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Investigation of the human monocyte receptors CD163 and CD64.

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Call No.	QH603.C43 S85 2003
Author	Sulahian, Timothy H.
Title	Investigation of the human monocyte receptors CD163 and CD64.
Date	2003
Abstract	Monocytes and macrophages play a key role in a number of immune functions including inflammation, antigen presentation and the clearance of microorganisms and free hemoglobin. Two surface receptors which are important for these functions are CD163 and CD64. In addition to being the scavenger receptor for free hemoglobin, CD163 has also been implicated in the regulation of inflammation. While it is known that CD163 expression is induced by glucocorticoids, the regulation of this molecule by other monocyte stimuli is not completely understood. The majority of this thesis addresses the issue of CD163 regulation by exposing monocytes to different stimuli and measuring CD163 levels by flow cytometry. Data is presented which demonstrates that surface CD163 is upregulated in response to the anti-inflammatory cytokine, IL-10. The development of an ELISA to measure soluble CD163 and the first evidence that soluble CD163 is present in normal plasma are included. In addition, the novel finding that crosslinking Fc gamma receptors induces the shedding of CD163 from the cell surface is reported. Taken together, these findings support the hypothesis that CD163 is associated with an anti-inflammatory phenotype and raise intriguing questions about the possible functions of soluble CD163. CD64 is an Fc receptor, i.e. a receptor for the Fc portion of IgG. Crosslinking CD64 results in the phagocytosis of opsonized particles and release of a number of cytokines and anti-microbial substances. In addition, CD64 targeted antigens are presented to T cells with extremely high efficiency, leading to the development of CD64 targeted vaccine strategies. However, the effect of targeting antigens to CD64 on the Th1/Th2 balance of the response has not yet been established. The second part of this thesis describes the generation of CD64 targeted antigenic fusion proteins. Three proteins were generated and purified using transfected NSO cells and affinity chromatography and were intended to (i) monomerically target, (ii) trimerize or (iii) multimerize the receptor while targeting ovalbumin to CD64. While aggregation of the monomeric construct compromises its use in addressing questions of CD64 targeting valency, the constructs are still useful for investigating the effect of CD64 targeting on the Th1/Th2 balance of the immune response.
Description	xiv, 150 leaves : ill. 28 cm.
Format	application/pdf
Advisor	Paul M. Guyre.
Subject	Cell receptors. Fc receptors. Monocytes. Immune response -- Regulation. Proteins -- Immunology. Receptors
College	Dartmouth Medical School -- Department of Physiology.
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D.)--Dartmouth College.
Copyright	All rights reserved. No part of this book may be reproduced in any form by any electronic or mechanical means without permission in writing from the author.
DOI	http://dx.doi.org/10.1349/ddlp.21

Title	Page 1
Format	application/pdf
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D.)--Dartmouth College.
Full Text	INVESTIGATION OF THE HUMAN MONOCYTE RECEPTORS CD163 AND CD64 A Thesis Submitted to the Faculty in partial fulfillment of the requirements for the "..... y--~ -, ,....... Carol L. FoIt, Ph.D. Dean of Graduate Studies degree of Doctor of Philosophy In Physiology by Timothy H. Sulahian DARTMOUTH COLLEGE Hanover, New Hampshire May 29, 2003 Examining Committee: Ei.ul M. ciuyr~, Ph.D.2(Chairmanl ~ ). Donald Bartlett, Jr., M.D. i J <;pqIel R. Wira, Ph.D. /funes D. ctorham, M.D., Ph.D. Robert F. Grliano, Ph.D.

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