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Chapter 137 : Evaluation of the Humoral Response in Transplantation

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Abstract:

This chapter focuses on HLA-specific antibodies. The term donor-specific antibody (DSA) is used to refer to an antibody (ies) specific for donor HLA. The chapter deals with the technical aspects of obtaining the information necessary to evaluate humoral sensitization to HLA antigens in the transplant patient. There are two categories of test used to assess a humoral response, the crossmatch and the antibody screen, the latter of which may be further subdivided into screens that test simply for the presence of antibody and those designed to provide antibody characterization. The advantages and disadvantages of the various techniques for testing antibodies are discussed in the chapter. The major disadvantages of the antibody screen are that it neither tests donor cells nor defines antibody directly. Additional specialized techniques discussed in the chapter may be useful in cases in which it may be crucial to confirm the presence or absence of a particular antibody. The increasing understanding of the role of the humoral response in transplantation indicates the need for certain types of information: (i) does the patient have antibodies; (ii) if so, what are the specificities, titers, and Ig classes; (iii) are the antibodies increasing or decreasing in strength or breadth; (iv) has the patient had antibodies in the past; and (v) has the patient experienced potentially sensitizing events.

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137

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Enzyme-Linked Immunosorbent Assay
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Flow Cytometric Assay
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Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
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Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
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FIGURE 1

Determination of specificity: serum 1. Reactions and presence of antigen are indicated by X.

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
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FIGURE 2

Determination of specificity: serum 2, first iteration. Reactions and presence of antigen are indicated by X. Shading of boxes representing serum reactions corresponds to antigen specificity indicated by similar shading. False-positive reactions for assigned specificities are indicated by boldfaced X’s. False-negative reactions for assigned specificities are indicated by O.

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
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Image of FIGURE 3
FIGURE 3

Determination of specificity: serum 2, A9 CREGs. Reactions and presence of antigen indicated by X. False-negative reactions for assigned specificities are indicated by boldfaced X’s. False-negative reactions for assigned specificities are indicated by O.

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
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Image of FIGURE 4
FIGURE 4

Determination of specificity: serum 2, B35 CREGs. Reactions and presence of antigen are indicated by X. Shading of boxes representing serum reactions corresponds to antigen specificity indicated by similar shading. False-positive reactions for assigned specificities are indicated by boldfaced X’s. False-negative reactions for assigned specificities are indicated by O.

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
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References

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26. Zachary, A. A., and , J. M. Hart. 1997. Relevance of antibody screening and crossmatching in solid organ transplantation, p. 477519. In M. S. Leffell,, A. D. Donnenberg, and , N. R. Rose (ed.), Handbook of Human Immunology. CRC Press, Boca Raton, Fla.
27. Zachary, A. A., and , A. G. Steinberg. 1997. Statistical analysis and applications of HLA population data, p. 132140. In N. R. Rose,, E. C. de Macario,, J. D. Folds,, H. C. Lane, and , R. M. Nakamura (ed.), Manual of Clinical Laboratory Immunology, 5th ed. ASM Press, Washington, DC.

Tables

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TABLE 1

Patient information

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table
TABLE 2

Assay characteristics

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table
TABLE 3

Advantages and disadvantages of different assays

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table
TABLE 4

Quality control factors

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
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Antigen

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table

Serum versus antigen

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table

Serum versus antigen

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table

HLA-A2

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table

Serum versus antigen

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137
Generic image for table

Serum versus antigen

Citation: Zachary A, Houp J, Vega R, Chesterton K, Lucas D. 2006. Evaluation of the Humoral Response in Transplantation, p 1215-1227. 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.ch137

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