Lecture 6: B-Lymphocytes, etc. 1. B cells arise in the bone marrow from progenitor lymphoid cells. a.

Refer to the hematopoiesis chart b. Remember B cells contain antigen-recognizing receptors for interaction with antigen. 2. B-cell receptor complex (BCR) a. Also known as membrane immunoglobulin (mIg) i. Expressed in association with CD79a/CD79b. b. Like antibodies it has two identical light/heavy chains linked by a disulfide bond. i. Variable regions 1. Light chain contains only V and J segments. 2. Heavy chain contains V, D, and J segments. 3. The generation of the heavy and light chain variable regions creates a unique antigen-binding site for each B-cell. The ways in which these regions are randomized is through: a. Multiple copies of V, D, and J. b. Random selection and combination of V, D, and J. i. Random selection is termed combinatorial diversity. c. Junctional diversity generated by addition/deletion of bases i. Junctional diversity is the deletion or insertion of V, D, and J segments to maintain an open reading frame downstream. 1. Mediated by terminal deoxynucleotidyl transferase (Tdt) d. Random assortment of light/heavy chains 4. Somatic recombination a. This is a process in which the DNA that encodes for the variable regions in the light and heavy chains is cut and recombined to form an intact gene. b. Recombination is triggered by RAG-1 and RAG-2. i. RAG = Recombination Activating Gene 5. B-cell repertoire a. A term that simply means all the possible combinations/forms of B cells and their variable regions. 6. Allelic exclusion a. Remember a chromosome has two possible outcomes b. The successful rearrangement of heavy chain variable regions from one chromosome, thus inhibiting the other outcome on the chromosome is termed allelic exclusion. 7. B-Cell Developmental Stages [Goal to create a B-cell repertoire that is non autoreactive] a. Pro-B cell i. This is a preparation stage. ii. Transcription of multiple genes: RAG-1/2, Tdt, CD79a/b, and CD19 [the Pan B cell marker!] b. Pre-B cell

i. Pre-BCR is expressed with CD79a/b 1. Pre-BCR is composed of: a. Two rearranged heavy chains b. Two pseudo light chains ii. Somatic recombination occurs in both the heavy/light chains. iii. CD20 is expressed and continues to be expressed from here on. c. Immature B cell i. Full BCR is expressed with two full heavy/light chains. ii. Tolerance induction is completed. 1. Apoptosis and Anergy is used to inactivate/destroy autoreactive B-cells. d. Mature B cell i. Characterized by co-expression of IgD and IgM. 1. These are formed from alternate splicing. e. Entrance into the blood i. Mature nave B cells, i.e. have not encountered antigen, enter the blood and circulate looking for antigen. f. Exit from the blood i. Exit is mediated by adhesive molecules (L-selectin) that interacts with High Endothelial Venules (HEV) in the lymph nodes. 8. X-linked Agammaglobulinemia (Brutons) a. Is as a result of mutations in Btk Kinase. b. Btk Kinase must be present in the Pro-B cell stage. i. If not the cells fail to move into the Pre-B cell stage. 1. As a result there are no mature B-cells in the system. c. Cumulative result: reduced B cell numbers and low serum Ig levels.