Characterization of B cell response alterations resulting from ablation of IκBNS expression
Author: Khoenkhoen, Sharesta
Date: 2020-08-28
Location: Ragnar Granit, room C0325, Biomedicum, Solna
Time: 9.30
Department: Inst för mikrobiologi, tumör- och cellbiologi / Dept of Microbiology, Tumor and Cell Biology
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Thesis (2.646Mb)
Abstract
The immune system is a sophisticated organization of cells and tissues that cooperate in safeguarding the integrity of our body by eliminating internal and external threats. Vaccines are the most impactful medical intervention and have contributed tremendously to lowering mortality rates of infectious diseases. The protective immunity induced by vaccination is mediated by eliciting neutralizing antibodies that are sustained for years or even for a lifetime. Antibodies are produced by plasma cells, which are generated from activated and terminally differentiated B cells. Many processes regulating B cell development and function are under the regulation of the NF-κB signaling pathway. Defects in NF-κB signaling have been shown, using mouse models, to be detrimental to the development of distinct B cell subsets as well as their ability to participate in humoral responses. In humans, mutations of components in the NF-κB pathway are increasingly associated with common variable immunodeficiencies. In this thesis, the role of IκBNS, a nuclear regulator of the NF-κB pathway, was investigated to gain a more comprehensive understanding of B cell development and function.
In paper I, we addressed the absence of B-1a cells in IκBNS-deficient bumble mice. We identified a precursor population in wildtype mice of IgM+CD93+CD19+CD5+ transitional B1a cells (TrB-1a), which exhibited other indicators of B-1 lineage commitment, such as natural IgM secretion and increased Igλ expression. We did not detect TrB-1a cells in bumble mice whereas the neonatal B-1 progenitor cell (B-1p) population was intact. These results indicate that IκBNS is required for the transition from B-1p to the TrB-1a stage. In paper II, we investigated potential reasons for the impaired T cell-independent (TI) responses in bumble mice. We found impaired expression of the surface receptor TACI, which is essential in responses to TI antigens, and reduced responsiveness to its ligands in bumble mice. In addition, bumble B cells did not fully silence Pax5 expression and exhibited rapid upregulation of Blimp1 during early division cycles. Thus, these results suggest that IκBNS is involved in TACI expression and function as well as in transcriptional regulation of PC differentiation. In paper III, we used nfkbidB- mice in which IκBNS was selectively depleted from B cells to distinguish between B cell intrinsic and extrinsic defects in T cell-independent (TD) responses. NfkbidB- mice exhibited normal GC formation, but antigen-specific antibodies of the IgG2c and IgG3 isotype were reduced. Hence, IκBNS appears to be important for class switching during TD responses. In paper IV, we developed a protocol for evaluation of plasma cell differentiation from human primary B cells. Cells that exhibited a phenotype resembling plasma cells were identified as CD38+IRF4hiPax5lo and CD38+IRF4intPax5lo. Both populations were generated when starting from fresh or cryopreserved samples, or isolated memory and naïve B cells. Application of the methods developed in this paper to patient samples could provide more insight into mechanisms underlying immune disorders.
Collectively, the results of this thesis contribute to our understanding of processes that are fundamental to establishing adequate humoral responses and are of direct relevance to immunodeficiency disorders.
In paper I, we addressed the absence of B-1a cells in IκBNS-deficient bumble mice. We identified a precursor population in wildtype mice of IgM+CD93+CD19+CD5+ transitional B1a cells (TrB-1a), which exhibited other indicators of B-1 lineage commitment, such as natural IgM secretion and increased Igλ expression. We did not detect TrB-1a cells in bumble mice whereas the neonatal B-1 progenitor cell (B-1p) population was intact. These results indicate that IκBNS is required for the transition from B-1p to the TrB-1a stage. In paper II, we investigated potential reasons for the impaired T cell-independent (TI) responses in bumble mice. We found impaired expression of the surface receptor TACI, which is essential in responses to TI antigens, and reduced responsiveness to its ligands in bumble mice. In addition, bumble B cells did not fully silence Pax5 expression and exhibited rapid upregulation of Blimp1 during early division cycles. Thus, these results suggest that IκBNS is involved in TACI expression and function as well as in transcriptional regulation of PC differentiation. In paper III, we used nfkbidB- mice in which IκBNS was selectively depleted from B cells to distinguish between B cell intrinsic and extrinsic defects in T cell-independent (TD) responses. NfkbidB- mice exhibited normal GC formation, but antigen-specific antibodies of the IgG2c and IgG3 isotype were reduced. Hence, IκBNS appears to be important for class switching during TD responses. In paper IV, we developed a protocol for evaluation of plasma cell differentiation from human primary B cells. Cells that exhibited a phenotype resembling plasma cells were identified as CD38+IRF4hiPax5lo and CD38+IRF4intPax5lo. Both populations were generated when starting from fresh or cryopreserved samples, or isolated memory and naïve B cells. Application of the methods developed in this paper to patient samples could provide more insight into mechanisms underlying immune disorders.
Collectively, the results of this thesis contribute to our understanding of processes that are fundamental to establishing adequate humoral responses and are of direct relevance to immunodeficiency disorders.
List of papers:
I. Gabriel K. Pedersen, Monika Ádori, Sharesta Khoenkhoen, Pia Dosenovic, Bruce Beutler, Gunilla B. Karlsson Hedestam. B-1a transitional cells are phenotypically distinct and are lacking in mice deficient in IκBNS. Proc Natl Acad Sci. 2014. 111(39): E41119-26.
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II. Sharesta Khoenkhoen, Elina Erikson, Monika Ádori, Jean M. Scholz, Michael P. Cancro, Gabriel K. Pedersen, Gunilla B. Karlsson Hedestam. TACI expression and plasma cell differentiation are impaired in the absence of functional IκBNS. Immunol Cell Biol. 2019 97(5): 485-497.
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III. Sharesta Khoenkhoen, Monika Ádori, Juan Darío Solis Sayago, Juliette Soulier, Jamie Russel, Bruce Beutler, Gabriel K. Pedersen, Gunilla B. Karlsson Hedestam. IκBNS expression in B cells is dispensable for germinal center formation but required for optimal antibody responses to T-dependent antigens. [Manuscript]
IV. Sharesta Khoenkhoen, Monika Ádori, Gabriel K. Pedersen, Gunilla B. Karlsson Hedestam. Flow cytometry-based protocols for the analysis of human plasma cell differentiation. [Manuscript]
I. Gabriel K. Pedersen, Monika Ádori, Sharesta Khoenkhoen, Pia Dosenovic, Bruce Beutler, Gunilla B. Karlsson Hedestam. B-1a transitional cells are phenotypically distinct and are lacking in mice deficient in IκBNS. Proc Natl Acad Sci. 2014. 111(39): E41119-26.
Fulltext (DOI)
Pubmed
View record in Web of Science®
II. Sharesta Khoenkhoen, Elina Erikson, Monika Ádori, Jean M. Scholz, Michael P. Cancro, Gabriel K. Pedersen, Gunilla B. Karlsson Hedestam. TACI expression and plasma cell differentiation are impaired in the absence of functional IκBNS. Immunol Cell Biol. 2019 97(5): 485-497.
Fulltext (DOI)
Pubmed
View record in Web of Science®
III. Sharesta Khoenkhoen, Monika Ádori, Juan Darío Solis Sayago, Juliette Soulier, Jamie Russel, Bruce Beutler, Gabriel K. Pedersen, Gunilla B. Karlsson Hedestam. IκBNS expression in B cells is dispensable for germinal center formation but required for optimal antibody responses to T-dependent antigens. [Manuscript]
IV. Sharesta Khoenkhoen, Monika Ádori, Gabriel K. Pedersen, Gunilla B. Karlsson Hedestam. Flow cytometry-based protocols for the analysis of human plasma cell differentiation. [Manuscript]
Institution: Karolinska Institutet
Supervisor: Karlsson Hedestam, Gunilla
Co-supervisor: Pedersen, Gabriel; Beutler, Bruce
Issue date: 2020-08-07
Rights:
Publication year: 2020
ISBN: 978-91-7831-915-2
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