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Document Type

Thesis - University Access Only

Award Date

2014

Degree Name

Master of Science (MS)

Department

Veterinary and Biomedical Sciences

First Advisor

Alan Young

Abstract

The production of antibodies against antigens is achieved through adaptive immunity. A key component of the adaptive immune response is the germinal center (GC) reaction that occurs in the secondary follicle of a lymph node. Activated GCs contain B cells that undergo antibody affinity maturation events. A crucial component of the GC is follicular dendritic cells (FDCs) which induce B cells to undergo immense proliferation and apoptosis, somatic hypermutation, positive selection, isotype switching and differentiation into memory cells or long-lived plasma cells. The current immunotherapeutic strategies to generate antigen-specific antibodies are unable to fulfill the entire need for these critically important antibodies. Therefore, an in vitro antibody production method is needed. Dr. Young’s laboratory has developed several methods to mimic active germinal centers in cell culture, which can be used as a technique for production of high-affinity antibodies in vitro. The goal of the project was to further investigate and optimize the in vitro system. The proliferation of human PBMCs using human or sheep FDCs was tested using a BrdU cell proliferation ELISA to assay the productivity of in vitro GCs. Using these in vitro GCs, the production of antigen-specific (influenza) responses of human PBMCs was investigated and analyzed via flow cytometry. The BrdU assay indicated that there was a trend toward higher proliferation of PBMCs when grown in culture with FDCs, although individual variations occurred. There was also a trend toward higher antigen-specific responses when cells were grown in culture with FDCs. Although these results are encouraging, further studies are needed to better optimize the system. This in vitro GC technology would help meet the highdemand for specific antibodies and may remove the need for in vivo development and production of polyclonal and monoclonal antibodies.

Library of Congress Subject Headings

Antigens
Immunity
Germinal center

Description

Includes bibliographical references (pages 63-67)

Format

application/pdf

Number of Pages

79

Publisher

South Dakota State University

Rights

In Copyright - Non-Commercial Use Permitted
http://rightsstatements.org/vocab/InC-NC/1.0/

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