulin precipitation, Sephacryl S300 gel filtration, and IgG-Sepharose affinity chromatography. Purified material, which contained no detectable IgG or IgA, was used in an enzyme-linked immunosorbent assay (ELISA) to quantitate cellular elaboration of IgM RF. Excellent standard curves (r2 = 0.98) were obtained without nonspecific binding of samples or antisera to IgG-coated microtiter plates and without cross-reactivity of standards with antisera other than anti-IgM. We found RA blood MNC (11 patients) spontaneously averaged 15 ng/ml IgM RF (6% of total IgM produced), but elaborated 254 ng/ml IgM RF following pokeweed mitogen (PWM) stimulation (22 patients), exceeding that of 13 normal controls. Bone marrow MNC spontaneously (4 patients) produced 71 ng/ml IgM RF and secreted 78 ng/ml IgM RF with PWM stimulation (9 patients). In contrast synovial fluid MNC (5 patients) spontaneously elaborated 6652 ng/ml IgM RF, significantly (P less than 0.05) more than blood or bone marrow MNC; PWM-stimulated synovial fluid MNC (5 patients) produced 5472 ng/ml IgM RF. These observations confirm selective localization of activated, IgM RF-producing cells to the rheumatoid synovial space.     

T-Cell Helper Activity and B-Cell Function of Synovial and Blood Lymphocytes from Patients with Rheumatoid Arthritis or Other Forms of Chronic Arthritis

The helper effect of T cells on B-cell immunoglobulin (Ig) responses induced by pokeweed mitogen (PWM) or purified protein derivative of tuberculin (PPD) was studied in lymphocytes from synovial fluid (SF) and blood of nine patients with rheumatoid arthritis (RA) and eight patients with other forms of chronic arthritis. In PWM cultures the helper effect of SF T cells on Ig responses (IgG, IgM, IgA) of autologous and allogeneic blood B cells was lower than that of blood T cells (P less than 0.01). This decrease was more pronounced in patients with RA than in patients with non-RA. In PPD cultures no significant difference was found between the helper effect of SF T cells and blood T cells on the Ig responses of allogeneic blood B cells or on the IgG response of autologous blood B cells, whereas the helper effect of SF T cells on the IgM and IgA responses of autologous blood B cells was decreased. The Ig responses to PWM or PPD in cocultures of autologous blood B and T cells were not significantly different between patients and healthy controls. The PWM- and PPD-induced Ig responses of SF B cells were lower than those of blood B cells when cocultured with autologous blood T cells. SF B cells produced IgG but usually little IgM and IgA. Thus there was a dysfunction of SF B cells and of SF T cells in a PWM-driven system, but a fairly good helper function of SF T cells in a PPD-driven system.     

LPS augments human B-cell differentiation by direct stimulation of PWM-responsive B cells

Bacterial lipopolysaccharide (LPS) augments production of IgM and IgG by two- to seven-fold in cultures of peripheral blood lymphocytes (PBL) stimulated by pokeweed mitogen (PWM), but only if monocytes are rigorously depleted. When PBL were separated into adherent cell (AC), B-cell-enriched, and T-cell-enriched fractions, pulsed with LPS, and recombined in culture with PWM, increased generation of plasma cells was seen only in cultures containing LPS-treated B cells. This effect of LPS appears to be independent of soluble factors. Supernatants from LPS-stimulated B cells or AC did not consistently increase PWM responses when cultured with fresh B cells in the presence of polymyxin B. Furthermore, pulsing of B cells with purified interleukin 1 from two different commercial sources failed to augment PWM-induced differentiation. When B cells were depleted of surface IgD (sIgD)-bearing cells by panning, no effect on LPS-mediated augmentation of PWM-driven differentiation was seen. B cells were also fractionated by rosetting with mouse erythrocytes. Treatment of BMR+ cells with LPS did not induce them to respond to PWM, while treatment of BMR- cells with LPS augmented generation of plasma cells. These results indicate that LPS acts directly to augment differentiation of PWM-responsive B cells, rather than recruiting sIgD+, BMR+ cells to become PWM responsive.     

Alteration of the immunoglobulin G subclass responses in mice to lipopolysaccharide: effects of nonbacterial proteins and bacterial membrane phospholipids or outer membrane proteins of Proteus mirabilis

The immunoglobulin M (IgM) and the IgG1, IgG2ab, and IgG3 subclasses of plaque-forming cells (PFC) specific for lipopolysaccharide (LPS) were measured after immunization of mice with LPS alone and compared with the responses to LPS in combination with nonbacterial proteins and with bacterial membrane phospholipid vesicles or two major outer membrane proteins from Proteus mirabilis. The relative numbers of IgG PFC belonging to the IgG1, IgG2, or IgG3 subclasses induced by immunization with LPS alone depended upon the type of LPS administered. Phospholipids and the proteins effected characteristic alterations in not only the strength but also the subclass of the IgG responses to LPS. The results suggest that the hydrophobic-hydrophilic nature or state of aggregation of the preparations plays a role in the induction of IgG1 and IgG2 subclasses of PFC specific for LPS. Complex formation with LPS and adjuvant was apparently necessary to obtain these effects.     

Suppression of polyclonal immunoglobulin biosynthesis by a soluble factor: T-cell dependent and T-cell independent mitogens

When normal human spleen cells are pulsed with concanavalin A (Con A), a portion become suppressor cells which in co-culture can inhibit immunoglobulin synthesis by other normal spleen cells stimulated by pokeweed mitogen (PWM). One mechanism whereby these Con-A activated spleen cells suppress Ig synthesis appears to be by the secretion of a soluble suppressor factor(s) since supernatants of Con-A stimulated splenocytes also suppress the polyclonal synthesis of immunoglobulin by human spleen cells. In this study, we report that supernatants of Con-A activated spleen cells suppress the in vitro synthesis of IgG, IgM and IgA by human spleen cells cultured with PWM. Our results indicate that the soluble suppressor factor(s) blocks an early stage in the differentiation of B lymphocytes into plasma cells without affecting the synthesis and secretion of immunoglobulin by more mature lymphocytes which appear to be irreversibly committed toward the pathway of synthesizing immunoglobulin. In addition, we studied the ability of normal human spleen cells to synthesize polyclonal immunoglobulin when cultured with either the T-cell dependent PWM or T-cell independent mitogens lipopolysaccharide (LPS) and Nocardia. Our results demonstrate that normal human splenic mononuclear cells cultured with either Nocardia, LPS or PWM are significantly stimulated to synthesize polyclonal IgG, IgM and IgA. Furthermore, supernatants of Con-A activated human spleen cells suppressed the polyclonal synthesis of these three antibody classes by human spleen cells responding to either T-cell dependent or independent mitogens.     

The optimum conditions for antibody production in vitro by Peyer's patch lymphocytes and a comparison between total immunoglobulins and antigen (TNP)-specific antibody synthesis following mitogen stimulation

Using optimised conditions in terms of cell concentration (2 X 10(6)/ml), duration of culture (7 days) and pH (7.0-7.1) and the highly sensitive enzyme-linked immunosorbent assay for antibody quantitation, we assessed the effects of lipopolysaccharide (LPS) and concanavalin A (Con A) on the levels of both total immunoglobulins and TNP-specific antibodies and compared the proportion of IgA anti-TNP to total IgA synthesised following the in vivo priming of mice, 5 days prior to culture, with trinitrophenylated sheep red blood cells (TNP-SRBC). LPS stimulation resulted in an increase in total IgA (ninefold), IgM (sixfold) and IgG (eightfold) synthesis over unstimulated cultures and similar increases were seen in IgA anti-TNP (eightfold), IgM anti-TNP (fivefold) and IgG anti-TNP (twelvefold). Con A stimulation resulted in a slight increase in total IgA (threefold), IgG (twofold) but a decrease in IgM (twofold) but did not appear to affect TNP-specific antibody levels. In unstimulated cultures, IgA anti-TNP comprised 7.8% of the total IgA synthesised. In LPS and Con A stimulated cultures the proportion was 6.6% and 3.2%, respectively. Inclusion of 10% fetal calf serum in the culture medium resulted in a substantial increase in the synthesis of IgG, IgM and IgA in unstimulated cultures but increased IgA levels only in LPS stimulated cultures. Our results emphasise the importance of optimising culture conditions when studying in vitro antibody synthesis and demonstrate that Peyer's patches respond quite substantially to parenterally administered antigen and that these responses can subsequently be monitored in vitro.     

Immunoglobulin G subclasses secreted by human B cells in vitro in response to interleukin-2 and polyclonal activators

The IgG subclasses secreted by human B cells in vitro in response to IL-2 have been analysed. B cells were prepared from tonsil, blood and spleen, and cultured with recombinant IL-2 in the presence or absence of two polyclonal activators: Staphylococcus aureus Cowan 1 (SAC) and bacterial lipopolysaccharide (LPS). Secretion of all four subclasses and of IgM was stimulated by IL-2, but the relative amounts varied according to (i) the tissue source of the B cells, and (ii) which polyclonal activator was used. The amount of IgG1 tended to be higher and IgG2 tended to be lower when SAC was the polyclonal activator (compared to LPS). This difference was most marked for tonsil B cells, and it was found that SAC had a negative effect on secretion of IgM and IgG2 in these cultures, whilst synergizing with IL-2 to stimulate the production of IgG1, 3 and 4. When the degree of stimulation of different pairs of isotypes was analysed, several interesting positive correlations emerged. In tonsil B-cell cultures, stimulation of IgM and IgG2 was linked with each other, but not with IgG1, whilst in blood B-cell cultures all isotypes appeared to be stimulated co-ordinately. Stimulation of IgG1 and IgG3 were positively correlated in cultures of B cells from all tissues. The results emphasize that the effects of a single cytokine on immunoglobulin isotype production can be influenced by the source of the B cells, and by other signals delivered to the cells.     

Are cord blood B cells functionally mature?

Very low immunoglobulin secretion occurs in pokeweed mitogen (PWM) stimulated cord blood mononuclear cells (MNC) and has been attributed to an 'immaturity' of both T and B cells of the newborn. The cord blood T cells are phenotypically 'naive' cells, in which suppressor activity for B cell function appears to dominate over helper activity. The cord blood B cells, in spite of their expression of different membrane immunoglobulin isotypes, secrete almost no IgG and IgA in the various B cell assays so far compared. We found that cord blood B cells are as competent as B cells from adults to generate clonal IgM, IgG and IgA responses in a culture system in which a cell contact with mutant EL-4 thymoma cells in conjunction with T cell supernatant leads to strong B cell activity. As regarding the possible causes of the low cord blood PWM response, we studied the role of transforming growth factor-beta 1 (TGF-beta 1), a potent inhibitor of lymphocyte functions. TGF-beta 1 sensitivity of B cells and TGF-beta 1 mRNA levels in MNC were found to be similar for adult and cord blood cells. A neutralizing anti-TGF-beta 1 antibody enhance the adult PWM response, but the immunoglobulin secretion in cord MNC remained very low. We conclude that suppression by endogenous TGF-beta 1 occurs in the PWM system but is not responsible for the low immunoglobulin response of cord blood MNC and that the newborn's B cell 'immaturity' can be overcome with potent T cell signals in vitro. This is consistent with the newborn's capacity to generate a T-dependent B cell response in vivo.     

Quantitation of Cells Secreting Immunoglobulins after Elution from Rheumatoid Synovial Tissue

Mononuclear cells (MNC) eluted from rheumatoid synovial tissue of 16 patients with rheumatoid arthritis were examined for immunoglobulin-secreting cells (ISC). Immediately after elution and separation synovial tissue MNC contained considerable numbers of ISC. IgG and IgA ISC were more abundant than IgM ISC. At the same time there were low numbers of ISC in blood. Synovial tissue ISC were lost during incubation with pokeweek mitogen (PWM), possibly because tissue MNC were already maximally triggered in vivo. This was in contrast to blood MNC, in which the number of ISC increased significantly during incubation with PWM.     

IL-2和PWM对人淋巴细胞产生免疫球蛋白的影响

本文初步观察了IL-2和PWM 对人淋巴细胞产生免疫球蛋白的影响。将PWM、MLA-144细血培养上清(富含IL-2)分别加入人扁桃体和外周血淋巴细胞体外培养。用酶联免疫吸附试验双抗体夹心法,分别检测第七天培养上清液中IgG、IgM和IgA的含量。加入PWM(10μg/ml)或 MLA-144 细胞培养上清(1:5稀释)的刺激组,IgG、IgM、IgA 的含量均较对照组明显升高,三种免疫球蛋白比较,IgG、IgM 产量高于IgA。相同条件下,扁桃体淋巴细胞产生的IgA,高于外周血淋巴细胞所产生者。同时加入PWM和MLA-144细胞上清联合刺激组较加PWM 或MLA-144 细胞上清单独刺激组IgG、IgM、IgA的产量增高。结果说明:适当刺激浓度的 IL-2和PWM 对人淋巴细胞免疫球蛋白的产生有明显促进作用。一定条件下,PWM和 IL-2联合刺激诱生免疫球蛋白有相加作用。