Intrinsic and Extrinsic Factors in the Maintenance of Long Lived Plasma Cells Pubblico

Derakhshan, Arsalan (2012)

Permanent URL: https://etd.library.emory.edu/concern/etds/cc08hg516?locale=it
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Abstract





Abstract

Intrinsic and Extrinsic Factors in the Maintenance of Long Lived Plasma Cells



Some vaccines that are received during childhood have been shown to elicit a humoral response that can be measured for the duration of an individual's life. The mechanism by which long-term humoral immunity can be maintained is not fully understood. Studies have described long-term bone marrow (BM) resident antibody secreting cells (ASCs) that have come to be known as long-lived plasma cells (LLPCs). Great strides have been made in understanding how LLPC populations are maintained and how they differ from plasma cell populations that are short-lived. However, we are far from understanding the complete picture. It is thought that LLPCs have been preprogrammed during B cell development to receive and process environmental signals as to render extended longevity. In this study, we investigate some of the intrinsic and environmental factors that govern how LLPCs are maintained and whether environmental factors differentially affect B lineage subsets. The role of CD59, a gene that is differentially over-expressed in LLPCs, is briefly explored in the context of longevity and affinity maturation. In this study, we also review some of the survival factors that are essential in LLPC maintenance and analyze various bone marrow cells and subsets that provide those survival factors. Through in vitro experiments, we identified many BM subsets capable of maintaining LLPCs, which casts doubts on the role of any single BM subset. Furthermore, our in vivo experiments suggest that eosinophils may not be required for the maintenance of LLPCs as was recently reported.






Intrinsic and Extrinsic Factors in the Maintenance of Long Lived Plasma Cells
By
Arsalan Derakhshan

Advisor: Dr. Rafi Ahmed, PhD

A thesis submitted to the Faculty of the James T. Laney School of Graduate Studies of Emory
University in partial fulfillment of the requirements for the degree of Bachelor of
Science/Master of Science in Biology
2012

Table of Contents



Table of Contents
INTRODUCTION .......................................................................................................................................... 1
MATERIALS AND METHODS ....................................................................................................................... 5

Harvesting and Prepping Bone Marrow Cells ....................................................................... 5

Staining Cells for Flowcytometry ......................................................................................... 5

In Vivo Depletion of Eosinophils .......................................................................................... 6

Gata-1 KO Vs WT Serial Antibody Titers .............................................................................. 7

Quantifying Antibody Titers- Enzyme Linked Immunosorbent Assay (ELISA) ...................... 7

Quantifying Affinity Maturation Using ELISA in NP-Hapten System ..................................... 9

CD55/CD59 Double Knockout Vs WT Serial Antibody Response to NP-Hapten & Alum .... 10

CD55/CD59 Double Knockout Vs WT Serial Antibody Response to NP-Hapten & CpG ...... 11

Antibody Response to NP-Hapten & Alum Vs NP-Hapten & CpG in CD55/CD59 KO Mice 11

Enumeration of Direct Ex-vivo Antibody Secreting Cells (ASC) From Mouse Tissue .......... 12

Co-culture Experiment 1 .................................................................................................... 14

Co-culture Experiment 2 ..................................................................................................... 15

Co-culture Experiment 3 ..................................................................................................... 17
RESULTS ........................................................................................................................................... 18

Figure 1 ............................................................................................................................... 18

Figure 2 ............................................................................................................................... 20

Figure 3 ............................................................................................................................... 21

Figure 4 ..........................................................................................................................23-24

Figure 5 ............................................................................................................................... 25

Figure 6 ............................................................................................................................... 26

Figure 7 ............................................................................................................................... 28

Figure 8 ............................................................................................................................... 29

Figure 9 ............................................................................................................................... 30


Figure 10 ............................................................................................................................. 31

Figure 11 ............................................................................................................................. 32
DISCUSSION ..................................................................................................................................... 33
REFERENCES ..................................................................................................................................... 40

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