Date of Award
2019
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Biological Sciences
Committee Chair
Kyung Ho Roh
Committee Member
Luis Cruz-Vera
Committee Member
Tanya Sysoeva
Subject(s)
B cells--Differentiation--Molecular aspects., Bioconjugates., Immunology.
Abstract
B cells mature within the microenvironment called germinal center and undergo affinity maturation through rounds of selective clonal expansion and so-matic hypermutation, isotype class switching, and differentiation to plasma cells and memory B cells. Induction of such complex germinal center reactions artifi-cially in vitro has not been achieved. Here we first develop and evaluate the use of reversible bioconjugation chemistry for surface modification of magnetic mi-crobeads with model proteins using desthiobiotin-streptavidin pair. Second, we employed the same reversible bioconjugation chemistry and magnetic sorting for isolation of antigen-specific B cells with a minimal amount of potentially toxic magnetic beads for the subsequent culture. Third, the reversible bioconjugation was employed to artificially present the CD40L signal on the surface of the mi-crobeads which mimics the follicular helper T cell (Tfh) and temporally control the surface presentation of CD40L signals mimicking the intrazonal dynamics within the germinal center. With further studies and development, the protocols presented here can be utilized to eventually yield production of antigen-specific effector B cells in larger quantities for various therapeutic purposes.
Recommended Citation
Vinod, Veena, "Isolation and activation of B lymphocytes using magnetic microbeads-assisted reversible bioconjugation" (2019). Theses. 278.
https://louis.uah.edu/uah-theses/278