Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays
Allergic rhinitis (AR) is an IgE-mediated nasal inflammatory disorder, commonly triggered by house dust mite, Dermatophagoides pteronyssinus. Der p 1 is a dominant allergenic protein derived from this species. The highly polymorphic human leukocyte antigen (HLA) genes were linked to AR predisposi...
| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2022
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/116689/ http://psasir.upm.edu.my/id/eprint/116689/1/116689%20-%20edited.pdf |
| _version_ | 1848867737340739584 |
|---|---|
| author | Jana Money, Vivek Prasad |
| author_facet | Jana Money, Vivek Prasad |
| author_sort | Jana Money, Vivek Prasad |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Allergic rhinitis (AR) is an IgE-mediated nasal inflammatory disorder, commonly
triggered by house dust mite, Dermatophagoides pteronyssinus. Der p 1 is a dominant
allergenic protein derived from this species. The highly polymorphic human leukocyte
antigen (HLA) genes were linked to AR predisposition/protection in different ethnicities.
Variation in this gene alters epitope-binding affinities for HLA molecule, causing
polarization of immune responses due to differential cytokines and antibodies secretions.
The relationship between HLA and Der p 1 epitopes in the induction of specific cytokines
and antibodies is unclear. During allergies, B cells perform various effector and
regulatory functions. Dysregulation of B cells with Der p 1 stimulation under HLA
regulation, remains unknown. This study aimed to use in silico approaches to predict
relationship between Der p 1 epitopes and HLA-DR serotypes in cytokines/antibodies
induction and evaluate in vitro changes in B and T cells of Der p 1-stimulated peripheral
blood mononuclear cells (PBMCs) from AR patients and controls with selected HLADRB1
alleles. Full amino acid sequence of Der p 1 (NCBI protein database) was
submitted into TepiTool, NetMHCIIpan 4.0, PREDIVAC and ABCpred, BCPREDS,
BepiPred 2.0 to predict T and B cell epitopes, respectively. AllerTOP 2.0, AllergenFP
1.0 and AlgPred determined allergenicities of predicted peptides. IL4Pred, IL-10Pred
and IFNepitope assessed T cell epitopes’ cytokine-inducing properties, while Bcipep and
IgPred evaluated B cell epitopes’ antibody isotype secretions. Needleman-Wunsch
alignment tool (NCBI BLAST) identified overlapping sequences between T and B cell
epitopes. For in vitro study, Malay adults were recruited and screened for HLA-DRB1
alleles via polymerase chain reaction with sequence-specific primers using buccal DNA
samples. Three subjects per HLA group for patients (DR7 and DR13) and controls
(DR15) were selected for skin prick testing and blood sample collection. Der p 1-
stimulated PBMCs were analyzed for B and T cells via flow cytometry. In silico analysis
identified 32 CD4+ T cell epitopes, mostly IL-4 inducers (n = 11) and associated with
allergenicity (p = 0.049). DR13 preferentially bound IL-4-inducing epitopes (75.00%)
whereas DR7 mostly bound IL-10/IFN-γ-inducing epitopes (70.59%). DR15 bound
almost equally IL-4 (42.86%) and IL-10/IFN-γ-inducing peptides (57.14%). From 13
predicted B cell epitopes, majority induced either IgE only (n = 4) or both IgE and IgA
(n = 4). DR13-associated epitopes, cross-matched between T and B cell peptides for at
least 60% similarities, were mostly IgE-inducing. Day 3 Der p 1-stimulated PBMCs
showed lower percentages of IL-10-secreting T cells in AR patients (DR7+ and DR13+)
compared to controls (DR15+) (27.08% vs 43.97%; p = 0.025). Memory B cell
percentages were higher in DR13+ than DR7+ group (63.00% vs 10.77%; p = 0.022)
whereas regulatory B cells were greater in DR7+ than DR13+ group (88.63% vs 32.53%;
p = 0.022). In conclusion, in silico and in vitro studies supported HLA-DR13 as AR
predisposing due to increased bindings of IL-4 and IgE-inducing epitopes, causing
proliferation of memory B cells. Conversely, HLA-DR7 protected against AR via
selective presentation of IL-10/IFN-γ-inducers, triggering differentiation of regulatory B
cells. The DR15 controls maintained a healthy immune profile, suggesting a neutral
outcome. |
| first_indexed | 2025-11-15T14:41:15Z |
| format | Thesis |
| id | upm-116689 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T14:41:15Z |
| publishDate | 2022 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1166892025-10-13T00:59:08Z http://psasir.upm.edu.my/id/eprint/116689/ Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays Jana Money, Vivek Prasad Allergic rhinitis (AR) is an IgE-mediated nasal inflammatory disorder, commonly triggered by house dust mite, Dermatophagoides pteronyssinus. Der p 1 is a dominant allergenic protein derived from this species. The highly polymorphic human leukocyte antigen (HLA) genes were linked to AR predisposition/protection in different ethnicities. Variation in this gene alters epitope-binding affinities for HLA molecule, causing polarization of immune responses due to differential cytokines and antibodies secretions. The relationship between HLA and Der p 1 epitopes in the induction of specific cytokines and antibodies is unclear. During allergies, B cells perform various effector and regulatory functions. Dysregulation of B cells with Der p 1 stimulation under HLA regulation, remains unknown. This study aimed to use in silico approaches to predict relationship between Der p 1 epitopes and HLA-DR serotypes in cytokines/antibodies induction and evaluate in vitro changes in B and T cells of Der p 1-stimulated peripheral blood mononuclear cells (PBMCs) from AR patients and controls with selected HLADRB1 alleles. Full amino acid sequence of Der p 1 (NCBI protein database) was submitted into TepiTool, NetMHCIIpan 4.0, PREDIVAC and ABCpred, BCPREDS, BepiPred 2.0 to predict T and B cell epitopes, respectively. AllerTOP 2.0, AllergenFP 1.0 and AlgPred determined allergenicities of predicted peptides. IL4Pred, IL-10Pred and IFNepitope assessed T cell epitopes’ cytokine-inducing properties, while Bcipep and IgPred evaluated B cell epitopes’ antibody isotype secretions. Needleman-Wunsch alignment tool (NCBI BLAST) identified overlapping sequences between T and B cell epitopes. For in vitro study, Malay adults were recruited and screened for HLA-DRB1 alleles via polymerase chain reaction with sequence-specific primers using buccal DNA samples. Three subjects per HLA group for patients (DR7 and DR13) and controls (DR15) were selected for skin prick testing and blood sample collection. Der p 1- stimulated PBMCs were analyzed for B and T cells via flow cytometry. In silico analysis identified 32 CD4+ T cell epitopes, mostly IL-4 inducers (n = 11) and associated with allergenicity (p = 0.049). DR13 preferentially bound IL-4-inducing epitopes (75.00%) whereas DR7 mostly bound IL-10/IFN-γ-inducing epitopes (70.59%). DR15 bound almost equally IL-4 (42.86%) and IL-10/IFN-γ-inducing peptides (57.14%). From 13 predicted B cell epitopes, majority induced either IgE only (n = 4) or both IgE and IgA (n = 4). DR13-associated epitopes, cross-matched between T and B cell peptides for at least 60% similarities, were mostly IgE-inducing. Day 3 Der p 1-stimulated PBMCs showed lower percentages of IL-10-secreting T cells in AR patients (DR7+ and DR13+) compared to controls (DR15+) (27.08% vs 43.97%; p = 0.025). Memory B cell percentages were higher in DR13+ than DR7+ group (63.00% vs 10.77%; p = 0.022) whereas regulatory B cells were greater in DR7+ than DR13+ group (88.63% vs 32.53%; p = 0.022). In conclusion, in silico and in vitro studies supported HLA-DR13 as AR predisposing due to increased bindings of IL-4 and IgE-inducing epitopes, causing proliferation of memory B cells. Conversely, HLA-DR7 protected against AR via selective presentation of IL-10/IFN-γ-inducers, triggering differentiation of regulatory B cells. The DR15 controls maintained a healthy immune profile, suggesting a neutral outcome. 2022-03 Thesis NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/116689/1/116689%20-%20edited.pdf Jana Money, Vivek Prasad (2022) Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays. Masters thesis, Universiti Putra Malaysia. http://ethesis.upm.edu.my/id/eprint/18292 Dermatophagoides pteronyssinus |
| spellingShingle | Dermatophagoides pteronyssinus Jana Money, Vivek Prasad Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays |
| title | Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays |
| title_full | Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays |
| title_fullStr | Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays |
| title_full_unstemmed | Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays |
| title_short | Role of HLA-DR in dysregulation of B cell homeostasis in Dermatophagoides pteronyssinus-induced allergic rhinitis among Malays |
| title_sort | role of hla-dr in dysregulation of b cell homeostasis in dermatophagoides pteronyssinus-induced allergic rhinitis among malays |
| topic | Dermatophagoides pteronyssinus |
| url | http://psasir.upm.edu.my/id/eprint/116689/ http://psasir.upm.edu.my/id/eprint/116689/ http://psasir.upm.edu.my/id/eprint/116689/1/116689%20-%20edited.pdf |