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Chapter 17 : Introduction

Author: Maurice R. G. O’Gorman
Content Type: Reference
Publication Year: 2006

Category: Immunology

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Introduction, Page 1 of 2

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Abstract:

This is the introductory chapter to the section . In an effort to capture the burgeoning field of clinical flow cytometry, the chapters presented in this section are divided into two parts. The first part focuses on current application, which encompasses updates in established procedures as well as new procedures that have been recently adopted. The second part is more innovative, with applications combining molecular methodologies and flow cytometry. This section provides a quick review of clinical flow cytometry applications. Stem cell transplant is no longer a rare procedure performed only at highly specialized institutions. With the advent of cord blood banking, new safer stem cell transplant protocols, and numerous new clinical trials, accurate enumeration of stem cells in a variety of products has become an essential procedure for an increasing number of institutions. The X-linked hyper immunoglobulin M (IgM) syndrome is a combined immune deficiency resulting from mutations in the genes encoding the CD40 ligand (CD154). A diagnostic flow cytometry screening test has been developed for the rapid detection of CD40 ligand expression abnormalities. Currently, the most commonly utilized laboratory methods for laboratory diagnosis of allergy involve the detection of allergen-specific IgE. Improved standardization of the allergens used for in vitro testing combined with improvements in the procedures will lead to an increased adoption of this testing modality in the near future.

Citation: O’Gorman M. 2006. Introduction, p 143-146. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch17
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Introduction
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Citation: O’Gorman M. 2006. Introduction, p 143-146. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch17
/content/10.1128/9781555815905.ch17.ch17fig01
FIGURE 1
Measurement of CD40 expression on CD4 T cells. CD4 T cells are gated indirectly by first gating (R1) on CD3 T cells (in the histogram of right-angle light scatter [RALS] versus CD3 and then gating (R2) on CD3CD8 T cells. The level of expression of CD40 ligand is then measured on the events that satisfy both of these gates (R1 and R2) as percent positive (number of events expressing fluorescence above an isotype control antibody). Shown below are the results on CD40 ligand expression levels of resting CD4 T cells in whole blood from a healthy control and on vitro-activated CD4 T cells in whole blood obtained from a healthy control, a patient diagnosed with the hyper IgM (XHIM) syndrome, and an XHIM carrier mother as labeled above.
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10.1128/9781555815905/f0144-01.gif
Image of FIGURE 1
FIGURE 1

Measurement of CD40 expression on CD4 T cells. CD4 T cells are gated indirectly by first gating (R1) on CD3 T cells (in the histogram of right-angle light scatter [RALS] versus CD3 and then gating (R2) on CD3CD8 T cells. The level of expression of CD40 ligand is then measured on the events that satisfy both of these gates (R1 and R2) as percent positive (number of events expressing fluorescence above an isotype control antibody). Shown below are the results on CD40 ligand expression levels of resting CD4 T cells in whole blood from a healthy control and on vitro-activated CD4 T cells in whole blood obtained from a healthy control, a patient diagnosed with the hyper IgM (XHIM) syndrome, and an XHIM carrier mother as labeled above.

Citation: O’Gorman M. 2006. Introduction, p 143-146. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch17
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/content/10.1128/9781555815905.ch17.ch17fig02
FIGURE 2
CD40 ligand expression on CD4 T cells, illustrating functional immune reconstitution in an XHIM patient transplanted with peripheral blood stem cells from an HLA identical donor sibling. Transplant date was 14 October 2004. Gating was performed as illustrated in Fig. 1 .
10.1128/9781555815905/f0145-01_thmb.gif
10.1128/9781555815905/f0145-01.gif
Image of FIGURE 2
FIGURE 2

CD40 ligand expression on CD4 T cells, illustrating functional immune reconstitution in an XHIM patient transplanted with peripheral blood stem cells from an HLA identical donor sibling. Transplant date was 14 October 2004. Gating was performed as illustrated in Fig. 1 .

Citation: O’Gorman M. 2006. Introduction, p 143-146. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch17
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References

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1. Boumiza, R.,, A. L. Debard, and , G. Monneret. 2005. The basophil activation test by flow cytometry: recent developments in clinical studies, standardization and emerging perspectives. Clin. Mol. Allergy 3: 9.
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7. Jacobsohn, D. A.,, K. M. Emerick,, P. Scholl,, H. Melin-Aldana,, M. O’Gorman,, R. Duerst, and , M. Kletzel. 2004. Non-myeloablative hematopoietic stem cell transplant for X-linked hyper-immunoglobulin M syndrome with cholangiopathy. J. Pediatr. 113: e122– 127.
8. O’Gorman, M. R.,, B. DuChateau,, M. Paniagua,, J. Hunt,, N. Bensen, and , R. Yogev. 2001. Abnormal CD40 ligand (CD154) expression in human immunodeficiency virusinfected children. Clin. Diagn. Lab. Immunol. 8: 1104– 1109.
9. O’Gorman, M. R.,, D. Zaas,, M. Paniagua,, V. Corrochano,, P. R. Scholl, and , L. M. Pachman. 1997. Development of a rapid whole blood flow cytometry procedure for the diagnosis of X-linked hyper-IgM syndrome patients and carriers. Clin. Immunol. Immunopathol. 85: 172– 181.
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