Lecture 7: B Cell Activation 1. Antigen-dependent B cell differentiation/activation a. Involves an initial antigen encounter in a secondary lymphoid tissue. b.

This results in initiation of the primary immune response. c. B cells undergo i. Isotype switching ii. Affinity maturation (somatic mutation) iii. Differentiation to plasma cells iv. Formation of memory B cells 2. Remember when the B-cell Receptor (BCR) encounters antigen, by crosslinking the antigen binding sites, the antigen is endocytosed and taken through exogenous processing eventually presenting via Class II MHC receptors on the cell surface. 3. CD4+ T-cells detect the Class II MHC receptors with antigen a. The B and T cells form a complex that is stabilized by adhesion molecules such as ICAM-1/LFA1 and LFA-3/CD2. 4. When B cells are fully activated they undergo clonal expansion. a. This large formation of B-cell clones creates germinal centers. 5. Plasma cell differentiation a. B-cells expresses antibodies on the cell surface, plasma cells secrete these antibodies. b. Differentiation occurs four days after immunization. i. IgM is the first secreted. 1. Later Isotype switching in Nave B-cells occurs so other antibodies can be secreted. ii. IgD is also present but it is not secreted. 6. Isotype switching, aka switch recombination a. This is a process by which cells expressing IgM and IgD are changed to produce antibodies of different isotypes (G, A, and E). b. Heavy chain portion of the antibody is modified to change isotypes. c. Specificity of the antigen-binding site is maintained because only the heavy chain portion is altered. d. Occurs roughly a week into B-cell activation. e. Cytokine note: disrupting IL-4 inhibits switching to IgE. 7. Affinity maturation, aka somatic mutation a. Occurs 7-10 days following B-cell activation. b. Random mutations in the VDJ gene region of light and heavy chains. i. Gives rise to point mutations ii. Results in gradual accumulation of higher affinity antibodies for immunizing agents. 8. Generation of memory B-cells a. Occurs roughly a week after B-cell activation i. Runs concurrently with isotype switching/affinity maturation. 1. As a result expresses isotypes other than IgM and high affinity immunoglobulins.

b. Some cells colonize secondary lymphoid organs while others join general circulation. c. Entrance into lymph nodes, a secondary lymphoid tissue, is different from other molecules. i. Most molecules use HEV to enter. ii. Memory B-cells lack L-selectin so they cannot use HEVs. 1. They enter via afferent lymphatics. d. Memory B-cells present for weeks or months without stimulation. 9. Negative signaling refers to inhibition of B-cells following antigen induced activation. This is basically a negative feedback loop to keep the system in check. 10. Secondary immune response a. Occurs when B-cells are activated a second time by an antigen. b. The response is quicker and stronger than before. i. It can also be initiated outside secondary lymphoid tissue. 11. Monoclonal activators a. Lead to cloning of one cell. b. T-independent antigens antigens that induce nave B-cells to activate. Leads mainly to IgM production. i. Example: Lipopolysaccharide (LPS) at low levels c. To enhance this response you can use an adjuvant. 12. Polyclonal activators (Mitogens) a. Leads to cloning of many different cells. i. Examples: 1. B-cell mitogens pokeweed mitogen and high levels of LPS. 2. T-cell mitogens concanavalin A, phytohemagglutinin, and pokeweed mitogen. 13. Oligoclonal activators a. Leads to cloning of a specific subset of T-cells i. Activates a very large and inspecific subset! b. Binds to a specific portion of T-cells called the V. i. There are only 200 of these portions shared by all T-cells. 1. As a result if you activate one you get a storm of T-cells.