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Color Plates

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Figures

Image of COLOR PLATE 1 (chapter 23)

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COLOR PLATE 1 (chapter 23)

A standard approach to analyzing flow cytometry data from tetramer-stained whole blood. Whole blood from an individual who was HLA-B7 and seropositive for EBV was stained with anti-CD3, anti-CD8, anti-CD11a, and an HLA tetramer composed of the HLA-B*0701 heavy chain and a well-characterized B7-restricted CD8 T-cell epitope from EBV (RPP for the first three amino acids in its peptide sequence). The same hierarchical gating procedure is shown in panels A and B. (A) Data displayed in pseudo-color dot plot format; (B) contour plot format, with the show-outliers option turned on for the third panel to show the low-frequency tetramer-positive population.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 2 (chapter 23)

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COLOR PLATE 2 (chapter 23)

Use of overlay plots to show the phenotype of a tetramer-positive population with two markers. In this example, whole blood from the same donor as in Color Plate 1 was stained with anti-CD8, anti-CD38, anti-HLA-DR, and the B7/EBV. RPP tetramer. After gating on lymphocytes and CD8 cells (two left-hand panels), CD8 cells are plotted as forward scatter versus tetramer, and a gate on the tetramer-positive cells is set. Finally, an overlay plot is created where all CD8 cells are shown in orange (as a zebra-plot format), while the tetramer-positive cells are presented as an overlay of blue dots.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 3 (chapter 36)

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COLOR PLATE 3 (chapter 36)

Blue light emission versus violet light emission as measured by INDO-1-loaded cells or control blue beads. Control positions of blue beads used in the alignment and UV detection of the calcium method can produce either a 45° angle or a 10° angle depending on the light filters used. The expected positions of simulated (free calcium release) and resting cells at baseline are indicated.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 4 (chapter 48)

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COLOR PLATE 4 (chapter 48)

Direct immunofluorescence of a frozen section from a renal biopsy specimen from a patient with IgA nephropathy, stained with FITC-rabbit antihuman IgA and showing diffuse granular staining in the glomerular mesangium.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 5 (chapter 48)

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COLOR PLATE 5 (chapter 48)

Direct immunofluorescence of a frozen section from a renal biopsy specimen from a patient with membranous glomerulonephritis, stained with FITC-rabbit antihuman IgG and showing diffuse granular deposits of IgG along the glomerular basement membrane.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 6 (chapter 48)

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COLOR PLATE 6 (chapter 48)

Direct immunofluorescence of a frozen section from a renal biopsy specimen from a patient with anti-GBM disease, stained with FITC-rabbit antihuman IgG and showing diffuse linear staining along the glomerular basement membrane.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 7 (chapter 58)

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COLOR PLATE 7 (chapter 58)

Mantoux test. Intradermal injection of tuberculin raises a weal with a characteristic peau d’orange appearance.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 8 (chapter 58)

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COLOR PLATE 8 (chapter 58)

Multipuncture Heaf test. This grade 1 reaction is commonly seen in healthy subjects who have received BCG vaccination.

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 9 (chapter 68)

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COLOR PLATE 9 (chapter 68)

Detection of influenza A virus in columnar respiratory epithelial cell by using a respiratory screen reagent. (Immunofluorescent staining; magnification, ×400.)

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 10 (chapter 68)

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COLOR PLATE 10 (chapter 68)

Detection of influenza A virus infection in R-mix cells by immunofluorescent staining at 24 h postinfection. (Courtesy Caroline Fong; magnification, ×200.)

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 11 (chapter 68)

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COLOR PLATE 11 (chapter 68)

Detection of CMV pp65 antigen in PBL nuclei by immunofluorescent staining. (Magnification, ×400.)

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 12 (chapter 97)

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COLOR PLATE 12 (chapter 97)

Whole blood from an individual with untreated latent infection was analyzed to determine the frequency of immediate cytokine-producing -specific CD4 T cells. Whole blood was stimulated with PPD for 20 h, the final 4 h in the presence of brefeldin A. CD4 T cells were analyzed on a flow cytometer (gating strategies not shown) for expression of various intracellular cytokines after stimulation with medium control, CMV, PPD, and SEB antigens (as shown across the top panel). In the first row of dot plots, with CD4 T cells expressing both CD69 (a marker of short-term activation) and IFN-γ, it is shown that 1.2% of CD4 T cells express both markers after stimulation with PPD antigen. By subtracting the 0.06% background frequency measured after stimulation with medium control antigen, the proportion of (really PPD)-specific CD4 T cells is 1.14%. In the second row of dot plots, parallel tubes demonstrate the frequency of CD4 T cells expressing both TNF-α and IL-2 under the same conditions. In the third row of dot plots, the events (cells) enumerated in the second row are visualized but color coded (gating strategy not shown) to indicate cells expressing all three cytokines of interest (i.e., the blue dots indicate CD4 T cells expressing IFN-γ, with TNF-α and/or IL-2 or neither, while the red dots indicate expression of TNF-α and/or IL-2 or neither).

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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Image of COLOR PLATE 13 (chapter 97)

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COLOR PLATE 13 (chapter 97)

PBMCs from an HIV-infected long-term nonprogressor were surface labeled with MAb to cell surface molecules and an HLA class I tetramer (HLA57 tetramer loaded with a peptide from the p24 protein). Cell frequencies from each of four quadrants from each of the eight dot plots representing the surface molecules tested are on the axis (CD25, HLA-DR, CD27, CD28, CD45RO, CD57, CD62L, and CCR7), while tetramer staining is shown on the axis. The data show that the frequency of antigen-specific CD8 T cells in this individual was ˜0.59% of all CD3 T cells (gating strategy not shown). In addition, the data show that these antigen-specific cells found in the peripheral blood were nearly all CD25, HLADR, CD45RO, CCR7, and heterogenous for the other markers measured. These data illustrate how HLA tetramers can help tease apart antigen-specific T-cell responses, particularly if immunophenotyping (shown here) is coupled with functional assays (e.g., CFSE proliferation assays and/or intracellular cytokine responses).

Citation: Detrick B, Hamilton R, Folds J. 2006. Color Plates, In Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC.
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