D031 LONGITUDINAL ANALYSIS OF SINGLE B CELLS, IGE ANTIBODIES, AND PLASMA IN A PEANUT ALLERGIC SUBJECT

The increased incidence of allergies and asthma has sparked interest in IgE, the central player in the allergic response. Interaction with its high-affinity receptor FcεRI leads to sensitization and allergen presentation, extracellular membrane-proximal domain in membrane IgE can act as an antigen receptor on B cells, and the interaction with low-affinity IgE receptor CD23 additionally influences its homeostatic range. Therapeutic anti-IgE antibodies act by the inhibition of IgE functions by interfering with its receptor binding or by the obliteration of IgE-B cells, causing a reduction of serum IgE levels. Fusion proteins of antibody fragments that can act as bispecific T-cell engagers have proven very potent in eliciting cytotoxic T-lymphocyte-mediated killing. We have tested five anti-IgE Fc antibodies, recognizing different epitopes on the membrane-expressed IgE, for the ability to elicit specific T-cell activation when expressed as single-chain Fv fragments fused with anti-CD3ε single-chain antibody. All candidates could specifically stain the cell line, expressing the membrane-bound IgE-Fc and bind to CD3-positive Jurkat cells, and the specific activation of engineered CD3-overexpressing Jurkat cells and non-stimulated CD8-positive cells was demonstrated for 8D6- and ligelizumab-based bispecific antibodies. Thus, such anti-IgE antibodies have the potential to be developed into agents that reduce the serum IgE concentration by lowering the numbers of IgE-secreting cells.

Download Full-text

Immunological responsiveness of neonatal A/J mice to isotypic determinants of syngeneic IgE.

Journal of Experimental Medicine ◽

10.1084/jem.168.2.713 ◽

1988 ◽

Vol 168 (2) ◽

pp. 713-724 ◽

Cited By ~ 11

Author(s):

S Haba ◽

A Nisonoff

Keyword(s):

T Cells ◽

B Cells ◽

Adoptive Transfer ◽

Time Dependent ◽

Spleen Cells ◽

Ige Antibodies ◽

Neonatal Mice ◽

Adult Mice ◽

Range Of Values ◽

Tolerant State

We have previously shown that adult A/J mice produce high titers of anti-IgE with isotypic or idiotypic specificities in response to challenge with a conjugate of KLH with syngeneic monoclonal IgE. Thus, B cells that can synthesize anti-IgE are present in the mice. Adult mice are unresponsive to unconjugated IgE in CFA, suggesting that tolerance exists at the level of T cells. The present study shows that neonatal mice produce anti-IgE antibodies in response to unconjugated IgE in CFA, but that this capacity is lost after the age of 2-3 wk. The loss of responsiveness corresponds closely with the appearance of detectable IgE in serum, suggesting that the IgE may induce tolerance. The affinities of anti-IgE antibodies produced by neonatal mice fall in the range of values obtained with KLH-IgE in adult mice. Tolerance to unconjugated IgE in CFA can be induced in neonatal mice by administration of IgE in saline. In addition, the tolerant state can be induced by adoptive transfer of spleen cells from adult mice. The time-dependent acquisition of tolerance provides a useful model for studying mechanisms of tolerance and autoimmunity.

Download Full-text

A Longitudinal Analysis Reveals Early Activation and Late Alterations in B Cells During Primary HIV Infection in Mozambican Adults

Frontiers in Immunology ◽

10.3389/fimmu.2020.614319 ◽

2021 ◽

Vol 11 ◽

Author(s):

Montse Jiménez ◽

Lucía Pastor ◽

Victor Urrea ◽

María Luisa Rodríguez de la Concepción ◽

Erica Parker ◽

...

Keyword(s):

B Cells ◽

Hiv Infection ◽

B Cell ◽

Longitudinal Analysis ◽

Chronic Infection ◽

Cell Activation ◽

Humoral Response ◽

Memory Cells ◽

Cell Compartment ◽

Primary Hiv Infection

Primary HIV infection (PHI) and subsequent chronic infection alter B-cell compartment. However, longitudinal analysis defining the dynamics of B-cell alterations are still limited. We longitudinally studied B-cell subsets in individuals followed for 1 year after PHI (n = 40). Treated and untreated chronic HIV infected (n = 56) and HIV-uninfected individuals (n = 58) were recruited as reference groups at the Manhiça District in Mozambique. B cells were analyzed by multicolor flow-cytometry. Anti-HIV humoral response and plasma cytokines were assessed by ELISA or Luminex-based technology. A generalized activation of B cells induced by HIV occurs early after infection and is characterized by increases in Activated and Tissue-like memory cells, decreases in IgM-IgD- (switched) and IgM-only B cells. These alterations remain mostly stable until chronic infection and are reverted in part by ART. In contrast, other parameters followed particular dynamics: PD-1 expression in memory cells decreases progressively during the first year of infection, Transitional B cells expand at month 3–4 after infection, and Marginal zone-like B cells show a late depletion. Plasmablasts expand 2 months after infection linked to plasma viral load and anti-p24 IgG3 responses. Most of well-defined changes induced by HIV in B-cell activation and memory subsets are readily observed after PHI, lasting until ART initiation. However, subsequent changes occur after sustained viral infection. These data indicate that HIV infection impacts B cells in several waves over time, and highlight that early treatment would result in beneficial effects on the B-cell compartment.

Download Full-text

Human IgE producing B cells have a unique transcriptional program and generate high affinity, allergen-specific antibodies

10.1101/327866 ◽

2018 ◽

Cited By ~ 1

Author(s):

Derek Croote ◽

Spyros Darmanis ◽

Kari C. Nadeau ◽

Stephen R. Quake

Keyword(s):

B Cells ◽

B Cell ◽

Single Cell ◽

Recombinant Expression ◽

Single Cells ◽

Food Allergies ◽

Model Organisms ◽

Biallelic Expression ◽

Ige Antibodies ◽

Helminth Infections

AbstractIgE antibodies provide defense against helminth infections, but can also cause life-threatening allergic reactions. Despite their importance to human health, these antibodies and the cells that produce them remain enigmatic due to their scarcity in humans; much of our knowledge of their properties is derived from model organisms. Here we describe the isolation of IgE producing B cells from the blood of individuals with food allergies, followed by a detailed study of their properties by single cell RNA sequencing (scRNA-seq). We discovered that IgE B cells are deficient in membrane immunoglobulin expression and that the IgE plasmablast state is more immature than that of other antibody producing cells. Through recombinant expression of monoclonal antibodies derived from single cells, we identified IgE antibodies which had unexpected cross-reactive specificity for major peanut allergens Ara h 2 and Ara h 3; not only are these among the highest affinity native human antibodies discovered to date, they represent a surprising example of convergent evolution in unrelated individuals who independently evolved nearly identical antibodies. Finally, we discovered that splicing within B cells of all isotypes reveals polarized germline transcription of the IgE, but not IgG4, isotype as well as several examples of biallelic expression of germline transcripts. Our results offer insights into IgE B cell transcriptomics, clonality and regulation, provide a striking example of adaptive immune convergence, and offer an approach for accelerating mechanistic disease understanding by characterizing a rare B cell population underlying IgE-mediated disease at single cell resolution.

Download Full-text

Anti-IgE therapy for IgE-mediated allergic diseases: from neutralizing IgE antibodies to eliminating IgE+ B cells

Clinical and Translational Allergy ◽

10.1186/s13601-018-0213-z ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 14

Author(s):

Jiayun Hu ◽

Jiajie Chen ◽

Lanlan Ye ◽

Zelang Cai ◽

Jinlu Sun ◽

...

Keyword(s):

B Cells ◽

Allergic Diseases ◽

Ige Antibodies ◽

Ige Mediated

Download Full-text

Longitudinal analysis of FcRL5 expression and clonal relationships among classical and atypical memory B cells following malaria

Malaria Journal ◽

10.1186/s12936-021-03970-1 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

S. Jake Gonzales ◽

Sebastiaan Bol ◽

Ashley E. Braddom ◽

Richard Sullivan ◽

Raphael A. Reyes ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Longitudinal Analysis ◽

Cell Receptor ◽

Memory B Cells ◽

B Cell Receptor ◽

Developmental Pathways ◽

Malaria Episode ◽

Transmission Region ◽

B Cell Subsets

Abstract Background Chronic and frequently recurring infectious diseases, such as malaria, are associated with expanded populations of atypical memory B cells (MBCs). These cells are different from classical MBCs by the lack of surface markers CD21 and CD27 and increased expression of inhibitory receptors, such as FcRL5. While the phenotype and conditions leading to neogenesis of atypical MBCs in malaria-experienced individuals have been studied extensively, the origin of these cells remains equivocal. Functional similarities between FcRL5+ atypical MBCs and FcRL5+ classical MBCs have been reported, suggesting that these cells may be developmentally related. Methods Here, a longitudinal analysis of FcRL5 expression in various B cell subsets was performed in two children from a high transmission region in Uganda over a 6-month period in which both children experienced a malaria episode. Using B-cell receptor (BCR)-sequencing to track clonally related cells, the connections between IgM+ and IgG+ atypical MBCs and other B cell subsets were studied. Results The highest expression of FcRL5 was found among IgG+ atypical MBCs, but FcRL5+ cells were present in all MBC subsets. Following malaria, FcRL5 expression increased in all IgM+ MBC subsets analysed here: classical, activated, and atypical MBCs, while results for IgG+ MBC subsets were inconclusive. IgM+ atypical MBCs showed few connections with other B cell subsets, higher turnover than IgG+ atypical MBCs, and were predominantly derived from naïve B cells and FcRL5− IgM+ classical MBCs. In contrast, IgG+ atypical MBCs were clonally expanded and connected with classical MBCs. IgG+ atypical MBCs present after a malaria episode mainly originated from FcRL5+ IgG+ classical MBCs. Conclusions Collectively, these results suggest fundamental differences between unswitched and class-switched B cell populations and provide clues about the primary developmental pathways of atypical MBCs in malaria-experienced individuals.

Download Full-text

Plasmalemma localisation of inorganic phosphate in B cells pancreas

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100083515 ◽

1978 ◽

Vol 36 (2) ◽

pp. 528-529

Author(s):

F. B. P. Wooding ◽

K. Pedley ◽

N. Freinkel ◽

R. M. C. Dawson

Keyword(s):

Electron Microscope ◽

B Cells ◽

B Cell ◽

Pancreatic Islets ◽

High Glucose ◽

Lead Acetate ◽

Inorganic Phosphate ◽

Slight Modification ◽

Uranyl Acetate ◽

Lead Phosphate

Freinkel et al (1974) demonstrated that isolated perifused rat pancreatic islets reproduceably release up to 50% of their total inorganic phosphate when the concentration of glucose in the perifusion medium is raised.Using a slight modification of the Libanati and Tandler (1969) method for localising inorganic phosphate by fixation-precipitation with glutaraldehyde-lead acetate we can demonstrate there is a significant deposition of lead phosphate (identified by energy dispersive electron microscope microanalysis) at or on the plasmalemma of the B cell of the islets (Fig 1, 3). Islets after incubation in high glucose show very little precipitate at this or any other site (Fig 2). At higher magnification the precipitate seems to be intracellular (Fig 4) but since any use of osmium or uranyl acetate to increase membrane contrast removes the precipitate of lead phosphate it has not been possible to verify this as yet.

Download Full-text

Control of Secretory Production in the Isopod Hepatopancreas

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094085 ◽

1972 ◽

Vol 30 ◽

pp. 166-167

Author(s):

John W. Roberts ◽

E. R. Witkus

Keyword(s):

B Cells ◽

Morphological Data ◽

Secretory Product ◽

Secretory Function ◽

Secretory Material ◽

Blind Ending ◽

Secretory Production ◽

Regenerative Cells ◽

Immature Cells ◽

And Storage

The isopod hepatopancreas, as exemplified by Oniscus ascellus. is comprised of four blind-ending diverticula. The regenerative cells at the tip of each diverticula differentiate into either club-shaped B-cells, which serve a secretory function, or into conoid S-cells, which serve in the absorption and storage of nutrients.The glandular B-cells begin producing secretory material with the development of rough endoplasmic reticulum during their process of maturation from the undifferentiated regenerative cells. Cytochemical and morphological data indicate that the hepatopancreas sequentially produces two types of secretory material within the large club-shaped cells. The production of the carbohydrate-like secretory product in immature cells seems to be phased out as the production of the osmiophilic secretion was phased in as the cell matured.

Download Full-text

Lymphoproliferative disorders (LPD) involving lungs: T and B cell subsets within pulmonary infiltrates and in peripheral blood

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010011338x ◽

1984 ◽

Vol 42 ◽

pp. 700-701

Author(s):

Irene Stachura ◽

Milton H. Dalbow ◽

Michael J. Niemiec ◽

Matias Pardo ◽

Gurmukh Singh ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Lymphoproliferative Disorders ◽

Mixed Population ◽

Lymphoid Cells ◽

Lymphomatoid Granulomatosis ◽

Cell Type ◽

Cell Surface Antigens ◽

Pulmonary Infiltrates ◽

B Cell Subsets

Lymphoid cells were analyzed within pulmonary infiltrates of six patients with lymphoproliferative disorders involving lungs by immunofluorescence and immunoperoxidase techniques utilizing monoclonal antibodies to cell surface antigens T11 (total T), T4 (inducer/helper T), T8 (cytotoxic/suppressor T) and B1 (B cells) and the antisera against heavy (G,A,M) and light (kappa, lambda) immunoglobulin chains. Three patients had pseudolymphoma, two patients had lymphoma and one patient had lymphomatoid granulomatosis.A mixed population of cells was present in tissue infiltrates from the three patients with pseudolymphoma, IgM-kappa producing cells constituted the main B cell type in one patient. In two patients with lymphoma pattern the infiltrates were composed exclusively of T4+ cells and IgG-lambda B cells predominated slightly in the patient with lymphomatoid granulomatosis.

Download Full-text

Ca++-ATPase activity in the hepatopancreas cells of Oniscus asellus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100124501 ◽

1992 ◽

Vol 50 (1) ◽

pp. 820-821

Author(s):

G.M. Vernon ◽

A. Surace ◽

R. Witkus

Keyword(s):

B Cells ◽

Epithelial Cell ◽

Atpase Activity ◽

Calcium Transport ◽

Carcinus Maenas ◽

Cell Types ◽

Molt Cycle ◽

Copper And Zinc

The hepatopancreas consists of a pair of bilobed tubules comprised of two epithelial cell types. S cells are absorptive and accumulate metals such as copper and zinc. Ca++ concentrations vary between the S and B cells and during the molt cycle. Roer and Dillaman implicated Ca++-ATPase in calcium transport during molting in Carcinus maenas. This study was undertaken to compare the localization of Ca++-ATPase activity in the S and B cells during intermolt.

Download Full-text