Biomedical subjects

G Delespesse

Publications and source records attributed to G Delespesse.

At least 91 records · Page 5Linked to original sources

PubMed1988

Elevation of IgE-binding factors in serum of patients with B cell-derived chronic lymphocytic leukemia.

One hundred nineteen sera from patients with various lymphoproliferative diseases and normal sera were tested by a solid-phase radioimmunoassay (RIA) for their content in IgE-binding factor (IgE-BF), a soluble glycoprotein binding to IgE and derived from low-affinity IgE receptors (Fc epsilon R). Fc epsilon R, recently identified as CD23, are known to be expressed on the surface of B cells at the intermediate stage of differentiation. The results indicate that in all cases of chronic lymphocytic leukemia (CLL) tested (n = 40), IgE-BF levels (45 to 8,656 U/mL) were 3-fold to 500-fold higher than in 24 controls (15.5 +/- 2 U/mL). With a few exceptions, serum IgE-BF levels could differentiate patients with CLL from those with other leukemias or lymphomas. In vitro studies indicated that B lymphocytes isolated from CLL patients produced 8 to 50 times more IgE-BF than did normal B cells (P less than 0.001). IgE-BF level was correlated with the Rai stage of the disease (P = 0.002) and the lymphocyte count (P = 0.041). IgE-BF was purified to homogeneity from one CLL serum sample by a combination of affinity chromatography and reverse-phase high-performance liquid chromatography (HPLC). this IgE-BF proved identical to IgE-BF isolated from the culture supernatant of RPMI 8866 cells, a B lymphoblastoid cell line bearing Fc epsilon R and secreting IgE-BF. Indeed, the two molecules had the same mol wt (25 to 27 kd), the same isoelectric point, and the same tryptic map. We suggest that determination of serum IgE-BF might prove useful for clinical monitoring of CLL.

B-Lymphocytes↗

PubMed1987

T-cell-derived IgE-binding factors. I. Cloned and transformed T cells producing IgE-binding factors.

In order to derive T cell clones that can produce factors that specifically regulate the IgE response, Fc epsilon receptor (Fc epsilon R)-positive T cells were isolated from two patients with hyper-IgE states (either the tropical pulmonary eosinophilia syndrome or the hyper-IgE syndrome) and transformed using the human T cell leukemia/lymphoma virus. After infection, these T4-positive, Fc epsilon R-positive T cells manifested a fourfold increase in the surface interleukin 2 receptor, a 10-fold increase in Fc epsilon R, and acquired viral markers not present in the uninfected cells. Cloning soon after transformation allowed for the isolation of 17 distinct T cell clones producing IgE-binding factors. The levels of these binding factors released in supernatant fluid from these clones ranged from 10 to 98 U (negative less than 1 U) using a radioimmunoassay. Furthermore, the presence and kinetics of production of these binding factors also could be demonstrated by metabolic labeling studies. These cells should prove potent immunologic tools for dissecting the various regulatory mechanisms involved in IgE production both in the normal state and in pathologic conditions.

Antigens, Viral↗

PubMed1987

T-cell-derived IgE-binding factors. II. Purification and characterization of IgE-binding factors produced by human T cell leukemia/lymphoma virus-1-transformed T lymphocytes.

We previously described human T cell leukemia/lymphoma virus-1-transformed T cell clones expressing surface molecules binding to monoclonal antibodies against lymphocyte Fc epsilon receptor (Fc epsilon R) and releasing soluble factors binding to both IgE and to monoclonal antibody to lymphocyte Fc epsilon R. In this study, one such clone (HE1-11) was tested for the presence of mRNA hybridizing with a cDNA probe coding for Fc epsilon R on B cells. Northern blot analysis revealed the presence of the same 1.7 kb Fc epsilon R mRNA in HE1-11 and in RPMI 8866 cells as well as in the macrophage cell line U937, expressing Fc epsilon R. The IgE binding factors (BF) released by HE1-11 cells appeared to be identical to those released by RPMI 8866 cells as shown by 1) one- and two-dimensional sodium dodecyl sulfate/polyacrylamide gel electrophoresis analysis and 2) tryptic peptides analysis. IgE-BF are microheterogeneous containing molecules with molecular mass of 25 to 27 kDa and with isoelectric point ranging from 4.5 to 5. Some preparations further contained 16-kDa fragments of IgE-BF. These findings suggest that the gene coding for both Fc epsilon R and IgE-BF may also be expressed on some human T cells.

Antibodies, Monoclonal↗

PubMed1987

Relationship between human IgE-binding factors (IgE-BF) and lymphocyte receptors for IgE.

This study indicates that human IgE-binding factors (IgE-BF) found in the cellfree culture supernatant (CSN) of Fc epsilon R-bearing B cells are breakdown products of the surface Fc epsilon R. This conclusion is suggested by the following observations. 1) Fc epsilon R and IgE-BF share several antigenic determinants as shown by immunoprecipitation with several Mab to Fc epsilon R (MabER) and SDS-PAGE analysis of the precipitates. 2) Upon incubation at 37 degrees C, normal tonsillar lymphocytes lose their Fc epsilon R and this is associated in a time-related manner with the release in the CSN of molecules reacting with two MabER. 3) Surface radioiodinated tonsillar lymphocytes or RPMI 8866 cells release labeled IgE-binding molecules displaying the same antigenic composition and the same migration on SDS-PAGE as purified IgE-BF. 4) Peptide mapping of highly purified IgE-BF and Fc epsilon R reveals the presence of several identical fragments after digestion with either alpha-chymotrypsin, trypsin, or papain. Moreover, papain digestion of the 25-27 kD IgE-BF and of the affinity-purified Fc epsilon R, generated a 15 kD fragment reacting with two MabER and that is known to bind IgE. Although these data strongly suggest that IgE-BF may be directly derived from cell surface IgE receptors, they do not exclude the possibility that some IgE-BF may also be secreted without being first anchored in the cell membrane.

Antibodies, Monoclonal↗

PubMed1987

Cloning and expression of the cDNA coding for a human lymphocyte IgE receptor.

Low-affinity receptors (Fc epsilon R) and secreted factors (IgE-BF) which bind to immunoglobulins of the IgE isotype play a key role in the regulation of human IgE synthesis. We report here the cloning of a cDNA coding for the Fc epsilon R of the human B-lymphoblast cell line RPMI 8866. The nucleotide sequence of this cDNA predicts a polypeptide with 321 amino acids and a mol. wt of 36,281 daltons. A functional Fc epsilon R capable of binding IgE was expressed in Chinese hamster ovary cells after stable transformation with the cDNA which had been cloned into a mammalian expression vector. Amino acid sequence analysis of IgE-BF purified from RPMI 8866 cells revealed an amino-terminal sequence of 19 residues which coincides with the predicted amino acid sequence of the Fc epsilon R, starting at residues 148 and 150. A computer search with the translated amino acid sequence of the Fc epsilon R revealed a domain of 120 amino acids having striking homology to the human asialoglycoprotein receptors.

Amino Acid Sequence↗

PubMed1987

Enhancement of IgE synthesis and histamine release by T cell factors derived from atopic patients with bronchial asthma.

Culture supernatants of unstimulated T cells (TCS) derived from normal donors or from atopic patients with bronchial asthma were tested for their ability to regulate the spontaneous IgE synthesis by B cells of normal and atopic subjects. The same TCS were also tested for their influence on the histamine release from leukocytes of house dust mites-sensitive patients. Addition of TCS to B cell cultures from allergic donors induced a dose-dependent increase of the spontaneous IgE production without affecting the synthesis of IgG, IgM, and IgA. The potentiating activity of TCS was observed only in B cell cultures spontaneously producing IgE; TCS were still active on irradiated B cells. The maximal IgE-enhancing activity was observed when TCS were added at the onset of B cell cultures. The supernatants of T cells lysed at day 0 did not contain IgE-potentiating factors. The antigen-induced but not the spontaneous histamine release from leukocytes of house dust mite-sensitive patients was enhanced by pretreatment with TCS from allergic donors. The enhancing activities of TCS on IgE synthesis and on histamine release could be removed by absorption with IgE-Sepharose and subsequently recovered by elution with glycine buffer. The results indicate that T cells of patients with asthma spontaneously release IgE-binding factors capable of increasing both the spontaneous IgE synthesis by B cells and the antigen-induced histamine release.

Asthma↗

PubMed1987

Modulation of IgE synthesis by IgE-binding factors released by T cells of asthmatic patients with elevated serum IgE.

The culture supernatants of unstimulated T cells (TCS) from asthmatic patients with elevated serum IgE were tested for IgE-binding factors (IgE-BFs) displaying the IgE-potentiating activity. The IgE-BFs were detected by their ability to inhibit the rosetting of RPMI 8866 cells with ox erythrocytes coupled with mouse monoclonal antibody (E-Mab) specific to Fc receptors for IgE (Fc epsilon R). TCS showing the rosette-inhibiting activity significantly enhanced the spontaneous IgE synthesis by B cells of allergic individuals. Interestingly, rosette-inhibiting factors could be removed by absorption with IgE-Sepharose from which they were subsequently eluated with acid buffer, indicating that the rosette inhibition was indeed mediated by IgE-BFs. In addition, such IgE-BFs had affinity for concanavalin A and lost their IgE-potentiating activity after treatment with trypsin and neuraminidase. In contrast, T cells treated with tunicamycin released IgE-suppressing factors capable of inhibiting the IgE-potentiating activity of TCS derived from untreated T cells. On the other hand, the culture supernatants from subpopulations depleted of Fc epsilon R+ T cells but not of Fc gamma R+ T cells contained neither rosette-inhibiting factors nor IgE-potentiating factors, suggesting that IgE-BFs were released by in vivo pre-activated Fc epsilon R+ T cells. With regard to circulating Fc epsilon R+ T cells determined by E-Mab, they were significantly higher in asthmatic patients with elevated serum IgE (0.77 +/- 0.15%) than in normal subjects (0.17 +/- 0.07%) in spite of a very small proportion of T cells bearing Fc epsilon R.

Asthma↗

PubMed1987

In vitro production of IgE-binding factors by human mononuclear cells.

This study documents the production of IgE-binding factors (IgE-BFs) by unstimulated and by mitogen-activated human mononuclear cells. IgE-BFs were detected by a sensitive radioimmunoassay employing monoclonal antibodies to lymphocyte Fc epsilon R (MabER). IgE-BFs were found in the 24-hr CSN of unfractionated tonsillar lymphocytes and of their B-cell but not of their T-cell enriched fractions. When cultured for 1 week, PBMC spontaneously synthesized and released IgE-BFs in the CSN; this was significantly reduced by IgE (10 micrograms/ml). PWM, PHA and Con A significantly increased the production of IgE-BFs by PBMC, and this was not influenced by IgE. The production of IgE-BFs in response to mitogens required interactions between T and non-T cells, and IgE-BFs seemed to be derived mainly from non-T cells. However, low levels of IgE-BFs could be detected in the CSN of highly purified T cells cultured for 1 week in the presence of PHA. The production of IgE-BFs by non-T cells was T-cell dependent and it was mediated by soluble factors released from mitogen-activated T cells. T-cell factors increased the secretion of IgE-BFs by: the macrophage cell line U937, adherent cells, and adherent cell-depleted B-cell preparations. It is concluded that the majority of IgE-BFs produced by cultured human mononuclear cells are derived from B cells and monocytes, and that their production is regulated by T lymphocytes.

B-Lymphocytes↗

PubMed1987

Purification and partial biochemical characterization of IgE-binding factors secreted by a human B lymphoblastoid cell line.

IgE-binding factors (IgE-BFs) were purified from the culture supernatant of RPMI-8866 cells, a human lymphoblastoid B-cell line expressing IgE receptors. The material, purified by affinity-chromatography on immunoadsorbents coupled to IgE or to monoclonal antibody against IgE receptor, was comprised of two major components with apparent molecular weight (MW) of 25,000-27,000 and 12,000, as determined by SDS-PAGE and silver staining. Only the 25,000-27,000 MW molecules were identified as IgE-BFs, as demonstrated by their reactivity with MabER in the Western blot and the immunoprecipitation assays, and their ability to inhibit rosette formation of U937 cells with IgE- but not with IgG-coated erythrocytes. IgE-BFs were purified to homogeneity by combining affinity-chromatography and either DEAE-ion exchange or reverse-phase chromatography on an HPLC system. Chromatofocusing analysis demonstrated the microheterogeneity of IgE-BFs that were comprised of molecules with isoelectric points ranging from 5.0 to 4.4. IgE-BFs were sensitive to treatment with O-glycosidase but not with N-glycanase. These molecules were resistant to heat and to pH ranging from 2 to 9; their immunoreactivity was lost after treatment with trypsin and pepsin. Papain digestion of purified IgE-BFs generated 14,000-16,000 MW molecules that were still binding to IgE and to MabER.

B-Lymphocytes↗

PubMed1986

Production and characterization of a monoclonal antibody to biotin.

Monoclonal antibodies to biotin have been prepared by using biotin linked to keyhole limpet haemocyanin (KLH) as the antigen. Spleen cells obtained from mice immunized with biotin-KLH were fused with the myeloma cell line NS-1. The resulting hybridomas were screened for the production of antibodies to biotin using an enzyme-linked immunosorbent assay. Clones producing antibodies to biotin were isolated by limiting dilution methods. Four cell lines, each derived originally from a different fusion, were chosen for the production of monoclonal antibodies. The monoclonal antibodies obtained have been characterized with respect to their ability to interact with biotin, biotin-bovine serum albumin, biotin-KLH and biocytin as well as to inhibit biotin-dependent enzymes. They have been used to produce cellular biotin deficiency in vitro for studies of biotin function.

Animals↗

PubMed1986

Production and characterization of rabbit anti-idiotypic antibodies specific to mouse monoclonal anti-human IgE and reacting with IgE-binding factors and lymphocyte receptors for IgE.

A mouse monoclonal antibody specific to human IgE (mAb 75) was employed to immunize a rabbit to obtain anti-idiotypes (aId) bearing the internal image of human IgE determinants and reacting with IgE-binding factors (IgE-BF) and/or lymphocyte receptors for IgE (Fc epsilon R). mAb 75 was selected on the basis of inhibition assays where the binding of mAb 75 to radiolabeled IgE was blocked by IgE-BF. The latter were produced by a lymphoblastoid cell line (RPMI 8866) expressing Fc epsilon R. Sequential samples of rabbit serum, collected during the immunization period, were extensively absorbed on mouse and human Ig-Sepharose 4B. The IgG fractions of the rabbit serum displayed the following activities: (a) they reacted with 125I-labeled mAb 75 but not with other labeled mouse Ig including mAb-aIgE, (b) this binding was inhibited in a dose-dependent fashion by human IgE but not by other human Ig classes nor by heat-inactivated IgE, (c) they reacted with a polyclonal rabbit anti-human IgE and (d) they blocked the binding of 125I-labeled IgE to mAb 75. It was concluded that the rabbit IgG contained aId (RaId) bearing the internal image of heat labile determinants of human IgE. The rosetting of IgE-coated bovine erythrocytes with Fc epsilon R-bearing cells was inhibited by preincubating the receptor-bearing cells with IgG RaId or its F(ab')2 but not with normal rabbit IgG. The ability of RaId to react with IgE-BF as well as with Fc epsilon R was also shown in inhibition experiments where IgE-BF and solubilized Fc epsilon R blocked the binding of mAb 75 to RaId. Finally, Western blot analysis of human colostrum, known to contain IgE-BF, indicated that radioiodinated RaId and IgE identified the same 12-16-kDa molecules corresponding to IgE-BF. It is concluded that RaId expresses the internal image of a heat-labile determinant of IgE which is involved in the binding of IgE to IgE-BF and Fc epsilon R. An alternative interpretation is that RaId reacts with an idiotypic determinant of mAb 75 which is shared by IgE-BF and Fc epsilon R.

Animals↗

PubMed1986

IgE receptors on human lymphocytes. I. Identification of the molecules binding to monoclonal anti-Fc epsilon receptor antibodies.

The present study indicates that the surface molecules from Fc epsilon receptor-bearing human lymphoblastoid cells binding to monoclonal antibodies to Fc epsilon receptor (mAbER) are identical to those binding to IgE. mAbER identify three components in the Nonidet-P40 cell lysate of surface-iodinated RPMI 8866 cells with approximately 65-95 kDa, 45 kDa and 37 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions and autoradiography. The same pattern is observed under nonreducing conditions, except for the low mol. wt component which displays an apparent molecular mass 31 kDa. When the labeled and solubilized membranes are adsorbed on IgE-Sepharose the 45-kDa component is always found in the eluate, whereas the 65-95-kDa component is occasionally detected and the low mol. wt component is rarely found. The binding of these molecules to IgE-Sepharose is inhibited by soluble IgE, mAbER but not by an unrelated mAb; reciprocally, the binding to mAbER-Sepharose is prevented by mAbER and to some extent by IgE. To improve the stability of IgE-Fc epsilon R complexes, biotinylated IgE was cross-linked on surface iodinated intact cells, which were then solubilized and adsorbed on avidin-Sepharose. Under these conditions, it was clearly shown that IgE binds to the same three surface molecules as mAbER. The isoelectric point of the high mol. wt component ranged from 4.2 to 4.4 as compared to 5.1-5.2 for the 45-kDa and the 37-kDa components. The observation in the Western blot assay that both the 65-95-kDa and the 45-kDa molecules react with 9 different mAbER indicates that these molecules have a polypeptide sequence in common.

Antibodies, Monoclonal↗

PubMed1986

IgE receptors on human lymphocytes. II. Detection of cells bearing IgE receptors in unstimulated mononuclear cells by means of a monoclonal antibody.

A monoclonal antibody (mAb) specific for lymphocyte IgE receptors (ER) was employed in a rosette assay for the detection of cells bearing IgE receptors (Fc epsilon R). The specificity of the assay was documented by inhibition studies with soluble immunoglobulins (Ig) and anti-Ig antibodies. Moreover, similar results were obtained by employing the F(ab')2 fragment of mAbER instead of intact molecule. Circulating mononuclear cells isolated from normal or allergic adults and from umbilical cord blood contained approximately 8% of Fc epsilon R-bearing cells with values ranging from 0.3 to 17%. Tonsillar lymphocytes contained about 30% of Fc epsilon R+ cells. After the removal of adherent cells, there was a small but significant reduction of the proportion of Fc epsilon R+ cells. When mononuclear cells were separated into T and B cell fractions by two-cycle rosetting with 2-aminoethylisothiouronium bromide hydrobromide-treated sheep red blood cells, most of the Fc epsilon R+ cells were in the B cell fraction; however, a small proportion of Fc epsilon R+ was also found in the enriched T cells and double-labeling experiments confirmed that these cells were indeed T lymphocytes. Fc epsilon R+ cells were purified by rosetting with mAbER-coated erythrocytes and their phenotype was compared to that of Fc epsilon R- cells; Fc epsilon R+ cells contained about 90% of B cells (B1+) together with a small proportion of OKT3+, Leu 7+ and Mo2+ cells. The bulk of T cells, macrophages and natural killer (NK) cells was found in the Fc epsilon R- cells which contained fewer B cells than the fraction of Fc epsilon R+ cells. These data thus indicated that the great majority of Fc epsilon R-bearing cells are B cells but that a small proportion of NK cells, macrophages and T lymphocytes also express Fc epsilon R. Upon incubation at 37 degrees C, B cells lost their Fc epsilon R and this phenomenon was selectively inhibited by IgE; however, purified T cells seemed to express more Fc epsilon R after overnight incubation at 37 degrees C and this was not influenced by IgE. It is finally shown that the expression of Fc epsilon R is cyclic and that Fc epsilon R-bearing B cells do not represent a functionally distinct subpopulation of B lymphocytes.

Adult↗

PubMed1986

Presence of IgE suppressor factors in human colostrum.

In spite of intensive investigations, the ability of breast feeding to delay and to attenuate atopic diseases in children remains debatable. This study documents a mechanism whereby breast feeding might interfere with the synthesis of IgE by breast-fed infants. Indeed, we show that colostrum contains IgE-binding factors (IgE-BF) capable of suppressing the in vitro synthesis of human IgE. Colostrum obtained from 15 donors was successively depleted of lipids and casein, filtered through Amicon XM50 membrane (mol. mass cut-off 50 kDa) and lyophilized. IgE-BF was demonstrated in such preparations by two different approaches, i.e. a classical rosette inhibition assay and Western blot analysis. In the first instance, lyophilized preparations of colostrum inhibited the binding of IgE-coated bovine erythrocytes to IgE recovered on the surface of RPMI 8866 lymphoblastoid cells. The rosette-inhibiting activity could be absorbed on IgE- but not on IgG-Sepharose 4B and it could be recovered in the eluate of IgE-Sepharose 4B. The molecular mass of IgE-BF was comprised between 10 to 20 kDa as estimated by gel filtration through a calibrated Sephadex G-75 column. After fractionation on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transfer to nitrocellulose membrane, colostrum displayed one band of 14 kDa and reacted with radiolabeled IgE but not with IgG nor IgM. This 14-kDa band could be removed by absorbing colostrum with IgE- but not with IgG-Sepharose 4B. Most importantly, the colostrum IgE-BF suppressed the spontaneous in vitro synthesis of IgE by B lymphocytes derived from allergic donors without altering the production of IgM.

Colostrum↗

PubMed1986

IgE receptors on human lymphocytes. III. Expression of IgE receptors on mitogen-stimulated human mononuclear cells.

Human peripheral blood mononuclear cells (PBMC) were tested for the expression of Fc epsilon-receptor (Fc epsilon R) after stimulation with various mitogens in the absence of IgE. Fc epsilon R were found on virtually all the cells from 19 Epstein-Barr virus-transformed B cell lines including those derived from cord blood, from one agamma-globulinemic patient and VDS-0 pre-B cells. Hence, the data clearly indicate that Fc epsilon R may be expressed on very immature B cells. PBMC cultures stimulated with either pokeweed mitogen (PWM), phytohemagglutinin (PHA) or concanavalin A displayed an early increase of their content in Fc epsilon R-bearing cells followed by a decrease to levels below those of control cultures. After fractionation of the PWM-stimulated cultures into T and B cell-enriched preparations, most of the Fc epsilon R+ cells were in the in the B cell fractions and the same low levels of Fc epsilon R+ cells were found in the T cell fractions isolated from the PWM-stimulated and from the control cultures. Double-labeling experiments, employing biotinylated F(ab')2 monoclonal antibody to FcR and either fluorescein isothiocyanate-conjugated B1 or Mo2 monoclonal antibodies, indicated that PWM mainly exerted its effect on B cells and on monocytes. This effect was T cell dependent and it was mediated by soluble factors of T cell origin. At the peak of the PHA or concanavalin A response, most of the Fc epsilon R-bearing cells were found in the B cell fraction but the T cells isolated from mitogen-stimulated cultures contained significant more Fc epsilon R+ cells than those from the control cultures, suggesting that T cell mitogens had increased the expression of Fc epsilon R on some T cells. This view was supported by the finding of a higher proportion of Fc epsilon R+ cells in PHA-stimulated than in control cultures of highly purified T cells with a maximum response at the end of the culture period. Double-labeling experiments at the peak (day 2) of the peripheral blood mononuclear cell response indicated that the expression of Fc epsilon R was increased on B cells (B1+) and on monocytes (Mo2+). By using the same approach at the peak of the T cell response (day 7), it was found that T cells isolated from PHA-stimulated cultures expressed more Fc epsilon R than those isolated from control cultures.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell Line↗

PubMed1986

Evaluation of the functional maturity of newborn T8+ suppressor cells and the resistance of newborn lymphocytes to suppression.

Concanavalin A (Con A) was used to study the suppressor function of human umbilical cord blood T cells. The suppressor activity and the phenotype of Con A-activated adult and cord T cells were compared using allogeneic adult mononuclear cell (MNC) response to pokeweed mitogen (PWM), phytohemagglutinin (PHA), and alloantigen as indicator culture systems. It was found that Con A-activated adult suppressor T cells were predominantly T8+ cells and suppressed PWM-induced IgG production by adult lymphocytes, as well as PHA- and alloantigen-induced proliferation of allogeneic adult MNC. Con A-activated cord T cells, however, exhibited no significant suppression in any of the three indicator systems. Furthermore, using Con A-activated adult T8+ cells as a source of suppression, the proliferative response of cord lymphocytes to PHA and alloantigen were as susceptible to suppression as those of adult lymphocytes. These results, together with previous findings that PWM-induced cord suppressor T cells were predominantly T4+ cell-mediated suppression, suggest that fetus/newborn lymphocytes achieved their selective inhibition on adult lymphocytes through the T4+ suppressor cell circuit.

Adult↗

PubMed1986

Human cord blood suppressor T lymphocytes. II. Characterization of inducer of suppressor cells.

Previously, we reported an antigen nonspecific inducer of T suppressor cell factor (TisF) produced by cord blood mononuclear cells (MNC) in 48-hr, two-way mixed lymphocyte cultures (MLC). The target of this factor was a radiosensitive, T4+ (T8-) adult suppressor T cell subset. The cellular origin of this TisF was examined in the present study. IgG production by pokeweed mitogen (PWM)-stimulated adult MNC was used as an assay for TisF activity. It was found that TisF-producing cells formed rosettes with sheep erythrocytes (E+) and were independent of adherent cells (AC) in the production of TisF. They were resistant to irradiation (2500 rads) and phenotypic characterization with T cell reactive monoclonal antibodies indicated that they resided in the T8- (T4+) population. Furthermore, both TQ1- and TQ1+ cells were required for the production of TisF activity and such activity could not be reconstituted by supernatants from TQ1- MLC and TQ1+ MLC. These results indicate that the production of TisF is dependent upon interactions between radioresistant E+, T8-, TQ1- and radioresistant E+, T8-, TQ1+ cells.

Antibodies, Monoclonal↗

PubMed1986

Production and characterization of mouse monoclonal antibodies to allergenic epitopes on LolpI (Rye I).

This study describes the production and characterization of mouse monoclonal antibodies (Mab) to LolpI (Rye I), the major allergen of rye-grass pollen. Hybridoma cell lines were screened for the secretion of Mab, the binding of which to labelled LolpI could be inhibited by pooled human sera containing IgE and IgG anti-LolpI antibodies. Twenty monoclonal hybridoma cell lines were expanded, and their product was purified and characterized. Each Mab bound to LolpI but not to unrelated antigens. Cross-inhibition studies allowed the grouping of these 20 Mab into five families differing by their epitope specificity. One representative Mab of each family was tested for its ability to inhibit human IgE/IgG anti-Lolp-I from 15 allergic donors as well as for its inhibition by affinity-purified anti-LolpI antibodies isolated from 14 donors. The results indicated that each family of Mab was specific for an epitope that was identical or close to that recognized by some human anti-LolpI antibodies. It is further shown that human IgE/IgG anti-LolpI are heterogeneous with some patients reacting preferentially with one or two epitopes defined by the Mab LolpI, and others reacting with epitopes different from those identified by the Mab LolpI.

Allergens↗