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> Anatomy > Tissue > Germinal Center

Germinal Center

The Germinal Center is a specialized microenvironment within secondary lymphoid organs, where B lymphocytes undergo affinity maturation and differentiation.
This dynamic process plays a crucial role in the generation of high-affinity antibodies and long-lived memory B cells, which are essential for effective humoral immune responses.
The Germinal Center is characterized by the proliferation and selection of B cells with increasingly refined antigen receptors, facilitated by interactions with follicular helper T cells and follicular dendritic cells.
Dysregulation of Germinal Center processes has been implicated in various immunological disorders and lymphoproliferative diseases.
Undertsanding the complex biology of the Germinal Center is crucial for advancing our knowledge of humoral immunity and developing targeted therapeutic interventions.

Most cited protocols related to «Germinal Center»

1
Survival Model Development for Lymphoma Subtypes
All aspects of identification of the gene-expression signatures and development of the survival model were based solely on the data from the CHOP training group and are outlined in detail in the Supplementary Appendix. No previous survival analysis or subgroup analysis was performed with the validation groups (i.e., the MMMLNP CHOP and the R-CHOP cohorts). A Cox model was used to identify genes associated with survival and to build multivariate survival models. The models and their associated scaling coefficients were fixed, based on the CHOP training group, and then evaluated in the validation groups. All reported P values are two-sided, except those in the validation groups, which are one-sided P values in the direction of the observed effect on the training group. P values reported for survival associations were based on single-hypothesis testing, except those for testing of multivariate models involving the germinal-center B-cell, stromal-1, proliferation, and MHC class II signatures in the R-CHOP cohort, which were not adjusted for multiple testing.
To discover new signatures associated with survival, we selected individual genes with expression patterns that contributed significantly (P<0.01) to the survival association in the CHOP training group, in a model containing that gene and the germinal-center B-cell and stromal-1 signatures. We organized these genes by hierarchical clustering according to their expression levels in the CHOP training group, and we identified five clusters of coordinately expressed genes (r>0.6). For each of these five candidate signatures, we averaged the expression levels of the component genes and tested whether the average for the signature added to the predictive significance of the bivariate survival model for the CHOP training group. One signature was clearly superior to the others with respect to its predictive contribution to the survival model and was therefore chosen for further analysis. This signature also added to the predictive significance of the bivariate model for the R-CHOP cohort (P = 0.001) and for the MMMLNP CHOP cohort (P = 0.011) (Fig. 8B and 8C in the Supplementary Appendix). In these survival models, this new signature was associated with reduced survival, whereas the stromal-1 signature was associated with increased survival, even though these two signatures were correlated with one another (r>0.8). Therefore, to refine this new signature, we identified genes that were more closely correlated with it than with the stromal-1 signature (P<0.02) in the CHOP training group, and we organized these genes into three signatures by hierarchical clustering, as described above. The signature that most improved the survival model (stromal-2) was chosen for subsequent analyses.
Lenz G., Wright G., Dave S.S., Xiao W., Powell J., Zhao H., Xu W., Tan B., Goldschmidt N., Iqbal J., Vose J., Bast M., Fu K., Weisenburger D.D., Greiner T.C., Armitage J.O., Kyle A., May L., Gascoyne R.D., Connors J.M., Troen G., Holte H., Kvaloy S., Dierickx D., Verhoef G., Delabie J., Smeland E.B., Jares P., Martinez A., Lopez-Guillermo A., Montserrat E., Campo E., Braziel R.M., Miller T.P., Rimsza L.M., Cook J.R., Pohlman B., Sweetenham J., Tubbs R.R., Fisher R.I., Hartmann E., Rosenwald A., Ott G., Muller-Hermelink H.K., Wrench D., Lister T.A., Jaffe E.S., Wilson W.H., Chan W.C, & Staudt L.M. (2008). Stromal Gene Signatures in Large-B-Cell Lymphomas. The New England journal of medicine, 359(22), 2313-2323.
Publication 2008
B-Lymphocytes DDIT3 protein, human Gene Clusters Gene Expression Genes Genes, MHC Class II Germinal Center
2
Generation and Characterization of IL-15 Deficient Mice
C57BL/6 mice genetically deficient in IL-15 were generated by homologous recombination in embryonic stem (ES) cells. The structure of the murine IL-15 genomic locus has been described 21. A gene targeting vector was constructed in which a 7.5-kb SpeI-EcoRV fragment containing IL-15 exons 3–5, encoding amino acids 1–65 of the primary translation product, was replaced with a PGK-neo cassette. A thymidine kinase cassette (MC-TK) was inserted into the 5′ end of the vector. C57BL/6-derived ES cells were electroporated with the IL-15 targeting vector and selected in G418 and ganciclovir as described 22. ES cell clones carrying a targeted IL-15 gene were identified by a combination of PCR and genomic Southern blot analyses and were injected into BALB/c blastocysts. Resulting male chimeras were bred to C57BL/6 females. The offspring were analyzed for germline transmission of the mutant IL-15 allele by PCR and genomic Southern blot analyses. Mice heterozygous for the IL-15 mutation (IL-15+/−) were intercrossed to generate IL-15–deficient (IL-15−/−) mice.
The IL-15−/− mice used throughout these studies were bred at Immunex and maintained on a C57BL/6 genetic background. All mice were housed under specific pathogen-free conditions and were used between 9 and 20 wk of age. Age- and sex-matched littermates (IL-15+/+ or IL-15+/−) or C57BL/6 mice (Taconic Farms, Inc.) were used as controls as indicated. Initial studies revealed no apparent difference between IL-15+/+ and IL-15+/− mice in cellularity of their secondary lymphoid tissues, phenotype of spleen or LN cells, or splenic NK cell responses.
Kennedy M.K., Glaccum M., Brown S.N., Butz E.A., Viney J.L., Embers M., Matsuki N., Charrier K., Sedger L., Willis C.R., Brasel K., Morrissey P.J., Stocking K., Schuh J.C., Joyce S, & Peschon J.J. (2000). Reversible Defects in Natural Killer and Memory Cd8 T Cell Lineages in Interleukin 15–Deficient Mice. The Journal of Experimental Medicine, 191(5), 771-780.
Publication 2000
Alleles Amino Acids antibiotic G 418 Blastocyst Cells Chimera Clone Cells Cloning Vectors Embryonic Stem Cells Exons Females Ganciclovir Genes Genetic Background Genome Germinal Center Germ Line Heterozygote Homologous Recombination Interleukin-15 Males Mice, Inbred C57BL Mus Mutation Natural Killer Cells Phenotype Southern Blotting Specific Pathogen Free Spleen Thymidine Kinase Transmission, Communicable Disease
3
Adoptive Transfer and Imaging of Germinal Center B Cells

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Publication 2010
Adoptive Transfer alum, potassium B-Lymphocytes Cells Cone-Rod Dystrophy 2 Escherichia coli Flow Cytometry FTY-720 Germinal Center Glutathione Institutional Animal Care and Use Committees Light Lycopersicon esculentum Lymphocyte Macrophage Medulla Oblongata Motility, Cell Mus NP 10 Pertussis Toxin Phycoerythrin PRDM1 protein, human Spleen SPN protein, human T-Lymphocyte Tissue Donors Transgenes Vaccination
4
Chimeric Antibody Production and Injection

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Publication 2010
Antibodies Chimera Germinal Center HEK293 Cells Mus
5
B Cell Switching Protocols for CSR
Spontaneous switching human IgM+ IgD+ 4B6 B cells, inducible switching human IgM+ IgD+ 2E2 B cells and inducible switching mouse IgM+ CH12F3 B cells were as we described12 (link). Human IgD− CD38+ CD19+ germinal center B cells and IgD+ CD38− CD19+ naïve B cells from tonsil and mouse B220+ primary B cells were prepared as we described12 (link), 68 (link). To analyze spontaneous CSR in human 4B6 B cells, IgG− IgA− cells were cultured at 5 × 104 cell ml−1 in RPMI-1640 medium (Invitrogen Corp.) supplemented with 10% fetal calf serum (Hyclone) and 1% penicillin-streptomycin (FCS-RPMI). To induce CSR to IgG1, human 2E2 B cells were cultured at 5 × 104 cell ml−1 and stimulated with agonistic mAb to hCD40 (IgG1 mAb G28-5, ATCC) plus recombinant hIL-4 (Genzyme Co.) for 4 d. Mouse B cells were labeled with CFSE (Invitrogen) and induced to undergo CSR by mCD154 or LPS (E. coli serotype 055:B5; Sigma-Aldrich Co.) plus appropriate cytokines, as we reported12 (link). All protocols involving human tissues and/or cells were in accordance to the rules and regulations of the IRB of UC Irvine.
Xu Z., Fulop Z., Wu G., Pone E.J., Zhang J., Mai T., Thomas L.M., Al-Qahtani A., White C.A., Park S.R., Steinacker P., Li Z., Yates J I.I.I., Herron B., Otto M., Zan H., Fu H, & Casali P. (2010). 14-3-3 adaptor proteins recruit AID to 5′-AGCT-3′-rich switch regions for class switch recombination. Nature structural & molecular biology, 17(9), 1124-1135.
Publication 2010
5-(6)-carboxyfluorescein diacetate succinimidyl ester agonists B-Lymphocytes Cells Cultured Cells Cytokine Escherichia coli Germinal Center Homo sapiens IgG1 Mus Palatine Tonsil Penicillins Streptomycin Tissues

Most recents protocols related to «Germinal Center»

1
Isolation and Staining of Lymphoid Cells
Secondary lymphoid organs (spleen and lymph nodes-axillary, brachial, inguinal, cervical, mesenteric, para-aortic) were isolated from mice and were pressed through a 70-μm nylon screen to generate a single-cell suspension. Single-cell suspensions were stained for 1 h at room temperature with allophycocyanin- or phycoerythrin-conjugated tetramers. Samples underwent enrichment for tetramer-binding cells as previously described (57 (link)). Cre61–71:I-Ab APC and PE tetramers were a generous gift from James Moon.
Martinez R.J., Breed E.R., Worota Y., Ashby K.M., Vobořil M., Mathes T., Salgado O.C., O’Connor C.H., Kotenko S.V, & Hogquist K.A. (2023). Type III interferon drives thymic B cell activation and regulatory T cell generation. Proceedings of the National Academy of Sciences of the United States of America, 120(9), e2220120120.
Publication 2023
allophycocyanin Aorta Arm, Upper Axilla Cells Germinal Center Groin Mesentery Mus Neck Nodes, Lymph Nylons Phycoerythrin Spleen Tetrameres
2
DLBCL Molecular Subtyping using NanoString
To determine the molecular subtype according to the cell-of-origin (COO) classification, extracted RNA was analyzed by the NanoString nCounter FLEX gene expression profiling (GEP) system (NanoString, Seattle, Washington, USA) as previously described [18 ]. The NanoString Lymphoma Subtyping Test (LST) algorithm allows the assignment of each analyzed sample to the germinal center B-cell like (GCB) subtype, the activated B-cell like (ABC) subtype, or to be unclassified (Supplementary Table 1) [19 , 20 (link)]. In brief, the LST CodeSet consists of capture and reporter probes for 20 genes: 7 genes overexpressed in GCB DLBCL (ASB13, ITPKB, MAML3, MME, MYBL1, S1PR2, SERPINA9), 8 genes overexpressed in ABC DLBCL (CCDC50, CREB3L2, CYB5R2, IRF4, LIMD1, PIM2, RAB7L1, TNFRSF13B), and 5 housekeeping genes (ISY1, R3HDM1, TRIM56, UBXN4, WDR55). Quality and quantity of used RNA was determined by spectrophotometer (Nanodrop, Thermo Scientific). The LST CodeSet was hybridized to 500 ng of total RNA for 18 h at 65 °C. Hybridized RNA samples were loaded into the nCounter Prep Station and expression of target mRNA was finally assessed by the nCounter Digital Analyzer.
Frontzek F., Staiger A.M., Wullenkord R., Grau M., Zapukhlyak M., Kurz K.S., Horn H., Erdmann T., Fend F., Richter J., Klapper W., Lenz P., Hailfinger S., Tasidou A., Trautmann M., Hartmann W., Rosenwald A., Quintanilla-Martinez L., Ott G., Anagnostopoulos I, & Lenz G. (2023). Molecular profiling of EBV associated diffuse large B-cell lymphoma. Leukemia, 37(3), 670-679.
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Publication 2023
B-Lymphocytes Cells Genes Genes, Housekeeping Genitalia Germinal Center interferon regulatory factor 4, human LIMD1 protein, human Lymphoma MAML3 protein, human MYBL1 protein, human PIM2 protein, human RNA, Messenger TNFRSF13B protein, human
3
Immune Cell Abundance Estimation Tool
The Immune Cell Abundance Identifier for the mouse (ImmuCellAI-mouse) is an online tool (http://bioinfo.life.hust.edu.cn/ImmuCellAI-mouse/#!/) to accurately estimate the abundance of 36 immune cell (sub)types using mouse transcriptome datasets.22 ImmuCellAI-mouse adopted a hierarchical strategy to divide 36 cell types into 3 layers. Layer 1 was composed of 7 major immune cell types: monocyte, macrophage, granulocyte, natural killer (NK) cell, dendritic cell (DC), B cell, and T cell. Second layer cells were mainly subtypes of the first layer immune cells, including macrophage subtypes (M1 and M2 macrophage), granulocyte subtypes (basophil, eosinophil, mast cell, and neutrophil), DC subtypes (conventional DC 1 [cDC1], conventional DC 2 [cDC2], monocyte-derived [MoDC], and plasmacytoid [pDC] cell), B cell subtypes (B1, follicular B, germinal center B, marginal zone B, memory B, and plasma B cell), and T cell subtypes (CD4+ T, CD8+ T, NKT, and γδ T cells). Finally, immune cells in layer 3 were subtypes of CD4+ T and CD8+ T cells, including naïve CD4+ T, CD4+ T memory (Tm), regulatory T cells (Treg), T helper, naïve CD8+ T, cytotoxic CD8+ T (Tc), CD8+ T central memory (Tcm), CD8+ T effector memory (Tem), and exhausted CD8+ T cells (Tex).
Xu X., Wei Y., Pang J., Wei Z., Wang L., Chen Q., Wang Z., Zhang Y., Chen K., Peng Y., Zhang Z., Liu J., Zhang Y., Jin Z.B, & Liang Q. (2023). Time-Course Transcriptomic Analysis Reveals the Crucial Roles of PANoptosis in Fungal Keratitis. Investigative Ophthalmology & Visual Science, 64(3), 6.
Publication 2023
Antigen-Presenting Cells B-Lymphocytes Basophils CD8-Positive T-Lymphocytes CDK1 protein, human Cells Eosinophil Germinal Center Granulocyte Macrophage Mast Cell Memory Monocytes Mus Natural Killer Cells Neutrophil Plasma Cells Regulatory T-Lymphocytes T-Lymphocyte Transcriptome
4
Retrospective Study of Kidney Lymphoma
This retrospective study was carried out at King Abdulaziz University Hospital and King Faisal Specialist Hospital and Research Centre, Jeddah, Saudi Arabia, which are 2 main referral hospitals in the western region of Saudi Arabia. Inclusion criteria included all cases diagnosed as kidney lymphoma between January 2002 and April 2022. The project started in January 2022 and was completed in August 2022. Exclusion criteria included cases with no available pathology slides and paraffin blocks. The definition proposed by Krol et al11 (link)
 was used to define primary extranodal NHL, which includes all patients who present with NHL originating at the kidney, even in the presence of disseminated disease, if the kidney component is clinically dominant.
The collected clinical data included age at presentation, gender, clinical features, and treatment. The immunohistochemistry slides were reviewed, and more immunohistochemistry markers were added in selected cases. The minimum immunohistochemistry panel included CD45, CD20, CD3, BCL-2, BCL-6, CD10, MUM-1, and KI-67. Additional panels were added in selected cases and included CD21, CD23, CD30, CD79a, CD38, CD138, Fascin, PAX-5, CD15, Epstein-Barr virus (EBV), and pankeratin. The additional panel was carried out to confirm subtype, to rule out Hodgkin lymphoma and viral inclusions. Histopathological classification of lymphomas was according to the 2017 World Health Organization (WHO) criteria.12
Diffuse large B-cell lymphomas were subclassified according to Hans algorithm to germinal center B-cells (GCB) and non-GCB which depends on the pattern of immunohistochemical expression for CD10, MUM-1, and BCL-6.13 (link)
 The study was approved by the Research Committee of the Biomedical Ethics Unit at King Abdulaziz University, Jeddah, Saudi Arabia (reference no.: 34-22). The study was carried out according to the principles of Helsinki Declaration. A review of morphology and additional immunohistochemistry markers allowed the re-classification of older cases into currently accepted diagnostic categories.
, & Al-Maghrabi J.A. (2023). Primary lymphoma of the kidney: Pathology experience from 2 tertiary hospitals in Western Saudi Arabia. Saudi Medical Journal, 44(1), 29-37.
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Publication 2023
B-Lymphocytes BCL2 protein, human BCL6 protein, human CD79A protein, human Diagnosis Diffuse Large B-Cell Lymphoma Epstein-Barr Virus Ethics Committees, Research fascin Germinal Center Hodgkin Disease Hospital Referral Immunohistochemistry Inclusion Bodies Kidney Lymphoma Paraffin Patients SDC1 protein, human
5
Flow Cytometric Analysis of Tfh, GCB, and Inflammatory Cells
Draining lymph nodes were collected at day 28, day 60, and at 4, 16, 24, and 72 hours post-first immunization to analyze Tfh cells and germinal center B (GCB) cells, and inflammatory cells, respectively. Draining lymph node cells were then washed and incubated with anti-CD16/32 (clone 2.4G2; BD Pharmingen, San Diego, CA, USA), before being stained for viability with the Zombie NIR™ Fixable Viability Kit (BioLegend). The cells were then immunostained with the antibodies outlined in Supplementary Table 1, before being examined using the BD LSR II flow cytometer (BD Bioscience, San Jose, CA, USA) or the NovoCyte 3000 flow cytometer (Agilent Technologies, Inc., Santa Clara, CA, USA). Data were analyzed by FlowJo software (Tree Star, Ashland, OR, USA) or NovoExpress 1.4.0 software (Agilent Technologies, Inc.).
Yoshioka Y., Kobiyama K., Hayashi T., Onishi M., Yanagida Y., Nakagawa T., Hashimoto M., Nishinaka A., Hirose J., Asaoka Y., Tajiri M., Hayata A., Ishida S., Omoto S., Nagira M, & Ishii K.J. (2023). A-910823, a squalene-based emulsion adjuvant, induces T follicular helper cells and humoral immune responses via α-tocopherol component. Frontiers in Immunology, 14, 1116238.
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Publication 2023
Antibodies B-Lymphocytes Cells Clone Cells Germinal Center Inflammation Lymphocyte Nodes, Lymph T Follicular Helper Cells Trees Vaccination

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More about "Germinal Center"

The Germinal Center (GC) is a specialized microenvironment within secondary lymphoid organs, such as lymph nodes and the spleen, where B lymphocytes undergo a dynamic process of affinity maturation and differentiation.
This crucial process plays a vital role in the generation of high-affinity antibodies and long-lived memory B cells, which are essential for effective humoral immune responses.
The GC is characterized by the proliferation and selection of B cells with increasingly refined antigen receptors, facilitated by interactions with follicular helper T (Tfh) cells and follicular dendritic cells (FDCs).
This complex interplay within the GC is essential for the development of robust and long-lasting antibody-mediated immunity.
Techniques such as flow cytometry, using instruments like the FACSAria II, BD Accuri C6, and FACSCanto II, are commonly employed to study the cellular dynamics and phenotypes of B cells and Tfh cells within the GC.
Markers like CD4-FITC and B220-FITC are often used to identify and isolate these key cell populations.
Dysregulation of GC processes has been implicated in various immunological disorders, such as autoimmune diseases and lymphoproliferative diseases.
Understanding the intricate biology of the GC is crucial for advancing our knowledge of humoral immunity and developing targeted therapeutic interventions.
PubCompare.ai, an AI-powered research optimization platform, can help researchers enhance their GC studies by locating the best protocols from literature, preprints, and patents.
With real-time comparisons and the integration of tools like the LSRFortessa and C6 Analysis software, researchers can improve the reproducibility and accuracy of their GC-related experiments.
By incorporating these insights and techniques, researchers can gain a deeper understanding of the GC and its role in the immune system, ultimately contributing to the development of more effective treatments and therapies.